KR20190136460A - Method and system for protein solubilization - Google Patents

Abstract

The present invention relates to a protein solubilization method, which comprises the following steps: adding alkali, for example lime, and heating the same to produce a reaction solution; and separating the solids from the reaction solution and then neutralizing the reaction solution to produce a neutralized liquid. In addition, the method comprises the steps of concentrating the neutralized liquid and returning water to the reaction solution. The present invention also comprises a system for performing the method.

Description

Method and system for protein solubilization

The present invention relates to a method for solubilizing a protein from a protein, in particular a source from which the protein is not readily solubilized.

Sea Sun provides a way to destroy prions in solubilized proteins

Growing global population has led to a sharp increase in food demand over the last few decades, increasing the need for protein sources for livestock.

Population growth has also produced increasing amounts of waste, which may be a useful raw material for producing animal feed.

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Processes for solubilizing proteins from biological sources are useful for converting proteins in waste into useful protein sources.

Thus, many of these methods have been previously developed. Some methods function only on proteins that are readily solubilized.

It is designed to improve solubilization of proteins from raw materials that are not easily solubilized, for example chicken feathers.

Thermo-chemical treatments promote the hydrolysis of protein-rich substances, breaking up complex proteins into smaller molecules and reducing their digestibility

Improve and use less feed to produce products that meet their needs for maintenance, growth and production

One previous method for solubilizing proteins in chicken feathers involves steaming. In this method, feathers are feather meal.

The process is only steamed to produce a solution that only slightly increases the solubility or digestibility of the protein in the feathers.

to be

Other previous methods involve acid treatment of protein sources. Acid treatment hydrolyzes amino acids, but the condition is that many amino acids

It is generally severe enough to break. Acid conditions also promote the formation of these bonds rather than the breakdown of disulfide bonds that aid in solubility.

All

In addition, the conditions of the prior art system may not be suitable for the destruction of prions in the original protein source.

<Overview of invention>

The present invention includes a novel method for solubilizing proteins. The method generally comprises alkali, for example lime

Feed to the raw material to produce a slurry. Proteins in the slurry are hydrolyzed to produce a liquid product.

The slurry may be heated to aid in hydrolysis. A solid residue may also be produced. The residue is a further process of the present invention.

Can be applied to

According to one specific embodiment, the present invention includes a protein solubilization method which comprises adding alkali to a protein source to produce a slurry.

Forming; Heating the slurry to a temperature sufficient to hydrolyze the protein in the protein source to obtain a reaction solution; Solid from the reaction solution

To separate; Neutralizing the reaction solution with an acid or acid source to produce a neutralized liquid; Concentrated liquid by concentrating the neutralized liquid and

To produce water; Conveying water to the slurry prior to or during the heating step.

According to another specific embodiment, the present invention includes a system for solubilizing a protein, wherein the system comprises a protein source and an alkali

And a heated reactor capable of producing a reaction solution by reacting the system.

Solid / liquid separators may also be included. The system may also allow the addition of acid to the reaction liquid to produce a neutralized liquid.

And a concentration tank capable of concentrating the neutralized liquid and producing concentrated liquid and water.

Additionally a conduit capable of passing from the concentration tank to the heated reactor and at least one heat exchanger capable of exchanging process heat

Can contain

The present invention relates to a method of solubilizing a protein from a biological raw material through hydrolysis.

And apparatus for solubilization and solubilization systems

Specific embodiments described hereinafter relate to solubilization of proteins from three different groups of biological sources.

The group includes resistant or keratinous protein sources such as chicken feathers and animal hairs. The second group includes labile or animal tissue proteins.

Raw, such as chicken ground and shrimp heads. The third group includes plant protein sources such as soybean hay and alfalfa.

Additional groups of white matter and examples included in those three groups will be apparent to those skilled in the art.

The method involves adding alkali, for example lime (Ca (OH) 2 or calcium hydroxide) to the protein source at a certain temperature.

The liquid product is obtained using a solid residue. In the specific embodiments described in Table 1, suitable for each of the three stock groups

Process conditions are provided

Additional advantages of some embodiments of the invention include

Mixtures of labile and resistant proteins can be processed simultaneously

The existing plug flow reactors can be used

Waste reduction is combined with food or protein supplement production

Solubilization greatly increases protein digestibility

The method of the present invention is simple and several components and heat recovery are possible.

Destruction of prions improves food safety

Polishing increases the reaction rate of protein digestion, allowing for increased product concentration and reduced product degradation

Unreacted components can be removed

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Protein product can be concentrated to dryness

Can destroy microorganisms

The present invention also includes a reactor system suitable for the house process of the present invention.

For easier understanding of the invention and its advantages, reference may be made to the description of the following exemplary embodiments and the accompanying drawings.

Suitable Processing Conditions for Protein Solubilization

Protein source resistant labile plant

In certain embodiments of the invention, different, using well-insulated, stirred reactors are used to obtain liquid products rich in amino acids.

Perform proteolytic (solubilization) for hours

Although lime is used in certain embodiments of the present invention, other alkalis such as magnesium oxide, magnesium hydroxide, sodium hydroxide,

Sodium carbonate, potassium hydroxide and ammonia can also be used in the present invention, however, most of these alkalis are recovered by carbonate.

Cannot be

In addition, lime offers advantages over some other alkalis because of its low solubility in water.

Sufficient lime is suspended in solution to maintain a relatively constant pH (~ 12) for the solution.

Ensures a constant pH during weaker hydrolysis conditions (compared to sodium hydroxide and other strong bases), whereby

Can reduce decomposition

Thermo-chemical treatment of high protein materials results in a mixture of small peptides and free amino acids

The newly formed carboxylic acid terminal of reacts with the alkaline medium to produce carboxylate ions, in which

Consumes other alkalis

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During proteolysis, a plurality of side reactions occur. FIG. 1 shows the hydrolysis of protein rich material under alkaline conditions.

Shows a sequential diagram Ammonia is the product of amino acid degradation (eg aspartate and glutamate as products

Deamidation of the resulting asparagine and glutamine). In some embodiments, the ammonia is trapped.

Can be neutralized with an acid, for example sulfuric acid, to produce an ammonium salt. The salt can then be used for fertilizer or other purposes.

Can

Arginine, threonine and serine are also sensitive to alkaline conditions. Sensitivity to degradation of arginine and threonine are both essential

It is nutritionally important because it is a minoic acid, reducing the contact time between the soluble peptide and the amino acid and the alkaline medium

Reduces degradation of compounds and increases nutrient quality. Also, the use of low temperatures (~ 100 ° C) can also reduce degradation.

Sequential treatment of protein-rich materials is used when long processing times are required for high solubilization efficiency (animal hair and chicken feathers)

A higher quality initial product is obtained during the initial treatment, while a lower quality product is produced later.

For example, a series of lime treatments can be used to obtain products with different properties when the initial waste is a mixture.

For example, in the leftover + feather mixture, the initial treatment can use low temperature and short time to target the hydrolysis of the grounded chicken.

However, the second lime treatment (longer time and higher temperature) can digest the feathers.

Table 2 shows the effects of suitable conditions and different processing parameters (temperature, concentration, lime loading and time) on the proteolysis of different materials.

It is a summary

Suitable Conditions for Thermo-Chemical Treatment of the Materials Studyed

Substance Note Recommended Conditions

Alfalfa hay

(158% protein)

Hydrolysis depends on temperature, and alfalfa hay concentration (up to 60 g / L)

D increases lime loading with minimal significant effects,

Converting Proteins to Small Peptides and Free Amino Acids

Required for ruminants

0075 g Ca (OH) 2 / g alfalfa, 100 ° C., 60

min, 60 g / L

Soybean hay

(19% protein)

Hydrolysis increases with lime loading and temperature (up to 100 ° C)

100 ° C is recommended due to high and lower energy requirements

Soybean hay concentration has no significant effect

Hum provides significantly lower hydrolysis conversion Rumin

Suitable for animals

005 g Ca (OH) 2 / g soybean,

100 ° C., 150 minutes

Shrimp head waste

The reaction is complete after 30 minutes. The temperature has no significant effect.

Hydrolysis increases with lime loading (005 g Ca (OH) 2 / g gun)

Suitable for unit animals)

005 Ca (OH) 2 / g dry shrimp,

At least 75 ° C, at least 15 minutes

leftover

(15% protein)

No significant change in conversion after 30 minutes

Concentration has no significant effect Hydrolysis depends on lime loading

Increase (up to 01 g Ca (OH) 2 / g dry residue) in unit animals

Fit

0075 g Ca (OH) 2 / g dry residue,

75 ° C, at least 15 minutes

Remnants + Feathers

A two-step method was studied: step 1 was used to determine the hydrolysis of the residue.

Produces a targeted, high quality amino acid mixture

Targets hydrolysis of feathers and produces ruminant food

Step 1: 0075 g Ca (OH) 2 / g dry residue,

50-100 ℃, 30 minutes

Step 2: 005 g Ca (OH) 2 / g feather, 100 ° C.,

2-4 h

feather

(96% protein)

Hydrolysis occurs faster than in hair, 70% before 6 h

Ring obtained. Suitable for ruminant.

01 g Ca (OH) 2 / g feathers,

100 ℃, 4-8 h

hair

(92% protein)

Prolonged treatment was required for high protein hydrolysis

A two-step process is recommended to reduce minnows acid degradation

Suitable for autumn animals

Step 1: 025 g Ca (OH) 2 / g hair,

100 ° C., 8 h

Step 2: 025 g Ca (OH) 2 / g hair,

100 ° C., 8 h

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The use of calcium hydroxide as an alkaline substance in the process of the present invention is a liquid product obtained in the reaction (in some embodiments the "centrifuged

Solution, which is also called "solution", and because some calcium salts have low solubility, calcium is CaCO3, Ca

It can be recovered by precipitation as (HCO3) 2 or CaSO4. Calcium carbonate has low solubility (00093 g / L, solubility in CaCO3).

(solubility product is 87 x 10-9). In contrast, the solubility of CaSO4 is 106 g / L,

The bandit is 61 x 10-5, the solubility of Ca (HCO3) 2 is 166 g / L, and the solubility level is 108. Also from CaCO3 than CaSO4

It is easier to regenerate Ca (OH) 2

50 to 70% of calcium is recovered by precipitation of calcium carbonate into the reaction liquid product by bubbling CO2.

High pH of the reaction product prior to calcium recovery may be recommended so that calcium carbonate is formed during the process.

May be sufficient The final pH after recovery may be between 88 and 90

Proteins produced from the process of the present invention may have many uses, including as animal feed.

Soluble proteins from star and plant protein sources do not have a well-balanced amino acid profile.

It is best used as a ruminant feed. In unstable proteins, the amino acid profile is well balanced, so that solubilized proteins

Thus, the end use of the protein solubilized by the method of the present invention is

It may be presented by the original source of the protein. An additional advantage in animal feed applications is that produced by some methods of the invention

May be lack of prion in the protein The lime treatment conditions are in many ways severe enough to substantially destroy the prion,

This can improve the safety of any feed produced using solubilized protein.

In addition, in some embodiments, the present invention provides for a period of time to destroy all or significant amounts of prion that may be present in the liquid.

And a holding step of heating the reaction liquid at an elevated temperature. For example, the liquid may be heated to a temperature of 125-250 ° C. for 1 second to 5 hours.

Can be heated

Protein-rich substances often found in wastes can be divided into three categories: keratinous, animal tissue and plant material

Each have different properties

Animal hair and chicken feathers have a high protein content (~ 92% and ~ 96%, respectively), and some contaminants that occur during the slaughter process, such as minine

Has Lal, Blood and Lipids The main components of animal hair and chicken feathers are keratin.Keratin is mechanically durable and chemically

As a nonreactive protein, this property is consistent with its physiological function and is a strong fibrous matrix for the tissue where keratin is found.

In mammalian hair, hooves, horns and wool, keratin is present as α-keratin and in bird feathers it is zoned as β-keratin.

Keratin has very low nutritional value, contains large amounts of cysteine, and is difficult to digest by most proteolytic enzymes

Has a very stable structure

The behavior of chicken feathers and animal hairs during the thermo-chemical treatment process of the present invention is shown in FIGS. 2 and 3. FIG. 22 shows chicken feathers rather than animal hairs.

Shows faster hydrolysis rates and greater final conversion to digestible proteins.

Easier access to lime for spear shape, or different large structures present in animal hair compared to chicken feathers (fibrillated spheres)

Crude, porosity, etc.) 01 g Ca (OH) 2 / g dry material for high hair conversion at 100 ° C. using lime loading

At least 8 hours are recommended, but for feathers a 70% conversion can be achieved within 4 hours

A linear relationship between reaction rate and conversion was found in both materials (FIG. 3), which is the first order for alkaline hydrolysis of proteins.

A pseudo equilibrium of hydrolysis versus degradation was found at high conversions

Animal tissue is less difficult to digest than keratinous materials Fluid matrices in which cells in animal tissue are maintained by a simple plasma membrane

Contains the nucleus and other organelles in the cytoplasm. The plasma membrane is easily broken down for glycogen, for digestion by enzymes or chemicals.

Releases Proteins and Other Ingredients

Animal tissues (waste and shrimp heads) are well hydrolyzed in less than 15 minutes (FIG. 4), do not require strong processing conditions,

It is suitable for lime loading and short time. Lipids and other substances present in animal tissues can be reacted through side reactions, eg lipid saponification.

Lime consumption is consumed more quickly, reducing the pH of the liquid product at the end of the process and making the liquid product sensitive to fermentation.

All

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Shrimp head and chicken ground are all animal protein by-products of the food industry, because they are animal tissue,

The acid distribution is expected to be similar to the animal requirements, but the quality may be different because these materials differ from batch to batch.

Dean can be a restriction amino acid in the liquid product

Another particular use for the method of the invention involves the treatment of dead birds in poultry farming. For example, about 5% of chickens in a slaughterhouse

However, typical chicken coops are processed for processing because they cause too many dead chickens to be processed on site.

There is a need for a method for storing dead chickens while waiting for a load. Using the method of the present invention, dead chickens are suitable equipment, for example

It can be crushed with a hammer mill and sprayed with lime to raise the chicken's pH to prevent decay. The lime concentration is about 01 g Ca (OH) 2 / dry

g (dead chicken) When lime-treated chicken is collected and transferred to a central processing plant, to complete the protein solubilization process

Can be heated

Finally, plants contain more difficult to digest lignocellulosic matrices in complex cell walls, making them more difficult to digest than animal tissue.

However, the presence of highly water soluble components allows for a high initial conversion of the protein into liquid during some of the processes of the present invention.

Figure 5 compares the rate of proteolysis for soybean and alfalfa hay. Figure 5 shows more soybean hay than alfalfa hay.

Shows a large soluble fraction and similar hydrolysis rates of both materials

Lime treatment of these plant materials produces products with low levels of lysine and threonine and increases the nutritional value of liquid products for unit animals.

Will reduce

In some embodiments of the invention used to solubilize proteins from plants, the resulting fiber in the solid residue is also ligated.

Digestion is easier because the nin and acetyl groups are removed Lime treatment of plant material is achieved by two products: protein (alkaline hydrolysis).

Liquid products rich in small peptides and amino acids), and to reduce their crystallinity and increase their degradability

Produces a solid residue that is rich in holocellulose that can be treated. Thus, some of the processes of the present invention may be combined with a plant digestion process.

When unexpected synergies exist

6 shows a method of solubilizing a protein in a protein containing material which does not include lime recovery.

White matter-containing material and lime are added to the reactor In a specific embodiment, the heat of reaction produces a hydrated form and calcined lime (Ca

Quicklime (CaO) is added so that (OH) 2) reduces the additional heating requirements of the reaction. Unreacted solids are trapped in unreacted solids.

Countercurrent wash to recover the solubilized protein. The liquid product exiting the reactor contains the solubilized protein.

The evaporator removes almost all of the water to concentrate the solubilized protein. Preferably, the evaporator can continue pumping the concentrated protein.

Sufficient water may remain

Suitable evaporators include multiple-effect evaporators or steam compressed evaporators. Steam compression allows mechanical compressors or jet ejectors to be used.

Because the pH is alkaline, any ammonia produced by proteolysis is volatilized and returned to the reactor.

Ultimately, ammonia levels can accumulate to unacceptable levels.

Excess ammonia can be removed. The removed ammonia can be neutralized with an acid.

Acid, propionic acid or butyric acid), neutralized ammonia can be fed to ruminants as a non-protein nitrogen source.

If gasic acid is added, neutralized ammonia can be used as fertilizer.

The concentrated protein slurry exiting the evaporator may be carbonate to react with excess lime. In some applications, the

Concentrated slurries can be added directly to the feed at short travel distances. However, long travel distances and stable storage are required.

If desired, the neutralized concentrated slurry can be spray dried to form a dry product which contains high calcium concentrations.

Because many animals need calcium for their food, calcium within solubilized protein is convenient to provide their calcium requirements

It may be one way

In Fig. 7, a similar process divided into two steps is shown, which process has a mixture of proteins suitable for ruminants and unit feed.

For example, dead birds include feathers (suitable for ruminants) and tailings (suitable for unit animals).

Step 1 uses mild conditions to solubilize unstable proteins that are subsequently concentrated, neutralized and dried.

The second step uses more stringent conditions to solubilize resistant proteins that can be concentrated, neutralized and dried.

The protein can be fed to ruminants

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FIG. 8 shows a process similar to FIG. 6 with an additional calcium recovery step to produce low calcium product.

For this purpose, the evaporation step is carried out in two stages. In the first evaporator, the protein of the outlet stream is retained in the solution.

To add carbon dioxide. During this step, the pH is preferably about 9.

Favorable pH Drops for Calcium Formation Calcium bicarbonate is much more soluble than calcium carbonate,

Reduced calcium recovery Calcium carbonate is recovered using a filter Calcium carbonate can be backwashed to recover soluble protein

The second evaporator then removes most of the remaining water. Sufficient water can be left to pump the slurry out.

Finally, the slurry can be spray dried to form a stable product upon storage.

FIG. 9 shows the two-step form of FIG. 8 that can be used to process a protein source having a mixture of labile and resistant proteins.

The first step is to solubilize the labile protein suitable for the unit animal and the second step to solubilize the protein suitable for the ruminant.

All

FIG. 10 shows a one stage continuous stirred tank reactor (CSTR) suitable for unstable protein processing Solids squeezed liquid from solids

To be discharged from the reactor using a screw conveyor

FIG. 11 shows a multistage CSTR. Four stages similar to the plug reactor are shown.

It is well suited for the use of protein sources. Plugs reduce the amount of reaction feed discharged with the solids consumed.

In, the liquid flow countercurrent to the solid flow

12 shows a multistage CSTR where the liquid flow co-currents with the solid flow.

13 shows a multi-stage CSTR where the liquid flow crosses into the solid flow.

14 shows a real plug reactor that is well suited for resistance and plant protein sources. Proteins are suitable solid equipment, for example

It is fed into the reactor using the screw conveyor shown in FIG. 14 or a V-ram pump not shown.

And a central shaft for rotating the "finger". The stationary phase "finger" refers to an unproductive rotation of the reactor contents.

It is attached to the reactor wall to prevent it. Water passes in countercurrent to the solid flow. Water exiting the top of the reactor is solubilized

Contains protein products which are discharged through the screen to block solids. Some sources of protein, eg chicken feathers, fur

And the fiber properties of the plant facilitate the filtration. The unreacted solid at the base of the reactor is a screw that squeezes liquid from the solid.

Removed using a conveyor In this embodiment, the compressed liquid is half rather than through a screen on the side of the screw conveyor.

The purpose of the arrangement is to preferentially allow the water to be added to the base of the reactor to flow upwards rather than downwards.

So that the solid is discharged as a gas tight plug.

No need to backwash these solids

FIG. 15 shows a plate similar to that shown in FIG. 14 except that the discharge screw conveyor is not connected to the central shaft of the reactor.

Shows the lug reactor This enables independent control of mixing speed and conveyor speed

FIG. 16 shows plugs similar to those shown in FIG. 14 except that the solid is discharged through the lock hopper rather than the screw conveyor

The reactor shows that the rock hopper can be vented between cycles to prevent gas from being introduced into the reactor.

FIG. 53 shows a method of solubilizing a protein in a protein containing material. First, in any polishing step, the protein source is determined by its surface area

To increase the reaction rate in the reaction step. Once the protein is solubilized in the reactor,

Can be reduced, thus reducing the amount of decomposition of the faster reaction stages.

Can be increased, thereby reducing the cost of recovery. If a polishing step is used, it may be a hammer mill, an in-line homogenizer, or

Can be achieved using other suitable equipment.

Next, the protein reacts with alkali at elevated temperature and high pH. The pH can be lowered to about 10 to 13, for example about 12 days.

Although any base may be used in the reaction step, in selected embodiments the base may be calcium oxide, calcium hydroxide, magnesium oxide.

Nesium, magnesium hydroxide, sodium hydroxide, sodium carbonate, potassium hydroxide or ammonia Calcium oxide and calcium hydroxide are in water

Low solubility and therefore easier to recover. They also buffer the pH up to about 12. In addition, calcium is a nutrient and the final

Other nutrient alkalins may remain in the final protein product.

The conditions may be as described herein, for example for different protein sources.

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The reactor can be a stirred tank, which can operate at 1 atm, but at higher pressures to achieve faster reaction rates.

It may be used in particular at higher temperatures. For example steam can be used directly into the reactor using steam from other parts of the process.

Can be purged to maintain reactor temperature

During the reaction, some amino acids are broken down into ammonia. The ammonia will generally be introduced into the gas phase.

For example, it may be neutralized with sulfuric acid to form ammonia salts. The ammonia salts may then be used for fertilizers or other purposes.

All

Next, the solid and the liquid are separated in the stream exiting the reaction. This can be achieved using a solid / liquid separator.

Recovered solids may include reactive solids, such as unsolubilized proteins, and inert solids such as both bone and stone.

Partial inert solids are denser than reactive solids and this property can be used to aid separation.

Enables repeated recycling of the sieve to improve the overall yield of the process. It is also possible that the presence of

Enables removal of inert solids that can reduce the rate

Density separators that can be used to separate reactive and inert solids include precipitators and hydroclone

Next, any maintenance step can be carried out. In this step, the liquid from the reaction step comprising the solubilized protein is constant

After heating to elevated temperature for a time, it may be cooled. After the reaction step, the liquid may comprise a complete prion.

It may later be detrimental to the health of any animal and human that consumes the solubilized protein.

May be sufficient to destroy all or a substantial portion of any prion present. The holding step may be similar to pasteurization.

For different kinds of prions, suitable temperatures and holding times may be different in most cases to achieve prion breakdown.

There will be sufficient various combinations of temperature and holding time. In specific embodiments, the holding step conditions may be at the required level of free.

May be selected to achieve warm destruction and at the same time limit amino acid degradation. For example, the holding step temperature may be 125-250 ° C.

The holding time can be from 1 second to 5 hours. To select the most suitable holding step conditions, the likelihood of occurrence in the protein source

I can confirm this prion beforehand

The holding step may be heated by steam. The system may heat the liquid to be used to help warm the liquid introduced.

It may include a heat exchange member that allows to escape this holding step.

The liquid can then be neutralized with an acid to reduce the pH to 2-9. The acid used in this step is almost all acid or

May be an acid source. In specific embodiments, the acid is carbon dioxide, phosphoric acid, carboxylic acid, such as acetic acid, propionic acid, and minor

It can be tyric acid, lactic acid, sulfuric acid, nitric acid and hydrochloric acid.

Carbon dioxide can be used as an acid source, especially when alkalis contain calcium. Carbon dioxide is a cheap, calcium-containing reaction

Calcium carbonate or calcium bicarbonate is produced depending on the pH during the neutralization of the liquid. Both calcium carbonate and calcium bicarbonate are prepared using a lime kiln.

Can be converted back to filtration. The lime can be reused in the reaction stage.

Since carbon dioxide is a gas, it is possible to foam the liquid during neutralization. To prevent the above problem, carbon dioxide is microporous.

Hydrophobic membranes, such as those manufactured by Celgard LLC, NC, USA, to be transferred into the liquid phase.

Can be reached

Phosphoric acid is used in certain other embodiments in which the reaction solution contains calcium because the calcium phosphate formed is an important mineral in bone formation.

Therefore, it is useful to add phosphoric acid to the final protein product.

In another embodiment, organic acids such as carboxylic acid and lactic acid are used to neutralize liquids containing any alkali

Since organic acids are an animal's energy source, it is useful to add them to the final protein product.

After neutralization, a selective solid / liquid separation can be carried out. This step allows acid neutralization to be insoluble salts such as calcium carbonate, calcium bicarbonate,

May be most useful when producing calcium sulfate or calcium phosphate. Some of these materials may be desirable for the final product and some may

It may be undesirable or it may be desirable to reduce its concentration in the final product.

Suitable solid / liquid separators are filter presses, rotary drums.

May include filters and hydroclones

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In one particular embodiment, neutralization via carbonation of the reaction liquid comprising calcium occurs at a pH of about 9 which is a solid / liquid fraction.

It is possible to substantially remove in the form of highly insoluble calcium carbonate through the backing group. After a significant amount of calcium carbonate has been removed,

Carbonation or other neutralization may continue to further lower the pH

After neutralization and any solid separation, the neutralized liquid can be concentrated. The reaction solution will generally comprise 2-6% of solubilized protein.

The concentration will not be significantly affected by the maintenance, neutralization and solid recovery steps. After concentration, the concentrated liquid is

May contain 65% solubilized protein

Concentration can be achieved by evaporation, for example using multi-effect, mechanical vapor-compression and jet ejector vapor compression evaporation.

Water can be removed from the neutralized liquid. Generally, dilute protein solutions tend to foam, whereas concentrated solutions

As a result, foam formation is reduced when the evaporator is operated with a liquid containing at least 15% of solubilized protein.

In addition, antifoaming agents may be added to the liquid, especially for thinner liquids. Vegetable oil is an effective antifoaming agent,

Add minutes to the final protein product

Filtration may also be used to concentrate the neutralized liquid. Specifically, dilute solutions may be suitable membranes, such as reverse osmosis membranes or

Can be concentrated by permeation of water through the hermetic nanofiltration membrane. To minimize concentration deflection, a vibrating disc filter (eg

VESP) can be used to achieve high permeation rates and high product concentrations.

The neutralized liquid can also be concentrated by freezing. As ice crystals form, proteins are largely excluded, whereby almost pure

Frozen water and concentrated amino acid / polypeptide solution are separated. Ice crystals can be used to remove concentrated product from its surface.

For example can be washed by reflux

Water can be prepared using a variety of immiscible amines, for example diisopropyl amine, trimethyl amine, methyl diethyl amine and other amines.

Can also be extracted from the liquefied liquid

The water removed during the concentration step can be returned to the reaction step. The water can be warmed from the process steam or other parts of the process.

It may be heated before its return via heat exchange with a fluid. If the water from the concentration step is too hot for the reaction step,

It may also be heat exchanged using a cooling fluid to bring it to an appropriate temperature before it is added to the reaction liquid.

Concentrated liquids may be optionally dried. Drying may be performed using standard equipment, for example spray dryers or scraped drum dryers.

Friction drum dryers can produce bulky final solids.

Steam may be recovered and used for process heat, for example reactor heating.

Thus, the method of FIG. 53 includes a selective grinder, reactor, ammonia collector, solid / liquid separator, optional density separator, optional beam

It can be carried out in a system with oil tank, neutralization tank, other optional solid / liquid separator, concentration tank and optional dryer.

These components can be linked together to allow processing of protein sources into liquid concentrates or dry products.

A return loop may be included to allow further processing and / or reuse. Adjusting the temperature and allowing the reuse of process heat

Can include heat exchanger for

The conditions, instruments, and other components of the systems and processes of the present invention are mutually compatible to produce modified protein solubilization methods and systems.

It will be readily understood that the components can be replaced. For example, a component described for one system or method can be used to digest a particular protein.

To achieve the desired product composition, to aid in the reuse and recovery of heat, and to facilitate interchangeability between different systems.

To other systems or methods.

Example

The following examples are provided to illustrate and further illustrate selected embodiments of the present invention.

It is not intended to be self-expressing. Modifications to these embodiments will be apparent to those skilled in the art and are included in the present invention.

In these examples, equations and experiment numbers are intended to represent only the equations and experiments of the indicated embodiments.

Are not numbered consecutively or similarly between

Example 1 General Methods and Equipment

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In this example the following general methods and formulas were used:

The concentrations of different compounds in the liquid product and the raw materials were determined by two different procedures: the amino acid composition was determined by HPLC

Water at the Laboratory of Protein Chemistry at Texas A & M University

Done); Total Kjeldahl Nitrogen and Mineral Crystals Expanded Soil, Water, and Forage Testing Laboratory at A & M University, Texas

(Extension Soil, Water and Forage Testing Laboratory) using standard methodology

Determination of digestibility of lignocellulosic material was performed by 3-d digestibility test using DNS method.

Polished to a suitable size Several locations at the Forest Science Research Center

Thomas-Wiley experimental wheat with sieve size was used

Lignin, cellulose, hemicellulose (holocellulose), ash and moisture content of the material were determined using the NREL method.

Water baths and shake air baths with thermocouples for temperature measurement and maintenance were used where required.

Achieved by heater Water and ice baths were used as cooling system

In general, the experiments in these embodiments include a 1-L auto with a thermostat and mixer powered by a variable speed motor.

Performed in a clave reactor (FIG. 17) The reactor was pressurized with N 2 to obtain a sample through the sampling port.

High mixing ratios (~ 1000 rpm) were used to induce good contact between the solid and the liquid.

Process variables for protein hydrolysis, ie temperature, time, raw material concentration (g dry matter / L) and calcium hydroxide loading (g Ca (OH) 2 / g dry water

Treatment conditions (for some organic materials) were deliberately changed to study the effects of quality).

The liquid phase was separated from the residual solid material by taking in and centrifugation.

Equation 1 was used to determine the conversion of centrifuged samples based on the initial total Kjeldahl nitrogen (TKN) of the organic material:

Conv1 = (V water x TKN centrifuged liquid) / (m dry sample x TKN dry sample) (1)

The liquid product was analyzed using two different methods to obtain amino acid concentrations and conversion of the reaction.

The total nitrogen content of the liquid sample was determined using the micro-Kjeldahl method Nitrogen content (TKN) x 625 was estimated crude protein content

In the second method, the concentration of individual amino acids present in the sample was obtained using HPLC. In this procedure, the sample was subjected to hydrochloric acid.

(150 ° C., 15 h or 100 ° C., 24 h) to convert proteins and polypeptides to amino acids; The measurement is total army

We call with acid composition We determine free amino acid composition

Additional measurements include: final pH of the liquid product, mass of soluble material in the liquid centrifuged after evaporation of water at 45 ° C.,

And the mass of residual solids after drying at 105 ° C. The mass of the residual solids, the final measurement, is the final horn without further washing with water.

The mixture was determined by filtration through a screen. The retained solid was dried at 105 ° C. The dry weight was not only insoluble solids,

Also included were soluble solids dissolved and retained in the residual solids.

Example 2: Protein Solubilization in Alfalfa Hay

Alfalfa hay is commonly used in ruminant nutrition

Higher feed digestibility ensures that animal demands are met with less feed, resulting in two distinct productions of alfalfa hay

Produces water, ie highly digestible soluble fractions found in liquid products, and delignification residual solids

Alfalfa hay was treated with calcium hydroxide on a minimum cost basis on the market. Table 3 summarizes the composition of alfalfa in different states.

All

TABLE 3

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Composition of Alfalfa in Different States (McDonald et al, 1995)

Alfalfa (% of dry mass) Soluble crude protein lignin cellulose hemi-cellulose

New early flowering 60 19 7 23 29

Medium bloom 54 183 9 26 26

Full bloom 48 14 10 27 21

Hay, Dried In The Sun, Early Bloom 58 18 8 24 27

Medium bloom 54 17 9 26 26

Late flowering 48 14 12 26 22

Mature flowering 42 129 14 29 22

Sun-dried alfalfa hay from Producers Cooperative, Brian, Texas

After obtaining; Thomas-Willy Experiment Mill (Arthur H Thomas Company, Philadelphia, Pennsylvania, USA)

Pia) and sieved through a 40-mesh screen Water content, total Kjeldahl nitrogen (estimated protein fraction), and Army

The acid content was determined to characterize the starting material.

Raw alfalfa hay was 8992% dry matter and 1008% moisture (Table 4) TKN was 2534% and about 1584% dry alfalfa

Corresponds to the crude protein concentration (Table 5). The remaining 8416% corresponds to fiber, sugar, minerals, etc.Amino to raw alfalfa hay

The acid compositions are shown in Table 6. Starting materials have relatively well balanced amino acid content (Table 6) with low levels of tyrosine.

Contained

TABLE 4

Moisture Content of Raw Alfalfa Hay

Sample solids (g) Dry solids (g) Dry solids (%)

1 71436 64248 8994

2 59935 53884 8990

Average 8992

TABLE 5

Protein and Mineral Content in Raw Alfalfa Hay

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

1 25492 02 227 18383 04591 6508 16 90 6 45

2 25 181 02 216 17 865 04 321 6176 16 94 5 42

Average 25 336 02 2 215 18 124 04 456 6342 16 92 55 435

TABLE 6

Amino Acid Composition of Air Dried Alfalfa Hay

Amino acid measured amino acid measured

ASP 1444 TYR 294

GLU 1185 VAL 561

SER 613 MEt 101

HIS 139 PHE 559

GLY 530 ILE 440

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THR 495 LEU 1006

ALA 563 LYS 577

CYS ND TRP ND

ARG 558 PRO 935

ND: Not measured Value is g AA / 100g total amino acids

Experiment 1 temperature effect

To determine the effect of temperature on solubilizing proteins in alfalfa hay, experiments were carried out using lime loading and alfalfa concentrations (each

0075 g lime / g alfalfa and 60 g dry alfalfa / L) were run at different temperatures while keeping the experimental conditions studied and

The measured parameters are summarized in Table 7.

TABLE 7

Experimental Conditions and Measured Parameters to Determine the Effect of Temperature on Protein Solubilization of Alfalfa Hay

Temperature (℃) 50 75 90 100 115

Mass of alfalfa (g) 567 534 567 567 567

Water Volume (mL) 850 800 850 850 850

Mass of lime (g) 43 40 43 43 43

Initial Temperature (℃) 503 732 941 931 105

pH final 111 113 107 99 985

Residual Solid (g) 395 349 37 368 35

Dissolved solid in 100 mL (g) 26 024 3549 34995 36248 31551

Protein in 100 mL (g) 0346 0390 0355 0338 0328

Protein Concentration (%) 133 110 101 93 104

Table 8 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different temperatures.

Based on fungal TKN (253%), proteolytic conversion was estimated (Table 9).

TABLE 8

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 1 (alfalfa hay)

Temperature

Time (min) 50 ℃ 75 ℃ 90 ℃ 100 ℃ 115 ℃

0 00506 00503 00526 00576 00474

5 00520 00669 00609 00641 00620

10 --- 00640 --- --- ---

15 00609 00653 00637 00713 00756

30 00665 00655 00679 00813 00813

45 00692 00771 00719 00958 00955

60 00679 00771 00761 01039 00927

120 --- 00778 --- --- ---

150 00554 --- 00568 00540 00525

180 --- 00624 --- --- ---

TKN is g nitrogen / 100 g liquid sample

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TABLE 9

Conversion of Total TKN to Soluble TKN for Experiment 1 (Alfalfa Hay)

Temperature

Time (min) 50 ℃ 75 ℃ 90 ℃ 100 ℃ 115 ℃

0 335 333 348 382 314

5 344 443 403 425 411

10 --- 424 --- --- ---

15 403 432 422 472 501

30 440 434 450 539 539

45 458 510 476 635 633

60 450 510 504 688 614

120 --- 515 --- --- ---

150 367 --- 376 358 348

180 --- 413 --- --- ---

The final product of proteolysis is individual amino acids, which react with hydroxyls, consume lime and reduce pH

Explain lower pH obtained for high protein conversion (Tables 7 and 9)

Similar initial conversions for all temperatures can be explained by the high fraction of soluble components in alfalfa (about 50%, see Table 3).

Final conversion lower than the merge is explained by different sampling methods All initial samples are sampled from the reactor at internal temperature

The sample was taken through a ring port. For the final sample, the fluid was cooled to 35 ° C., the nitrogen pressure was released, the solid was filtered and the sample was

The sampling procedure for the final sample was modified to measure more parameters. The same procedure was repeated for other experiments.

Followed

Highly soluble alfalfa components are present in the dissolved solids. Table 7 shows approximately 11% protein concentration in the remaining solids after liquid evaporation at 75 ° C.

This is actually lower than the protein content in the raw alfalfa, but the processing step converts the protein into a highly digestible amino acid.

, And these amino acids are mixed with other highly digestible alfalfa ingredients to increase the nutritional value of the final product

18 shows proteolysis (% conversion) as a function of time for the different temperatures studied. Conversion is increased at higher temperatures.

The conversion to 100 ° C. is similar to that obtained at 115 ° C .; Therefore, less amino acids will degrade at lower temperatures.

And the energy required is smaller and the working pressure is lower.

Experiment 2 lime loading effect

To determine the effect of lime loading on protein solubilization of alfalfa hay, experiments were carried out at temperature and alfalfa concentrations (75 ° C. and 40 g, respectively).

The dry alfalfa / L) was run at different lime / alfalfa ratios while maintaining a constant Table 10 shows the experimental conditions and measured parameters.

To summarize

TABLE 10

Experimental Conditions and Measured Parameters for Determining Lime Loading Effect in Protein Solubilization of Alfalfa Hay

Lime loading (g lime / g alfalfa) 0 005 0075 01 02 04

Mass of alfalfa (g) 378 378 378 378 378 378

Water volume (mL) 850 850 850 850 850 850

Mass of lime (g) 0 19 29 38 76 152

Temperature (℃) 75 75 75 75 75 75

Initial Temperature (℃) 781 712 782 583 803 815

pH final 57 10 107 --- 114 112

Residual Solid (g) 235 241 228 203 237 295

Dissolved solid in 100 mL (g) 13489 18645 20201 19289 19215 21651

Protein in 100 mL (g) 0286 0249 0231 0267 0264 0251

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Table 11 shows the total nitrogen content in the centrifuged liquid samples as a function of time for different temperatures.

TABLE 11

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 2 (alfalfa hay)

Lime loading

Time (min) 0 g / g 005 g / g 0075 g / g 01 g / g 02 g / g 04 g / g

0 00360 00364 00353 00370 00319 00345

5 00401 00394 00370 00392 00394 00373

15 00457 00423 00377 00427 00423 00401

30 00457 00452 00451 00441 00423 00450

45 00485 00466 00488 00462 00481 00457

60 00485 00511 00510 00478 00481 00498

150 00457 00394 00370 00427 00554 00401

TKN is g nitrogen / 100 g liquid sample

Based on the average TKN (253%) for dry alfalfa hay, proteolytic conversion was estimated and presented in Table 12.

TABLE 12

Conversion of Total TKN to Soluble TKN for Experiment 2 (Alfalfa Hay)

Lime loading

Time (min) 0 g / g 005 g / g 0075 g / g 01 g / g 02 g / g 04 g / g

0 357 361 350 367 316 342

5 398 391 367 389 391 370

15 453 419 374 423 419 398

30 453 448 447 437 419 446

45 481 462 484 458 477 453

60 481 507 506 474 477 494

150 453 391 367 423 549 398

Again, the initial conversion is similar for all lime loadings because of the highly soluble components present in alfalfa (about 50%, see Table 3).

The final conversion to the experiment (150 min) for 02 g lime / g alfalfa increased while the others decreased and differed from each other 02

In the case of g lime / g alfalfa, the final sample was taken through the sampling port while the final sample for other loading opened the reactor and the sample

Was taken out and drunk

19 shows solubilized protein (% conversion) as a function of time for the different lime loadings studied.

For experiments without using lime. The behavior is related to the highly soluble content of alfalfa hay.

In the limeless experiment, the soluble protein is present in the aqueous phase; The hydroxyl group is dilute to react at the solid or solid-liquid interface

The hydrolysis reaction will be slower under these conditions and therefore less free amino acids are present.

Was 57; Likewise, due to the acid released from the biomass (eg acetyl group) and the amino acid released from the protein

pH became acidic Since no lime was used, the concentration of dissolved solids was lower. In all other cases in Table 10, lime

Was part of the dissolved solids

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19 shows that lime loading has no significant effect on protein solubilization of alfalfa hay.

It may be recommended to avoid acid hydrolysis of proteins that tend to be more damaging than hydrolysis.

Will produce higher concentrations of free amino acids in water

Experiment 3 Alfalfa Concentration Effect

To determine the effect of the initial alfalfa concentration on protein solubilization of alfalfa hay, experiments were carried out with temperature and lime loading (75 ° C. and

0075 g lime / g alfalfa) were run at different alfalfa concentrations while maintaining a constant experimental conditions and measured parameters.

Summarize in Table 13

TABLE 13

Experimental conditions and measured parameters to determine the effect of initial alfalfa concentration on protein solubilization

Alfalfa Concentration (g Dry Alfalfa / L) 20 40 60 80

Mass of alfalfa (g) 189 378 534 756

Water volume (mL) 850 850 800 850

Mass of lime (g) 15 29 40 57

Temperature (℃) 75 75 75 75

Initial temperature (℃) 781 782 732 821

pH final 107 107 113 11

Residual Solid (g) 97 228 349 533

Dissolved solid in 100 mL (g) 10072 20201 3549 41349

Protein in 100 mL (g) 0154 0231 0390 0450

Table 14 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different alfalfa concentrations.

Based on one mean TKN (253%), proteolytic conversion was estimated and presented in Table 15.

TABLE 14

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 3 (alfalfa hay)

Alfalfa concentration

Time (min) 20 g / L 40 g / L 60 g / L 80 g / L

0 00175 00353 00503 00514

5 00182 00370 00669 00571

10 --- --- 00640 ---

15 00204 00377 00653 00770

30 00211 00451 00655 00727

45 00218 00488 0,0771 00946

60 00218 00510 00771 00883

120 --- --- 00778 ---

150 00247 00370 --- 00720

180 --- --- 00624 ---

TKN is g nitrogen / 100 g liquid sample

TABLE 15

Conversion of Total TKN to Soluble TKN for Experiment 3 (Alfalfa Hay)

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Alfalfa concentration

Time (min) 20 g / L 40 g / L 60 g / L 80 g / L

0 346 350 333 256

5 360 367 443 284

10 --- --- 424 ---

15 404 374 432 383

30 418 447 434 362

45 431 484 510 471

60 431 506 510 440

120 --- --- 515 ---

150 489 367 --- 358

180 --- --- 413 ---

The final conversion to the experiment (150 minutes) at 20 g alfalfa / L increased while the others decreased, which was different from the others 20 g

In the case of alfalfa / L, the final sample was taken through the sampling port, while the final sample for the other concentration opened the reactor and pulled out the sample.

Drunken

20 shows protein solubilization (% conversion) as a function of time for different alfalfa concentrations studied.

Increase with addition until reaching a maximum of 60-80 g / L; At this point, the mass of lime and alfalfa is very large

, It is difficult for alfalfa to contact the liquid phase, which reduced the conversion to 80 g / L as compared to that obtained for 20 g / L.

Similar In addition, the conversions for 40 and 60 g / L are similar. As shown in Table 13, the dissolved solids are at higher alfalfa concentrations.

More about

Experiment 4 statistical analysis

To determine if there is a correlation between the variables studied in protein solubilization of alfalfa hay, temperature, lime loading as variables

And alfalfa loading, and additional 23 factorial experiments using TKN solubilization (conversion) at 60 min as response variables.

The studied conditions are summarized in Table 16 together with the conversions obtained for each experiment.

TABLE 16

23 Experimental conditions studied with factorial design

Condition

Var 1

Temperature

(℃)

Var 2

Lime loading

(g lime / g solid)

Var 3

Alfalfa concentration

(g / L)

Y

transform

(%)

1 75 0075 40 506

2 100 0075 40 539

3 75 01 40 474

4 100 01 40 586

5 75 0075 60 510

6 100 0075 60 688

7 75 01 60 604

8 100 01 60 673

Using response variables, Yates using conversion values to obtain interactions between means, variable effects, and studied variables

The algorithm was performed. The information is summarized in Table 17. To determine the variability of the measurements, repeat conditions I and 5 in triplicates.

(Table 18)

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TABLE 17

Yates algorithm results (Milton and Arnold, 1990)

Column 1 Column 2 Column 3 Yates Results Interpretation of Yates Results

10449 21048 45800 5725 Average

10598 24752 3932 983 E1 (Effect of Variable 1)

11987 1458 927 232 E2 (Effect of Variable 2)

12765 2474 -300 -075 I12 (interaction of variables 1 and 2)

337 149 3704 926 E3 (Effect of Variable 3)

1120 778 1016 254 I13 (Interactions of Variables 1 and 3)

1779 783 629 157 I23 (Interaction of Variables 2 and 3)

696 -1083 -1866 -467 I123 (interaction of variables 1, 2 and 3)

TABLE 18

Standard Deviation Calculations and Results

Condition 1 repetition 2 repetitions 3 repetitions Average

5 5468 4766 5104 5113

1 5195 5056 5512 5255

S2 8891 SE 1491

In Table 18, the variance (S2) was calculated as the mean variance of the two conditions studied, followed by SE (standard deviation of the variable effect) versus four values.

Estimated using one mean variance (effects and interactions at 23 factorials are the average of four calculations) 4 degrees of freedom and 99% confidence

Given a degree, the t-student value is 3747. Then, multiplying the t-value by SE (1491) gives the result in the Yates result column.

Provides a limit of unintended effects (-559 and 559)

From Table 17, the only significant effects were from variable 1 (temperature, E1 = 983> 559) and variable 3 (alfalfa concentration, E3 = 926> 559).

This is consistent with the observations in Experiments 1 and 3 From the values obtained in factorial design, the presence of insignificant variable interactions

Means that the effects of temperature and alfalfa concentration are additive, providing the highest conversion when both variables are high.

Because different phenomena can occur at the above, the assay is more likely to be subjected to higher temperatures and concentrations (as seen in Experiment 3).

Cannot be shoveled

There was no significant effect of lime loading on the solubilization of protein from alfalfa hay (E2 = 232 <559), which was in contrast to other variables.

Without interaction (I12 and I23 <559); Thus, lime loading results in acid addition of the protein entirely to amino acids instead of protein solubilization

Conversion may be based on the prevention of hydrolysis of nitrogen (protein) in the liquid product rather than the individual hydrolyzed amino acids.

Only present

Comparing the composition of raw and residual solids provides information on the effectiveness of lime treated alfalfa on protein solubilization

The compositions for are shown in Table 19. These results are shown for condition 5 (75 ° C., 0075 g lime / g alfalfa and 60 g alfalfa / L) of factorial design.

Got

TABLE 19

Comparison of Protein and Mineral Contents in Residual Solids after Raw Alfalfa Hay and Lime Treatment

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(%)

Zn

(%)

Fe

(%)

Cu

(%)

Mn

(%)

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dry

Alfalfa

25336 020 221 18124 04456 5342 16 92 55 435

Residual Solid 22383 018 142 33554 04166 3969 71 137 17 37

Table 19 shows that the calcium concentration of residual solids is greater than that of raw alfalfa.

Increases due to added lime The values for potassium and sodium decrease during lime treatment due to the solubility of these salts

Nitrogen present in the sieve is similar to the value obtained for the raw material prior to lime treatment.

Means similar

The fraction of alfalfa solubilized under condition 5 was calculated as follows:

Soluble fraction = 1-{325 g residual solid-[(355 g dissolved solid / 100 mL liquid) * 200 mL moisture]} / 534 g initial alfalfa = 0524

g solubilized / g alfalfa

The calculation is corrected for dissolved solids contained in 200 mL of liquid. The above values (0524 g solubilized / g alfalfa) are reported in Table 20.

do

TABLE 20

Variable measured for condition 5

Alfalfa Mass (g) 534 pH Final 113

Volume of Water (mL) 800 Residual Solid (g) 325

Mass of lime (g) 40 Dissolved solids in 100 mL (g) 355

Temperature (℃) 75 Soluble Fraction of Alfalfa 0524

Experiment 5 Amino Acid Analysis

Alfalfa hay was limed for 60 min and 24 h with recommended conditions: 100 ° C., 0075 g lime / g alfalfa and 60 g alfalfa / L.

Amino acid analysis was performed in three different ways:

1) Centrifuged liquid product-free amino acid assay The assay was performed without extra HCl hydrolysis of the sample.

Although not destroyed by the procedure, soluble polypeptides may be lost in the assay

2) Centrifuged liquid product-total amino acid analysis The analysis consisted of 24-h HCl hydrolysis of the liquid sample.

Was destroyed by a stone procedure or converted to another amino acid; Soluble polypeptide is measured in the assay

3) Dry product after evaporation of water from the centrifuged liquid Since the sample is a solid, HCl hydrolysis was required.

Minosan (asparagine, glutamine and tryptophan) was destroyed by acid and could not be measured

Tables 21 and 22 show the free and total amino acid concentrations for alfalfa treated with lime at 60 min and 24 h, respectively.

23 shows the protein and mineral content for both samples

TABLE 21

Free and total amino acid concentrations for centrifuged liquid product of lime-hydrolyzed alfalfa hay at 60 min

amino acid

Hydrolyzed-Free Amino Acids Hydrolyzed-Total Amino Acids

Concentration (mg / L) Percentage (%) Concentration (mg / L) Percentage (%)

ASN 16587 1717 000 000

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GLN 000 000 000 000

ASP 5430 562 33481 2304

GLU 10911 1129 15535 1069

SER 4487 464 7872 542

HIS 000 000 000 000

GLY 4450 461 8683 598

THR 1897 196 4365 300

ALA 3734 387 7642 526

ARG 7727 800 11028 759

TYR 000 000 1868 129

CYS 3657 379 ND 000

VAL 3931 407 7103 489

MET 468 048 000 000

PHE 920 095 4782 329

ILE 2262 234 3962 273

LEU 2735 283 6406 441

LYS 558 058 3122 215

TRP 1881 195 ND 000

PRO 24978 2585 29447 2027

Total 96 615 100 145295 100

Table 22

Free and total amino acid concentrations for liquid product centrifuged from lime-hydrolyzed alfalfa hay at 24 h

amino acid

Hydrolyzed-Free Amino Acids Hydrolyzed-Total Amino Acids

Concentration (mg / L) Percentage (%) Concentration (mg / L) Percentage (%)

ASN 7610 807 000 000

GLN 000 000 000 000

ASP 7026 745 23979 1751

GLU 11633 1233 15716 1147

SER 3893 413 7664 559

HIS 000 000 000 000

GLY 9601 1018 14165 1034

THR 948 100 3728 272

ALA 3719 394 7406 541

ARG 7525 798 9355 683

TYR 000 000 843 062

CYS 3566 378 ND 000

VAL 3889 412 6617 483

MET 000 000 000 000

PHE 1048 111 4845 354

ILE 2190 232 3984 291

LEU 2595 275 6090 445

LYS 000 000 2676 195

TRP 1756 186 ND 000

PRO 27328 2897 29916 2184

Total 94 324 10000 136982 10000

TABLE 23

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Comparison of Protein and Mineral Contents in the Lime-treated Centrifuged Liquid of Alfalfa Hay

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

60 min 00742 00062 0149 02342 0027 538 2 4 0 2

24 h 00926 00082 0155 02342 0031 518 2 6 0 2

For all experiments, the centrifuged liquid was measured in Kjeldahl crystals but very high concentrations which may not be measured in amino acid analysis

Suspended particulate matter, which is the difference between the estimated protein concentration using amino acid determination and Kjeldahl analysis (145).

Vs. 464 and 137 vs. 579 g protein / L)

Comparing Table 21-23, the nitrogen concentration increases from 60 min to 24 h, but the total amino acid concentration remains relatively constant, thus

Hydrolysis of Alfalfa Hay Shows No Long-term Treatment Required

Finally, the amino acid composition of the product was compared to the necessary essential amino acids of various livestock

Table 24 shows the amino acid compositions of the dry and liquid products (both free amino acids and total amino acids-Table 21) 60

The amino acid composition of lime-hydrolyzed alfalfa hay at min balances well with the essential amino acid requirements of different unit livestock.

Does not achieve particularly low values for histidine, threonine, methionine and lysine; Some other amino acids are not all

Lime hydrolysis of alfalfa hay is enough for proline and asparagine.

But these are not essential amino acids in livestock feed

TABLE 24

Amino Acid Analysis of Products and Essential Amino Acid Requirements for Various Livestocks (Alfalfa Hay)

Amino Acid Catfish Dog Cat Chicken Pig

dry

product

Liquid

(FAA)

Raw material

Alfalfa

ASN 1717

GLN 000

ASP 752 562 1444

GLU 1140 1129 1185

SER 532 464 613

HIS 131 100 103 140 125 071 000 139

GLY 650 461 530

THR 175 264 243 350 250 253 196 495

ALA 455 387 563

ARG 375 282 417 550 000 636 800 558

VAL 263 218 207 415 267 900 407 561

CYS 200 * 241 * 367 * 400 * 192 * 636 379 ND

MET 200 * 241 * 207 225 192 * 095 048 101

TYR 438+ 405+ 293+ 585+ 375+ 278 000 294

PHE 438+ 405+ 140 315 375+ 553 095 559

ILE 228 205 173 365 250 554 234 440

LEU 306 327 417 525 250 1077 283 1006

LYS 447 350 400 575 358 149 058 577

TRP 044 091 083 105 075 ND 195 ND

PRO 1270 2585 935

* Cysteine + Methionine + Tyrosine + Phenylalanine FAA Free Amino Acid

All values are g amino acids / 100 g protein

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The difference between the two liquid samples (free versus total amino acids) is due to some amino acids (especially tryptophan, asparagine)

And glutamine), some proteins in the centrifuged liquid are not hydrolyzed by lime.

Can be present as a soluble polypeptide that is not detected by HPLC analysis.

The difference in amino acids is explained by the high concentration of suspended material (centrifuged for 5 minutes at 3500 rpm) present in the liquid sample.

The suspended material is not determined during the total amino acid measurement since the first step before HCl hydrolysis is centrifugation at 15000 rpm.

Suspended material forms an important part of the dry product, which accounts for very different results on the amino acid composition

The highest protein solubilization (68%) for alfalfa was obtained using 60 min, 0075 g Ca (OH) 2 / g alfalfa, 100 ° C., and 60 g dry alfalfa / L.

Protein solubilization increases with temperature; Initial concentrations of higher alfalfa resulted in a conversion of 60-80 g alfalfa / L.

Increase to the limit

Due to the solubility of the alfalfa component, solubilized protein is high and did not change dramatically in all cases studied (43% to 68%)

Lime loading has the least effect of four variables studied, but some limes contain amino acids that are naturally present in alfalfa.

To prevent damage and to obtain higher proportions of free amino acids in the final product

Finally, the amino acid composition of the product is poor compared to the essential amino acid requirements for various unit livestock.

Low in styrene (underestimated in analysis), threonine, methionine and lysine are particularly rich in asparagine and proline, but they

Not required for water feed Protein products are best suited for ruminants

Lime treatment increases the digestibility of the holocellulose fraction (Chang et al, 1998), thereby reducing the value of residual solids from thermochemical treatment.

Increased use of both products as ruminant food ensures more efficient digestion than the initial material

Example 3: Protein Solubilization in Soybean Hay

Soybeans are usually harvested for some food production. During the harvesting process, large amounts of unused waste products are produced.

In addition, some special climatic conditions (e.g. long dry season, long rainy season) hinder soybean growth.

Leads to the production of animal feed (soybean hay) instead of aquaculture

Treatment of soybean hay will produce two distinct products: highly digestible soluble fraction and delignification residual solids.

Feed digestibility ensures that animal needs are met with less feed

Sun-dried soybean hay (ie, leaves, stems and beans of a cut soybean plant) was obtained from the Terrabon Company;

Was ground using a Thomas-Willy experimental mill (Arthur H. Thomas Company) and sieved through a 40-mesh screen.

Total nitrogen (estimated protein fraction), and amino acid content were determined to characterize the starting material.

In Table 25, the composition of soybeans in different states is summarized.

TABLE 25

Composition of Soybeans in Different States (McDonald et al, 1995)

Big head

Crude fiber

(g / kg)

Crude protein

(g / kg)

Digestive Crude Protein

(g / kg)

Starch and sugar

Soybean meal 58 503-124

Soy flour, batteries 48 415-91

Hay, dried in the sunlight 366 156 101-

Soybean hay was 9131% dry matter and 869% moisture (Table 26). TKN was 302% and crude protein in about 19% dry soybean hay.

Corresponds to the quality concentration (Table 27). The remaining 81% corresponds to fiber, sugar, minerals, etc.

We present at 28

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TABLE 26

Moisture Content of Air Dried Soybean Hay

Sample

solid

(g)

Dry solid

(g)

Dry solid

(%)

1 51781 47297 9134

2 55 824 50967 9130

3 54826 50048 9129

Average 9131

TABLE 27

Protein and Mineral Content of Air Dried Soybean Hay

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

Raw soybean 30 183 037 224 16 477 03 606 1399 34 280 13 53

TABLE 28

Amino Acid Composition of Air Dried Soybean Hay

Amino acid measured amino acid measured

ASP 1679 TYR 282

GLU 1510 VAL 485

SER 565 MET 088

HIS 255 PHE 536

GLY 446 ILE 427

THR 423 LEU 932

ALA 482 LYS 593

CYS ND TRP ND

ARG 775 PRO 521

ND: not measured Value is g AA / 100 g total amino acids

Repeatability of Experiment 1 Results

To determine the repeatability of the results for solubilizing proteins in soybean hay, experiments were run under the same conditions: temperature, lime loading

And soybean hay concentrations (100 ° C., 005 g lime / g soy hay and 60 g dry soybean hay / L, respectively).

The measured parameters are summarized in Table 29.

TABLE 29

Experimental Conditions and Measured Variables to Determine Repeatability of Results in Protein Solubilization of Soybean Hay

Experiment B E J K

Mass of soybean hay (g) 559 559 559 559

Water volume (mL) 850 850 850 850

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Mass of lime (g) 28 28 28 28

Initial Temperature (℃) 93 935 105 981

pH final 86 89 86 89

Residual Solid (g) 353 368 37 354

Dissolved solids in 100 mL (g) 25706 23927 27449 27116

Protein in 100 mL (g) 0770 0799 0837 0779

Table 30 shows the total nitrogen content in the centrifuged liquid sample as a function of time for the same conditions of temperature, lime loading and soybean hay concentration.

Based on the average TKN (302%) for dry soybean hay, proteolytic conversion was estimated (Table 31).

TABLE 30

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 1 (soybean hay)

Time (min) B E J K

0 00808 00741 00799 00831

5 00768 00837 00837 00876

15 00916 00876 00965 00996

30 01 002 00939 01028 01078

45 01068 00977 01084 01203

60 01008 01009 01239 01222

150 01231 01277 01338 01246

TKN is g nitrogen / 100 g liquid sample

Table 31

Conversion of Total TKN to Soluble TKN for Experiment 1 (Soybean Hay)

Time (min) B E J K Average

0 446 409 441 458 438

5 424 462 462 483 458

15 505 483 532 550 518

30 553 518 567 595 558

45 589 539 598 664 598

60 556 557 684 674 618

150 679 705 738 687 702

21 shows proteolysis of soybean hay as a function of time for four different runs under the same experimental conditions.

Relatively little change every time; Dispersion tends to increase at the median and is smaller at the extreme From time behavior, at 150 minutes

The value is close to the maximum conversion-because the rate of change is relatively small for all cases

Experiment 2 temperature effect

To determine the effect of temperature on solubilizing proteins in soybean hay, experiments were carried out using lime loading and soybean hay concentration (each

005 g lime / g soybean hay and 60 g dry soybean hay / L) were run at different temperatures while keeping constant.

The key and measured parameters are summarized in Table 32.

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Table 32

Experimental Conditions and Measured Parameters to Determine the Effect of Temperature on Protein Solubilization of Soybean Hay

Temperature (℃) 75 100 115

Mass of soybean hay (g) 559 559 559

Water volume (mL) 850 850 850

Mass of lime (g) 28 28 28

Initial Temperature (℃) 753 93 1002

pH final 95 86 8

Residual Solid (g) 362 353 346

Dissolved solid in 100 mL (g) 27593 25706 26568

Protein in 100 mL (g) 0647 0770 0823

Table 33 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different temperatures.

Based on average TKN (302%), proteolytic conversion was estimated (Table 34).

Table 33

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 2 (soybean hay)

Temperature

Time (min) 75 ℃ 100 ℃ * 115 ℃

0 00822 00795 00781

5 00869 00830 00856

15 00889 00938 0093

30 00916 01012 01008

45 00969 01083 01094

60 00982 01120 01140

150 01035 01273 01315

* Average TKN for 4 experiment runs is g nitrogen / 100 g liquid sample

Table 34

Conversion of Total TKN to Soluble TKN for Experiment 2 (Soybean Hay)

Temperature

Time (min) 75 ℃ 100 ℃ * 115 ℃

0 454 438 431

5 479 458 472

15 490 518 513

30 505 558 556

45 535 598 604

60 542 618 629

150 571 702 726

* Average of 4 experiment runs

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22 shows proteolysis (% conversion) as a function of time for the different temperatures studied. Conversion is increased at higher temperatures.

The conversion to 100 ° C. is similar to that obtained at 115 ° C .; Therefore, less amino acids are degraded at lower temperatures.

And less energy required and lower working pressure.

The analysis in Table 32 again shows that as more lime reacts with the amino acid product, the protein ratio of the product increases with increasing conversion.

Increase in protein solubility shows a decrease in pH as protein solubilization increases

Conversion at 75 ° C. is statistically different from conversion at 100 and 115 ° C. In all cases, the reaction rate decreases at 150 minutes.

Have incense

Experiment 3 lime loading effect

To determine the effect of lime loading on protein solubilization of soybean hay, temperature and soybean hay concentrations (100 ° C. and 60 g drying, respectively)

The experiments were run at different lime / soybean hay ratios while maintaining soybean hay / L) constant.

The numbers are summarized in Table 35.

Table 35

Experimental Conditions and Measured Parameters for Determining Lime Loading Effect in Protein Solubilization of Soybean Hay

Lime loading (g lime / g soybean hay) 0 005 01

Mass of soybean hay (g) 559 559 559

Water volume (mL) 850 850 850

Mass of lime (g) 0 28 56

Temperature (℃) 100 100 100

Initial Temperature (℃) 935 981 905

pH final 59 89 108

Residual Solid (g) 361 354 344

Dissolved solids in 100 mL (g) 21803 27116 34937

Protein in 100 mL (g) 0560 0779 0906

Table 36 shows the total nitrogen content in liquid samples centrifuged as a function of time for different lime loadings.

Based on the average TKN (302%) for the proteolytic conversions are estimated and shown in Table 37. Soluble present in soybean hay

Due to the sex component the initial conversion is similar for all lime loadings

TABLE 36

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 3 (soybean hay)

Lime loading

Time (min) 0 (g / g) 005 (g / g) * 01 (g / g)

0 00787 00795 00761

5 00850 00830 00811

15 00908 00938 01147

30 00895 01012 00965

45 00914 01083 01128

60 00888 01120 01178

150 00895 01273 01448

* Average TKN for 4 experiment runs is g nitrogen / 100 g liquid sample

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TABLE 37

Conversion of Total TKN to Soluble TKN for Experiment 3 (Soybean Hay)

Lime loading

Time (min) 0 (g / g) 005 (g / g) * 01 (g / g)

0 434 438 420

5 469 458 447

15 501 518 633

30 494 558 532

45 504 598 622

60 490 618 650

150 494 702 799

* Average of 4 experiment runs

Figure 23 shows solubilized protein (conversion) as a function of time for the different lime loadings studied.

Increased with addition, giving maximum effect when changing to 005 g / g lime loading in lime-free experiments.

Achieved at 15 minutes and further processing at 100 ° C. does not produce additional protein solubilization

Minimum lime loading is required for protein solubilization The difference between lime loadings of 005 and 01 g / g is statistically only for 150 minutes

Keep in mind

In the limeless experiments, the final pH was 59. Similarly, from acids (eg, acetyl groups) and proteins released from the biomass

The pH became acidic due to released amino acids. Since no lime was used, the concentration of dissolved solids was lower.

In all other cases reported, lime was part of the dissolved solids

Experiment 4 Soybean Hay Concentration Effect

To determine the effect of initial soybean hay concentration on protein solubilization, experiments were carried out with temperature and lime loading (100 ° C. and 005 g lime, respectively).

Ash / g soybean hay) was run at different soybean hay concentrations while maintaining a constant.

To summarize

Table 38

Experimental Conditions and Measured Variables to Determine the Effect of Early Soybean Hay Concentration on Protein Solubilization

Soybean Hay Concentration (g Dry Soybean Hay / L) 40 60 80

Mass of soybean hay (g) 378 534 756

Water volume (mL) 850 800 850

Mass of lime (g) 29 40 57

Temperature (℃) 75 75 75

Initial Temperature (℃) 782 732 821

pH final 107 113 11

Residual Solid (g) 228 349 533

Dissolved solid in 100 mL (g) 20 201 3549 41349

Protein in 100 mL (g) 0231 0390 0450

Table 39 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different soybean hay concentrations.

Based on average TKN (302%) for seconds, proteolytic conversion was estimated and presented in Table 40

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TABLE 39

Total Kjeldahl nitrogen content in centrifuged liquid phase as a function of time for Experiment 4 (soybean hay)

Soybean Hay Concentration

Time (min) 40 g / L 60 g / L 80 g / L

0 00531 00741 01065

5 00503 00837 01215

15 00592 00876 01264

30 00639 00939 01399

45 00681 00977 01514

60 00701 01009 01472

150 01028 01277 01221

TKN is g nitrogen / 100 g liquid sample

TABLE 40

Conversion of Total TKN to Soluble TKN for Experiment 4 (Soybean Hay)

Soybean Hay Concentration

Time (min) 40 g / L 60 g / L 80 g / L

0 440 438 441

5 417 458 503

15 491 518 523

30 530 558 579

45 565 598 627

60 581 618 609

150 852 702 505

24 shows protein solubilization (conversion) as a function of time for the different soybean hay concentrations studied.

For times less than min, it does not change with soybean hay concentration. Values at 150 minutes correspond to the previous values.

Probably have some sampling problems. From Table 38, the dissolved solids and proteins present in the final product

Increases with increasing hay concentration

Comparing the composition of raw and residual solids provides information on the effectiveness of lime-treated soybean hay on protein solubilization.

The compositions for the materials are shown in Table 41. These results were obtained for 100 ° C., 005 g lime / g soybean hay and 60 g soybean hay / L.

All

Table 41

Comparison of protein and mineral content in raw soybean hay with residual solids and centrifuged liquid after lime treatment

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na (ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

Raw soybean 30 183 037 224 16 477 03 606 1399 34 280 13 53

Residual Solid 19824 033 078 31171 01845 1326 19 158 9 35

Centrifuged liquid 01176 00104 0155 02114 00146 104 2 10 0 2

* About 150 minutes

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Table 41 shows that the calcium concentration of residual solids is greater than that of raw soybean hay.

The value of other minerals decreases during the lime treatment due to the solubility of these salts.

Nitrogen present in the sieve is 33% less than the value obtained for the raw material before lime treatment

Centrifuged liquids have a very high concentration of calcium due to lime, which leads to calcium concentration in the final product (after evaporation of the centrifuged liquid).

The degree will be higher than the nitrogen content. The ratio of protein to calcium in the final product is the ratio = (01176 x 625) / 02114 =

348 g protein / g Ca

The fraction of solubilized soybean hay is calculated as follows:

Soluble fraction = 1-{262 g residual solid-[(156 g dissolved solid / 572 mL liquid) * 200 mL moisture]} / 559 g initial soybean hay =

0450 g solubilized / g soybean hay

The calculation corrects for dissolved solids contained in 200 mL of liquid. The solids are not washed, so the retained liquid contains dissolved solids.

Record this value (0450 g solubilized / g soybean hay) in Table 42

Table 42

Parameters measured for 100 ° C., 005 g lime / g soybean hay, and 60 g soybean hay / L

Mass of soybean hay (g) 559 pH Final 97

Water Volume (mL) 850 Residual Solid (g) 362

Mass of lime (g) 28 Dissolved solids in 572 mL (g) 156

Temperature (℃) 100 Soluble fraction of soybean hay 045

Experiment 5 Amino Acid Analysis

Soybean hay was limed at 150 ° C. and 24 h with recommended conditions: 100 ° C., 005 g lime / g soy hay, and 60 g soy hay / L.

Multi-amino acid analysis was performed in three different ways:

1) Centrifuged liquid product-free amino acid assay The assay was performed without extra HCl hydrolysis of the sample.

Although not destroyed by the procedure, soluble polypeptides may be lost in the assay

2) Centrifuged liquid product-total amino acid analysis The analysis consisted of 24-h HCl hydrolysis of the sample.

Destroyed by tea or converted to another amino acid; Soluble polypeptide is measured in the assay

3) Dry product after evaporation of water from the centrifuged liquid Since the sample is a solid, HCl hydrolysis was required.

Minosan (asparagine, glutamine and tryptophan) was destroyed by acid and could not be measured

Tables 43 and 44 show the free and total amino acid concentrations for soybean hay treated with lime at 150 minutes and 24 h, respectively.

Table 45 shows the protein and mineral contents for the two samples.

Table 43

Free and total amino acid concentrations for centrifuged liquid product of lime-hydrolyzed soybean hay at 150 min

amino acid

Hydrolyzed-Free Amino Acids Hydrolyzed-Total Amino Acids

Concentration (mg / L) Percentage (%) Concentration (mg / L) Percentage (%)

ASN 21348 3064 000 000

GLN 000 000 000 000

ASP 6949 997 44 776 3301

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GLU 4646 667 17272 1273

SER 912 131 5272 389

HIS 1451 208 3529 260

GLY 6158 884 10668 787

THR 636 091 3701 273

ALA 2063 296 5807 428

ARG 9744 1398 14270 1052

TYR 000 000 1678 124

CYS 3645 523 000 000

VAL 2071 297 4820 355

MET 000 000 000 000

PHE 2563 368 5538 408

ILE 1035 148 3489 257

LEU 1321 190 5462 403

LYS 000 000 3777 278

TRP 2586 371 000 000

PRO 2558 367 5572 411

Total 69685 100 135633 100

Table 44

Free and total amino acid concentrations for centrifuged liquid product of lime-hydrolyzed soybean hay at 24 h

amino acid

Hydrolyzed-Free Amino Acids Hydrolyzed-Total Amino Acids

Concentration (mg / L) Percentage (%) Concentration (mg / L) Percentage (%)

ASN 9837 1704 000 000

GLN 000 000 000 000

ASP 8254 1430 33684 2565

GLU 4562 790 19613 1493

SER 644 112 5293 403

HIS 000 000 2571 196

GLY 9790 1696 15013 1143

THR 000 000 3385 258

ALA 2650 459 6922 527

ARG 8184 1418 12209 930

TYR 000 000 2091 159

CYS 3426 594 000 000

VAL 1919 333 5005 381

MET 000 000 000 000

PHE 2172 376 5420 413

ILE 1079 187 3779 288

LEU 783 136 6064 462

LYS 000 000 3550 270

TRP 2327 403 000 000

PRO 2088 362 6749 514

Total 57 716 100 131 348 100

TABLE 45

Comparison of Protein and Mineral Contents in the Lime-treated Centrifuged Liquid of Soybean Hay

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Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

150 min 01 176 00 104 0155 02 114 00146 104 2 10 0 2

24 h 01562 00146 0149 02716 00186 104 2 16 0 2

For both cases, the total hydrolyzed amino acid concentration is approximately twice the free amino acid concentration, which is 50% less amino acids

Shows what exists in de form

For all experiments, the centrifuged liquid was measured in Kjeldahl crystals but very high concentrations which may not be measured in amino acid analysis

It contains suspended particulate matter, which is the difference between protein concentrations estimated from amino acid determination and Kjeldahl analysis (136 vs.

735 and 131 vs. 976 g protein / L)

Comparing Tables 43-45, the nitrogen concentration increased from 150 minutes to 24 h, but the total amino acid concentration remained relatively constant, thus

Hydrolysis of Soybean Hay Shows No Long-term Treatment Required

Finally, the amino acid composition of the protein product was compared to the essential amino acid needs of various livestock.

Table 46 shows that the amino acid products from hydrolysis of soybean hay are not well balanced with respect to the requirements of different unit livestock.

Particularly low values for histidine, threonine, methionine and lysine; Some other amino acids (tyrosine, valine) are all ah

Although enough for most animals, lime hydrolysis of soybean hay produces a product that is very rich in asparagine, but

Not essential in the best feed for vertebrates

TABLE 46

Amino Acid Analysis of Products and Essential Amino Acid Requirements for Various Livestocks (Soybean Hay)

Amino Acid Catfish Dog Cat Chicken Pig Dry Product Liquid (FAA) Raw Material

ASN 3064

GLN 000

ASP 668 997 1679

GLU 956 667 1510

SER 711 131 784

HIS 131 100 103 140 125 000 208 255

GLY 1069 884 446

THR 175 264 243 350 250 180 091 423

ALA 505 296 482

ARG 375 282 417 550 000 619 1398 775

VAL 263 218 207 415 267 708 297 485

CYS 200 * 241 * 367 * 400 * 192 * 922 523 ND

MET 200 * 241 * 207 225 192 * 087 000 088

TYR 438+ 405+ 293+ 585+ 375+ 271 000 282

PHE 438+ 405+ 140 315 375+ 526 368 590

ILE 228 205 173 365 250 515 148 427

LEU 306 327 417 525 250 981 190 932

LYS 447 350 400 575 358 110 000 593

TRP 044 091 083 105 075 ND 371 ND

PRO 1170 367 521

* Cysteine + Methionine + Tyrosine + Phenylalanine FAA Free Amino Acid

All values are g amino acids / 100 g protein

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The difference between the two liquid samples (free versus total amino acids-Table 43 and Table 45) can be attributed to some amino acids (especially trips)

(Tophan, asparagine and glutamine) can also be explained by the acid decomposition of some proteins in the centrifuged liquid

May not be hydrolyzed and may be present as a soluble polypeptide that is not detected by HPLC analysis

The difference in total amino acids in the product is dependent on the high concentration of suspended material (centrifuged at 3500 rpm for 5 minutes) present in the liquid sample.

The suspended material is the total amino acid side since the first step before HCl hydrolysis is centrifugation at 15000 rpm.

Suspended material forms an important part of the dry product, which is a very different texture for amino acid composition.

Explain and

Highest protein solubilization (85%) was achieved using 005 g Ca (OH) 2 / g soybean hay, 150 minutes, 100 ° C, and 40 g dry soybean hay / L.

The effects of the variables studied in this experiment can be summarized as follows:

Protein solubilization increases with temperature and gives the same result as 115 ° C at 100 ° C.

100 ° C because less and no pressure vessel is required. The initial concentration of soybean hay is protein soluble at less than 60 min.

Minimal lime loading (at least 005 g Ca (OH) 2 / g soybean hay) can be used to efficiently solubilize proteins

In all cases, protein solubilization increases with time and the maximum value is obtained for 150 minutes.

Secondary concentration has the least significant effect of the four variables studied

A well balanced biometric analysis of amino acid analyzes for hydrolysis products and essential amino acid requirements for various unit livestock

It is not a holy substance. It has a high concentration of asparagine (which is a non-essential amino acid).

As in the case of alfalfa hay, protein products are best suited for ruminants. Lime treatment reduces the digestibility of the holocellulose fraction.

Increasing (Chang et al, 1998), increasing the value of residual solids from thermo-chemical treatment.

Use ensures more efficient digestion than the initial material

Example 4: Protein Solubilization in Chicken Ground

Chicken grounds are available from the Texas A & M Poultry Science Department.

Generally, residues may include bones, heads, beaks and feet, but in the present case only the intestines (eg heart, lungs, intestines, liver)

The residue was blended in an industrial blender for 10 minutes, collected in a plastic bottle, and finally -4 ° C for later use.

Water content, total nitrogen (estimated protein fraction), ash (mineral fraction) using a sample of the blended material

And the amino acid content was obtained to characterize the starting material.

Equation 1 defines the conversion of centrifuged samples based on the initial total Kjeldahl nitrogen (TKN) of the residue:

Conv1 = {V water x TKN centrifuged liquid} / {m dry dregs x TKN dregs} (1)

Equation 2 defines the conversion of uncentrifuged samples based on the initial total Kjeldahl nitrogen (TKN) of the residue:

Conv2 = {V water x TKN uncentrifuged liquid} / {m dry dregs x TKN dregs} (2)

Equation 3 uses the mass balance to estimate the fractional loss TKN of the initial residue nitrogen:

LTKN = 1- [{Vwater x TKN uncentrifuged liquid} / {m dry residue x TKN dry residue}] (2)

The raw residue was 333% dry matter and 667% moisture (see Table 47). The crude protein concentration of the dry residue was about 45% and ashed.

The amount is about 1%; The remaining 54% were fiber and fat

TABLE 47

Water content of raw waste (g)

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Crucible residue (g) Dry matter (g)% Dry weight

J 322197 106402 33024

A 308807 104 548 33855

4 286961 9512 33147

Average 33342

Dry material (105 ℃ oven)

Experiment 1 Effect of Process Variables

Experiment 1 included eight runs labeled A to H. Runs A, B and C were 20 g dry residue / L and 01 g Ca at 100 ° C.

(OH) 2 / g residues were used. These conditions were the optimal results of previous experiments that studied the same kind of response to chicken feathers.

(Chang and Holtzapple, 1999) The remaining runs (D to H) were obtained for different operating conditions as shown in Table 48.

Was performed

TABLE 48

Experimental conditions used in Experiment 1 (chicken ground)

Execution

Temperature

(℃)

Ca (OH) 2 Mass

(g)

Wet residue

Mass (g)

Water volume

(mL)

Ca (OH) 2 loading

(g / g dry dregs)

Dry residue sea bass

Degree

(g / L)

Final pH

A 100 170 515 850 0099 2020 950

B 100 170 512 850 0100 2008 965

C 100 170 515 850 0099 2020 950

D 100 340 1023 850 0100 4013 955

E 100 510 1533 850 0100 6013 950

F 100 255 1025 850 0075 4021 890

G 100 170 1024 850 0050 4017 910

H 75 340 1024 850 0100 4017 1010

Table 49 shows the total nitrogen content in the centrifuged liquid sample as a function of time for eight runs. Average for dry residues

Based on TKN (7132%), proteolytic conversion was estimated and shown in Table 50. Conversions in Table 50 are shown in FIGS. 25-28 V4.

Pro city

Table 49

Total Kjeldahl nitrogen content in the liquid phase centrifuged as a function of time for Experiment 1 (chicken ground)

Experiment

Time (min) A B C D E F G H

5 00698 00520 00635 01332 02112 01438 00862 01191

10 00721 00543 00658 01354 02112 01461 00851 01191

15 00721 00543 00647 01366 02134 01473 00851 01213

25 00721 00554 00658 01388 02156 01495 00874 01179

35 00721 00566 00647 01388 02145 01517 00874 01191

45 00721 00554 00635 01388 02168 01495 00874 01179

60 00721 00600 00658 01399 02156 --- --- ---

90 00721 00600 00669 01445 02156 --- --- ---

120 00721 00589 00669 01433 02168 01507 00918 01202

180 00765 00623 00681 01433 02179 --- --- ---

TKN is g nitrogen / 100 g liquid sample

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TABLE 50

Fraction Conversion of Total TKN to Soluble TKN for Experiment 1 (Chicken Ground-Equation 1)

Experiment

Time (min) A B C D E F G H

5 0467 0350 0425 0466 0511 0502 0301 0416

10 0482 0365 0440 0473 0511 0510 0297 0416

15 0482 0365 0433 0478 0516 0514 0297 0424

25 0482 0373 0440 0485 0522 0522 0305 0412

35 0482 0381 0433 0485 0519 0529 0305 0416

45 0482 0373 0425 0485 0525 0522 0305 0412

60 0482 0404 0440 0489 0522 --- --- ---

90 0482 0404 0447 0505 0522 --- --- ---

120 0482 0396 0447 0501 0525 0526 0321 0420

180 0512 0419 0456 0501 0527 --- --- ---

25-28 show that under these conditions, the conversion of nitrogen in the solid phase to the liquid phase is not efficient (45-55%)

This means that many proteins in the solid phase do not react with the hydroxide or the formed amino acids precipitate back into the solid phase.

Matter is the presence of fats in the raw materials that consume hydroxide to slow the proteolysis

25-28 show that the reaction occurs during the first 10 or 15 minutes of contact time and then conversion (concentration) remains constant.

25 shows that results from different runs using the same experimental conditions provide similar conversions.

It is similar for different initial concentrations of materials. This is true for residue starting concentrations with higher amino acid concentrations in the liquid phase.

Means higher

27 shows that low lime loading has low conversion; Thus, the reaction requires minimal loading 0075 and 01 lime

Since similar results are obtained for loading, a minimum of 0075 g Ca (OH) 2 / g dry residue will be used. FIG. 28 shows the reaction at 75 ° C.

This shows as fast as almost 100 ° C, which is advantageous because at lower temperatures amino acids will be less degraded

Experiment 2 Process Optimization

In Experiment 2, the goal was to find more (more efficient) conditions for conversion. Experiment 2 was a total of eight threads labeled I to P.

The reaction is fast and the conversion is constant after 15 minutes, so only one sample is needed to obtain representative conditions of the reaction.

Table 51 shows the experimental conditions and the TKN concentration in the liquid sample.

Table 51

Experimental conditions and results for Experiment 2 (chicken ground-2 samples for each run)

Execution

Temperature

(℃)

Ca (OH) 2 concentration

(g / g dry dregs)

Dry residue concentration

(g / L)

Final pH

time

Sample

TKN TKN

I 50 0 100 40 835 15 h 02067 02067

J 100 0075 40 845 30 min 0169 02209 (a)

K 100 0075 40 845 2 h 01722 02296 (a)

L 75 0075 40-30 min 02046 0234 (a)

M 75 0075 40 2 h 02 231 02 318 (a)

N 100 0400 40 1205 1 h 01 116 01094

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O 100 0 300 40 120 1-2 h 01 203 01289

P 75 0 300 40 120 1-2 h 0143 01463

(a) Uncentrifuged liquid sample TKN is g nitrogen / 100 g liquid sample

Table 52 shows that for Runs I to M, the conversion is 63% to 84% using Equation 1 (ie liquid TKN / TKN added in solids).

For Runs J through M, conversion was performed using Equation 2 (ie, liquid TKN / TKN added in solids in uncentrifuged samples).

From 83% to 87% Equation 3 shows 13% loss of initial residue nitrogen at 75 ° C. and initial residue at 100 ° C. for Runs J through M

15% loss of nitrogen is unknown where the lost nitrogen goes

Table 51 and Table 52 show that for the run with the highest conversion, the final pH is different for Experiment 1 and in Experiment 2

Lower than all obtained for other runs. From Experiment 2, we have a lime loading of 0075 gCa (OH) 2 / g dry residue.

Can recommend a temperature of 75 ℃

Table 52

Fraction Conversion of Total TKN to Soluble TKN for Experiment 2 (Chicken Ground)

Run Conversion Sample 1 Conversion Sample 2 Fraction Loss of TKN

I 0781 (1) 0781 (1)

J 0634 (1) 0829 (2) 0171 (3)

K 0646 (1) 0861 (2) 0139 (3)

L 0768 (1) 0879 (2) 0121 (3)

M 0838 (1) 0870 (2) 0130 (3)

N 0436 (1) 0411 (1)

O 0452 (1) 0484 (1)

P 0536 (1) 0548 (1)

(1) Conversion calculated using Equation 1

(2) Conversion calculated using Equation 2

(3) Loss Nitrogen Calculated Using Equation 3

Experiment 3 Analysis of the Final Product

FIG. 29 shows the conditions of Experiment 2 (lime loading 0075 g Ca (OH) 2 / g dry residue, temperature 75 ° C., residue 40 g dry residue / L and time

Shows amino acid spectra for two centrifuged liquid samples obtained under 1 h).

The amino acid composition in the centrifuged liquid sample was determined by HPLC analysis. Second, the centrifuged liquid sample was treated with 6-N HCl.

Treated for 1 h, which hydrolyzed the protein to its corresponding amino acid. By comparing the two results,

It can be concluded that hydrolysis is to individual amino acids; The results in both cases are essentially the same

Figure 30 compares the amino acid spectra for raw residues and solid residues remaining after lime treatment.

Was dried at 105 ° C. for 24 h and samples were taken for protein determination. The water content of the solid residue was about 80%,

The defined protein originates from the liquid and solid phases. The amino acid content in the residual solids causes the amino acids to dissolve into the liquid phase.

Much less than raw waste

Using the mass balance and the data shown in Figure V6, the amount of each amino acid "extracted" from the raw materials is 50% to 75%.

B, which includes the protein in the liquid attached to the solid. Subtracting the protein dissolved in the attached liquid, the extraction for each amino acid

52% to 76% of the crude protein, which is similar to the result obtained in Experiment 2

Another important problem is to determine the decomposition of the individual amino acids at the reactor operating conditions.

It is necessary to obtain the amino acid concentration in time. FIG. 31 shows the amino acid present in the liquid phase centrifuged at 30 min at 2 h.

Almost identical to; This means that amino acids are stable under operating conditions. FIG. 32 shows residues of different starting concentrations.

Using; Again, it shows that amino acids are the same concentration at 30 min and 2 h

Patent Publication 10-2007-0020094

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33 compares the results of three different initial debris concentrations for the same time, temperature and lime loading. These results are centrifuged.

Amino acid concentration in the liquid phase, as expected, is higher for raw materials with higher initial concentrations

Figure 34 examines amino acid concentration as a function of time during the first 10 minutes of the reaction. The concentration is stable for all amino acids after 10 minutes.

And the 30-min value is also similar because as concluded in Experiment 1, the reaction occurs during the first 10-30 min of contact.

it means

From experiments conducted using HPLC and Kjeldahl methods, the nitrogen content was similar in both cases (see Table 53).

Means that the main contribution to the total nitrogen content is amino acids (i.e. protein content of chicken grounds)

Table 53

Comparison of results for nitrogen content (g nitrogen / 100 g liquid sample) using HPLC and Kjeldahl methods for experiments in FIG. V10

2 min 3 min 5 min 10 min

HPLC 0065 0072 0211 0216

Kjeldahl 011 011 018 017

Table 54 compares the various requirements for essential amino acids to the needs of the various livestock shown in Table 55.

The composition of the phosphorus animal feed is shown and can also be compared to Table 54.

TABLE 54

(a) at 75 ° C., 0075 g Ca (OH) 2 / g dry residue, 60 g dry residue / L and 30 min; And (b) 0100 g Ca (OH) 2 / g at 50 ° C

Amino acids present in the liquid phase of the two experiments, crude residue, 40 g dry residue / L and 90 min, were compared to the dietary requirements of different animals.

School

Amino Acid Catfish Dog Cat Chicken Pig

Solubilization residue

(a)

Solubilization residue

(b)

ASN 214 082

ASP 362 636

GLU 1056 870

SER 454 721

HIS 131 100 103 140 125 292 223

GLY 489 535

THR 175 264 243 350 250 574 647

ALA 847 666

ARG 375 282 417 550 000 795 522

VAL 263 218 207 415 267 753 660

CYS 200 * 241 * 367 * 400 * 192 * 07 ND

MET 200 * 241 * 207 225 192 * 383 423

TYR 438+ 405+ 293+ 585+ 375+ 168 436

PHE 438+ 405+ 140 315 375+ 542 465

ILE 228 205 173 365 250 636 519

LEU 306 327 417 525 250 1091 937

LYS 447 350 400 575 358 327 742

TRP 044 091 083 105 075 226 ND

PRO 611 698

* Cysteine + Methionine + Tyrosine + Phenylalanine ND Not measured

Values are expressed in g individual amino acids / 100 g total amino acids

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TABLE 55

Nutritional Needs for Livestock During Growth (Ponder, 1995)

Catfish dog cat

chicken

Broiler

pig

Crude protein (%) 320 220 300 200 120

Arginine (%) 120 062 125 110 000

Methionine (%) 064 * 053 * 062 045 023 *

Cysteine (%) 064 * 053 * 110 * 080 * 023 *

Histidine (%) 042 022 031 028 015

Isoleucine (%) 073 045 052 073 030

Leucine (%) 098 072 125 105 030

Lysine (%) 143 077 120 115 043

Tyrosine (%) 140 ** 089 ** 088 ** 117 ** 045 **

Phenylalanine (%) 140 ** 089 ** 042 063 045 **

Threonine (%) 056 058 073 070 030

Tryptophan (%) 014 020 025 021 009

Valine (%) 084 048 062 083 032

Note: 1) * cysteine + methionine 2) ** tyrosine + phenylalanine

3) All values are expressed as a percentage of the feed (g / 100 g feed)

TABLE 56

Composition of Different Feeds Used in Animal Feed (Pond et al, 1995)

Blood meal Fish meal ** Soy flour Gluten flour Corn flour Milo

Sex and bone

minute

Feather

Dry Matter (%) 910 920 89 910 930 890 94 910

Crude fiber (%) 10 09 60 40 120 20 24 47

Crude protein (%) 799 612 458 429 180 110 509 854

Digestibility (%) * 623 564 417 357 148 78 450 602

Arginine (%) 350 374 320 140 120 036 305 533

Cysteine (%) 140 058 067 060 032 018 046 321

Glycine (%) 340-210 150-040--

Histidine (%) 420 144 110 100-027 096 047

Isoleucine (%) 100 285 250 230-053 147 351

Leucine (%) 1030 448 340 760 170 142 302 042

Lysine (%) 690 474 290 080 090 027 289 167

Methionine (%) 090 175 060 100 035 009 008 054

Phenylalanine (%) 610 246 220 290 080 045 165 359

Threonine (%) 370 251 170 140 090 027 160 363

Tryptophan (%) 110 065 060 020 030 009 028 052

Tyrosine (%) 180 193 140 100 150 036 079 235

Valine (%) 650 319 240 220 130 053 214 585

Note: 1) * as-fed basis for ruminants 2) ** There are three types of fishmeal: anchovies, menhaden and herring

Values given for fish 3) The value of amino acid is the feed basal percentage (g amino acid / 100 g food)

Patent Document 10-2007-0020094-40

The tabulated results indicate that the solubilized protein meets or exceeds the essential amino acid requirements of the animals during the growth phase for run at 50 ° C.

On the other hand, the values for tyrosine and lysine at 75 ° C. (optimal conversion conditions) are lower than required

Chicken grounds containing 15% protein (wet basis) or 45% protein (dry basis) are treated with Ca (OH) 2 at a temperature below 100 ° C.

Can be used to obtain amino acid-rich products.

Can be used

For all conditions of temperature studied, lime loading and residue concentration, no significant change occurred in conversion after 30 minutes of reaction.

To maximize protein conversion (up to 80%), the optimum conditions are 0075 g Ca (OH) 2 / g dried at 75 ° C. for at least 15 min.

The initial residue concentration does not have a significant effect on the conversion or amino acid spectrum of the product.

However, in order to obtain a highly concentrated product, to reduce the energy requirements for concentrating the final product,

Degree is recommended

Almost no amino acid degradation was observed for all experiments performed below 100 ° C. and up to 2 hours.

Evaporation at near temperatures will result in little decomposition

At 50 ° C., the spectrum of essential amino acids obtained meets or exceeds the requirements for many livestock during the growing season.

The amino acid rich solid product obtained by lime treatment of residues can serve as protein supplement for these animals

The product obtained at 75 ° C. has less lysine and tyrosine than required and therefore will not be efficient

Example 5: Protein Solubilization in Chicken Ground and Feathers

Disposal of animal organs by slaughter is an important environmental problem. Poultry farming has developed technologies to treat these wastes.

Generates a large amount of waste (waste, feathers, and blood) that is concentrated in the slaughterhouse in a volume large enough during the period.

Can be processed into liquid fractions (heat-dried blood used as feed supplement), hydrolyzed feather powders, poultry dry powders and fats

5% of poultry weight is feathers Due to their high protein content (897% of dry weight, Table 57), feathers are a potential protein for food

Although a vaginal source, complete destruction of hard keratin structures is required (Dalev, 1994)

Table 57

Composition of poultry ground and chicken feathers (Wisman et al, 1957, and Dalev, 1994)

% Gross Weight New Ground Dry Matter Feather (Dry Matter)

Moisture 695--

Crude protein 172 565 897

Ether Extract (Fat) 80 262 14

Crude fiber 01 04 ND

Ash 37 121 63

Nitrogen free extract 15 48 ND

Calcium (Ca) 05 17 035

Inn (P) 06 20 013

Sodium (Na) ND-04

Potassium (K) ND-09

Poultry ground contains much more histidine, isoleucine, lysine and methionine than chicken feathers (characterizing chicken ground and feathers

Thus, poultry residue and feather meal will have better balanced amino acids together (EI

Boushy and Van der Poel, 1994) Feather / Drag processes are more difficult than feathers to degrade or hydrolyze.

I can adjust it

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Table 58

Amount of surviving microorganisms in poultry waste (Acker et al, 1959)

Unwashed washed used agar

Total Aerobic Bacteria 280000 90000 Trypticase Soi

Total Anaerobic Bacteria 98000 28000 Linden Thioglycolate

Spore-forming anaerobes

Clostridium botulinum

(Clostridium botulinum))

4500 2000 Linden Thioglycolate

Coliform Group (Salmonella) 20000 9000

Violet Red Vail

(Violet red bile)

Lactobacilli 270000 97000 Tomato juice

Yeast 28000 26000 Litman oxgall

Courtney fungi <100 <100 Litman Oxgal

Modulus / g wet weight

Table 59

Composition of Poultry Waste (Acker et al, 1959)

Unwashed Washed Unit

Crude Protein 205 177 g / 100 g Wet Substance

Digestive Protein 912 915 g / 100 g Protein

Ether Extract 84 76 g / 100 g Wet Substance

Crude fiber 11 10 g / 100 g Wet Substance

Moisture 685 721 g / 100 g Wet Substance

Ash 40 43 g / 100 g Wet Substance

Loss on combustion 275 235 g / 100 g Dry matter

Calcium 14 18 g / 100 g Wet Substance

Phosphorus 11 13 g / 100 g Wet Substance

Riboflavin 38 31 mg / 100 g Dry Substance

Niacin 48 63 mg / 100 g Dry Substance

Ca pantothenate 23 11 mg / 100 g dry matter

Pyrodoxin 011 009 mg / 100 g dry matter

B12 526 95 μg / 100 g dry matter

Vitamin A 8068 11639 USP Unit / 100 g Dry Substance

Carotene 3562 6568 international unit / 100 g dry matter

Total Vitamin A 11630 18207 International Units / 100 g Dry Substance

Total Vitamin C 479 269 mg / 100 g Dry Substance

Vitamin E 34 77 International Units / 100 g Dry Substance

Inositol 2181 1315 mg / 100 g dry matter

Thiamine 013 007 mg / 100 g dry matter

Folic Acid 011 004 mg / 100 g Dry Substance

Arginine 66 71 g / 100 g Protein

Histidine 12 14 g / 100 g protein

Isoleucine 105 110 g / 100 g protein

Leucine 89 100 g / 100 g Protein

Lysine 133 136 g / 100 g Protein

Methionine 27 28 g / 100 g Protein

Phenylalanine 55 50 g / 100 g Protein

Threonine 25 32 g / 100 g Protein

Tryptophan 09 07 g / 100 g Protein

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Valine 29 34 g / 100 g Protein

The addition of excreta to animal feed can adversely affect growth (Acker et al, 1959) and also public health considerations.

Because of this, residues used for feeding can be treated to reduce bacterial loads (Table 58).

Content (calcium and phosphorus) and vitamins are present (Table 59) Poultry residue is important for vitamins, minerals, and possibly unidentified growth factors

Seems to be a source (Acker et al, 1959)

One way of dealing with poultry by-products is by rendering, which includes five phases:

Raw material storage

Cooking and drying (sterilization)

·congelation

Fat extraction

Powder handling

Poultry blood, feathers and debris, slaughter waste, and dead birds reach the cooker in different ways. Hydrolysis and sterilization

The material takes place in a reactor where it is heated for a given time at an established temperature and pressure. The material then preserves the quality of the product.

In order to dry at the lowest possible temperature, condensation of water vapor is required according to environmental regulations.

Finally, the product prepared in this way may have a fat content of greater than 16%; Thus, lower than 10-12%

Fat extraction (eg lard discharge through the perforated bottom to the adjacent tank) is required to ensure fat content.

The released fat can be used as an additive to feed and for other purposes (El Boushy and Van der Poel, 1994).

Sterilization takes place during cooking Drying is achieved in separate dryers Two different kinds of dryers have been used: discs

Dryers and flash dryers Flash dryers provide smaller floor space, heating by oil or gas, and high quality final

Most commonly used because of the same benefits as products (El Boushy and Van der Poel, 1994)

The rendering process can be used to treat different wastes or produce different products as follows:

Feathers using only chicken feathers (FM)

Poultry by-product powder or dregs powder from dregs (intestines, head, feet and blood)

Mixed poultry by-product powder (PBM) from a mixture of poultry ground and chicken feathers

Compositions and nutritional values for feather powder and poultry by-product flour using different treatment conditions are shown in Tables 60-63.

TABLE 60

Composition of Poultry By-Product Powder

% Gross weight New dry matter

Moisture 61-

Crude protein 546 581

Ether extract 149 159

Crude fiber 08 09

Ash 170 181

Nitrogen free extract 66 70

Calcium 80 85

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Phosphorus 30 32

TABLE 61

Residue Powder Composition Using Rendering Process in Different Industrial Plants (McNaughton et al, 1977)

Factory 1 Factory 2 Factory 3

Crude protein 5399 5310 5401

Crude fat 2534 2520 2470

Ash 552 596 606

Moisture 1115 1101 998

Crude fiber 400 473 525

Calcium 146 165 178

Inn 100 108 110

Value is a percentage of gross weight

Table 62

Amino Acid Content of Food from Different Poultry Waste Processes (El Boushy and Van der Poel, 1994)

Amino acid FM (batch) FM (continuous) PBM (batch) PBM (continuous)

ASP 590 575 520 517

THR 405 435 240 233

SER 750 925 270 270

GLU 1010 1035 983 970

PRO 955 885 643 650

GLY 675 685 787 740

ALA 535 475 443 493

VAL 540 580 287 303

CYS 260 300 063 060

MET 050 040 107 143

ILE 415 425 223 230

LEU 700 725 420 437

TYR 235 240 180 200

PHE 430 410 240 253

LYS 180 190 370 380

HIS 060 055 110 120

ARG 665 660 477 477

Crude protein 8455 8640 6363 6476

FM Feather Powder (Batch) 30-60 min, 207-690 kPa, ~ 150 ℃

(Continuous) 6-15 min, 483-690 kPa, ~ 150 ℃

PBM poultry by-product flour (blood, feathers and dregs),

Batch or continuous, 30-40 min, 380 kPa, 142 ° C

TABLE 63

Amino Acid Content and Availability of Different Poultry Wastes (El Boushy and Van der Poel, 1994)

FM Usability PBM Usability

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ASP 502 56 546 67

GLU 796 62 800 77

SER 673 64 609 81

HIS 055 59 108 72

GLY 447-659-

THR 336 62 322 76

ALA 485 78 435 78

ARG 544 77 545 84

TYR 223 65 252 77

VAL 641 75 481 77

MET 079 65 114 77

PHE 389 77 363 79

ILE 415 78 325 79

LEU 619 73 578 78

LYS 157 64 281 77

PRO 939 71 613 77

CYS 426 65 243 62

Fecal flour contains about 85% crude protein; It is rich in cysteine, threonine, and arginine, but methionine, lysine, histidine

And lack of tryptophan (El Boushy and Roodbeen, 1980).

The addition will improve the quality of the product. At high pressures, chicken feathers tend to "gum" and thus have a non-free flow

Provide flour

Leftovers and feathers were obtained from Texas A & M Fowltree Science Department. Used leftovers are bones, heads, beaks and feet.

And intestines (eg, heart, lung, intestine, liver). The residue is blended in an industrial blender for 10 minutes and collected in a plastic bottle.

And finally frozen at −4 ° C. for later use with a sample of the blended material using the water content, total nitrogen (protein

Estimates of vaginal fractions) and amino acid content were obtained to characterize the starting material. The feathers were washed several times with water and air at ambient temperature.

Dried, dried at 105 ° C. and finally ground using a Thomas-Willy experimental mill (Arthur H. Thomas Company)

Sieved through 40-mesh screen

Experiments were carried out in two autoclave reactors (12-L and 1-L) with a thermostat and mixer powered by a variable speed motor.

The conditions studied were established from previous experiments using both chicken feathers and chicken grounds.

Concentration (dry debris + feathers / L), calcium hydroxide loading (g Ca (OH) 2 / g dry debris + feathers) and time.

The liquid phase was taken from the reactor and then centrifuged to separate the liquid phase from the residual solid material.

A group of steps was followed to collect data for different intermediate products for the process shown in FIG. 35.

Raw residue was 334% dry matter and 666% moisture The crude protein concentration of dry residue was -34% (TKN 540% residue),

Ash content is -10%; The remaining 56% were fiber and fat Amino Acid Analysis of Solid Raw Waste (Table 64)

Shows good balance against total protein content from amino acid analysis is 26 g protein / 100 g dry residue (Table 65)

Some of the amino acids were destroyed during the acid hydrolysis used in the HPLC determination, and the Kjeldahl (TKN) value was found to approximate the protein content.

In this regard, these two values are similar.

Table 64

Amino Acid Analysis for Dry Raw Waste

Amino Acid Concentration (mg / L)

percentage

(g amino acid / 100 g protein)

ASP 29565 9900

GLU 50559 16930

SER 12453 4170

HIS 5826 1951

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GLY 22557 7553

THR 12409 4155

ALA 20943 7013

ARG 22753 7619

TYR 10015 3354

VAL 15172 5080

MET 6894 2309

PHE 13456 4506

ILE 13100 4387

LEU 28257 9462

LYS 20266 6786

PRO 14409 4825

TABLE 65

Determination of Amino Acid Content for Dry Raw Waste Samples

Variable value

Total Amino Acid Concentration (mg / L) 29863

Total mass of amino acids in solid samples (mg) 2389

Mass of solid sample for analysis (mg) 92

Proportion of Amino Acids in Dry Samples 26

Chicken feathers were 92% dry matter and 8% moisture The crude protein concentration of dry feathers was about 957% (feather TKN 153%); Remainder

43% were fiber and ash

Experiment 1 Complete Residue Hydrolysis

Experiment 1 consisted of a sample using only the gut (chapter V) previously performed for protein solubilization of a complete debris sample (bone, head, beak, foot and gut).

The conditions used in Experiment 1 were 75 ° C., 010 g lime / g residue and 40 g dry residue / L.

Summarized variables in Table 66

TABLE 66

Experimental conditions and measured parameters to determine protein solubilization of debris samples with bones, heads, beaks, feet and intestines

Variable value

Temperature (℃) 75

Mass of Ca (OH) 2 (g) 35

Mass of residue (g) 1021

Volume of Water (mL) 850

Lime loading (g Ca (OH) 2 / g dry residue) 0103

Dry residue concentration (g Dry residue / L) 4005

Residual Solid (g) 142

Table 67 shows the total nitrogen content in the centrifuged liquid sample as a function of time for this experiment.

Based on TKN (540%), proteolytic conversion was estimated and shown in Table 68.

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TABLE 67

Protein and mineral content of the product after raw waste and lime hydrolysis

Condition

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

Dry residue 53995 06269 09181 03845 00622 3150 59 493 46 10

Liquid 30 min 01189 00041 00311 00539 0001 104 0 11 0 0

Liquid 90 min (*) 01 925 00187 00321 02 00031 104 2 9 2 0

Liquid 90 min 01 145 00041 00311 00487 0001 104 0 3 0 0

Dry Residual Solid 25867 05606 01005 41793 01078 560 97 187 58 15

(*) Uncentrifuged samples

TABLE 68

Total TKN Conversion to Availability TKN

Sample conversion

Centrifuged Liquid 30 min 594

Uncentrifuged liquid 90 min 962

Centrifuged Liquid 90 min 572

Under the conditions studied, the conversion of nitrogen in the solid phase to the liquid phase was 60% efficient.

Although lower than that obtained for this, this can be explained by the presence of bones, heads, beaks and feet that did not exist before.

Powder contains higher proportions of ash, minerals, and insoluble ingredients that reduce the efficiency of the hydrolysis process.

unchanged from min to 90 min (Table 68), similar to previous results; To avoid possible degradation of heat-sensitive amino acids

30 min is the recommended time No significant loss of nitrogen occurred during hydrolysis (962% for samples not centrifuged)

Is explained)

A significant reduction (about 50%) of protein in the solid was achieved, from 337% in raw waste to 162% in residual solids after lime treatment

(Similar to the 133% value obtained from the analysis of amino acids, Table 69) .Amino acids and other solubles present in the raw waste

The solubilization of minutes results in a 58% reduction in the weight of the dry solids. The residual solids are stable without severe odors and may be

Have a well balanced amino acid content (Table 70) that meets or exceeds essential amino acid requirements

TABLE 69

Determination of Amino Acid Content on Residual Solids After Lime Treatment

Variable value

Total Amino Acid Concentration (mg / L) 18050

Total mass of amino acids in solid samples (mg) 1354

Mass of solid sample for analysis (mg) 102

Proportion of Amino Acids in Dry Samples 1327

TABLE 70

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Amino Acid Analysis for Residual Solids After Lime Treatment

Amino Acid Concentration (mg / L)

percentage

(g amino acid / 100 g protein)

ASP 19289 10686

GLU 25776 14280

SER 8512 4716

HIS 4314 2390

GLY 9178 5085

THR 8314 4606

ALA 10392 5757

ARG 12771 7075

TYR 7805 4324

VAL 10546 5843

MET 4967 2752

PHE 10376 5749

ILE 9545 5288

LEU 20762 11502

LYS 9858 5462

PRO 8096 4485

Treatment of chicken residues with lime allows hydrolysis of existing proteins into small peptides and free amino acids that are soluble in water.

Thus, 60% TKN conversion from solid to liquid phase shows the efficiency of protein recovery in the liquid phase.

Show amino acid balance for deeply separated liquids

TABLE 71

Amino Acid Analysis on Centrifuged Liquid Samples (3 min)

amino acid

density

(mg / L)

percentage

(g amino acid / 100 g protein)

ASP 69983 3530

GLU 129448 6529

ASN 3937 0199

SER 98378 4962

GLN 26346 1329

HIS 25379 1280

GLY 69551 3508

THR 73033 3684

CIT 54309 2739

B-ALA 4170 0210

ALA 147275 7428

TAU 200813 10129

ARG 162465 8195

TYR 93992 4741

CYS-CYS 102601 5175

VAL 80385 4055

MET 51049 2575

TRP 36910 1862

PHE 86256 4351

ILE 74689 3767

LEU 179141 9036

LYS 136399 6880

PRO 76073 3837

Total Amino Acid Concentration 19826 mg / L

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Comparing the amino acid content of the raw debris, centrifuged liquid product, and residual solids (Table 72) in the centrifuged liquid and residual solids

Shows that the amino acid content of is similar to that of the raw waste, so that solubilization of all amino acids occurs in a similar ratio,

Means little destruction of specific amino acids

TABLE 72

Comparison of Amino Acid Contents for Different Substances During Lime Treatment of Chicken Ground

Amino Acid Residue Solid Solid Centrifuged Liquid *

ASP 990 1069 450

GLU 1693 1428 833

SER 4,17 472 633

HIS 195 239 163

GLY 755 508 448

THR 416 461 470

ALA 701 576 948

ARG 762 708 1046

TYR 335 432 605

VAL 508 584 517

MET 231 275 329

PHE 451 575 555

ILE 439 529 481

LEU 946 1150 1153

LYS 679 546 878

PRO 483 449 490

* Consider only amino acids present in solid assays

Treating chicken residues with lime at medium temperature and time reduces the amount of microorganisms present in the liquid phase.

Rapid evaporation of liquids is essential as it contains all the nutritional requirements for the intestines

Amino acid analysis of the sample (Table 73) again balances very well to meet or exceed the essential amino acid requirements of the animal during the growth phase.

Shows slightly lower values for histidine

TABLE 73

Amino acid analysis of raw materials and products compared to essential amino acid requirements for various livestock (whole residue)

Amino Acid Catfish Dog Cat Chicken Pig

Centrifuged

Liquid

solid

leftover

Residual solid

ASN 020

GLN 133

ASP 353 990 1069

GLU 653 1693 1428

SER 496 417 472

HIS 131 100 103 140 125 128 195 239

GLY 351 755 508

THR 175 264 243 350 250 368 416 461

ALA 743 701 576

ARG 375 282 417 550 000 819 762 708

VAL 263 218 207 415 267 405 508 584

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CYS 200+ 241+ 367+ 400+ 192+ 518 ND ND

MET 200+ 241+ 207 225 192+ 257 231 275

TYR 438 * 405 * 293 * 585 * 375 * 474 335 432

PHE 438 * 405 * 140 315 375 * 435 451 575

ILE 228 205 173 365 250 377 439 529

LEU 306 327 417 525 250 904 946 1150

LYS 447 350 400 575 358 688 679 546

TRIP 044 091 083 105 075 186 ND ND

PRO 384 483 449

* Cysteine + Methionine + Tyrosine + Phenylalanine ND Not measured

Values are expressed in g individual amino acids / 100 g total amino acids

Experiment 2 Waste and Feather Treatment

Chicken feathers and grounds have different compositions and their main components behave differently during proteolysis using lime

Tin proteins are more difficult to hydrolyze than proteins in debris and require longer times or higher temperatures and lime concentrations.

Residual waste from the slaughterhouse often contains a mixture of debris and feathers, which can lead to protein-rich

It is possible to obtain the sacred product. Two products can be produced: capable of meeting the amino acid requirements for various unit livestock.

One with well-balanced amino acid content (from waste), and one for ruminant (from feathers)

Hydrolysis of the chicken feather / flour mixture was studied using the process shown in FIG. 35. Initial treatment of the mixture was mainly in the dregs.

Hydrolysis of the protein present in the product to obtain a liquid product and a residual solid.

Precipitate CaCO3 (which can be converted back to lime) and reduce the concentration of Ca in the liquid phase.

Obtain a First Solid Amino Acid Rich Product

Residual solids in one phase may be further treated with lime for a longer time (different conditions) to promote hydrolysis of chicken feather proteins.

The second product will be followed by a similar step to the one phase which is returned to the reactor.

Condition 1 was used in Experiments A1, B1 and C1, while Condition 2 was used in Experiments A2, B2 and C2.

The experimental conditions and measured parameters studied during Experiment 2 are summarized in Table 74.

The ratio of 175 g wet residue / 7 g wet feather was used as the normal value in the waste products of the slaughterhouse.

TABLE 74

Experimental Conditions and Measured Parameters for Determining Protein Solubilization of the Leftover / Feather Mixture

Variable Experiment A1 Experiment A2 Experiment B1 Experiment B2 Experiment C1 Experiment C2

Temperature (℃) 50 75 75 75 75 100

Mass of Ca (OH) 2 36 414 207 207 48 27

Mass of residue (g) 685 343 913

Mass of feathers (g) 274 410 137 2118 365 487

Water volume (mL) 6000 3000 3000 2000 800 800

Ca (OH) 2 (g / d dry residue) 0075 0101 0086 0098 0075 0055

Dry substance (g / L) 8008 13653 8013 10579 8002 6081

Dry residue (g / L) 3806 3812 3805

Total TKN (g) 5094 2548 679

TKN (%) 1060 1060 1060

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Table 75 shows the total nitrogen content in the centrifuged liquid sample as a function of time for this experiment. Dry Ground (540%) and Chicken

Mean TKN for feathers (153%) gave a mixture initial TKN of 106%. Proteolytic conversion was estimated and Table 76.

And Table 77. Table 76 considers the conversions relating to debris (condition 1) and secondly to feather (condition 2), while Table 77 shows

Provides a conversion for the initial TKN of the composite. Under the conditions studied, the highest conversion of nitrogen in the solid phase to the liquid phase was 60%.

Table 75

Total Kjeldahl nitrogen content in the centrifuged liquid phase as a function of time for Experiment 2 (Drag / Feather Mixture)

Time (min) Experiment A1 Experiment A2 Experiment B1 Experiment B2 Experiment C1 Experiment C2

5 01 126 01015 --- 01 183 --- ---

10 01 210 --- 01 109 --- --- ---

15 01154 00973 01238 01262 --- ---

30 01182 01126 01182 01431 --- ---

60 --- 01514 01349 01723 02300 ---

120 --- 02188 --- 02299 --- 02600

TKN is g nitrogen / 100 g liquid sample

Table 76

Conversion of total TKN to soluble TKN for Experiment 2, with respect to debris (A1, B1 and C1) and feather (A2, B2 and C2) TKN, respectively

Time (min) Experiment A1 Experiment A2 Experiment B1 Experiment B2 Experiment C1 Experiment C2

5 592 79 --- 123 --- ---

10 636 --- 582 --- --- ---

15 606 76 649 131 --- ---

30 621 87 620 148 --- ---

60 --- 118 708 179 1209 ---

120 --- 170 --- 238 --- 269

Table 77

Conversion of Total TKN to Soluble TKN for Experiment 2 (Drag / Feather Mixture)

Time (min) Experiment A1 Experiment A2 Experiment B1 Experiment B2 Experiment C1 Experiment C2

5 143 60 --- 93 --- ---

10 154 --- 141 --- --- ---

15 147 57 157 99 --- ---

30 150 66 150 112 --- ---

60 --- 89 172 135 293 ---

120 --- 129 --- 180 --- 307

Total 279 352 60

Based on the data in Table 76, no significant effect on conversion occurred when the temperature was changed from 50 to 75 ° C. Experiments A1 and B1.

The results from show a higher conversion at 60 min compared to 30 min; This means that keratin proteins hydrolyze more slowly

It is expected to continue to react during contact with lime. In addition, comparing Table 68 and Table 76,

Similar results are obtained for the conversion as for the residue alone; Therefore, the residue present in the mixture is only residue

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Hydrolyze at the same rate at the temperatures studied in Experiments A1 and B1, the hydrolysis of chicken feathers is relatively slow compared to the residue

Multiprotein hydrolysis is significantly increased by changing the temperature from 75 to 100 ° C. for Condition 1 (Experiment C1).

This is explained by the larger conversion expected for chicken feathers under these conditions (60% for chicken feather hydrolysis at 2 h) (Chang

and Holtzapple, 1999)

The results from experiments A2 and B2 indicate that the initial "pretreatment" of chicken feathers in a mixture with chicken residues results in a hydrolysis conversion to feathers (at 17%).

Slightly higher) and higher temperatures or longer times may be required to fully hydrolyze the chicken feathers.

Results from experiment C2 show higher conversion at 100 ° C. compared to 75 ° C. [Chang and

From Holtzapple's study, much higher temperatures or longer reaction times may be used to further increase proteolysis.

Can

Table 78-80 shows the total nitrogen and mineral content of the samples from different stages of the lime treatment process of the residue / feather mixture.

A slight decrease in the calcium content (8%) is obtained after bubbling the liquid with CO 2 until a pH ˜6 is achieved.

Achievement by Dead Reduction (Table 78) The results show that calcium precipitation with CO2 is a very inefficient process for the studied conditions.

Shows

Table 78

Protein and mineral content of the product after lime hydrolysis for experiments A1 & A2

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

With solid (30 min) 04257

Liquid 1 (30 min) 01 182 00093 00404 00746 0001 259 0 3 1 0

After bubbling 01098 00083 00352 00684 0 207 0 2 1 0

With solid (2h) 05420

Liquid 2 (2 h) 02 188 00041 00197 01523 0 155 1 6 1 0

After bubbling 02 108 00031 00176 01503 0 145 1 2 1 0

Residual Solid 1 90 254 0571 03 119 40 974 00756 3264 104 210 35 13

Residual Solid 2 79 002 02974 01492 56684 01109 2694 104 301 31 16

Table 79

Protein and mineral content of the product after lime hydrolysis for experiments B1 & B2

TKN (%) P (%) K (%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

With solid

(60 min)

04257

Liquid 1 (60 min) 01349 00104 00383 00984 0001 259 1 5 1 0

With solid

(2 h)

05926

Liquid 2 (2 h) 02299 00031 00166 01668 0 135 1 2 1 0

Residual Solid 1 87163

Residual Solid 2 80 355 0313 00705 59482 00839 2518 77 166 20 9

TABLE 80

Protein and mineral content of the product after lime hydrolysis for experiments C1 & C2

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TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

Liquid 1 (60 min) 023 0 004 01 0 228 2 1 0 0

Liquid 2 (2 h) 026 0 001 014 0 83 1 1 0 0

Residual Solid 1 1279 03 032 292 005 1617 73 152 19 5

Residual Solid 2 977 053 009 429 009 819 95 269 24 9

Final Product 1171 012 055 517 001 2912 38 21 11 8

Table 79 shows that, after the second lime treatment, the protein content in the solid is from 106% (TKN) in the raw mixture to 79% (TKN) in the final residual solid.

In addition, the total dry weight (soluble material) is reduced by about 35%.

Calcium is relatively high (~ 6% in all cases), and the amino acid content in some of the amino acids required for animal growth

Poor; Similar to the residue obtained for chicken feathers alone

Since calcium concentration is high in residual solid # 1, in all cases, less lime is similar for proteolytic conversion.

Can be added to the second lime treatment with one result

The concentrations of all minerals are compared for all cases studied (Table 78-80). The nitrogen content in the centrifuged liquids # 1 and # 2 is the highest.

Increase in temperature The mineral content (phosphorus, potassium and sodium) is increased in liquid # 1 as more salts are solubilized with temperature and time.

Decreases with liquid # 2

Tables 81-83 show amino acid contents for different liquid products obtained under the conditions studied. For Experiments A2 and B2, the samples

Hydrolyzed with HCl for 24 h, followed by amino acid analysis to determine total amino acid concentration from chicken feather hydrolysis.

In experiment C2, hydrolysis was not performed for comparison purposes

TABLE 81

Amino Acid Analysis on Liquid Samples Centrifuged in Experiments A1 and A2

Experiment A1 Experiment A2

amino acid

density

(mg / L)

percentage

(g amino acid /

100 g protein)

density

(mg / L)

percentage

(g amino acid /

100 g protein)

ASP 20570 512 41220 750

GLU 45438 1130 64967 1181

ASN 992 025 4051 074

SER 23514 585 35129 639

GLN 000 000 000 000

HIS 5093 127 000 000

GLY 17000 423 36521 664

THR 14934 372 13127 239

CIT 5303 132 9938 181

B-ALA 644 016 472 009

ALA 27672 688 44372 807

TAU 38912 968 10669 194

ARG 29898 744 25601 466

TYR 17899 445 37828 688

CYS-CYS 10961 273 12771 232

VAL 16471 410 49055 892

MET 11056 275 9993 182

TRP 6881 171 4619 084

PHE 16255 404 23689 431

ILE 14170 352 33424 608

LEU 35104 873 57880 1053

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LYS 30546 760 28356 516

PRO 12691 316 6232 113

Total Concentration 402004 549914

Table 82

Amino Acid Analysis on Liquid Samples Centrifuged in Experiments B1 and B2

Experiment B1 Experiment B2

amino acid

density

(mg / L)

percentage

(g amino acid /

100 g protein)

density

(mg / L)

percentage

(g amino acid /

100 g protein)

ASP 20838 488 60653 823

GLU 45589 1069 78825 1070

ASN 939 022 000 000

SER 24538 575 94375 1281

GLN 2055 048 000 000

HIS 5198 122 000 000

GLY 19449 456 95665 1298

THR 16133 378 16624 226

CIT 6751 158 000 000

B-ALA 957 022 000 000

ALA 30078 705 38708 525

TAU 39107 917 000 000

ARG 32920 772 54622 741

TYR 20469 480 27413 372

CYS-CYS 7444 174 000 000

VAL 17131 402 40103 544

MET 11850 278 10284 140

TRP 4172 098 000 000

PHE 16173 379 37028 503

ILE 13892 326 33031 448

LEU 36399 853 68405 928

LYS 34567 810 10663 145

PRO 19960 468 70417 956

Total concentration 426610 736815

TABLE 83

Amino Acid Analysis on Liquid Samples Centrifuged in Experiments C1 and C2

Experiment C1 Experiment C2

amino acid

density

m / L

percentage

(g amino acid /

100 g protein)

density

m / L

percentage

(g amino acid /

100 g protein)

ASP 28042 481 7339 695

GLU 67571 1159 14871 1408

ASN 1489 026 088 008

SER 24452 420 9968 944

GLN 000 000 000 000

HIS 8050 138 000 000

GLY 24911 427 9198 871

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THR 22713 390 641 061

CIT 23891 410 7504 710

B-ALA 661 011 000 000

ALA 43812 752 10695 1012

TAU 19922 342 2259 214

ARG 26288 451 3932 372

TYR 9779 168 1370 130

CYS-CYS 18 157 312 4773 452

VAL 29399 504 5611 531

MET 14891 255 1441 136

TRP 11375 195 000 000

PHE 25851 444 4800 454

ILE 27012 463 5445 515

LEU 59913 1028 10736 1016

LYS 40843 701 2554 242

PRO 53785 923 2420 229

Total Concentration 582807 105646

From Tables 81-83, a comparison of the results from Experiments A1, B1 and C1 shows similar amino acid content for all cases; Ta

Thus, the effect of temperature on the rate of hydrolysis is similar for different individual amino acids.

(100 ° C. vs. 75 ° C., Table 76 and Table 77), does not affect amino acid content in lime treatment of chicken feather / waste mixture

Comparing experiments A1, B1 and C1 with the amino acid content for chicken residues alone (Table 71), similar results are obtained in all cases chickens

The amino acid content and protein hydrolysis of the residues are not affected by the presence of chicken feathers in the mixture, and hydrolysis of these feathers

Is relatively low in the studied conditions. Proline increases for higher temperatures may be associated with connective tissue and

Can be explained by the hydrolysis of bone (in debris)

Comparing the results from experiments A2, B2 and C2 shows a greater difference in amino acid content than experiments A1, B1 and C1

For these temperatures different amounts of unhydrolyzed residue left in residual solid # 1 may account for these differences.

Table 84 and Table 85 compare essential amino acid requirements of various livestock with different products.

TABLE 84

Amino Acid Analysis of Raw Materials and Products Compared to Essential Amino Acid Requirements for Various Livestock (Fragment / Feather Mixture Condition 1)

Amino Acid Catfish Dog Cat Chicken Pig Experiment A1 Experiment B1 Experiment C1

ASN 025 022 026

GLN 000 048 000

ASP 512 488 481

GLU 1130 1069 1159

SER 585 575 420

HIS 131 100 103 140 125 127 122 138

GLY 423 456 427

THR 175 264 243 350 250 372 378 390

ALA 688 705 752

ARG 375 282 417 550 000 744 772 451

VAL 263 218 207 415 267 410 402 504

CYS 200+ 241+ 367+ 400+ 192+ 273 174 312

MET 200+ 241+ 207 225 192+ 275 278 255

TYR 438 * 405 * 293 * 585 * 375 * 445 480 168

PHE 438 * 405 * 140 315 375 * 404 379 444

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ILE 228 205 173 365 250 352 326 463

LEU 306 327 417 525 250 873 853 1028

LYS 447 350 400 575 358 760 810 701

TRP 044 091 083 105 075 171 098 195

PRO 316 468 923

* Phenylalanine + Tyrosine + Cysteine + Methionine

All values are g amino acids / 100 g protein

TABLE 85

Amino Acid Analysis of Raw Materials and Products Compared to Essential Amino Acid Requirements for Various Livestock (Degment / Feather Mixture Condition 2)

Amino Acid Catfish Dog Cat Chicken Pig Experiment A2 Experiment B2 Experiment C2

ASN 074 000 008

GLN 000 000 000

ASP 750 823 695

GLU 1181 1070 1408

SER 639 1281 944

HIS 131 100 103 140 125 000 000 000

GLY 664 1298 871

THR 175 264 243 350 250 239 226 061

ALA 807 525 1012

ARG 375 282 417 550 000 466 741 372

VAL 263 218 207 415 267 892 544 531

CYS 200+ 241+ 367+ 400+ 192+ 232 000 452

MET 200+ 241+ 207 225 192+ 182 140 136

TYR 438 * 405 * 293 * 585 * 375 * 688 372 130

PHE 438 * 405 * 140 315 375 * 431 503 454

ILE 228 205 173 365 250 608 448 515

LEU 306 327 417 525 250 1053 928 1016

LYS 447 350 400 575 358 516 145 242

TRP 044 091 083 105 075 084 000 000

PRO 113 956 229

* Phenylalanine + Tyrosine + Cysteine + Methionine

All values are g amino acids / 100 g protein

For the liquid product obtained after the first hydrolysis of the chicken feather / waste mixture, the results shown in the table indicate that the solubilized protein is

Means that the essential amino acid requirements of the animal are met or exceeded.

All

On the other hand, the value after the second hydrolysis (feather), ie threonine, cysteine + methionine, tryptophan, and especially the values for lysine and histidine

Lower than this requirement, which is a poor product for unit animal nutrition, but it is suitable for ruminants.

Experiment 3 calcium recovery and regeneration

The use of calcium hydroxide as alkaline material results in relatively high calcium concentrations in centrifuged liquid solutions.

Since is low, calcium can be recovered by precipitating it with calcium carbonate, calcium bicarbonate or calcium sulfate (gypsum).

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Calcium carbonate is preferred because of its low solubility (00093 g / L, solubility of CaCO3 is 87x10-9).

Its solubility is 106 g / L and its solubility is 61x10-5 It is also easier to regenerate Ca (OH) 2 from calcium carbonate than calcium sulfate

Since CaSO4 is more soluble and gypsum is more difficult to regenerate, it is more efficient to use CaCO3 as a precipitate.

Phosphorus is fair

When CO 2 is bubbled into the centrifuged solution, carbonic acid (H 2 CO 3) is formed.

Quantum acid H2CO3, HCO3

-And CO3

An equilibrium between 2- is produced and the fraction of each component in the mixture is a function of pH Ca (HCO3) 2

Since is water soluble (166 g / L water, solubility 108), the precipitation efficiency of the process is also a function of pH

To measure and study calcium recovery by CO 2 bubbling; Centrifuged Liquid from the Hydrolysis Process of Chicken Feathers and Grounds

The product was collected in a plastic bottle and kept at 4 ° C. for later use. Known volumes of centrifuged liquid material (400 mL)

Into an Erlenmeyer flask with a stir bar (constantly stirring), and CO 2 was bubbled from the pressurized vessel.

Depending on the liquid sample (~ 10 mL) was collected and centrifuged. Total nitrogen and calcium content in the clarified liquid was measured different

Samples of initial pH were used to study whether these parameters influence precipitation efficiency.

36 shows calcium and total nitrogen content as a function of pH for two different samples: one from chicken tailing hydrolysis

(C1), the other from chicken feather hydrolysis (C2) In both cases, the TKN concentration remains constant, which means that nitrogen during calcium precipitation

Means no loss

36 also shows that calcium concentration decreases to a minimum at pH ˜9 (calcium recovery 50-70%) and increases at lower pH.

Increased calcium concentration due to the high solubility of calcium bicarbonate and the conversion of calcium carbonate to calcium bicarbonate and carbonate at low pH (below 8)

Is expected the initial pH for the centrifuged liquid shown in FIG. 36 is relatively high (102 and 111, respectively); In both cases, carbonates

Equilibrium between exists in bands with relatively high carbonate concentrations (pKa2 = 1025)

On the other hand, Figure 37 shows the calcium and total nitrogen content of the sample with a relatively low initial pH (~ 92) collected samples are carbonic acid and bicarbonate

Since it exists in the equilibrium zone between salts, calcium cannot be recovered as a precipitate (calcium bicarbonate solubility)

Experiment 4 Preservation of Chicken Waste Under Alkaline Conditions

The chicken leftovers and feathers described above in this example were used as raw material for further experimental sets.

Performed without mixing at ambient temperature in the lask; To avoid unpleasant odors, the flask was placed in a hood

Calcium hydroxide loading (g Ca (OH) 2 / g dry residue + to determine the lime required to preserve the waste material mixture

The occurrence of severe odors (fermentation products) is considered the end point of the study

Two experiments were run under the same conditions. Samples were taken from the reactor at different times and centrifuged to remove liquid from the solid material.

Sieves were separated Total nitrogen content and pH were measured in centrifuged liquid samples.

To determine the lime required for the preservation of chicken waste mixtures and to study the protein solubilization of the waste material, different lime loadings were used.

Several experiments were carried out without mixing at ambient temperature. The experimental conditions and measured parameters are summarized in Table 86.

TABLE 86

Experimental Conditions During Preservation Studies of Chicken Feather and Ground Mixtures

Experiment G1 Experiment G2 Experiment H1 Experiment H2 Experiment I1 Experiment I2

Temperature (℃) 25 25 25 25 25 25

Mass of Ca (OH) 2 (g) 33 33 66 66 99 99

Mass of residue (g) 913 913 913 913 913 913

Mass of feathers (g) 365 365 365 365 365 365

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Water volume (ml) 800 800 800 800 800 800

Ca (OH) 2 (g / g dry matter) 0052 0052 0103 0103 0155 0155

Dry substance (g / L) 8002 8002 8002 8002 8002 8002

Dry residue (g / L) 3805 3805 3805 3805 3805 3805

Total TKN (g) 679 679 679 679 679 679

Total TKN (%) 1060 1060 1060 1060 1060 1060

Table 87 shows the pH variation as a function of time, while Table 88 shows the total nitrogen content of the centrifuged liquid.

TABLE 87

PH as a Function of Time During Preservation Studies of Chicken Ground and Feather Mixtures

Time (days) Experiment G1 Experiment G2 Experiment H1 Experiment H2 Experiment I1 Experiment I2

0 901 912 121 1214 121 1215

1 --- --- 1152 1156 1214 1217

2 --- --- 1116 1125 1208 1214

4 --- --- 1082 1103 1203 1206

7 --- --- 1065 1085 1205 1206

11 --- --- 905 101 1206 1209

14 --- --- --- --- 1206 121

17 --- --- --- --- 1204 1207

TABLE 88

Total Kjeldahl Nitrogen Content as a Function of Time During Preservation Studies of Chicken Ground and Feather Mixtures

Time (days) Experiment G1 Experiment G2 Experiment H1 Experiment H2 Experiment I1 Experiment I2

0 01438 01427 01002 01103 00924 00991

1 --- --- 01248 01314 01325 01381

2 --- --- 01337 01337 01460 01472

4 --- --- 01 348 01337 01596 01630

7 --- --- 01371 01416 01835 01824

11 --- --- 01472 01427 02099 02020

14 --- --- --- --- 02239 02251

17 --- --- --- --- 02297 02297

TKN is g nitrogen / 100 g liquid sample

Proteolytic conversion was estimated and presented in Table 89 and Table 90. Table 89 summarizes the conversion considerations with regard to the residue nitrogen content.

Cotton, Table 90, provides conversions for the initial TKN of the mixture. Under the conditions studied, the highest transition of nitrogen into the liquid phase in the solid phase

The ring was ~ 30%

Table 89

Conversion rate in the liquid phase as a function of time (preservation experiment)

Time (days) Experiment G1 Experiment G2 Experiment H1 Experiment H2 Experiment I1 Experiment I2

0 755692 749911 526567 579644 485577 520786

1 --- --- 655844 690528 696309 725738

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2 --- --- 702 615 702 615 767 253 773 560

4 --- --- 708396 702615 838724 856591

7 --- --- 720482 744131 964322 958541

11 --- --- 773 560 749 911 1103058 1061542

14 --- --- --- --- 1176630 1182937

17 --- --- --- --- 1207110 1207110

Table 90

Conversion in liquid phase relative to total nitrogen as a function of time (preservation experiment)

Time (days) Experiment G1 Experiment G2 Experiment H1 Experiment H2 Experiment I1 Experiment I2

0 183018 181618 127527 140382 117600 126127

1 --- --- 158836 167236 168636 175764

2 --- --- 170 164 170 164 185 818 187 345

4 --- --- 171 564 170 164 203 127 207 454

7 --- --- 174491 180218 233545 232145

11 --- --- 187345 181618 267 145 257091

14 --- --- --- --- 284963 286491

17 --- --- --- --- 292345 292345

In Table 89, values greater than 100% mean solubilization of chicken feather protein for long-term preservation studies.

If high protein hydrolysis is correlated with high pH pH reduction during the hydrolysis process (Table 87) leads to the production of new free amino acids

A value close to 9 was measured the day before severe odor occurrence

Monitoring pH during the preservation of chicken waste mixtures is a viable alternative for maintaining a stable (non-fermentable) solution

Based on the results obtained, a pH value of 105 can be used as a lower limit for excess lime addition to avoid bacterial growth.

Lime is a relatively water-insoluble base and, due to this low solubility, produces weak alkaline conditions (pH -12) in solid-liquid mixtures.

Relatively low pH reduces the likelihood of unwanted decomposition reactions compared to strong bases (eg sodium hydroxide)

Ash also promotes protein digestion and solubilization in the liquid phase while the chicken waste mixture is preserved (Table 90).

Chicken grounds and feathers can be used to obtain amino acid rich products by treatment with Ca (OH) 2 at temperatures below 100 ° C.

Simple low pressure vessels can be used for this process because of low temperature requirements.

Chicken feather / pile mixtures can be used to obtain two amino acid rich products: one well balanced (one) and some

Lack of amino acids but high in protein and minerals

For the first lime treatment of the mixture (running at 50-100 ° C.), the spectrum of essential amino acids obtained from the experiment was high during the growth phase.

Therefore, the amino acid rich solid product obtained by lime treatment of chicken residues

Can serve as a protein supplement for these animals

For the second lime treatment of the mixture (running at 75-100 ° C.), the spectrum of essential amino acids obtained from the experiment showed several amino acids.

Lack of acid Thus, the amino acid-rich solid product obtained by the second lime treatment of the chicken feather / waste mixture is present in ruminants.

Can serve as a source of nitrogen and minerals

Bubbling CO2 into centrifuged liquid product to precipitate calcium carbonate recovers between 50 and 70% of calcium High initial pH

Is recommended (> 10), while calcium carbonate is formed during the process and no calcium bicarbonate is formed; From final pH to 88-90

Ensures high calcium recovery for CaSO4 because CaSO4 is more soluble and gypsum is more difficult to regenerate,

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It is a more efficient process to use. Finally, the lime solution contains chicken processing waste containing keratinous substances in chicken feathers.

Hydrolysis and preservation during absence of decay odor, continuous protein hydrolysis to liquid phase, and preservation of chicken waste mixture

Due to the possibility of continuous monitoring of pH, the process maintains a stable (non-fermentable) solution and keeps the chicken waste mixture during storage on the farm

Become a viable alternative to preserve

Example 6: Protein Solubilization in Cow Hair

According to the USDA, 188 lbs of lean meat and poultry is consumed per person in the United States each year, with ~ 116 lbs of beef and pork

Animal slaughterers generate large amounts of waste, and animal hair represents 3 to 7% of the total weight. Better use of waste residues

And conversion of these into useful products is necessary and desired

The wet hairs were obtained from the Terrabonn company and then air dried. To characterize the starting material, the moisture content, total nitrogen (protein

Estimates of fractions), and amino acid content were determined

Air dried hair is used as starting material for these experiments. The dry matter content, chemical composition and amino acid balance are respectively listed.

91, Table 92 and Table 93.

TABLE 91

Dry matter content of air dried cattle

Sample Moisture Solid (g) Dry Solid (g) Dry Matter (%)

1 40883 38350 9380

2 37447 35163 9390

Average 9385

Table 92

Protein and Mineral Contents of Air Dried Wool

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

Hair 1473 00508 00197 01658 0029 5244 58 185 50 37

TABLE 93

Amino Acid Composition of Air Dried Cow Hair

Amino acid measured literature amino acid measured literature

ASP 663 30 TYR 24 4 34

GLU 1447 122 VAL 680 55

SER 891 72 MET 071 06

HIS 129 07 PHE 309 30

GLY 552 108 ILE 420 44

THR 748 66 LEU 977 77

ALA 450 10 LYS 553 21

CYS ND 139 TRP ND 14

ARG 1098 77 PRO 768 85

ND: not measured

Value is g AA / 100 g total amino acid

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Starting materials have relatively well-balanced amino acid content and low levels of histidine, methionine, tyrosine and phenylalanine

The content is very low (~ 1%), the crude protein content is high (~ 921%) and the starting moisture content is 615%.

Experiment 1 Hair Concentration Effect

To determine the effect of initial hair concentration on solubilization of proteins, temperature and lime loading (100 ° C. and 010 g lime / g air dried, respectively)

The experiments were run at different concentrations while keeping the hair constant. The experimental conditions and parameters measured were summarized in Table 94.

Table 94

Experimental Conditions and Measured Parameters to Determine the Effect of Initial Hair Concentration on Protein Solubilization of Cow Hair

Hair Concentration (g Hair / L) 40 60

Mass of hair (g) 34 51

Water volume (mL) 850 850

Mass of lime (g) 34 51

Temperature (℃) 100 100

Initial temperature (℃) 1014 871

pH final 92 98

Residual Solid (g) 288 449

Dissolved solids in 100 mL (g) 118 192

Protein in 100 mL (g) 081 104

Table 95 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different hair concentrations.

Based on one mean TKN (1473%), proteolytic conversion is estimated and presented in Table 96

Table 95

Total Kjeldahl nitrogen content in the liquid phase centrifuged as a function of time for Experiment 1 (Fur)

Air dried hair concentration

Time (h) 40 g / L 60 g / L

0 00160 00327

05 00185 00497

1 00435 00699

2 00718 01000

3 00754 01194

4 00868 01368

6 01088 01629

8 01298 01662

TKN is g nitrogen / 100 g liquid sample

TABLE 96

Conversion of Total TKN to Soluble TKN for Experiment 1 (Fur)

Air dried hair concentration

Time (h) 40 g / L 60 g / L

0 272 370

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05 314 562

1 738 791

2 1219 1131

3 1280 1351

4 1473 1548

6 1847 1843

8 2203 1881

38 shows protein solubilization (conversion) as a function of time for the different hair concentrations studied.

It does not have a significant effect on degradation (conversion) and obtains about 70% of the conversions achieved using chicken feathers, another keratin substance.

Shows that more lime loading or longer treatment periods are required

As shown in Table 94, the dissolved solids are more for higher hair concentrations as expected, the final pH for both cases.

Is lower than the initial 120, indicating that lime was consumed during hydrolysis and that lime was not present as a solid in the final mixture.

It's beautiful

Experiment 2 lime loading effect

To determine the effect of lime loading on protein solubilization of air dried hair, temperature and hair concentration (100 ° C. and 40 g air guns, respectively)

The experiments were run at different lime / hair ratios while keeping the coarse hair / L) constant.

To summarize

TABLE 97

Experimental Conditions and Measured Parameters for Determining Lime Loading Effect in Protein Solubilization of Cow Hair

Lime loading (g lime / g whisk) 010 020 025 035

Mass of hair (g) 34 34 34 34

Water volume (mL) 850 850 850 850

Mass of lime (g) 34 68 85 119

Temperature (℃) 100 100 100 100

Initial Temperature (℃) 1014 1023 756 902

pH final 92 103 114 112

Residual Solid (g) 288 1744 (*) 226 229

Dissolved solid in 100 mL (g) 118 292 (*) 296 299

Protein in 100 mL (g) 081 177 218 240

(*) Measured after 48 h rather than 8 h as in the other three conditions

Table 98 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different lime loadings.

Based on the mean TKN (1473%) for the proteolytic conversion is estimated and presented in Table 99

TABLE 98

Total Kjeldahl nitrogen content in the liquid phase centrifuged as a function of time for Experiment 2 (Fur)

Lime loading

Time (min) 010 g / g 020 g / g 025 g / g 035 g / g

0 00160 00144 00241 00133

05 00185 --- 00454 00637

1 00435 00845 00922 00822

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2 00718 01425 01350 01438

3 00754 --- 01549 01792

4 00868 02145 01951 02023

6 01088 --- 02699 02999

8 01298 02832 03487 03837

TKN is g nitrogen / 100 g liquid sample

TABLE 99

Conversion of Total TKN to Soluble TKN for Experiment 2 (Fur)

Lime loading

Time (min) 010 g / g 020 g / g 025 g / g 035 g / g

0 272 244 409 226

05 314 --- 771 1081

1 738 1434 1565 1395

2 1219 2419 2291 2441

3 1280 --- 2629 3041

4 1473 3641 3311 3433

6 1847 --- 4581 5090

8 2203 4807 5918 6512

39 shows solubilized protein (conversion) as a function of time for the different lime loadings studied, which is 01 g lime / g air

The conversion is similar for all lime loadings except for the dried hair. FIG. 38 shows that the conversion is higher at longer times.

Shows that this different and reaction does not slow at 8 h for any lime loading studied.

A minimum lime loading is required to increase the ring and make the process efficient

As shown in Table 97, dissolved solids are more for more lime loading as expected (more knives in solution)

Calcium salt and higher conversion) The final pH increases with increasing lime loading and in all cases lower than 120, which in turn is hydrolyzed

Mean that the lime consumption, and the final OH-concentration (pH) can be related again to the efficiency of the treatment.

The behavior shown in FIG. 39 may be related to the requirement for hydroxyl groups as catalysts for hydrolysis reactions.

Maintains a "constant" lime concentration in all treatments (02 to 035 g lime / g air dried hair), but is not affected by its consumption during the process.

Slower lime loading reactions or faster leveling

Effect of Long-Term Treatment Than Experiment 3

In order to establish the effect of long-term treatment on the solubilization of the protein, two different conditions, 100 ° C., 02 g lime / g air dried, respectively, were applied.

Whisk, 40 g air dried whisk / L; And 100 ° C., 035 g lime / g air dried hair, 40 g air dried hair / L

Experimental conditions and measured parameters are summarized in Table 100.

TABLE 100

Experimental conditions and measured parameters to determine the effect of longer treatment duration on protein solubilization of bovine hair

Lime loading (g lime / g air dried whisk) 02 035

Mass of hair (g) 34 34

Water volume (mL) 850 850

Mass of lime (g) 68 119

Temperature (℃) 100 100

pH final 103 1199

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Residual Solid (g) 1744 1074

Dissolved solids in 100 mL (g) 292 401

Protein in 100 mL at 48 h (g) 225 263

Table 101 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different lime loadings.

Based on the mean TKN (1473%) for the proteolytic conversion is estimated and presented in Table 102.

TABLE 101

Total Kjeldahl Nitrogen Content in the Liquid Phase Centrifuged as a Function of Time for Experiment 3 (Fur)

Lime loading

Hour (h) 020 g / g 035 g / g

0 00144 00133

1 00845 ---

2 01 425 ---

4 02145 02088

8 02832 02832

12 03089 ---

24 03319 03988

36 03617 04265

48 03597 04210

TKN is g nitrogen / 100 g liquid sample

Table 102

Total TKN Conversion to Soluble TKN for Experiment 3 (Fur)

Lime loading

Hour (h) 020 g / g 035 g / g

0 244 226

1 1434 ---

2 2419 ---

4 3641 3544

8 4807 4807

12 5243 ---

24 5633 6768

36 6139 7239

48 6105 7145

FIG. 40 shows protein solubilization (conversion) as a function of time for the two different conditions studied.

It is different for treatments and shows that the reaction reaches the highest conversion in 24 to 36 hours of treatment The tube between lime availability and conversion

Relevance is better understood in the long-term treatment studies.

Starts at 24 hours, an easily recognizable ammonia odor occurs, which suggests amino acid degradation over longer periods

One method of reducing the problem is to separate residual solids into liquid phase for further alkaline hydrolysis in subsequent processing steps.

To recover unhydrolyzed amino acids.

Experiment 4 Determination of Ammonia During Alkaline Hydrolysis of Air Dried Cattle (Amino Acid Degradation)

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The effect of long-term treatment on solubilization of proteins and degradation of soluble amino acids was determined by ammonia measurement.

For two experimental conditions of Experiment 3 and at 100 ° C., 02 g lime / g air dried hair, 40 g air dried hair / L for 5 hours

Further runs using centrifuged liquids of the experiments performed were determined as a function of time studied experimental conditions and measured

The variables are summarized in Table 103.

Table 103

Experimental conditions and measured parameters to determine the effect of longer treatment periods on amino acid degradation

Lime loading (g lime / g air dried hair) 02 (Experiment A1) 035 (Experiment A2) 035 (Experiment A3)

Mass of hair (g) 34 34 **

Water volume (mL) 850 850 850

Mass of lime (g) 68 119 85

Temperature (℃) 100 100 100

Initial temperature (℃) 1023 988 966

pH final 103 1199 1208

Residual Solid (g) 1744 1074 828

Dissolved solid in 100 mL (g) 292 401 250

Protein in 100 mL at 48 h (225 g) 225 263 141

** No solid material used, only centrifuged liquid from previous experiments

Table 104-106 and FIGS. 41-43 show the total nitrogen content and free arm in the centrifuged liquid sample as a function of time for different experimental conditions.

Shows moniker concentration

Table 104

Total Kjeldahl Nitrogen Content, Ammonia Concentration and Estimated Protein Nitrogen (Fleece) in the Centrifuged Liquid Phase as a Function of Time for Experiment A1

Time (h)

[ammonia]

(ppm)

TKN

(%)

TKN

(ppm)

Protein-N

(ppm)

0 34 00144 144 110

1 33 00845 845 812

2 41 01 425 1425 1384

4 76 02 145 2145 2069

8 175 02832 2832 2657

12 236 03089 3089 2853

24 274 03319 3319 3045

36 327 03617 3617 3290

48 316 03597 3597 3281

TKN is g nitrogen / 100 g liquid sample

Table 105

Total Kjeldahl Nitrogen Content, Ammonia Concentration and Estimated Protein Nitrogen (Fleece) in the Centrifuged Liquid Phase as a Function of Time for Experiment A2

Time (h)

[ammonia]

(ppm)

TKN

(%)

TKN

(ppm)

Protein-N

(ppm)

0 0 0 0 0

4 85 02088 2088 2003

8 115 02 832 2832 2717

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24 111 03988 3988 3877

36 141 04 265 4265 4124

48 110 04 210 4210 4100

TKN is g nitrogen / 100 g liquid sample

TABLE 106

Total Kjeldahl Nitrogen Content, Ammonia Concentration and Estimated Protein Nitrogen (Fleece) in the Centrifuged Liquid Phase as a Function of Time for Experiment A3

Time (h)

[ammonia]

(ppm)

TKN

(%)

TKN

(ppm)

Protein-N

(ppm)

0 50 02 332 2332 2282

1 50 02 426 2426 2376

2 51 02 449 2449 2398

4 60 02 449 2449 2389

8 90 02382 2382 2292

12 106 02393 2393 2287

24 86 02326 2326 2240

48 87 02 248 2248 2161

The ammonia concentration in the centrifuged liquid is determined by the Kjeldahl method, but there is no initial hydrolysis of the sample

TKN is g nitrogen / 100 g liquid sample

41 and 42 show that total protein-N concentration increases as a function of time until the maximum is reached at 24 to 36 h of treatment.

The free ammonia concentration also increases as a function of time, which suggests the decomposition of amino acids. In Experiments A1 and A2, into the liquid

Further hydrolysis of the hair provides a pure improvement of protein-N up to the 24-36 h period in excess of amino acid degradation.

In Experiment A3, there is no solid hair and therefore no protein source other than previously solubilized protein, in this case a reduction of protein-N

Occurs after 4 h and continues to 48 h, which means that some amino acids are sensitive to degradation under the conditions studied

Experiment 4A Amino Acid Degradation Study

For experiments A2 and A3, the amino acid composition of the liquid sample was analyzed to determine the stability of the individual amino acids in the protein hydrolysate.

Was

Two different amino acid analyzes of lime-hydrolyzed bovine were performed:

1) Free amino acid analysis in the centrifuged liquid was performed without excess HCl hydrolysis of the sample.

Although not destroyed, soluble polypeptides are lost in the assay

2) HCl hydrolysis was performed prior to total amino acid HPLC determination in the centrifuged liquid. Some amino acids (asparagine, glutamine,

Cysteine and tryptophan) were destroyed by acid and could not be measured

Tables 107 and 108 compare estimated amino acids using total amino acids (HCl hydrolysis), free amino acids, and TKN values.

Tables show that the hair protein is mainly hydrolyzed to small soluble peptides instead of free amino acids (to free amino acids total amino

Compared to acid columns)

Table 107

Protein Concentration Comparison for Experiment A2 (Fleece)

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Time (h) TKN (%) Protein (mg / L) Free AA (mg / L) Total AA (mg / L)

4 02088 130 500 3304 47835

8 02 832 177000 6845 93004

24 03988 249 250 14549 122084

36 04 265 266563 16992 136801

48 04 210 263 125 17426 139896

Table 108

Protein Concentration Comparison for Experiment A3 (Fleece)

Time (h) TKN (%) Protein (mg / L) Free AA (mg / L) Total AA (mg / L)

0 02 332 145 750 4136 73730

1 02 426 151625 8166 94906

2 02 449 153063 9894 110754

4 02 449 153063 11547 120404

8 02382 148875 13939 105491

12 02393 149563 15719 99884

24 02 326 145 375 22669 84648

48 02 248 140 500 22369 87823

Table 108 also shows an increase in total amino acid concentration from 0 to 4 h. This experiment (A3) shows that only centrifuged liquid (no solid hair)

Increasing values result in the presence of suspended polypeptide particles in solution which are further hydrolyzed in liquid.

In solid separation, the liquid was centrifuged at 3500 rpm, while 15000 rpm was used before HPLC analysis.

Table 108 shows that the total amino acid concentrations are very consistent at the estimated protein (TKN) and 4 h, where amino acid degradation is relatively

There is very little, and the conversion of "suspended substances" in the liquid phase is very high. In Table 107, the difference is not suspended in the amino acid analysis.

Can be explained by the presence of a substance

For Experiment A2, FIG. 44 shows the concentration of individual free amino acids present in the centrifuged liquid as a function of time,

45 shows the total concentration of individual amino acids as a function of time Histidine concentration elutes just before very high concentration of glycine;

Cannot be measured or underestimated because peaks cannot be separated

Figure 45 shows an increase in all amino acid concentrations up to 36 h except for arginine, threonine and serine Figure 44 shows in particular arginine

And similar behavior except for lower concentrations for threonine. At 36 hours, amino acid concentrations were leveled off.

(Except arginine, threonine and serine), suggests an equilibrium between solubilization and degradation processes

For experiment A3 (using only liquid centrifuged without solid hair added), FIG. 45 is in the centrifuged liquid as a function of time.

While showing the concentration of the individual free amino acids present, FIG. 46 shows the total concentration of the individual amino acids as a function of time.

In Figure 46, the concentration of free amino acids is increased up to 24 h and then leveled again, except for arginine, threonine and serine,

The first two have very low concentrations as free amino acids

47 shows an increase in all individual amino acid concentrations from 0 to 4 h which in turn is hydrolyzed to liquid phase from 0 to 4 h.

The presence of suspended particles in the previously centrifuged liquid. After this initial trend, the concentration of all amino acids decreases with time.

High, which suggests the degradation of all amino acids under the conditions studied for long-term treatment. Arginine (16% of the concentration obtained at 4 h)

Is present at 48 h), threonine (31%) and serine (31%) are more degraded than other amino acids

Increased concentrations of ornithine and citrulline, both not present in appreciable amounts in the hair, suggest that they are possible degradation products

do

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Table 109 shows the weight ratio of each amino acid as a function of time for Experiment A2. Similar content of arginine, threonine

For most amino acids except and serine. Some amino acid ratios increase due to greater resistance to degradation.

Other things decrease

Table 109

Individual amino acids present in Experiment A2 as a function of time compared to the initial material

amino acid

Time (h)

4 8 24 36 48 Hair

ASP 676 690 703 696 677 663

GLU 1331 1464 1596 1642 1637 1447

SER 668 376 153 111 100 891

HIS 111 000 000 000 000 129

GLY 933 948 850 825 829 552

THR 240 166 085 066 054 748

CIT 091 095 156 168 168 000

ALA 540 650 863 947 927 450

ARG 922 779 438 289 211 1098

TYR 535 543 578 587 574 244

VAL 674 713 745 740 725 680

MET 080 090 105 100 109 071

PHE 317 305 313 317 315 309

ILE 404 419 452 462 455 420

LEU 881 966 1092 1121 1125 977

LYS 209 271 389 408 414 553

PRO 1377 1507 1460 1502 1660 768

Value is g AA / 100 g total amino acid

Experiment 5 Two-stage Treatment of Material

Amino acid degradation observed in previous experiments undermines the overall efficiency of the hydrolysis process.

Separating the dissociated protein from subsequent solubilization (residual solids) of the protein in a series of processing steps.

Two conditions were studied to determine the effect of the two-step process on amino acid degradation of proteins in air and air dried hair.

The experimental conditions and measured parameters are summarized in Table 110.

TABLE 110

Experimental Conditions and Measured Variables for Determining Lime Loading Effect in Protein Solubilization (Fleece-Two Step Treatment)

Experiment Experiment C1 Experiment C2 Experiment D1 Experiment D2

Mass of hair (g) 34 20 34 20

Water volume (mL) 850 850 850 850

Mass of lime (g) 85 5 119 5

Temperature (℃) 100 100 100 100

Initial Temperature (℃) 756 965 902 105

pH final 114 112 112 112

Residual solid at 8 h (g) 226 127 229 124

Dissolved solid in 100 mL (g) 296 115 299 117

Protein in 100 mL at 8 h (g) 180 091 178 086

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Table 111 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different experimental conditions.

Based on the mean TKN (1473%) for the proteolytic conversions were estimated and shown in Table 112. FIG. 48 shows the empty time as a function of time.

Shows total conversion for tablets (step 1 + step 2)

Table 111

Total Kjeldahl nitrogen content in the liquid phase centrifuged as a function of time for Experiment 5 (Fur)

Time (h) Experiment C1 Experiment C2 Experiment D1 Experiment D2

0 00241 00363 00133 00365

05 00454 00553 00637 00481

1 00922 00560 00822 00571

2 01 350 00620 01438 00631

3 01 549 00756 01792 00704

4 01951 00745 02023 00798

6 02299 01135 02269 01042

8 02887 01450 02837 01383

TKN is g nitrogen / 100 g liquid sample

Table 112

Conversion of Total TKN to Soluble TKN for Experiment 5 (Fur)

Time (h) Experiment C1 Experiment C2 Experiment D1 Experiment D2

0 409 616 226 619

05 771 939 1081 816

1 1565 950 1395 969

2 2291 1052 2441 1071

3 2629 1283 3041 1195

4 3311 1264 3433 1854

6 3902 1926 3851 1768

8 4900 2461 4815 2347

FIG. 48 shows similar conversion for the two conditions studied. At 16 h of treatment, a total of 70% of the initial nitrogen was recovered in the liquid phase.

The total conversion increased during the second treatment, with lower concentrations of ammonia present than in the first stage treatment (Table 113),

Suggest less degradation Therefore, further treatment of the residual solid with lime will hydrolyze more hair, but in the second step

The concentration of bovine (protein / amino acid) is only 40% of that obtained in the initial treatment, which increases the energy required for water evaporation.

Since the initial concentration of kinda fur has no significant effect on the conversion, higher product concentrations can be obtained in semisolid reactions.

Table 113

Total Kjeldahl Nitrogen and Ammonia Concentrations for Two-Step and One-Step Processes

Step 1 (8 h) Step 2 (8 h) Step 1 (16 h)

TKN 02984 01154 03525

Ammonia 87 39 363

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The separation of the initial liquid at 8 h yields a ratio for sensitive amino acids (arginine, threonine and serine) with about 50% conversion of the initial protein.

Ensure high levels of maturation. The second step provides lower total concentrations of these amino acids while providing greater total conversion.

Unreacted residual solids after step 2 (approximately 30% of the initial hair using 7 g nitrogen / 100 g dry solids) were added in a total of 80%

Can be further processed to provide white matter recovery. The step will probably require 24 to 36 hours.

Experiment 6 Amino Acid Composition and Process Mass Balance of the Product

This section presents the amino acids of the product obtained using the total mass balance and two proposed 8-h step processes and one 16-h step treatment.

We suggest the furtherance

Table 113 compares the total Kjeldahl nitrogen and ammonia concentrations for the three centrifuged liquid products.

Shows solid composition (nitrogen and minerals) for FIG. 49 shows mass balance for two-step and one-step processes

Nonuniformity causes very large fluctuations in concentration

Table 114

Protein and mineral content of air dried hair and residual solids in the process

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

Hair 1473 00508 00197 01658 0029 5244 58 185 50 37

RS1 8h 10234 00622 00176 70083 01233 3005 108 457 61 17

RS2 8h 6974 00725 00155 101003 01938 2301 117 702 62 22

RS3 16h 5803 00642 00228 97181 01617 2404 79 472 56 18

Table 115 compares amino acid compositions for three different products and hairs. As expected from previous experiments, Step 1

Provides higher values for threonine, arginine, and serine. Except for the aforementioned amino acids, steps 1, 2, and 1

The concentration of product from the system process is very similar.

Table 115

Individual amino acids present in the solid product and starting material

Amino Acid Stage 1 (8 h) Stage 2 (8 h) Stage 1 (16 h) Hair

ASP 819 868 785 663

GLU 1746 1930 1751 1447

SER 301 110 157 891

HIS 106 083 094 129

GLY 1000 697 984 552

THR 132 083 076 748

ALA 734 780 864 450

ARG 795 494 525 1098

TYR 175 214 259 244

VAL 782 899 820 680

MET 073 099 075 071

PHE 337 339 338 309

ILE 462 521 482 420

LEU 1101 1304 1152 977

LYS 277 482 391 553

PRO 1162 1094 1245 768

Value is g AA / 100 g total amino acid

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Finally, in Table 116, the amino acid composition of the product was compared to the required essential amino acids of the various unit livestock.

Table 116

Amino Acid Analysis of Products and Essential Amino Acid Requirements for Various Livestock

amino acid

Step 1

(8h)

Step 2

(8h)

Stage 1

16h

Hairless catfish dog cat chicken pig

ASP 819 868 785 663

GLU 1746 1930 1751 1447

SER 301 110 157 891

HIS 106 083 094 129 131 1 103 14 125

GLY 1000 697 984 552

THR 132 083 076 748 175 264 243 35 25

ALA 734 780 864 450

ARG 795 494 525 1098 375 282 417 55 0

VAL 782 899 820 680 263 218 207 415 267

CYS ND ND ND ND 2+ 241+ 367+ 4+ 192+

MET 073 099 075 071 2+ 241+ 207 225 192+

TYR 175 214 259 244 438 * 405 * 293 * 585 * 375 *

PHE 337 339 338 309 438 * 405 * 14 315 375 *

ILE 462 521 482 420 228 205 173 365 25

LEU 1101 1304 1152 977 306 327 417 525 25

LYS 277 482 391 553 447 35 4 575 358

TRP ND ND ND ND 044 091 083 105 075

PRO 1162 1094 1245 768

+ Cysteine + methionine * tyrosine + phenylalanine ND not measured

All values are g amino acids / 100 g protein

As shown in Table 116, the amino acid composition of the lime-hydrolyzed cow's hair was based on the essential amino acid requirements of different livestock animals.

Not well balanced in relation to histidine (underestimated in the analysis), particularly low values for threonine, methionine and lysine

Minor other amino acids are sufficient for most, but not all, animals (tyrosine, phenylalanine).

Although glutamine + glutamate produce very rich products, they are not essential amino acids in feed for livestock units

Multi-amino acid products can be used for ruminants

Higher serine and threonine concentrations can be obtained by reducing the time in step 1

Air dried bovine hair containing 92% protein (wet basis) was treated with Ca (OH) 2 at 100 ° C. to obtain an amino acid rich product.

Because of the low temperature requirements, a simple unpressurized container may be used for this process.

While hair concentrations do not have a significant effect on proteolysis, they can also be obtained from other keratin substances, chicken feathers.

High lime loading (more than 01 g Ca (OH) 2 / g hair) and long term treatment period (t> 8 h) are required to achieve about 70% conversion.

Protein solubilization varies with lime loading only for prolonged treatment, which requires hydroxyl groups as a catalyst for the hydrolysis reaction.

Although shown, its consumption during the process slows down the smaller lime loading reaction or makes the leveling faster

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To maximize protein conversion (up to 70%) the optimum conditions were 035 g Ca (OH) 2 / g hair treated at 100 ° C. for at least 24 hours.

Easily recognizable ammonia odor starting at 24 hours multi offers amino acid decomposition Arginine, Threonine and Serine Eggs

More sensitive amino acids under kali hydrolysis

Decomposition of amino acids is already hydrolyzed into liquid phase by separating residual solids for further alkaline hydrolysis in subsequent processing steps

The recovery of the initial liquid (step 1) at 8 h has about 50% conversion of the initial protein.

While ensuring relatively high concentrations for sensitive amino acids (arginine, threonine, and serine).

Lower concentrations provide greater total conversion (approximately 70%)

The nitrogen concentration (protein / amino acid) in step 2 is only 40% of that obtained in the initial treatment, which is the energy required for water evaporation.

The initial concentration of the hair does not have a significant effect on the conversion, so more product concentrations are obtained by semisolid reactions.

Can lose

The amino acid composition of the composition is poor compared to essential amino acid requirements for different livestock unit animals.

Low histidine, methionine and lysine Asparagine and proline are particularly rich, but they are not required in animal feed

Products obtained by conventional processes are useful as ruminant foods, have very high digestibility, high nitrogen content and are highly soluble in water

to be

Example 7: Protein Solubilization in Shrimp Heads

Significant shrimp processing by-products are disposed of annually. In commercial shrimp processing, approximately 25% (w / w) of fresh shrimp is recovered as shrimp meat.

Sieve waste contains about 30-35% tissue protein; Calcium carbonate and chitin are other major fractions Chitin and chitosan production is currently

Based on waste from processing of keratin During chitosan production, about 3 kg of protein is discarded for every 1 kg of chitosan production

(Gildberg and Stenberg, 2001)

Chitin is a widely distributed, naturally rich amino polysaccharide, insoluble in water, alkalis and organic solvents, slightly soluble in strong acids

Ida chitin is a structural component of the crustacean exoskeleton consisting of ~ 15-20% chitin by dry weight.

Similar to cellulose in structure and biological function (Kumar, 2000)

At present, chitin-containing materials (crust, shrimp waste, etc.) are diluted salts after treatment for 1-3 h in boiling aqueous sodium hydroxide (4% w / w).

Decalcify in acid (1-2 N HCl) for 8-10 h (calcium carbonate removal) Subsequently, chitin is concentrated in sodium hydroxide (40-

Deacetylated in 50% w / w) to form chitosan

Frozen large whole white shrimp was obtained from the grocery store. The shrimp tail was removed and residual waste (head, stutter)

Etc.) in an industrial blender for 10 minutes, collected in plastic bottles and finally frozen at -4 ° C for later use.

Water content using the sample of the blended material, total nitrogen (estimated protein ~ 16% + chitin fraction by total weight)

164% is nitrogen), ash (mineral fraction) and amino acid content were obtained to characterize the starting material.

Shrimp head waste was 2146% dry matter and 172 g ash / 100 g dry weight (Table 117 and Table 118) TKN was approximately 1025%

Corresponds to 641% crude protein and chitin fraction (Table 119). The remaining 18% corresponds to lipids and other components.

The amino acid compositions for are shown in Table 120.

Table 117

Moisture content in shrimp head waste

Sample solids (g) Dry solids (g) Dry solids (%)

1 641091 137745 2149

2 585 237 125662 2147

3 617193 132126 2141

Average 2146

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Table 118

Ash content in shrimp head waste

Sample solids (g) Dry solids (g) Dry solids (%)

1 32902 05859 1781

2 3068 05148 1678

3 30486 05196 1704

Average 1721

Table 119

Protein and Mineral Content in Shrimp Head Wastes

Sample

TKN

(%)

P

(%)

K

(%)

Ca

(%)

Mg

(%)

Na

(ppm)

Zn

(ppm)

Fe

(ppm)

Cu

(ppm)

Mn

(ppm)

1 102 134 107 45 430 03 896 120 90 90 355 160 10

2 103 121 102 47 162 03586 11550 90 167 155 9

Average 1025 127 1045 46296 03781 11820 90 261 1575 95

TABLE 120

Amino Acid Composition of Shrimp Head Waste

Amino acid measured amino acid measured

ASP 1113 TYR 315

GLU 1583 VAL 577

SER 408 MET 184

HIS 178 PHE 493

GLY 694 ILE 454

THR 406 LIEU 830

ALA 683 LYS 563

OYS ND TRIP ND

ARG 725 PRO 796

ND: not measured Value is g AA / 100 g total amino acids

Starting material had a well balanced amino acid content (Table 120); Histidine and methionine levels are relatively low.

Calcium and potassium make substances a useful source for minerals in animal feed

Experiment 1 repeatability

To determine the repeatability of the solubilization method of proteins in shrimp head waste, the same conditions (100 ° C., 40 g dry shrimp / L, respectively)

And 010 g lime / g dried shrimp) were run. The experimental conditions and measured parameters are summarized in Table 121.

Table 121

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Experimental Conditions and Measured Variables to Determine Repeatability in Protein Solubilization of Shrimp Head Wastes

Experiment A B

Mass of Shrimp Head Waste (g) 149 149

Volume of Water (mL) 750 750

Mass of lime (g) 32 32

Initial Temperature (℃) 97 87

pH final 1064 102

Moist Residual Solid (g) 13719 1827

Dry residual solid (g) 1724 1974

Dissolved solid in 100 mL (g) 23757 24322

Table 122 shows the total nitrogen content in the centrifuged liquid sample as a function of time for two different runs.

Based on the average TKN (1025%) for the waste, proteolytic conversion was estimated and presented in Table 123.

The mean standard deviation is 113 or 15% of the mean result (793% conversion)

TABLE 122

Total Kjeldahl nitrogen content in the centrifuged liquid phase as a function of time for Experiment 1 (shrimp head waste)

Time (minutes) A B

0 02837 02934

10 03005 03017

20 03053 02981

30 03029 03005

60 03053 02969

120 03077 03005

TKN is g nitrogen / 100 g liquid sample

Table 123

Total TKN Conversion to Soluble TKN for Experiment 1 (Shrimp Head Waste)

Time (minutes) A B

0 751 776

10 795 798

20 808 789

30 801 795

60 808 786

120 814 795

FIG. 49 shows protein solubilization (conversion) as a function of time for two different runs, where conversion takes place after the initial 5-10 minutes.

Remain stable and show that the proteolytic process is clearly repeatable under the conditions studied.

The condenser is sealed and pressurized and then taken, the process takes 8 to 12 minutes

Experiment 2 temperature effect

To determine the effect of temperature on the solubilization of protein in shrimp head waste, lime loading and material concentration (010 g lime / g respectively)

The experiment was run at different temperatures while keeping the shrimp and 40 g dry shrimp / L) constant.

Summarizes at 124

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Table 124

Experimental Conditions and Measured Variables to Determine the Effect of Temperature on Protein Solubilization of Shrimp Head Wastes

Temperature (℃) 75 100 125

Mass of shrimp (g) 149 149 149

Volume of Water (mL) 750 750 750

Mass of lime (g) 32 32 32

Initial temperature (℃) 785 97 108

pH final 101 1064 988

Moisture Residual Solid (g) 13304 13719 13058

Dry Residual Solid (g) 1606 1724 1742

Dissolved solid in 100 mL (g) 26439 23757 26808

Table 125 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different temperatures. Dry shrimp head waste

Based on the average TKN (1025%) for, proteolytic conversion was estimated and presented in Table 126.

TABLE 125

Total Kjeldahl nitrogen content in the centrifuged liquid phase as a function of time for Experiment 2 (Shrimp Head Waste)

Temperature

Time (min) 75 ℃ 100 ℃ 125 ℃

0 03 160 02837 03053

10 03196 03005 03101

20 03101 03053 03101

30 03101 03029 03112

60 03101 03053 03101

120 03172 03077 03101

TKN is g nitrogen / 100 g liquid sample

Table 126

Total TKN Conversion to Soluble TKN for Experiment 2 (Shrimp Head Waste)

Temperature

Time (min) 75 ℃ 100 ℃ 125 ℃

0 836 751 808

10 846 795 821

20 821 808 821

30 821 801 823

60 821 808 821

120 839 814 821

51 shows proteolysis (conversion) as a function of time for the different temperatures studied. Conversion is not temperature dependent.

At the lower temperature (statistically the same value), less amino acids will decompose and are required to maintain the process at that temperature.

Advantageous because less energy is saved

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Experiment 3 lime loading effect I

To determine the effect of lime loading on protein solubilization of shrimp head waste, temperature and shrimp concentrations (100 ° C and 40 g, respectively)

The experiment was run at different lime / shrimp ratios while keeping the crude shrimp / L) constant. The experimental conditions and measured parameters are shown in Table 127.

Summarize

Table 127

Experimental Conditions and Measured Parameters for Determining Lime Loading Effect in Protein Solubilization of Shrimp Head Wastes

Lime loading (g lime / g shrimp) 0 005 01 02

Mass of shrimp head waste (g) 149 149 149 149

Volume of Water (mL) 750 750 750 750

Mass of lime (g) 0 16 32 64

Initial Temperature (℃) 96 95 97 103

pH final 81 920 1064 12

Moisture Residual Solid (g) 1794 1488 1372 1225

Dry Residual Solid (g) 1772 165 1724 1828

Dissolved solid in 100 mL (g) 23 576 25 146 23 757 24516

Table 128 shows the total nitrogen content in the centrifuged liquid sample as a function of time for different lime loadings.

Proteolytic conversion was estimated based on mean TKN (1025%) for the substance (Table 129).

Table 128

Total Kjeldahl nitrogen content in the centrifuged liquid phase as a function of time for Experiment 3 (Shrimp Head Waste)

Lime loading

Time (minutes) 0 g / g 005 g / g 01 g / g 02 g / g

0 02477 02890 02837 02573

10 02452 02978 03005 02573

20 0244 03035 03053 02621

30 02488 03035 03029 02669

60 02452 03051 03053 02766

120 02513 03035 03077 02897

TKN is g nitrogen / 100 g liquid sample

Table 129

Total TKN Conversion to Soluble TKN for Experiment 3 (Shrimp Head Waste)

Lime loading

Time (minutes) 0 g / g 005 g / g 01 g / g 02 g / g

0 655 765 764 681

10 649 788 797 681

20 646 803 798 694

30 658 803 798 706

60 649 807 797 732

120 665 803 805 767

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Figure 52 shows solubilized protein (conversion) as a function of time for the different lime loadings studied which do not use lime

Except that experiments (statistically different) show that the conversion is similar for all lime loadings

In the limeless experiment, soluble protein is present in the aqueous phase; Hydroxyl groups dilute, slowing the hydrolysis reaction and cell destruction

The final pH for the limeless experiment was 81 Similarly, the alkaline pH was calculated from calcium carbonate and bicarbonate released from shrimp waste.

Is caused by calcium

Lime addition is required to ensure rapid proteolysis in the liquid phase and likewise higher fractions of oil in the product

In addition, the lime treatment is considered a preliminary step to produce chitin and chitosan, so high protein

Recovery reduces the chemicals required for subsequent steps during processing and involves higher quality chitin or chitosan products.

All

The recovery of carotenoids (astaxanthin) from suspended solids is intended to produce additional useful products from the process.

Since calcium carbonate and chitin are structural components of crustaceans, the mixture is filtered and the suspended solids

Tenoids can be recovered (Gildberg and Stenberg, 2001)

Experiment 4 Amino Acid Analysis

Table 130 shows the total amino acid composition of the hydrolyzate for different process conditions. Serine and treo in high lime loading experiments.

Except for the relatively high fluctuations in the nin and cysteine contents, the composition of the final product does not change with treatment conditions.

As can be seen, the limeless experiment produces lower protein concentrations in the hydrolyzate

TABLE 130

Total Amino Acid Composition with Different Process Conditions in Protein Hydrolysis of Shrimp Head Wastes

Condition

100 ℃

60 min

01 lime

100 ℃

120 min

02 lime

100 ℃

120 min

01 lime

100 ℃

120 min

Wuxi Lime

75 ℃

120 min

01 lime

125 ℃

120 min

01 lime

ASP 966 1019 927 978 946 940

GLU 1568 1585 1550 1568 1503 1520

SER 457 392 * 433 446 441 438

HIS 000 000 000 000 000 000

GLY 777 831 732 726 705 742

THR 357 230 * 401 446 440 377

ALA 715 753 728 720 669 717

TAU 000 000 000 000 000 000

ARG 700 647 759 490 * 794 660

TYR 382 427 378 394 383 413

CYS-CYS 067 048 082 142 109 074

VAL 579 613 608 617 624 630

MET 219 215 221 225 215 214

TRP ND ND ND ND ND ND

PHE 443 490 443 467 457 481

ILE 401 432 431 430 433 451

LEU 860 894 875 902 883 897

LYS 779 731 734 752 753 759

PRO 730 692 697 697 645 685

ND: not measured Value is g AA / 100 g total amino acids

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Table 131 shows the free amino acid composition of the hydrolyzate for different process conditions Composition variability is greater than for total amino acids

Larger treatment conditions affect sensitive amino acids; Stronger conditions (eg, longer time, higher temperature, or

Many lime loadings) accelerate the degradation reaction and produce different compositions, especially in free amino acid crystals

Tryptophan represents about 2% of the free amino acid composition, while taurine is close to 4%.

Can be used as an estimate of their concentration

Table 131

Free Amino Acid Composition at Different Process Conditions for Protein Hydrolysis of Shrimp Head Wastes

Condition

100 ℃

60 min

01 lime

100 ℃

120 min

02 lime

100 ℃

120 min

01 lime

100 ℃

120 min

Wuxi Lime

75 ℃

120 min

01 lime

125 ℃

120 min

01 lime

ASP 161 385 209 293 216 275

GLU 349 554 386 446 408 420

ASN 187 083 215 240 253 212

SER 301 415 317 337 320 359

GLN 167 000 205 269 329 018

HIS 000 000 000 000 000 000

GLY 851 861 655 654 580 659

THR 244 138 300 338 325 291

CIT 052 113 058 038 067 036

B-ALA 050 025 009 002 000 015

ALA 871 921 841 845 785 898

TAU 651 563 431 384 348 395

ARG 1145 937 1163 653 1146 951

TYR 393 435 472 540 506 525

CYS-CYS ND ND ND ND ND ND

VAL 410 461 484 487 485 550

MET 278 322 322 336 301 289

TRP 278 257 232 217 216 186

PHE 455 474 517 615 587 556

ILE 386 392 482 432 445 572

LEU 763 815 890 982 960 975

LYS 1031 939 982 1098 932 982

PRO 978 910 828 795 791 837

ND: not measured Value is g AA / 100 g total free amino acids

An average of 40% of the total amino acids are present as free amino acids.

Obtained

Thermo-chemical treatment of shrimp waste produces a mixture of free amino acids and small soluble peptides, which are possible nutritional products

Hydrolyzate products contain high fractions of essential amino acids, making them high-quality nutrients for unit animals.

Shows a comparison between the total amino acid composition and the requirements for various livestock, since histidine is underestimated during the analysis.

Meet or exceed the essential amino acid requirements of animals during the growing phase, using the 178 g / 100 g value calculated for the waste material.

High quality protein supplements are produced

Table 132

Amino Acid Analysis of Products and Essential Amino Acid Requirements for Various Livestock (Shrimp Head Waste)

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ASN 215

GLN 205

ASP 927 209

GLU 1550 386

SER 433 317

HIS 131 100 103 140 126 000 000

GLY 732 655

THR 175 264 243 350 250 401 300

ALA 728 841

ARG 375 282 417 550 000 759 1163

VAL 263 218 207 415 267 608 448

CYS 200 * 241 * 367 * 400 * 192 * 082 ND

MET 200 * 241 * 207 225 192 * 221 322

TYR 438+ 405+ 293+ 585+ 375+ 378 472

PHE 438+ 405+ 140 315 375+ 443 517

ILE 228 205 173 365 250 431 482

LEU 306 327 417 525 250 875 890

LYS 447 350 400 575 358 734 992

TRP 044 091 083 105 075 ND 232

PRO 697 828

* Cysteine + Methionine + Tyrosine + Phenylalanine ND Not measured

All values are g amino acids / 100 g protein

In addition to ~ 20% ash, shrimp head waste contains 64% protein + chitin, both used to produce some useful products

Thermo-chemical treatment of the waste with lime can result in a well-balanced amino acid that can be used as an animal feed supplement.

Produces protein-rich material with a sufficient volume of carotenoids by recovering the treated mixture and centrifuging the liquid product.

Finally, residual solids rich in calcium carbonate and chitin are also used to produce chitin and chitosan via well known methods.

Can be

For all conditions of temperature studied, lime loading and time, no significant change in conversion occurred after 30 minutes of reaction.

Almost no amino acid degradation was observed for all conditions and treatments up to 2 h.

Lime addition is required during the treatment to obtain higher nitrogen conversion to the liquid phase. It is also necessary for the production of chitin and chitosan to be produced.

Will reduce the chemicals required for further treatment of the solids

Products obtained by lime treating shrimp waste materials are suitable, meeting or exceeding the essential amino acid requirements for the unit animal.

Become a protein supplement

While only exemplary embodiments of the invention have been described above in detail, they are intended to be within the spirit and scope of the invention.

It will be understood that variations and modifications of the invention are possible.

Claims (1)

Claim 1
Alkali is added to the protein source to form a slurry; The slurry is heated to a temperature sufficient to hydrolyze the protein in the protein source.
Obtaining a reaction solution; Separating solid from the reaction solution; Neutralizing the reaction solution with an acid or acid source to produce a neutralized liquid;
Concentrating the neutralized liquid to produce concentrated liquid and water; Comprising conveying water to the slurry before or during the heating step
White solubilization method
Claim 2
The method of claim 1 further comprising polishing the protein source.
Claim 3
The method of claim 1 wherein the alkali comprises calcium oxide or calcium hydroxide.
Claim 4
The method of claim 1 wherein the alkali is magnesium oxide, magnesium hydroxide, sodium hydroxide, sodium carbonate, potassium hydroxide, ammonia, and their
Comprising a compound selected from the group consisting of any combination
Claim 5
The method of claim 1, wherein the heating further comprises producing ammonia and neutralizing the ammonia with an acid.
Claim 6
The method of claim 1 further comprising returning the separated solid to a protein source.
Claim 7
The method of claim 1 further comprising separating the reactive solid from the inert solid in the separated solid.
Claim 8
The reaction solution of claim 1, wherein the reaction solution is held at an elevated temperature for a time sufficient to destroy all or substantially all prions in the reaction solution.
How to further include
Claim 9
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The method of claim 1 wherein the temperature rise is 125-250 ° C. and the time is 1 second to 5 hours.
Claim 10
The method of claim 1 further comprising separating the solid from the neutralized liquid.
Claim 11
The method of claim 10, wherein the separated solid comprises an alkali and further comprises adding the separated solid to a protein source.
Claim 12
The method of claim 1 wherein the concentration of the neutralized liquid further comprises evaporating the neutralized liquid.
Claim 13
The method of claim 1 wherein the concentration of the neutralized liquid further comprises filtering the neutralized liquid.
Claim 14
The method of claim 1 wherein the concentration of the neutralized liquid further comprises freezing the neutralized liquid.
Claim 15
The method of claim 1 wherein the concentration of the neutralized liquid further comprises adding an immiscible amine.
Claim 16
The method of claim 1 further comprising drying the concentrated liquid.
Claim 17
The method of claim 1 further comprising generating a process heat and reusing the process heat.
Claim 18
A heated reactor operable to react the protein source and the alkali to produce a reaction solution; Operate to separate solids from the reaction solution
Possible solid / liquid separators; A neutralization tank operable to allow addition of acid to the reaction liquid to produce a neutralized liquid; Neutralized
A concentration tank operable to concentrate the liquid and produce concentrated liquid and water; Water is passed from the concentration tank to the heated reactor
A conduit operable to cause; And at least one heat exchanger operable to exchange process heat.
System for raising
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Claim 19
19. The system of claim 18, further comprising a polisher operable to polish the protein source.
Claim 20
19. The system of claim 18, wherein the heated reactor comprises a stirred tank.
Claim 21
19. The arm of claim 18, wherein the arm is operable to collect ammonia from the heated reactor and to allow neutralization of the ammonia with acid.
System additionally contains Monia Collector
Claim 22
19. The system of claim 18, further comprising a conduit operable to return the separated solids to the heated reactor.
Claim 23
19. The system of claim 18, further comprising a density separator operable to separate the reactive solid from the inert solid.
Claim 24
24. The system of claim 23, wherein the density separator comprises a settler or a hydroclone.
Claim 25
19. The method of claim 18, wherein the reaction is carried out at a temperature and for a time sufficient to destroy all or substantially all prions in the reaction solution.
The system further comprising a holding tank operable to heat the liquid
Claim 26
19. The system of claim 18, further comprising a solid / liquid separator operable to separate the solid from the neutralized liquid.
Claim 27
19. The process of claim 18, wherein the concentration tank comprises a multi-effect evaporator, a mechanical vapor compression evaporator, a jet ejector vapor compression evaporator, a reverse osmosis membrane,
Further comprising a component selected from the group consisting of wheat nanofiltration membranes, freezers, amine recovery systems, and any combination thereof
System to include
Claim 28
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19. The system of claim 18, further comprising a dryer operable to dry the concentrated liquid.
Claim 29
The system of claim 28, wherein the dryer comprises a spray dryer or a scraped drum dryer.
Claim 30
Reaction means operable to react the protein source and the alkali to produce a reaction solution; Operable to separate solids from reaction solution
One solid / liquid separation means; Neutralizing means operable to allow addition of acid to the reaction liquid to produce a neutralized liquid; Neutralized liquid
Concentration means operable to concentrate the sieve and produce concentrated liquid and water; Passing water from the concentration tank to the heated reactor
Means operable to turn on; And at least one heat exchange means operable to exchange process heat.
System for raising
Claim 31
33. The system of claim 30, further comprising polishing means operable to polish the protein source.
Claim 32
31. The collection of claim 30, wherein the collection is operable to collect ammonia from the reaction means and to allow neutralization of the ammonia with acid.
System further comprising means
Claim 33
31. The method of claim 30, at a temperature and for a time sufficient to destroy all or substantially all prions in the reaction solution.
The system further comprises a holding means operable to heat the liquid
Claim 34
33. The system of claim 30, further comprising drying means operable to dry the concentrated liquid.

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