CN86106166A - The method for preparing amino acid composite liquid by cocoon chrysalis - Google Patents

Abstract

The present invention is to be raw material with the cocoon chrysalis, makes papoid directly act on pupa protein liquid, in 65 ℃ of reactions 80 minutes, adds veltol plus again and removes smelling removal under neutral, normal pressure, and the thing of gained is amino acid composite liquid.This law technology is simple, equipment need not corrosion-resistantly, does not produce the three wastes in the production.The amino acid that contains needed by human in the amino acid composite liquid that makes, A wide selection of colours and designs, quantity are many.Can be used for preparation height, middle-grade beverage and medicine.

Description

The method for preparing amino acid composite liquid by cocoon chrysalis

The present invention relates to adopt the biochemical method of papain enzymolysis tussah silkworm chrysalis protein.

Acid, the basic hydrolysis pupa albumen of adopting in producing at present more.(see: silkworm industry comprehensive utilization of resources technical information compilation) need to add strong acid (sulfuric acid, hydrochloric acid) and highly basic (sodium hydroxide) in the reaction, adopt technologies such as Zeo-karb and activated carbon decolorizing again, the corrosion of the necessary acid-and base-resisting of not only production process complexity, and equipment.Simultaneously with the reaction process of acid, alkali in, part effective constituent (as: tryptophane) is destroyed.Owing to adopt strong acid, highly basic, must carry out the three wastes and handle.This not only brings adverse factors economically to also giving on the technology.Also the someone utilizes pig trypsin digestion pupa albumen in recent years, produces amino acid composite liquid.(seeing 1984 the 6th phase P21 of Food science) because pig trypsinase is subjected to the restriction in Pancreas Sus domestica source, price is higher.The vigor that pancreas is put enzyme for a long time descends, and purified pig trypsinase costs an arm and a leg.The time of pig trypsin digestion is long, and the amino acid composite liquid that obtains behind the enzymolysis still has the inherent fishy smell of pupa own, so can't promote or eat.

The object of the present invention is to provide the processing method that a kind of production technique is simple, cost is low, do not have the three wastes to produce, make the product that makes to keep original aminoacid component not have fishy smell again, be suitable for eating.

The present invention is a method of producing amino acid composite liquid with the enzymolysis tussah silkworm chrysalis protein, it is characterized in that adopting papoid enzymolysis under the following conditions:

A, enzyme amount: bright pupa amount=1: 200～400(weight ratio)

B, bright pupa amount: the water yield=1: 3.0～4.5(weight ratio)

C, hydrolysis temperature: 60～70 ℃

D, enzymolysis time: 60～120 minutes

Reaction solution PH=7-8 when e, enzymolysis

The top condition of its enzymolysis is:

A, enzyme amount: bright pupa amount=1: the 270(weight ratio) (vigor of enzyme is in 600,000 units/gram)

B, bright pupa amount: the water yield=1: the 3.75(weight ratio)

C, hydrolysis temperature: 65 ℃

D, enzymolysis time: 80 minutes

Reaction solution PH=7.5 when e, enzymolysis

Add veltol plus behind the enzymolysis in the work in-process of amino acid composite liquid, its quantity is:

Bright pupa: veltol plus=5000: the 1(weight ratio)

Be the Production Flow Chart of producing amino acid composite liquid with papain enzymolysis albumen below:

Water cleans (adding water)/() colloidal mill and grinds (pump)/() reactor (adding papoid)/(heating.Transfer pH value) enzymolysis

(cooling)/() centrifuging (twice) (0 ℃)/() storage → centrifuging

(adding veltol plus)/(adding liquor) high-pressure filteration (adding Sodium Benzoate)/() ultraviolet disinfector → amino acid composite liquid.

In this production process, papain enzymolysis pupa albumen is the principal reaction part, so must be noted that the condition of control enzyme digestion reaction:

1. the consumption of enzyme

The quantity of the enzyme that adds directly influences the effect of enzymolysis.In bright pupa amount one regularly, the papoid consumption increases, and amino acid whose content also increases thereupon in the product, but is not proportional relation.As shown in Figure 1, when the consumption of enzyme reached some amount, even increase the consumption of enzyme, the amplitude that amino acid whose content increases in the product was also little.Consumption that consider to select suitable enzyme from economic angle is promptly so:

Enzyme amount: bright pupa amount=1: 200～400(weight ratio) comparatively suitable.And wherein with the enzyme amount: bright pupa amount=1: be good the 270(weight ratio).(vigor of papoid is in 600,000 units/gram).Every batch of the vigor of enzyme is different, has both made with a collection of enzyme, and along with the prolongation of storage period, the vigor of enzyme also descends thereupon.Therefore must measure the vigor (method is attached) of enzyme before using, converse the consumption of actual required enzyme according to the vigor of the enzyme that records by following formula:

The actual amount of enzyme=(consumption of the standard vigor * enzyme of enzyme (by the standard vigor))/(vigor of the enzyme that records)

The papoid that this law is used is the standard vigor with 600,000 units/gram.

2. concentration of reactants

When concentration of reactants lower, promptly the water yield of Jia Ruing more for a long time, not only amino acid whose total amount is lower in the product, and has reduced usage ratio of equipment.And concentration of reactants is when higher, though the plant factor height because the system thickness, mixes, conducting heat all encounters difficulties, so amino acid whose total amount is not high yet in the product.Bright pupa amount all is 40 grams in the table 1, except that the concentration of reactants difference, and the contrast of total amino acid content situation in products therefrom when other condition is all identical:

Table 1

The water yield: bright pupa amount	2.25	3	3.75	4.5	5.25
						Total amino acid content in the product (gram)	2.079	2.688	2.94	2.772	2.2785

As can be seen from the table when the water yield: bright pupa amount=3～4.5: effect is better in the time of 1, and works as the water yield: the best results of bright pupa amount=3.75: 1.

3. time of enzymolysis

The reaction needed regular hour between papoid and the pupa albumen.Along with the reaction times increases, amino acid whose content also increases thereupon in the product as can be seen from Figure 2.As can be seen from Figure 2 when reaction times during at 60～120 minutes, amino acid whose content is higher in the product.When surpassing 120 minutes, though increase the reaction times, also not die is inadvisable for the amplitude that aminoacids content increases in the product, get 60～120 minutes comparatively suitable, and wherein enzymolysis time adopt 80 minutes comparatively good.So not only aminoacids content is higher in the product, and reaction time, short energy consumption was low.

4. the temperature of enzymolysis

During papain enzymolysis pupa albumen, carry out at a certain temperature, temperature is crossed the activity of low enzyme and is not given full play to, and the too high part enzyme of temperature can loss of activity.When other reaction conditions was identical, the two all can cause, and amino acid whose content reduced in the product.As shown in table 2, be both 40 gram pupas when other condition is identical, aminoacids content situation contrast in the product under differing temps:

Table 2

Hydrolysis temperature ℃	50	60	70	80
					Aminoacids content in the product (grams per liter)	20.7	23.75	24.15	17.7

By last table can get temperature of reaction generally 60～70 ℃ comparatively reasonable, and papain is a best results at 65 ℃.

5. the pH value of enzymolysis

Bright pupa will carry out under suitable pH value when enzymolysis, and when other reaction conditions was identical, reaction solution was the acid or alkaline activity that all can influence enzyme, makes amino acid whose content reduction in the product, and is as shown in the table:

Table 3

When PH=7-8, being more favourable to enzyme digestion reaction as can be seen from the table, is optimum when PH=7.5 especially.The homogenate of bright pupa adds that pH value is between 6.5～6.8 behind the water, and available like this about 30% sodium bicarbonate is transferred to 7.5 with the pH value of reaction solution.

In the work in-process of amino acid composite liquid, adding veltol plus behind the enzyme digestion reaction, not only can eliminate the fishy smell of cocoon chrysalis, but also can improve mouthfeel.The quantity that adds is:

Bright pupa amount: veltol plus amount=5000: the 1(weight ratio)

The present invention adopts papoid to directly act on pupa protein liquid, technology is simple, enzymolysis efficiency is high, with short production cycle, do not produce the three wastes, equipment is not had corrosion-resistant requirement.The papoid source is sufficient, price is low, and the amino acid composite liquid that makes is essential amino acid A wide selection of colours and designs but also content height not only, does not have the peculiar smell mouthfeel good.

Embodiment 1:

Get 20 kilograms of bright cocoon chrysalis, with tap water flushing three times, boiling water is washed two times.Control water purification branch adds 75 kg of water (bright pupa is heavy: water is heavy=1: 3.75) then, makes it to wear in colloidal mill homogenate (5～20 microns of particle diameters), is squeezed in the reactor by beverage pump.Insulation jacket to reactor feeds steam, start agitator simultaneously, make slurries be warming up to 65 ℃, and the pH value of slurries is transferred to 7.5 with 30% sodium bicarbonate, add 74 gram papoids (vigor that records enzyme is 600,000 units/gram) this moment, keep reaction 80 minutes (during this period of time getting final product) with this understanding with intermittently stirring.Be cooled to room temperature then, filter (filter cloth: first pass woven dacron, second time with double-deck tussah silk silk) twice with whizzer (4000 rev/mins).About 0 ℃, preserve and spend the night.Next day is centrifuging again.The liquor and the 4 gram veltol pluss (1 gram veltol plus adds 500 gram water dissolution) that add 1.5 kilograms then, mix the back high-pressure filteration, add Sodium Benzoate 150 grams again, filtrate is passed through ultraviolet disinfector, promptly obtain 74 kilograms of amino acid composite liquid finished products, amino acid whose content is 18.5 grams per liters.

Embodiment 2:

Repeat operation and the condition of embodiment 1, the vigor that wherein records papoid is 550,000 units/gram, and the amount that then adds papoid is:

74 * 60/55=80.7(gram)

76 kilograms of the amino acid composite liquid finished products that obtains, amino acid whose content are 18.7 grams per liters.

Embodiment 3:

Get 30 kilograms of dried pupas (one kilogram of dried pupa is equivalent to 2.5～2.8 kilograms of bright pupas), with tap water flushing three times, boiling water is washed one time, logical steam to boiling was soaked 24 hours, add 285 kg of water then, make it in colloidal mill, to wear into homogenate (5～20 microns of particle diameters), squeeze in the reactor by beverage pump.Insulation jacket to reactor feeds steam, starts agitator simultaneously, makes slurries be warming up to 65 ℃ rapidly, and regulates PH to 7.5, adds 383 gram papoids (vigor of enzyme is 580,000 units/gram) this moment.Intermittently stir, reacted 80 minutes.Reflux cooling is to room temperature, twice of centrifuging.Filtrate is preserved about 0 ℃ and is spent the night, and next day is centrifuging again.The liquor and the 17.6 gram veltol pluss that add 7.5 kilograms then mix the back high-pressure filteration, add Sodium Benzoate 550 grams, and filtrate promptly obtains 272 kilograms of amino acid composite liquid finished products by ultraviolet disinfector, and amino acid whose content is 19.2 grams per liters.

The papoid vigour-testing method

One, reagent

1. enzyme liquid: enzyme 0.1 gram is put in the mortar, adds a small amount of frozen water and grinds 100 milliliters of frozen water constant volumes.

2. enzyme diluent: (face and use stylish preparation)

Get 0.4577 gram cysteine hydrochloride, get 0.223 gram EDTANa ₂Transfer PH to 5.0 with 4N sodium hydroxide or 4N hydrochloric acid before 100 milliliters of the constant volumes, constant volume.

3.4N sodium hydroxide.

4.4N hydrochloric acid.

5. casein solution: get 100 milliliters of 1.79 gram Sodium phosphate dibasic constant volumes, be 0.05M Sodium phosphate dibasic liquid, get 0.6 gram casein and add 80 milliliters of above-mentioned disodium phosphate solns, transfer behind the PH=7.0 100 milliliters of constant volumes with 1/10N hydrochloric acid after the stirring and dissolving.

6. trichloroacetic acid solution: get 1.896 milliliters of 99% glacial acetic acids, the back that is dissolved in water adds 1.805 gram sodium acetates, and the dissolving back that is mixed adds 1.797 gram Tricholroacetic Acids dissolving back constant volumes to 100 milliliter.

7. standard amino acid: (50.0 micrograms tyrosine/milliliter) gets 5 milligrams of tyrosine with 100 milliliters of 1/10N hydrochloric acid constant volumes.

Two, measuring method

1. get 5 milliliters of enzyme liquid and put in 10 milliliters of volumetric flasks, 10 milliliters of enzyme-added diluent constant volumes, this is for supplying examination enzyme liquid.

2. get for examination enzyme liquid and in test tube with ground stopper, put 37 ℃ of water bath heat preservations 10 minutes for 1 milliliter, add and give 5 milliliters of hot casein solutions, the reaction 10 minutes of clocking.

3. add 5 milliliters of Tricholroacetic Acids during by 10 minutes, shake all and filter.

4. get confession examination enzyme liquid and add 5 milliliters of Tricholroacetic Acids for 1 milliliter, add 5 milliliters of casein solutions after 10 minutes, filter, be blank solution.

5.275nm measure absorption value A.

6. the 275nm absorption value of the accurate tyrosine of mark (is with water blank) As.

Three, calculate

The vigor of papoid (units/gram)=A/As * 50 * (trial-product extension rate)/(trial-product is heavy)

Claims (3)

1, a kind of method of producing amino acid composite liquid by the enzymolysis tussah silkworm chrysalis protein is characterized in that, adopts papoid to carry out enzymolysis under the following conditions:

A, enzyme amount: bright pupa amount=1: 200～400 (weight ratios)

B, bright pupa amount: the water yield=1: 3.0～4.5 (weight ratio)

C, hydrolysis temperature: 60～70 ℃

D, enzymolysis time: 60～120 minutes

Reaction solution PH=7～8 when e, enzymolysis

2, the amino acid composite liquid according to claim 1 is characterized in that, adopts the condition of papain enzymolysis to be:

A, enzyme amount: bright pupa amount=1: the 270(weight ratio) (vigor of enzyme is in 600,000 units/gram)

B, bright pupa amount: the water yield=1: the 3.75(weight ratio)

C, hydrolysis temperature: 65 ℃

D, enzymolysis time: 80 minutes

Reaction solution PH=7.5 when e, enzymolysis

3, the amino acid composite liquid according to claim 1 or 2 is characterized in that, adds veltol plus in the work in-process of amino acid composite liquid, and its quantity is:

Bright pupa amount: veltol plus amount=5000: the 1(weight ratio)

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