CN117737169A - Process for preparing protein peptide by enzymolysis of corn steep liquor - Google Patents
Process for preparing protein peptide by enzymolysis of corn steep liquor Download PDFInfo
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- CN117737169A CN117737169A CN202311860463.0A CN202311860463A CN117737169A CN 117737169 A CN117737169 A CN 117737169A CN 202311860463 A CN202311860463 A CN 202311860463A CN 117737169 A CN117737169 A CN 117737169A
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- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 48
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- 108010059892 Cellulase Proteins 0.000 claims description 3
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 claims description 3
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- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims description 2
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 6
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- 229910052757 nitrogen Inorganic materials 0.000 abstract description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 2
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- LINOMUASTDIRTM-UHFFFAOYSA-N vomitoxin hydrate Natural products OCC12C(O)C(=O)C(C)=CC1OC1C(O)CC2(C)C11CO1 LINOMUASTDIRTM-UHFFFAOYSA-N 0.000 description 3
- 239000005019 zein Substances 0.000 description 3
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- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 2
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a process for preparing protein peptide by enzymolysis of corn steep liquor, belonging to the technical field of microorganism submerged fermentation. The preparation process comprises the following steps: the method comprises the steps of (1) plate activation of strains, (2) shake flask seed liquid preparation, (3) primary seed liquid preparation, (4) secondary seed liquid preparation, (5) corn steep liquor treatment, (6) fermentation production, (7) corn steep liquor enzymolysis treatment, (8) detoxification treatment, (9) evaporation concentration and (10) spray drying. The preparation process of the invention selects corn steep liquor as a substrate for strain fermentation, fully utilizes nutrient substances in the corn steep liquor, does not add carbon sources, nitrogen sources and other nutrient substances in the fermentation process, does not regulate and control pH value in the whole process, and monitors the change of lactic acid and reducing sugar by natural pH value fermentation, thereby effectively changing the viscosity of the corn steep liquor, and the obtained protein product has high protein content, high utilization rate, low moisture content and low vomit toxin content and is easy to digest.
Description
Technical Field
The invention relates to a process for preparing protein peptide by enzymolysis of corn steep liquor, belonging to the technical field of microorganism submerged fermentation.
Background
Corn steep liquor is a byproduct of corn deep processing enterprises. Corn steep liquor contains abundant proteins, growth factors, precursor substances and other nutrients, and contains 40% -50% of solid substances, so that the corn steep liquor can be applied to fermentation culture. Because of the abundant protein content, the microbial fermentation method can be used as a nitrogen source for fermentation, but part of amino acids exist in the form of macromolecular proteins, which is unfavorable for the utilization of the microbial fermentation method. In addition, phytate contained in corn steep liquor complexes proteins in corn into insoluble phytic acid-protein complex, which damages the structure of the proteins and also affects the digestibility of the proteins. In addition, corn steep liquor contains about 10% of lactic acid and 5-7% of reducing sugar, and the viscosity of corn steep liquor is obviously affected in the concentration and drying process due to the hygroscopicity of lactic acid and the extensibility of reducing sugar.
At present, the zein peptide is mainly prepared by an acidolysis method or an enzymolysis method, the acidolysis method is adopted to prepare the zein peptide, the reaction is relatively severe, the reaction process is difficult to control, partial amino acid is easy to damage, the taste of the product is poor, the cost is high, and the popularization and the use of the zein peptide are seriously influenced.
Therefore, the adverse factors in the corn steep liquor are reduced or even eliminated by utilizing a biological enzymolysis method, the quality of the corn steep liquor is improved, the existing resources are fully utilized, waste is changed into valuable, the production cost is reduced, and the wide utilization of the Yu Yu corn steep liquor is facilitated.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a process for preparing protein peptide by enzymolysis of corn steep liquor, which can effectively treat the corn steep liquor, and the obtained protein product has high protein content, high utilization rate, low moisture content, low vomit toxin content and easy digestion.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a process for preparing protein peptide by enzymolysis of corn steep liquor comprises the following steps:
(1) Plate activation of the strain: inoculating yeast to a plate culture medium under aseptic condition, and culturing in an incubator at 28-32deg.C for 22-24 hr to obtain plate activated strain;
(2) Preparing shake flask seed liquid: inoculating the plate activated strain into shake flask liquid culture medium, shake culturing at 30-33deg.C and rotation speed of 200r/min for 16-18 hr to OD 600 0.8-1.2 to obtain shake flask seed liquid;
(3) Preparing primary seed liquid: inoculating the shake flask seed liquid into the primary seed culture medium, wherein the adding amount of the shake flask seed liquid is 0.4-0.6% of the primary seed culture medium by volume, and culturing in a seed tank under stirring until OD 600 1.0 to 1.1 to obtain first-stage seed liquid;
(4) Preparing a secondary seed liquid: transferring the primary seed liquid into a secondary seed culture medium, wherein the adding amount of the primary seed liquid is 9-11% of that of the secondary seed culture medium by volume, and culturing in a seed tank under stirring until reaching OD 600 1.5 to 1.6 to obtain secondary seed liquid;
(5) Corn steep liquor treatment: beating fresh corn steep liquor with dry matter content of 40-45% into a sterilized fermentation tank, diluting with sterile hot water to dry matter content of 18-25%, continuously stirring for 1 hr, and cooling to 35-40deg.C to obtain sterile diluted corn steep liquor;
(6) Fermentation production: inoculating the second-level seed liquid into sterile diluted corn steep liquor, and fermenting in a fermentation tank to obtain fermentation liquor;
(7) Enzymolysis treatment of corn steep liquor: adding complex enzyme into the fermentation broth, and stirring for 2-4 hours at 58-62 ℃ to obtain corn steep liquor enzymatic hydrolysate;
(8) Detoxification treatment: adding detoxication agent into the corn steep liquor enzymolysis liquid, and uniformly stirring to obtain detoxified fermentation liquor;
(9) And (3) evaporating and concentrating: performing four-effect evaporation concentration on the detoxified fermentation liquor until the dry matter content of the concentrated liquor is 60-65%, thus obtaining concentrated liquor;
(10) Spray drying: and (3) carrying out spray drying on the concentrated solution to obtain the protein peptide particles.
Further, in the step (1), the preparation method of the plate culture medium comprises the following steps: preparing a YEPD agar medium, sterilizing with steam at 115 ℃ for 25min, and cooling to room temperature to obtain a flat-plate medium.
Further, in the step (2), the preparation method of the shake flask liquid culture medium comprises the following steps: 40g of peptone, 20g of yeast powder and 30g of glucose are weighed and dissolved in distilled water, steam sterilization is carried out for 25min at 115 ℃, and the shaking liquid culture medium is prepared after cooling to room temperature.
Further, in the step (3), the preparation method of the seed culture medium comprises the following steps: weighing 0.8kg of glucose, 4kg of corn steep liquor and 15mL of defoamer, sterilizing by steam in a seed tank at 125-126 ℃ for 20min, and cooling to room temperature to obtain the first-stage seed culture medium.
Further, in the step (3), the conditions of the stirred tank culture are as follows: 75-85% of dissolved oxygen is corrected before inoculation, the tank pressure is 0.1Mpa, and the initial air quantity is 2-3 m 3 Per min, the air quantity in the culture process is 2-4 m 3 And (3) per minute, dissolved oxygen is 10-20%, pH value is controlled to be 5.4-5.6 by liquid ammonia, culture temperature is 31-35 ℃ and time is 14-16 hours, and in the step (4), conditions of stirring culture in the seed tank are as follows: correcting dissolved oxygen before inoculation by 75-85%, tank pressure by 0.1Mpa, initial air volume of 31-33 m/min, air volume of 32-63 m/min in culture process, dissolved oxygen by 10-20%, liquid ammonia controlling pH value to 5.4-5.6, and culture temperature of 33-35 ℃ for 8-10h.
Further, in the step (4), the preparation method of the secondary seed culture medium comprises the following steps: weighing 8kg of glucose, 40kg of corn steep liquor and 150mL of defoamer, continuously sterilizing in a seeding tank by steam at 125-126 ℃ for 20min, and cooling to room temperature to obtain the secondary seed culture medium.
Before the OD values are measured in the step (3) and the step (4), the material is diluted by 10 times by distilled water, then the OD value is detected, the instrument detects the OD value to the maximum extent, and before the dilution, the OD value of the material is large and cannot be detected. And the materials in the step (2) can be directly detected without dilution.
Further, in step (5), the temperature of the sterile hot water is > 90 ℃. Sterile hot water is selected to kill a few of the miscellaneous bacteria in the fresh corn steep liquor at high temperature. Diluting with sterile hot water to dry matter content of 18-25%, and the corn steep liquor has high dry matter content, high viscosity, and can not be used by strain to inhibit its growth, and the diluted corn steep liquor has reduced viscosity, and the concentration ratio of nutrient components is suitable for strain growth metabolism.
Further, in the step (6), the culture conditions of the fermenter are as follows: the tank pressure is controlled at 0.10-0.12Mpa before inoculation, the corrected dissolved oxygen is 80%, the dissolved oxygen is controlled at 5-10% in the culture process, the air quantity is controlled at 300-500 m/min, and the culture time is 24-26h. The pH value is not regulated and controlled in the whole fermentation process, the pH change is concerned, when the pH value rises to above 6.3, the pH value does not rise within 30min, and the fermentation is finished.
Further, in the step (6), the protein content in the fermentation liquor is 52-54%, the lactic acid content is less than 0.1g/L, the reducing sugar content is less than 0.8%, and the viscosity of the fermentation liquor is 220-230mpa/s. The corn steep liquor contains reducing sugar and lactic acid, has high viscosity, is easy to ferment and deteriorate, is unfavorable for storage, and after fermentation, the lactic acid in fermentation liquor is basically consumed, the reducing sugar is mostly consumed, and the viscosity of the fermentation liquor is greatly reduced, thereby being favorable for subsequent spray drying.
In the step (7), the complex enzyme is one or more of phytase, papain and cellulase, and the adding amount of the complex enzyme is 1-2 per mill of the fermentation liquor by mass.
In the step (7), the protein content in the corn steep liquor enzymatic hydrolysate is 55-57%, the dry ammonia nitrogen content is 2.5-2.7g/100mL, and the dry matter content is 15-17%.
Further, in the step (8), the detoxicant is Na 2 SO 3 、NaHSO 3 The addition amount of the detoxicant is 0.07-0.1% of the corn steep liquor by mass.
Further, in the step (8), the vomit toxin content in the detoxified fermentation broth is less than 500ug/kg. The content of vomitoxin contained in different corn steep liquor is different, and the addition amount of the detoxication agent is regulated according to the content of vomitoxin in corn steep liquor.
Further, in the step (9), the temperatures of the four-effect evaporation concentration are respectively: one effect is 75-92 ℃, two effects are 65-88 ℃, three effects are 55-75 ℃, and four effects are 40-65 ℃. The evaporation concentration can be carried out synchronously after the detoxication agent is uniformly mixed, the temperature of the evaporation concentration has no influence on the detoxication effect, and the time can be saved.
Further, in the step (10), the temperature of the spray drying hot air is more than or equal to 100 ℃, and the mixing temperature of materials is 69-72 ℃.
Further, in the step (10), the protein content of the protein peptide particles is more than or equal to 56%, the amino nitrogen content is 2.8-3.2g/100mL, and the moisture content is less than 4.0%. The prepared protein peptide particles have the particle size of 12-30 meshes, and the content of the particles is more than or equal to 90 percent. The protein peptide particles are not easy to agglomerate, the product quality is good, and the protein peptide particles are not easy to deteriorate, thereby being beneficial to preservation.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention only uses the corn steep liquor as a substrate for strain fermentation, and fully utilizes the nutrient substances in the corn steep liquor. In the fermentation process, no carbon source, nitrogen source and other nutrient substances are added, the pH value is not regulated and controlled in the whole process, the fermentation is carried out at the natural pH value, the change of lactic acid and reducing sugar is monitored, the viscosity of corn steep liquor is effectively changed, and the protein content is increased. No solid waste is produced, the process is simple, the production is easy, the energy consumption is saved, and the social benefit is obvious.
(2) Unlike traditional corn steep liquor treatment, the present invention uses complex enzyme to treat corn steep liquor fermentation liquor, hydrolyzes macromolecular protein into micromolecular peptide, and hydrolyzes PO in phosphate in phytic acid structure 4 3+ Hydrolysis is carried out to decompose protein from the protein-phytic acid complex, so that the content of micromolecular protein is increased, the digestibility of the protein is improved, the index is also improved, the ash content of corn steep liquor is also reduced, and the relative protein resource is effectively relievedShortage problem.
(3) The invention can synchronously carry out detoxification reaction and evaporation concentration, and the detoxification effect can be satisfied, thereby effectively reducing vomitoxin in corn steep liquor, being easy to digest, not causing harm to animals, increasing the utilization rate of organic wastewater, improving the nutritive value and increasing the economic benefit of corn processing enterprises.
(4) The invention ferments the corn processing byproducts and carries out enzymolysis detoxification to produce protein peptide products, which can realize effective utilization of resources, bring economic benefit to enterprises, promote the development of the enterprises and reduce environmental pollution.
Drawings
FIG. 1 is a process flow diagram of the preparation of a protein peptide by enzymolysis of corn steep liquor.
Detailed Description
Example 1
A process for preparing protein peptide by enzymolysis of corn steep liquor and application thereof comprise the following steps:
(1) Plate activation of the strain: preparing a YEPD agar medium, sterilizing with steam at 115 ℃ for 25min, cooling to room temperature to obtain a plate medium, inoculating saccharomycetes to the plate medium under aseptic condition, and culturing in an incubator at 28 ℃ for 24h to obtain plate activated strain;
(2) Preparing shake flask seed liquid: weighing 40g of peptone, 20g of yeast powder and 30g of glucose, dissolving in distilled water, sterilizing with steam at 115 ℃ for 25min, cooling to room temperature to obtain shake flask liquid culture medium, inoculating plate activated strain into shake flask liquid culture medium, shake culturing at 30 ℃ and 200r/min for 18h until OD is reached 600 0.8, obtaining shake flask seed liquid;
(3) Preparing primary seed liquid: weighing glucose 0.8kg, corn steep liquor 4kg and defoamer 15mL, sterilizing with steam at 125 ℃ in a seed tank for 20min, cooling to room temperature to obtain a first-stage seed culture medium, inoculating shake flask seed liquid into the first-stage seed culture medium, wherein the adding amount of the shake flask seed liquid is 0.4% of the first-stage seed culture medium by volume, and stirring and culturing in the seed tank to OD 600 1.0 to obtain first-level seed liquid;
(4) Preparing a secondary seed liquid: weighing glucose 8kg, corn steep liquor 40kg and defoamer 150mL, sterilizing with steam at 125-126 ℃ in a seed tank for 20min, cooling to room temperature to obtain a secondary seed culture medium, transferring primary seed liquid into the secondary seed culture medium, wherein the addition amount of the primary seed liquid is 10% of that of the secondary seed culture medium by volume, and stirring and culturing in the seed tank to OD 600 1.5, obtaining a secondary seed solution;
(5) Corn steep liquor treatment: pumping fresh corn steep liquor with dry matter content of 40% into a sterilized fermentation tank, diluting with sterile hot water to dry matter content of 18%, continuously stirring for 1 hr, and cooling to 35deg.C to obtain sterile diluted corn steep liquor;
(6) Fermentation production: inoculating the second-stage seed solution into sterile diluted corn steep liquor, fermenting in a fermenter, controlling dissolved oxygen at 5% and air volume at 500m 3 And/min, culturing for 26h to obtain fermentation liquor;
(7) Enzymolysis treatment of corn steep liquor: adding cellulase with the addition amount of 1 per mill into the fermentation liquor, and stirring for 2 hours at the temperature of 58 ℃ to obtain corn steep liquor enzymatic hydrolysate;
(8) Detoxification treatment: adding detoxication agent NaHSO into corn steep liquor enzymolysis liquid 3 Stirring uniformly to obtain detoxified fermentation liquor;
(9) And (3) evaporating and concentrating: sequentially performing four-effect evaporation concentration on the detoxified fermentation liquor at the temperature of 75 ℃, 88 ℃, 75 ℃ and 40 ℃ until the dry matter content of the concentrated liquor is 65%, thus obtaining the concentrated liquor;
(10) Spray drying: and (3) spray drying the concentrated solution at the temperature of 100 ℃ to obtain the protein peptide particles.
Example 2
A process for preparing protein peptide by enzymolysis of corn steep liquor and application thereof comprise the following steps:
(1) Plate activation of the strain: preparing a YEPD agar medium, sterilizing with steam at 115 ℃ for 25min, cooling to room temperature to obtain a plate medium, inoculating saccharomycetes to the plate medium under aseptic condition, and culturing in an incubator at 32 ℃ for 22h to obtain plate activated strain;
(2) Seed shaking bottlePreparing a sub-liquid: weighing 40g of peptone, 20g of yeast powder and 30g of glucose, dissolving in distilled water, sterilizing with steam at 115 ℃ for 25min, cooling to room temperature to obtain shake flask liquid culture medium, inoculating plate activated strain into shake flask liquid culture medium, shake culturing at 33 ℃ and 200r/min for 16h until OD is reached 600 0.8, obtaining shake flask seed liquid;
(3) Preparing primary seed liquid: weighing glucose 0.8kg, corn steep liquor 4kg and defoamer 15mL, sterilizing with steam at 126 ℃ in a seed tank for 20min, cooling to room temperature to obtain a first-stage seed culture medium, inoculating shake flask seed liquid into the first-stage seed culture medium, wherein the adding amount of the shake flask seed liquid is 0.6% of the first-stage seed culture medium by volume, and stirring and culturing in the seed tank to OD 600 1.1, obtaining first-stage seed liquid;
(4) Preparing a secondary seed liquid: taking 8kg of glucose, 40kg of corn steep liquor and 150mL of defoamer, sterilizing by steam in a seed tank at 126 ℃ for 20min, cooling to room temperature to obtain a secondary seed culture medium, transferring a primary seed liquid into the secondary seed culture medium, wherein the adding amount of the primary seed liquid is 9% of that of the secondary seed culture medium by volume, and stirring and culturing in the seed tank until OD 600 1.6, obtaining a secondary seed solution;
(5) Corn steep liquor treatment: pumping fresh corn steep liquor with the dry matter content of 45% into a sterilized fermentation tank, diluting with sterile hot water until the dry matter content is 25%, continuously stirring for 1 hour, and cooling to 38 ℃ to obtain sterile thin corn steep liquor;
(6) Fermentation production: inoculating the second-stage seed solution into sterile diluted corn steep liquor, fermenting in a fermenter, controlling dissolved oxygen to 10% and air volume to 300m during culture 3 And/min, culturing for 26h to obtain fermentation liquor;
(7) Enzymolysis treatment of corn steep liquor: adding papain with an addition amount of 1 per mill into the fermentation liquor, and stirring for 3 hours at the temperature of 62 ℃ to obtain corn steep liquor enzymatic hydrolysate;
(8) Detoxification treatment: adding detoxication agent Na into corn steep liquor enzymolysis liquid 2 SO 3 Stirring uniformly to obtain detoxified fermentation liquor;
(9) And (3) evaporating and concentrating: sequentially performing four-effect evaporation concentration on the detoxified fermentation liquor at the temperature of 92 ℃, 65 ℃, 55 ℃ and 65 ℃ until the dry matter content of the concentrated liquor is 60%, thus obtaining the concentrated liquor;
(10) Spray drying: and (3) spray drying the concentrated solution at the temperature of 105 ℃ to obtain the protein peptide particles.
Example 3
A process for preparing protein peptide by enzymolysis of corn steep liquor and application thereof comprise the following steps:
(1) Plate activation of the strain: preparing a YEPD agar medium, sterilizing with steam at 115 ℃ for 25min, cooling to room temperature to obtain a plate medium, inoculating saccharomycetes to the plate medium under aseptic condition, and culturing in an incubator at 28 ℃ for 24h to obtain plate activated strain;
(2) Preparing shake flask seed liquid: weighing 40g of peptone, 20g of yeast powder and 30g of glucose, dissolving in distilled water, sterilizing with steam at 115 ℃ for 25min, cooling to room temperature to obtain shake flask liquid culture medium, inoculating plate activated strain into shake flask liquid culture medium, shake culturing at 30 ℃ and 200r/min for 18h until OD is reached 600 1.03, obtaining shake flask seed liquid;
(3) Preparing primary seed liquid: weighing glucose 0.8kg, corn steep liquor 4kg and defoamer 15mL, sterilizing with steam at 126 ℃ in a seed tank for 20min, cooling to room temperature to obtain a first-stage seed culture medium, inoculating shake flask seed liquid into the first-stage seed culture medium, wherein the adding amount of the shake flask seed liquid is 0.5% of the first-stage seed culture medium by volume, and stirring and culturing in the seed tank to OD 600 1.09 to obtain first-level seed liquid;
(4) Preparing a secondary seed liquid: taking 8kg of glucose, 40kg of corn steep liquor and 150mL of defoamer, sterilizing by steam in a seed tank at 126 ℃ for 20min, cooling to room temperature to obtain a secondary seed culture medium, transferring a primary seed liquid into the secondary seed culture medium, wherein the adding amount of the primary seed liquid is 11% of that of the secondary seed culture medium by volume, and stirring and culturing in the seed tank until OD 600 1.56, obtaining a secondary seed solution;
(5) Corn steep liquor treatment: feeding fresh corn steep liquor with the dry matter content of 43% into a sterilized fermentation tank, diluting with sterile hot water to the dry matter content of 20%, continuously stirring for 1 hour, and cooling to 40 ℃ to obtain sterile diluted corn steep liquor;
(6) Fermentation production: inoculating the second-stage seed solution into sterile diluted corn steep liquor, fermenting in a fermenter, controlling dissolved oxygen at 5% and air volume at 500m 3 And/min, culturing for 26h to obtain fermentation liquor;
(7) Enzymolysis treatment of corn steep liquor: adding phytase into the fermentation broth, and stirring for 4 hours at the temperature of 58 ℃ to obtain corn steep liquor enzymatic hydrolysate;
(8) Detoxification treatment: adding detoxication agent into the corn steep liquor enzymolysis liquid, and uniformly stirring to obtain detoxified fermentation liquor;
(9) And (3) evaporating and concentrating: sequentially performing four-effect evaporation concentration on the detoxified fermentation liquor at the temperature of 75 ℃, 65 ℃ and 75 ℃ and 65 ℃ until the dry matter content of the concentrated liquor is 65%, thus obtaining the concentrated liquor;
(10) Spray drying: and (3) carrying out spray drying on the concentrated solution to obtain the protein peptide particles.
Experimental example 1
The components and contents of the protein peptide particles prepared in examples 1 to 3 were examined. The results of the measurements are shown in Table 1.
TABLE 1 composition and content of protein peptides
As is clear from Table 1, the protein contents of the protein peptides obtained in examples 1 to 3 were 56.1%, 57.2% and 56.8%, the moisture contents were 3.95%, 3.81% and 3.89%, the amino nitrogen contents were 2.97g/100ml, 3.12/g/100 ml and 3.04/g/100 ml, respectively, and the vomit toxin contents were 496 ug/kg, 487ug/kg and 486 ug/kg, respectively, and it was found that the protein peptides prepared by the present invention were high in protein content, low in moisture content and low in vomit toxin content. The preparation process can effectively treat the corn steep liquor, and the obtained protein peptide product has high protein content, high utilization rate, easy digestion and no harm to animals.
Claims (10)
1. A process for preparing protein peptide by enzymolysis of corn steep liquor is characterized in that: the method comprises the following steps:
(1) Plate activation of the strain: inoculating yeast to a plate culture medium under aseptic condition, and culturing in an incubator at 28-32deg.C for 22-24 hr to obtain plate activated strain;
(2) Preparing shake flask seed liquid: inoculating the plate activated strain into shake flask liquid culture medium, shake culturing at 30-33deg.C and rotation speed of 200r/min for 16-18 hr to OD 600 0.8-1.2 to obtain shake flask seed liquid;
(3) Preparing primary seed liquid: inoculating the shake flask seed liquid into the primary seed culture medium, wherein the adding amount of the shake flask seed liquid is 0.4-0.6% of the primary seed culture medium by volume, and culturing in a seed tank under stirring until OD 600 1.0 to 1.1 to obtain first-stage seed liquid;
(4) Preparing a secondary seed liquid: transferring the primary seed liquid into a secondary seed culture medium, wherein the adding amount of the primary seed liquid is 9-11% of that of the secondary seed culture medium by volume, and culturing in a seed tank under stirring until reaching OD 600 1.5 to 1.6 to obtain secondary seed liquid;
(5) Corn steep liquor treatment: beating fresh corn steep liquor with dry matter content of 40-45% into a sterilized fermentation tank, diluting with sterile hot water to dry matter content of 18-25%, continuously stirring for 1 hr, and cooling to 35-40deg.C to obtain sterile diluted corn steep liquor;
(6) Fermentation production: inoculating the second-level seed liquid into sterile diluted corn steep liquor, and fermenting in a fermentation tank to obtain fermentation liquor;
(7) Enzymolysis treatment of corn steep liquor: adding complex enzyme into the fermentation broth, and stirring for 2-4 hours at 58-62 ℃ to obtain corn steep liquor enzymatic hydrolysate;
(8) Detoxification treatment: adding detoxication agent into the corn steep liquor enzymolysis liquid, and uniformly stirring to obtain detoxified fermentation liquor;
(9) And (3) evaporating and concentrating: performing four-effect evaporation concentration on the detoxified fermentation liquor until the dry matter content of the concentrated liquor is 60-65%, thus obtaining concentrated liquor;
(10) Spray drying: and (3) carrying out spray drying on the concentrated solution to obtain the protein peptide particles.
2. The process for preparing protein peptide by enzymolysis of corn steep liquor according to claim 1, wherein: in the step (3), the conditions of the stirred culture in the seed tank are as follows: 75-85% of dissolved oxygen is corrected before inoculation, the tank pressure is 0.1Mpa, and the initial air quantity is 2-3 m 3 Per min, the air quantity in the culture process is 2-4 m 3 And (3) per minute, dissolved oxygen is 10-20%, pH value is controlled to be 5.4-5.6 by liquid ammonia, culture temperature is 31-35 ℃ and time is 14-16 hours, and in the step (4), conditions of stirring culture in the seed tank are as follows: correcting dissolved oxygen before inoculation by 75-85%, tank pressure by 0.1Mpa, initial air volume of 31-33 m/min, air volume of 32-63 m/min in culture process, dissolved oxygen by 10-20%, liquid ammonia controlling pH value to 5.4-5.6, and culture temperature of 33-35 ℃ for 8-10h.
3. The process for preparing protein peptide by enzymolysis of corn steep liquor according to claim 2, wherein: in the step (6), the protein content in the fermentation liquor is 52-54%, the lactic acid content is less than 0.1g/L, the reducing sugar content is less than 0.8%, and the viscosity of the fermentation liquor is 220-230mpa/s.
4. A process for preparing a protein peptide by enzymolysis of corn steep liquor according to any one of claims 1-3, wherein: in the step (7), the complex enzyme is one or more of phytase, papain and cellulase, and the adding amount of the complex enzyme is 1-2 per mill of the fermentation liquor by mass.
5. The process for preparing protein peptide by enzymolysis of corn steep liquor as claimed in claim 4, wherein: in the step (7), the protein content in the corn steep liquor enzymatic hydrolysate is 55-57%, the dry ammonia nitrogen content is 2.5-2.7g/100mL, and the dry matter content is 15-17%.
6. The process for preparing protein peptide by enzymolysis of corn steep liquor according to claim 1, wherein: in the step (8), the detoxicant is Na 2 SO 3 、NaHSO 3 The addition amount of the detoxicant is 0.07-0.1% of the corn steep liquor by mass.
7. The process for preparing protein peptide by enzymolysis of corn steep liquor according to claim 5 or 6, wherein: in the step (8), the vomit toxin content in the detoxified fermentation broth is less than 500ug/kg.
8. The process for preparing protein peptide by enzymolysis of corn steep liquor as claimed in claim 7, wherein: in the step (9), the four-effect evaporation concentration temperatures are respectively as follows: one effect is 75-92 ℃, two effects are 65-88 ℃, three effects are 55-75 ℃, and four effects are 40-65 ℃.
9. The process for preparing protein peptide by enzymolysis of corn steep liquor according to claim 1, wherein: in the step (10), the temperature of the spray drying hot air is more than or equal to 100 ℃, and the mixing temperature of materials is 69-72 ℃.
10. The process for preparing protein peptide by enzymolysis of corn steep liquor according to claim 8 or 9, wherein: in the step (10), the protein content of the protein peptide particles is more than or equal to 56%, the amino nitrogen content is 2.8-3.2g/100mL, and the water content is less than 4.0%.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN118805843A (en) * | 2024-09-12 | 2024-10-22 | 山东福洋生物科技股份有限公司 | A corn gluten meal processing technology and its application in food |
| CN119385244A (en) * | 2024-11-04 | 2025-02-07 | 山东谊智合生物科技有限公司 | Production method of corn steep liquor fermented protein and corn steep liquor fermented protein |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN118805843A (en) * | 2024-09-12 | 2024-10-22 | 山东福洋生物科技股份有限公司 | A corn gluten meal processing technology and its application in food |
| CN119385244A (en) * | 2024-11-04 | 2025-02-07 | 山东谊智合生物科技有限公司 | Production method of corn steep liquor fermented protein and corn steep liquor fermented protein |
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