CN105166322A - Method for high yield production of aerobic single-cell protein by autolysis process - Google Patents
Method for high yield production of aerobic single-cell protein by autolysis process Download PDFInfo
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Abstract
The invention relates to the technical field of single-cell protein processing, in particular to a method for high yield production of aerobic single-cell protein by an autolysis process. The process is characterized in that in each pre-processing unit, the material obtained by a centrifugal dewatering step is transported to a post-processing unit to conduct centralized treatment, and the post-processing unit consists of an autolysis step, a drying step, a sterilization step and a crushing and packaging step. The technology and the prepared product involved in the invention employ the autolysis process, during autolysis, autolytic enzyme fully decomposes macromolecular nutrients and cell walls, autolysis can produce high content free amino acid and free nucleotide, and at the same time cell wall polysaccharides are decomposed, and cells can produce permeability so as to ensure that active substances can flow out of cells to as to be directly absorbed and utilized by animals. Autolysis is carried out after centrifugal dewatering, high efficiency of autolysis can be ensured, and the content of water needing to be removed in a drying process after autolysis can be reduced, thereby reducing the cost of drying.
Description
[technical field]
The present invention relates to single cell protein processing technique field, particularly relate to a kind of method that high yield uses self-dissolving explained hereafter aerobic single cell protein.
[background technology]
Single cell protein (Singlecellprotein is called for short SCP) is also known as microprotein or mycoprotein, utilize industrial wastewater, waste gas, natural gas, petroleum alkane class, agricultural and sideline converted products and organic waste etc. as culture medium, the unicellular microorganism such as culture yeasts, non-pathogenic bacterium, miniature bacterium, fungi, then making after purge drying process, is the important protein source of food industry and feed industry.Single cell protein product contains rich in protein, and amino acid whose A wide selection of colours and designs ratio is suitable.Single cell protein also containing abundant fat, carbohydrate, nucleic acid, vitamin and inorganic salts, and containing mineral matter elements such as calcium, phosphorus, potassium, iron, magnesium, sodium, manganese, also contains multiple enzyme.Yeast, yeast hydrolyate have been used successfully to the substituting of fish meal protein in animal feed, animal immune strengthens, the improvement etc. of feed mouthfeel.Single cell protein nucleotide and derivative thereof have very important biochemical functions, and main function has: as the raw material of RNA and DNA synthesis; As the Auto-regulator of metabolism; As the composition of coenzyme, as coacetylase; Activating the important intermediary of glycogen and glycoprotein synthesis, is also the intermediary of phosphatide synthesis, activation intermediate product; Participation hereditary information is transmitted; ATP is the direct energy of life process; AMP regulates the shrinkage of smooth muscle and adjustable CBF; Nucleotides can also as the donor etc. of methyl.
Due to the consideration that harmful microorganism, harmful chemical, heavy metal etc. can work the mischief to Product quality and safety.Or carry out single cell protein product of less types of industrialization development at present, mainly concentrate on that use safety risk is low, the product that comes from food industry or byproduct carry out cultivation and the production of single cell protein as raw material.If brewer's yeast and enzymolysis brewer's yeast are as the use of feedstuff on animal, beer needs to reach strict food security standard when producing, the organic accessory substance produced by it and the biology of Yeast protein, chemistry, physical hazard all reach minimum, thus provide reliable safety assurance for the application of feed industry.
Patent of invention WO2009059163A1 provides a kind of method of cultivating single cell protein, the single cell protein cultivated out can be used as the protein sources of high-quality in animal feed, the fluid nutrient medium that this technology uses the waste water of Foods or drinks processing factory as brewery produces to cultivate as thalline.There is in Wastewater From The Beverage Industry the nutriment such as abundant organic matter, nitrogen phosphorus, and there is no the pollution of harmful microorganism, toxic chemical, although be not worth for these waste water beverage plant, and process can pay larger cost containing waste water that is organic and nitrogen phosphorus, angle is cultivated from microorganism, nutriment in waste water can become nutritious culture medium, can be used for producing specific single cell protein.Patent of invention WO2009059163A1 by regulating the ratio of various nutriment in waste water, use the bacterium through improvement of genes to produce protein content is higher, be of high nutritive value single cell protein product, this single cell protein is the mixture of a kind of bacterium, mainly containing aerobic bacteriums such as micrococcus luteus, bacillus, nitrobacteria, alcaligenes.Simultaneously; Wastewater From The Beverage Industry nutrition content after cultivation microorganism can reduce; it reduce the degree of water pollution; this patented technology not only can realize the production of high quality protein but also organic concentration in waste water can be reduced to below statutory standards; thus there is the dual value of economy and environment protection, therefore this technology has very good industrialization prospect.Because the growth cycle of bacterium is short, nutriment utilization ratio is high, the output that single cell protein is cultivated is very high, and this provides guarantee economically for industrialization development, but due to the feature of bacterial growth, during cultivation, the content of solid content is very low, generally can lower than 4000mg/L.If single cell protein will be processed into product, just need to consider how to be extracted from water and the dry product being processed into low moisture content of low cost by single cell protein.Based on the characteristic of the single cell protein described in patent of invention WO2009059163A1, current process technology has following defect:
1) single cell protein is just used as general proteins raw material and is produced by processing technology, just concentrate on the dehydration of albumen, drying, sterilizing, be produce albumen for target, the bacterium reckoned without in patent of invention WO2009059163A1 produces high-quality product by further processing method;
2) single cell protein that patent of invention WO2009059163A1 technology is turned out finds, containing a large amount of endogenous enzymes, specifically to comprise nuclease, protease and lysozyme through research.These enzymes can decompose macro-nutrients, therefore use the endogenous enzymes of self-dissolving technology bacterium can go out multiple active material by enzymolysis, as free amino acid, nucleotides, monose etc.But existing technique is convection drying and microwave sterilization after dewatering, this just destroys most endogenous enzymes, therefore also would not produce active material;
3) protein digestibility of existing processing technology is still not high, still has the space promoted further;
4) single cell protein has the nucleic acid of high level, nucleic acid decomposition is not nucleotides and is still in cell by common process drying process, and by certain method by nucleic acid decomposition be nucleotides single cell protein can be made to be applied to animal feed time there are food calling and these two obvious effects of immunity, existing technique does not consider to be how the nucleotides of high value by nucleic acid decomposition.
5) although the output of the single cell protein cultivated is very high, but the production capacity of following process equipment still can be significantly higher than the production capacity of cultivating single cell protein, the production capacity of the process equipment of such as patent of invention CN2014100556582 is limited to the low yield of cultivation output and cannot reaches the highest production capacity, even can not reach process equipment average productivity.Which results in the idle of investment goods and waste.
The patent No. be 201410055658.2 Chinese patent disclose a kind of method utilizing biological clay standby single cell protein white powder, utilize biological mud to produce the resource recycle method of single cell protein (SCP), after the biological mud of biological cultivation system precipitation being discharged is concentrated, through centrifugal dehydration, hot blast is dried instantaneously, air-flow cools fast, after microwave sterilization, can obtain unicellular crude protein powder.This patent is the earlier application of this department, but owing to lacking self-dissolving operation, cell protein utilization ratio is lower.Self-dissolving refer to thalline utilize the endogenous enzymes (protease, nuclease, lysozyme etc.) of self endobacillary polymer substance is hydrolyzed to amino acid, nucleotides, when micromolecular processes such as sugar.Autolytic process is a more complicated process, by various factors, comprises bacterial species, moisture, temperature, pH value, time, endogenous enzymes kind etc.The more important thing is under field conditions (factors), self-dissolving is not single process, be the corrupt after death last process of organism, macromolecular substances is decomposed into the nutriment such as amino acid, nucleotides through self-dissolving, and this provides important source of nutrition for putrefactive microorganisms grows.Therefore, the result of self-dissolving is also because microbe species, Autolysis Condition, target product etc. have bigger difference, and self-dissolving unsuccessfully can cause spoilage product as generations such as ammonia, hydrogen sulfide, trimethylamines.
The patent No. is that 2008800240991 and 2011102810796 two Chinas patent discloses yeast and use self-dissolving technique to process, and produces yeast autolyzate, in order to improve product special flavour.But these two patents are also not suitable for this patent, the raw material that this patent application adopts is anaerobic bacteria, bacteria characteristic and aerobic bacteria difference larger, the mode of self-dissolving also has very big difference, such as the time of anaerobic bacteria self-dissolving is longer, and need the outer enzyme that adds to carry out promotion self-dissolving, anaerobic bacteria self-dissolving has more restriction for reaction vessel, moisture content, pH, temperature etc., and product is difficult to control, and easily produces the spoilage product of high level.
[summary of the invention]
The object of the invention is to make up the above-mentioned deficiency preparing single cell protein and defect, provide a kind of high-quality, be rich in the method for use self-dissolving explained hereafter aerobic single cell protein of active component.
To achieve these goals, design a kind of method that high yield uses self-dissolving explained hereafter aerobic single cell protein, comprise several preprocessing unit and an aft-loaded airfoil unit, preprocessing unit comprises precipitation concentration step, centrifugal dehydration step and trafficking step, the material of 78% ~ 85% moisture content will obtained after centrifugal dehydration step in each preprocessing unit, be transported to aft-loaded airfoil unit after cooling to focus on, described aft-loaded airfoil unit comprises autolysis process, drying steps, sterilization steps and crushing packing step, and described autolysis process is as follows:
A) from the mass transport of preprocessing unit to being incubated feed bin, be then delivered to self-dissolving equipment and carry out self-dissolving process;
B) self-dissolving equipment is by heating up to the heat supply of twin-screw hollow blade, reaches the temperature requirement of self-dissolving, by designing and regulating the cavity volume supporting with charging rate, reaches the time requirement of self-dissolving;
Before the twin-screw hollow blade of c) described self-dissolving equipment, 1/4 to 1/2 is segmented into bringing-up section, and object is that heating material is temperature required to self-dissolving, and remaining segment is soaking zone, keeps material from solubility temperature;
D) temperature requirement of self-dissolving is: the temperature of charge after heating remains on 65% ~ 95 DEG C;
E) time requirement of self-dissolving is: the time that material is warming up to target temperature by bringing-up section should be less than 30 minutes, and the self-dissolving time of soaking zone should at 30 minutes ~ 360 minutes;
The material moisture of described self-dissolving requires: after self-dissolving, moisture content of material should be not less than 70%;
Described sterilization steps adopts microwave sterilization.
Temperature of charge after described heating remains on 65% ~ 80 DEG C, and the self-dissolving time is 60 ~ 180 minutes.
Concrete grammar step is as follows:
(1) preprocessing unit:
A. precipitation concentration: the mycoprotein that bioreactor produces is entered precipitation concentration pond by pipeline, and mycoprotein is at the concentration basin bottom precipitation of taper, and bottoms material moisture content is reduced to less than 97%, can carry out follow-up dehydration procedure;
B. use mateiral pump that concentration basin bottoms material is delivered to centrifugal-dehydration device;
C. centrifugal dehydration: use screwed bonnet type centrifugation apparatus to carry out Separation of Solid and Liquid, centrifugal rear material water ratio is reduced to 78% ~ 85%, material is in wet pureed; Described centrifugal sloughed moisture is the ECW of mycoprotein, and in born of the same parents, moisture still remains in cell, and this creates condition for intracellular endogenous enzymes carries out self-dissolving; The material of described centrifugal rear 78% ~ 85% moisture content can carry out autolysis;
D. transport: be transported to aft-loaded airfoil unit after the material cooling after centrifugal and focus on; Described cooling is that material is cooled to less than 10 DEG C after centrifugation; Described transport is, by thermal insulated vehicle, the wet pureed material shipment of preprocessing unit output is carried out following process to aft-loaded airfoil unit; Time of described transport should in 0 ~ 12 hour, and the material temperature that material arrives aft-loaded airfoil unit is not higher than 5 DEG C of material temperature when leaving preprocessing unit;
(2) aft-loaded airfoil unit:
A. self-dissolving: from the mass transport of preprocessing unit to being incubated feed bin, being then delivered to self-dissolving equipment and carrying out self-dissolving process; Self-dissolving equipment, by heating up to the heat supply of twin-screw hollow blade, reaches the temperature requirement of self-dissolving, by designing and regulating the cavity volume supporting with charging rate, reaches the time requirement of self-dissolving; Before the twin-screw hollow blade of described self-dissolving equipment, 1/4 to 1/2 is segmented into bringing-up section, and object is that heating material is temperature required to self-dissolving, and remaining segment is soaking zone, keeps material from solubility temperature; The temperature requirement of self-dissolving is: the temperature of charge after heating remains on 65% ~ 95 DEG C; The time requirement of self-dissolving is: the time that material is warming up to target temperature by bringing-up section should be less than 30 minutes, and the self-dissolving time of soaking zone should at 30 minutes ~ 360 minutes; The material moisture of described self-dissolving requires: after self-dissolving, moisture content of material should be not less than 70%;
B. dry: the material after self-dissolving is delivered to air-flow flash drying equipment by flood dragon feeding machine and carries out drying; The material Contact Temperature of described expansion drying should lower than 100 DEG C, and the moisture content of described dried material should 10% ~ 40%;
C. sterilizing: material is after drying delivered to tunnel type micro wave drying sterilization equipment by gas lift equipment and carries out sterilizing; After described microwave sterilization, material water ratio is 5% ~ 10%;
D. crushing packing: the material after sterilizing is packaged into finished product after crushed.
Described self-dissolving equipment comprises: belt wheel, reductor, travelling gear, cover plate, charging aperture, heating hollow blade, insulation hollow blade, overflow plate, discharge gate, steam is imported and exported, motor, host shell and transmission main shaft, host shell top is provided with cover plate, transmission main shaft is provided with in host shell, transmission main shaft is provided with twin-screw hollow blade, cover plate is provided with charging aperture, discharging opening is provided with bottom host shell, overflow plate is provided with near discharging opening in host shell, transmission main shaft one end connection for transmission gear, travelling gear and output end of reducer gears meshing, reductor input connects motor output end by belt wheel, the transmission main shaft other end connects steam or conduction oil is imported and exported, before described twin-screw hollow blade, 1/4 to 1/2 is segmented into bringing-up section, remaining segment is soaking zone.Two transmission main shafts and two twin-screw hollow blades are provided with in host shell.
Adopt the aerobic single cell protein that above-mentioned method is obtained, employing protein content is the aerobic single cell protein water content 5.0%-10.0% that the mycoprotein of 33%-60% obtains; Protein content 33.0%-60.0%; The free nucleotide of unit-protein is not less than 1.40%; The free base of unit-protein is not higher than 0.90%; Free base and free nucleotide ratio range should at 0-0.60.
Compared with the existing technology, technique and the product made thereof have the following advantages in the present invention:
1) embody single cell protein and there is this situation of endogenous enzymes, take full advantage of the nuclease, protease, the lysozyme that have in mycoprotein of the prior art;
2) self-dissolving technique is employed, in autolytic process, autolytic enzyme fully decomposes large molecular nutrition thing and cell membrane, free amino acid, the free nucleotide of high level can be produced by self-dissolving, make cell wall polysaccharides be decomposed simultaneously, make cell produce permeability thus guarantee that active material can flow out extracellular, thus directly can be absorbed by animal;
3) self-dissolving technique can not need the ECW of mycoprotein, endogenous enzymes can play a role in intracellular microenvironment, therefore, self-dissolving is carried out after have employed centrifugal dehydration, the high efficiency of self-dissolving can be guaranteed, also can reduce the moisture removed needed for the drying process after self-dissolving, thus decrease drying cost;
4) quick heating of the self-dissolving equipment used, temperature are easy to control, the time of staying of material in equipment be controlled, can guarantee that mycoprotein carries out self-dissolving under the suitableeest Parameter Conditions;
5) use the digestibility of the product of self-dissolving technique to improve further, apparent digestibility of the dry matter can bring up to 76.9% from 57.7%, and albumen apparent digestibility can be increased to 89.1% from 66.4%;
6) product made not only albumen quality is high and have various active composition, have the functional protein feedstuff of different physiological roles, physiological function comprises: food calling and the improvement improving feed intake, mouthfeel, smell, immunologic function, liver protecting, resisting stress, prevent diarrhoea, the albumen of high-quality, efficient rate; Have to have than the mycoprotein of non-self-dissolving in animal experiment and better show performance.
7) use new production technology to reduce further the energy consumption of production, reason is: a) self-dissolving preheats the material before drying, improves the temperature before drying; The cell membrane permeability of the mycoprotein b) after self-dissolving strengthens, and moisture and intracellular organic matter more easily flow out outside born of the same parents, and dry difficulty reduces further.
8) production capacity that new technology take into account the technique such as self-dissolving, drying of aft-loaded airfoil is significantly higher than single cell protein and cultivates and this situation of dehydration production capacity.Whole processing technology is divided into two sections, under the assistance of the transportation resources of efficiently controlling temperature, connect whole technological process.Thus the production efficiency of all process equipments can be played, can production capacity be significantly improved, equipment overlapping investment can be reduced and then reduce costs.
[accompanying drawing explanation]
Fig. 1 is step schematic diagram of the present invention;
Fig. 2 is the chromatogram of free nucleotide standard items in embodiment 1 in the present invention;
Fig. 3 is the chromatogram of free base standard items in embodiment 1 in the present invention;
Fig. 4 is the front view of self-dissolving equipment in the present invention;
Fig. 5 is the side view of self-dissolving equipment in the present invention;
Fig. 6 is the top view of self-dissolving equipment in the present invention;
In figure: 1. belt wheel 2. reductor 3. travelling gear 4. cover plate 5. charging aperture 6. heats hollow blade 7. and is incubated hollow blade 8. overflow plate 9. discharge gate 10. steam and imports and exports 11. motor 12. host shell 13. transmission main shafts;
Specify Fig. 1 as Figure of abstract of the present invention.
[detailed description of the invention]
Below in conjunction with accompanying drawing, the invention will be further described, and the equipment of this side's ratio juris and employing is very clearly concerning the people of this specialty.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Embodiment 1: the foundation of free nucleotide and free base detection method
Use free nucleotide and free base as the leading indicator contrasted before and after self-dissolving technical arrangement plan and product self-dissolving.Principle is: the raw material such as yeast, mycoprotein can containing free nucleotides and base through self-dissolving or enzymolysis, sample after extraction with aqueous solution enters efficient liquid phase chromatographic analysis, through C18 post, UV-detector, buffer salt make mobile phase, and external standard method can be used to do quantitative assay to 5 kinds of nucleotides, 5 kinds of nucleosides and 5 kinds of bases.
Free nucleotide refers to the summation of 5 kinds of nucleotides and 5 kinds of nucleosides, comprises cytidine monophosphate, uridylic acid, guanylic acid, inosinicacid, adenylate, cytidine, uridine, inosine, guanosine, adenosine.Free base refers to the summation of 5 kinds of nucleic acid bases, specifically comprises cytimidine, uracil, guanine, thymidine, adenine.
Single cell protein product contains abundant nucleotides, and on market, the product of yeast class is the main nucleotides supplier of feed industry.Due to process characteristic, the products such as autolysing yeast, enzymatic yeast, hydrolyzed yeast contain the nucleic acid hydrolysis thing of more free state, and research finds existing a lot of yeast product both containing free nucleotide, the base also containing high level.Free nucleotide can be absorbed fast by animal, and play important physiological function, base then can cause harmful effect to animal's liver kidney.Base contents too much means has the nucleotides of important nutritive value by enzymolysis, hydrolysis or microbial metabolism in the middle of production process.This can cause the Quality Down of yeast product, and the nucleotide content provided reduces, and then the effect of immunologic function can be provided to reduce for animal, and the base simultaneously increased such as adenine then can cause toxicity to animal.Judge the destructiveness of production technology to nucleotides of single cell protein, use can reaction process condition quality again can for control of product quality provide science, reflection nutritive value appraisal procedure just become very important.Free nucleotide represents the degree of self-dissolving, and the higher explanation of numerical value represents endogenous enzymes and reduces nucleic acid as the degree of nucleotides is higher.Free base is the product of bacterial metabolism, and the growth of content higher explanation putrefactive microorganisms is more, as the index of checking product degree of spoilage after self-dissolving.
Reagent and solution:
Except as otherwise noted, it is pure that this law agents useful for same is analysis, and water is deionized water, meets the regulation of GB/T6682 secondary water.
Potassium dihydrogen phosphate: analyze pure.
Potassium hydroxide: analyze pure.
Methyl alcohol: chromatographically pure.
Acetonitrile: chromatographically pure.
20%w/v potassium hydroxide solution: take 100 grams, potassium hydroxide, be dissolved in 500ml water, be stored in volumetric flask.
Mobile phase A: 0.1M potassium dihydrogen phosphate, takes potassium dihydrogen phosphate 27.2 grams, is dissolved in 1000ml water, adds 20% potassium hydroxide solution 1.0ml, adds water and be settled to 2 liters.For subsequent use after 0.45um water system membrane filtration.
Mobile phase B: 25% methanol solution, measures methyl alcohol 250ml and is dissolved in 750ml water, for subsequent use after stirring and evenly mixing rear 0.45um organic system membrane filtration.
Adenylate AMP:sigma#A2252.
Cytidine monophosphate CMP:sigma#C1131.
Inosinicacid IMP:sigma#I2897.
Sodium guanylate GMP:sigma#G8377.
Uridylic acid UMP:sigma#U1752.
Adenosine A denosine:sigma#A9251.
Cytidine Cytidine:sigma#C122106.
Guanosine Guanosine:sigma#V900311.
Inosine Inosine:sigma#I4125.
Uridine Uridine:sigma#V900421.
Prepare nucleotides hybrid standard liquid: use uridylic acid, adenylate, Sodium guanylate, cytidine monophosphate, inosinicacid, guanosine, adenosine, uridine, inosine, cytidine to prepare hybrid standard liquid successively, each nucleotide concentration is followed successively by 20.000ug/ml, 20.000ug/ml, 20.000ug/ml, 20.000ug/ml, 20.00ug/ml, 10.000ug/ml, 10.000ug/ml, 10.000ug/ml, 10.000ug/ml, 10.000ug/ml and divides and be filled in 0.5ml cryopreservation tube,-20 DEG C of preservations, are valid for one year.
Cytimidine Cytosine:sigma#V900462.
Uracil Uracil:sigma#V900439.
Guanine Guanine:sigma#V900473.
Thymidine Thymine:sigma#V900437.
Adenine Adenine:sigma#V900471.
Preparation base hybrid standard liquid: use cytimidine, uracil, guanine, thymidine, adenine to prepare hybrid standard liquid successively, the concentration of hybrid standard liquid caustic soda base is 16.00ug/ml, 16.00ug/ml, 20.00ug/ml, 16.00ug/ml, 16.00ug/ml according to this.Dividing is filled in 0.5ml cryopreservation tube, and-20 DEG C of preservations, are valid for one year.
Instrument and equipment:
High performance liquid chromatograph system (gradient elution, UV detect, controlled column temperature); Chromatographic column: AthenaC18,4.6 × 150mm; Assay balance; Centrifuge; Ultrasonic cleaning machine; 1mL disposable syringe; The disposable aqueous phase filter of 0.45um; 0.5ml centrifuge tube; 100ml volumetric flask; 250ml volumetric flask; 40ml blind nut glass tube with cover; 1000ul pipettor; 5000ul pipettor; 15mL plastic centrifuge tube.
Determination step:
Sample treatment:
(1) 0.1000 gram of sample is taken as in 40ml glass tube.Each testing sample repeated sampling at least two group.
(2) add water 15ml (volume of dissolution, V), and whirlpool shakes 3 times, each 1min, every minor tick 20min.
(3) ultrasonic 20min after concussion.Whirlpool concussion again.
(4) get in glass tube in test solution 3ml to 5ml centrifuge tube.12000g high speed centrifugation 15min.
(5) get in supernatant 2ml to 5ml centrifuge tube, add water 2ml, dilutes one times.Preserve the above machine that filters after mixing to measure.
By high performance liquid chromatography, sample is measured, adopts following chromatographic condition:
| Environment temperature: | 25℃～30℃ |
| Chromatographic column: | Athena C18，4.6×150mm |
| Determined wavelength: | 260nm |
| Column temperature: | 30℃ |
| Mobile phase A: | 0.1mol/L potassium dihydrogen phosphate |
| Mobile phase B: | 25% methyl alcohol |
| Sample size: | 20ul |
| Flow velocity: | 0.5mL/min |
Elution program is as following table 1:
Table 1
As shown in Figure 2 and Figure 3, this several free nucleotides, base can well be separated by above-mentioned steps.
Result calculates:
External standard method is used to go out the content of free base, nucleotides in sample.
Base, nucleosides, nucleotide content ω in sample, with mass fraction mark, numerical value represents with %, by formulae discovery:
In formula:
The chromatographic peak area of A---sample oligonucleotide, base
A
st---the chromatographic peak area of standard items
C
st---the concentration of standard items
X---a certain nucleotides or nucleosides, or base
C---nucleotide concentration ug/ml;
V---volume of dissolution ml;
M---example weight (g);
ω
x---content %;
The measurement result arithmetic mean of instantaneous value of parallel determination represents, retains 2 position effective digitals.
Precision:
Repeat above detection, make the absolute difference of twice of acquisition independent measurement result must not exceed 10% of arithmetic mean of instantaneous value.
The free nucleotide of self-dissolving handicraft product using said determination method to record and the content of free base see the following form 2, can find out that product can produce the nucleotides of high level after self-dissolving, and be higher than commercially available yeast product in table.
Table 2
Embodiment 2: self-dissolving production technology and product
Self-dissolving production technology and the product of the present invention's exploitation are the single cell protein products utilizing the culture technique of patent of invention WO2009059163A1 to make, but are not limited thereto, and are also applicable to the processing of the aerobic single cell protein product similar with this invention.
A processing method for high yield self-dissolving single cell protein, the mode of realization is that process is divided into two large divisions, has been come respectively by different production units.Wherein Part I is preprocessing unit, contain single cell protein cultivated after from precipitation concentration to the process of centrifugal dehydration.In this part, multiple preprocessing unit can be set, as 2 ~ 10 are all fine.Part II is aft-loaded airfoil unit, contains the operation of self-dissolving, drying, sterilizing.The mode that Part I to Part II have employed transport realizes.In this part, an aft-loaded airfoil unit is set.
When preprocessing unit is 1, preprocessing unit and aft-loaded airfoil unit can unite two into one.
When base unit is more than 2 and 2, preprocessing unit separates separately with aft-loaded airfoil unit.
Preprocessing unit and aft-loaded airfoil cell distance are 0 ~ 500 kilometer.Preferably within 400 kilometers.
As shown in the figure, 3 preprocessing unit and an aft-loaded airfoil unit are set in this example.
Following operation is completed in preprocessing unit:
1) precipitation concentration: regularly the mycoprotein that bioreactor produces is entered precipitation concentration pond by pipeline, mycoprotein is at the concentration basin bottom precipitation of taper, and bottoms material moisture content is reduced to less than 97%, can carry out follow-up dehydration procedure.
A) the pond body diameter in the present embodiment: 15 meters, the degree of depth: 3.5 meters, dischargeable capacity: 477 cubic metres.
2) use positive displacement single-screw (single screw) pump that concentration basin bottoms material is delivered to centrifugal-dehydration device.
3) centrifugal dehydration: use screwed bonnet type centrifugation apparatus to carry out Separation of Solid and Liquid.Centrifugal rear material water ratio is reduced to 78% ~ 82%, and material is in wet pureed.
The material disposal ability of the centrifuge a) in the present embodiment is 10-30 cubic meter/hour, and working speed is 2000-3200 rev/min, and load is 1.5 tons/hour, and charging solid content is 3.0-5.0%.Solid content≤0.2% of supernatant, material recovery rate >=95%.
4) transport: be transported to aft-loaded airfoil unit after the material cooling after centrifugal and focus on.
A) before transport, material is cooled to less than 5 DEG C in the present embodiment.
B) by thermal insulated vehicle, the wet pureed material shipment of preprocessing unit output is carried out following process to aft-loaded airfoil unit in the present embodiment.
C) in the present embodiment, 3 preprocessing cell distance aft-loaded airfoil unit are all less than 300 kilometers, and the time of transport, the material temperature that material arrives aft-loaded airfoil unit was not higher than 4 DEG C of material temperature when leaving preprocessing unit in 4 hours.
Haulage time is determined by the test of different temperatures, different time processing sample.The material protein content of self-dissolving is 50%, moisture content 82%.Sample is deposited at different tests temperature and different holding time conditions, after carry out self-dissolving.The condition of self-dissolving is: temperature 70 C, and the time is 180 minutes.That tests the results are shown in following table:
Can finding out in table 3, carrying out lower than placing product in 15 hours when 10 DEG C the exception that self-dissolving can't cause nucleotides and base, illustrate that when preserving lower than 10 DEG C, the factor such as enzyme work, harmful bacteria of mycoprotein there is no great change.The transport carrying out mycoprotein under this temperature conditions can be described within a certain period of time can't impact product.
Table 3
Following operation is completed in aft-loaded airfoil unit:
5) self-dissolving: from the mass transport of 3 preprocessing unit to being incubated feed bin, being then delivered to self-dissolving equipment and carrying out self-dissolving process.
A) the present embodiment self-dissolving equipment is improved by twin-screw hollow blade dryer, by heating up to hollow blade heat supply, reach the temperature requirement of self-dissolving, the cavity volume of equipment is 7.5 cubic metres, dischargeable capacity is at 5.0 ~ 7.0 cubic metres, use overflow plate height regulable control dischargeable capacity, equipment can meet the material production of embodiment 3 preprocessing unit.
Before two twin-screw hollow blades of b) the present embodiment self-dissolving equipment, 1/3 is segmented into bringing-up section, uses the steam of 0.5-0.6Mpa pressure to heat up.Remaining segment is soaking zone, uses the steam condensate (SC) after 0.1Mpa low-pressure steam and leading portion heating to be incubated, to reach the effect maintained from solubility temperature.
C) temperature requirement of the present embodiment self-dissolving is: the temperature of charge after heating remains on 68% ~ 80 DEG C, from solubility temperature is preferably: 70 DEG C.
D) time requirement of the present embodiment self-dissolving is: the time that material is warming up to target temperature by bringing-up section should be less than 20 minutes.The self-dissolving time of soaking zone should at 60 ~ 90 minutes.
E) the material moisture requirement of the present embodiment self-dissolving is: after self-dissolving, moisture content of material should be not less than 70%.
6) dry: the material after self-dissolving is delivered to air-flow flash drying equipment by flood dragon feeding machine and carries out drying.
A) the material Contact Temperature of the present embodiment expansion drying should lower than 100 DEG C.
B) moisture content of the dried material of the present embodiment should 15% ~ 30%.
7) sterilizing: material is after drying delivered to tunnel type micro wave drying sterilization equipment by gas lift equipment and carries out sterilizing.
Total number of molds, the total number of bacteria of the material a) after the present embodiment microwave sterilization meet GB13078-2001 forage health standard.
B) after described microwave sterilization, material water ratio is 5% ~ 10%.
8) crushing packing: the material after sterilizing is packaged into finished product after crushed.
The single cell protein made through above-mentioned production technology should have following quality index simultaneously:
A) the free nucleotide content of unit-protein should higher than 1.40%
B) the free base contents of unit-protein should lower than 0.90%
C) ratio of free base and free nucleotide should in 0 ~ 0.60 scope.
The key index of the product that the mycoprotein that protein content is 50% obtains after the processing of above-mentioned processing method is in table 4.
Table 4
Embodiment 3: digestibility is tested
The mycoprotein and the difference of common mycoprotein on apparent digestibility that use self-dissolving technique is have evaluated by animal experiment.
Digestibility is that in the feed ingredient absorbed of animal, digestible nutrient accounts for the percentage eating feed nutrient.Indicator method is exactly with that exist in feed or the artificial indicator evenly mixed.The amount of contained unit indicator from unit feed and excrement of feeding after fish asks calculation digestibility.With the digested value of feedstuff nutrient for working out according to carrying out formula, significant for digestive utilization ratio, the pollution of minimizing feed substances to cultivation water environment improving mixed feed.
Testing program:
Assessment raw material: by protein content be 50% mycoprotein carry out self-dissolving production according to the method for embodiment two, common non-self-dissolving protein content is all the same batch of single cell protein of 50%.
Test feed is made up of " 70% basal feed+30% raw material to be measured ".Add 0.01% yittrium oxide in test feed as indicator, adopt step by step diffusion method Homogeneous phase mixing in feed powder.During test preparation feed, all raw materials all cross 40 orders, make the pellet of particle diameter 3mm with granulator.
Test is carried out in 145 liters of plastic aquaria, and water source is aeration running water, and aquarium uses oxygen charging pump inflation, and each aquarium uses heating rod temperature control.Change water 2/3 every day, keep water cleaning.Duration of test water temperature 15 ± 1 DEG C, pH6.5-7.5, dissolved oxygen >=4.0mg/l.Test fish is rainbow trout.
Basal feed and 2 kinds of test feeds totally 3 test group, often organize 3 repetitions, totally 9 aquariums.Every case puts 50 tail fishes.
After test fish input aquarium tames 7 days, test feed of throwing something and feeding, started formal test after 7 days.Duration of test, be satiated with food every day the 2 (8:30 that throw something and feed; 15:30), each bait throwing in after 0.5 hour with siphon pipe by residual bait sucking-off.Every day collects ight soil in test steck excrement peak period, by close diddle-net, ight soil is pulled out in time at every turn, then with tweezers, excrement particles complete for coating is chosen in culture dish, dry in 70 DEG C of baking ovens to constant weight, then ight soil is put into drier to wait to analyze, till ight soil enough detects.
Result calculates:
The computing formula of test feed and basal feed dry, protein apparent digestibility is:
Apparent digestibility of the dry matter (%)=100 × (1-B/B ')
The apparent digestibility of nutritional labeling (%)=[1-(A '/A) × (B/B ')] × 100
Wherein, A, B are respectively certain nutritional labeling and yttria levels in feed, and A ', B ' are respectively certain corresponding nutritional labeling and yttria levels in ight soil.
Raw material dry, protein and amino acid whose apparent digestibility computing formula are
[1,2,3]:
In formula: DT and DR is respectively the digestibility of test feed and basal feed nutritional labeling
Result:
As can be seen from following table 4, the dry of the single cell protein of self-dissolving technique and protein digestibility is used to be significantly improved, illustrate that the cell membrane of bacterium obtains abundant broken wall after self-dissolving, the nutriment such as nucleotides, albumen dissociates to extracellular, thus is more easily absorbed by animals.
Table 4
Embodiment five: feed intake is tested
Mycoprotein and the difference of common mycoprotein in food calling and feed intake of self-dissolving technique is used by the extra quality groove Preference test assessment of piglet.The extra quality groove model test of piglet can measure the preference of piglet to specified raw material local flavor, and the feed for its preference of searching for food piglet can improve feed intake, and then improves the speed of growth.
Testing program:
Assessment raw material: albumen is the self-dissolving single cell protein of 50%, and common non-self-dissolving protein content is all the single cell protein of 50%.Test feed, basal diet is commercially available weanling pig feed, and test feed proportioning sees the following form 5.
Experimental animal: select Weaning Age, body weight close, the weanling pig searching for food normal, healthy 45, is divided into 3 groups at random.
Table 5
Test method:
The material level hopper identical with specifications and models 2 (requiring that the material level of each hopper must be consistent with test pig quantity) is put in same hurdle, the test feed adding some respectively in hopper (requires that the quantity of feed must be sufficient, ensure the situation that there will not be feed inadequate in 2 hours), A hopper adds A material, B hopper adds B material, allow remaining feed in pig free choice feeding 2 h before harvest hopper measuring, then in A hopper, add B material, add A material in B hopper and carry out above-mentioned identical test.
Observe two hoppers at duration of test to search for food the quantity of feed pig, and to search for food situation and pig number change situation with video camera and camera track up two hopper pigs.Require: test carries out 3 ~ 4 times, each 3 ~ 4 repetitions continuously.Totally 3 ~ 5 days, or a certain feed search for food terminate till.
Result calculates:
Testing index: feed intake and partially addicted to index:
The total feed intake of A daily ration partial eclipse index=A group/total feed intake of B group
Result of the test shows, and under embodiment experimental condition, weanling pig has Preference for self-dissolving single cell protein, for higher containing its cost feed feed intake, shows phagostimulating effect from the free nucleotide of stripping.
Claims (4)
1. the method for a high yield use self-dissolving explained hereafter aerobic single cell protein, comprise several preprocessing unit and an aft-loaded airfoil unit, preprocessing unit comprises precipitation concentration step, centrifugal dehydration step and trafficking step, it is characterized in that the material of 78% ~ 85% moisture content will obtained after centrifugal dehydration step in each preprocessing unit, be transported to aft-loaded airfoil unit after cooling to focus on, described aft-loaded airfoil unit comprises autolysis process, drying steps, sterilization steps and crushing packing step, and described autolysis process is as follows:
A) from the mass transport of preprocessing unit to being incubated feed bin, be then delivered to self-dissolving equipment and carry out self-dissolving process;
B) self-dissolving equipment is by heating up to the heat supply of twin-screw hollow blade, reaches the temperature requirement of self-dissolving, by designing and regulating the cavity volume supporting with charging rate, reaches the time requirement of self-dissolving;
Before the twin-screw hollow blade of c) described self-dissolving equipment, 1/4 to 1/2 is segmented into bringing-up section, and object is that heating material is temperature required to self-dissolving, and remaining segment is soaking zone, keeps material from solubility temperature;
D) temperature requirement of self-dissolving is: the temperature of charge after heating remains on 65% ~ 95 DEG C;
E) time requirement of self-dissolving is: the time that material is warming up to target temperature by bringing-up section should be less than 30 minutes, and the self-dissolving time of soaking zone should at 30 minutes ~ 360 minutes;
The material moisture of described self-dissolving requires: after self-dissolving, moisture content of material should be not less than 70%;
Described sterilization steps adopts microwave sterilization.
2. a kind of high yield as claimed in claim 1 uses the method for self-dissolving explained hereafter aerobic single cell protein, and it is characterized in that the temperature of charge after described heating remains on 65% ~ 80 DEG C, the self-dissolving time is 60 ~ 180 minutes.
3. a kind of high yield as claimed in claim 1 uses the method for self-dissolving explained hereafter aerobic single cell protein, it is characterized in that the order of described concrete grammar step is as follows:
(1) preprocessing unit:
A. precipitation concentration: the mycoprotein that bioreactor produces is entered precipitation concentration pond by pipeline, and mycoprotein is at the concentration basin bottom precipitation of taper, and bottoms material moisture content is reduced to less than 97%, can carry out follow-up dehydration procedure;
B. use mateiral pump that concentration basin bottoms material is delivered to centrifugal-dehydration device;
C. centrifugal dehydration: use screwed bonnet type centrifugation apparatus to carry out Separation of Solid and Liquid, centrifugal rear material water ratio is reduced to 78% ~ 85%, material is in wet pureed; Described centrifugal sloughed moisture is the ECW of mycoprotein, and in born of the same parents, moisture still remains in cell, and this creates condition for intracellular endogenous enzymes carries out self-dissolving; The material of described centrifugal rear 78% ~ 85% moisture content can carry out autolysis;
D. transport: be transported to aft-loaded airfoil unit after the material cooling after centrifugal and focus on; Described cooling is that material is cooled to less than 10 DEG C after centrifugation; Described transport is, by thermal insulated vehicle, the wet pureed material shipment of preprocessing unit output is carried out following process to aft-loaded airfoil unit; Time of described transport should in 0 ~ 12 hour, and the material temperature that material arrives aft-loaded airfoil unit is not higher than 5 DEG C of material temperature when leaving preprocessing unit;
(2) aft-loaded airfoil unit:
A. self-dissolving: from the mass transport of preprocessing unit to being incubated feed bin, being then delivered to self-dissolving equipment and carrying out self-dissolving process; Self-dissolving equipment, by heating up to the heat supply of twin-screw hollow blade, reaches the temperature requirement of self-dissolving, by designing and regulating the cavity volume supporting with charging rate, reaches the time requirement of self-dissolving; Before the twin-screw hollow blade of described self-dissolving equipment, 1/4 to 1/2 is segmented into bringing-up section, and object is that heating material is temperature required to self-dissolving, and remaining segment is soaking zone, keeps material from solubility temperature; The temperature requirement of self-dissolving is: the temperature of charge after heating remains on 65% ~ 95 DEG C; The time requirement of self-dissolving is: the time that material is warming up to target temperature by bringing-up section should be less than 30 minutes, and the self-dissolving time of soaking zone should at 30 minutes ~ 360 minutes; The material moisture of described self-dissolving requires: after self-dissolving, moisture content of material should be not less than 70%;
B. dry: the material after self-dissolving is delivered to air-flow flash drying equipment by flood dragon feeding machine and carries out drying; The material Contact Temperature of described expansion drying should lower than 100 DEG C, and the moisture content of described dried material should 10% ~ 40%;
C. sterilizing: material is after drying delivered to tunnel type micro wave drying sterilization equipment by gas lift equipment and carries out sterilizing; After described microwave sterilization, material water ratio is 5% ~ 10%;
D. crushing packing: the material after sterilizing is packaged into finished product after crushed.
4. adopt the aerobic single cell protein that method according to claim 1 is obtained, it is characterized in that adopting protein content to be the obtained aerobic single cell protein of the mycoprotein of 33%-60%; Protein content 33.0%-60.0%; The free nucleotide content of unit-protein is not less than 1.40%; The free base contents of unit-protein is not higher than 0.90%; , free base and free nucleotide ratio range should at 0-0.60.
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