CN102948614A - Method for preparing peptide used for active feed by bacteria and enzyme synergistic fermentation bean pulp - Google Patents
Method for preparing peptide used for active feed by bacteria and enzyme synergistic fermentation bean pulp Download PDFInfo
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Abstract
The invention discloses a method for preparing peptide used for the active feed by the bacteria and enzyme synergistic fermentation bean pulp. The method comprises the following steps of: preparing complex enzyme used for synergistic fermentation; preparing complex strain used for the synergistic fermentation; mixing and adding the complex enzyme and the complex strain into the bean pulp; blending the bean pulp with warm water; carrying out grading temperature-control fermentation on the bean pulp containing water; and post-processing materials formed after fermentation to obtain a finished product. Compared with the prior art, the method has the advantages that trypsin inhibitor is almost removed, big-molecule protein in the bean pulp is effectively degraded, and the actions of the microorganism and the enzyme are furthest cooperated.
Description
Technical field
The present invention relates to microbial enzyme hydrolysis and fermentation technical field, especially relate to the method that a kind of bacterium enzyme cooperative fermentation dregs of beans prepares active feeding peptide.
Background technology
As large agricultural country, aquaculture is occupied critical role in China's agricultural, and feed is the important foundation of aquaculture.Usually, the content of protein and quality are the major criterion of its quality of evaluation in the feed, but also are directly connected to health and the level of production of animal.Feed can be divided into animal protein raw material and vegetable protein raw material two large classes with protein raw material.Wherein, the animal protein raw material comprises fish meal, meat meal tankage and SDPP etc.Because the amino acid similarity that animal protein amino acid composition is required with growth of animals or poultry, absorptivity is high, absorption is fast and produce the advantages such as pollution is few so animal protein has.Yet the safety issue of scarcity of resources and feed product is restricted the application of animal protein raw material.Therefore, people more and more pay close attention to the application of vegetable protein raw material.The vegetable protein raw material comprises Soybean Meal, rapeseed dregs, Cottonseed Meal, peanut meal etc.Compare the cheap and wide material sources of vegetable protein cost of material with the animal protein raw material.There are abundant and inexpensive vegetable protein raw material in China.Producing approximately 1,000 ten thousand tons, the dish dregs of rice per year such as: dregs of beans produces approximately 700 ~ 8,000,000 tons, cottonseed cake (dregs of rice) per year and approximately produces more than 600 ten thousand tons per year.But except dregs of beans, the utilization rate of the assorted dregs of rice such as Cottonseed Meal, leached tea oil slag is less than 30%, and effective exploitation vegetable protein raw material has good prospect.
Along with people to the deepening continuously of the feeding peptide of activity research, the active feeding peptide with various functions constantly is found, active feeding peptide has become the important form of protein performance nutrition physiology effect.Utilize the feeding peptide of bioactive activity that has of vegetable protein raw material production to have some advantages.Such as: can promote protein metabolism, improve the utilization rate of protein; Promote absorption and the utilization of mineral matter, strengthen immunity of organisms, improve the animal dis motility rate; Also has physiological regulatory action; Can be directly promote the secretion of enteron aisle hormone receptor and enzyme and play a role as neurotransmitter; Promote the growth of the digestive systems such as intestinal villi and pancreas; Strengthen the breeding of profitable strain, improve the synthetic of mycoprotein, stimulate the growth of enteron aisle, strengthen premunition, thereby improve growing of animal.
Than horn of plenty, the method for utilizing dregs of beans to prepare active feeding peptide mainly contains dregs of beans: acid hydrolyzation, based on alkaline hydrolysis, electrolysis, microbe fermentation method and enzymatic isolation method in the resource of China.Wherein, acid hydrolyzation, based on alkaline hydrolysis and electrolysis are multiplex in laboratory research, are not suitable for large-scale production.Although microbe fermentation method and enzymatic isolation method can large-scale productions, contain relatively large ANFs in the prepared active feeding peptide.Wherein, ANFs comprises SNSP, Soybean antigen protein and residual protease inhibitor etc.The existence of these ANFs or suppressed that some digestive ferment produces in the animal body perhaps becomes stodgy composition with the nutriment complexing, so that the absorptivity of the digestibility of dregs of beans material and animal descends.
Summary of the invention
The purpose of this invention is to provide the method that a kind of bacterium enzyme cooperative fermentation dregs of beans prepares active feeding peptide, it has the lower characteristics of ANFs content in the prepared active feeding peptide.
To achieve these goals, the technical solution adopted in the present invention is: bacterium enzyme cooperative fermentation dregs of beans prepares the method for active feeding peptide, comprises the steps:
A) the used complex enzyme of preparation cooperative fermentation;
B) the used composite bacteria of preparation cooperative fermentation;
C) with complex enzyme be added in the dregs of beans after composite bacteria mixes, regulate dregs of beans with warm water;
D) with step C) in contain moisture dregs of beans carry out stage by stage temperature controlled fermentation;
E) with step D) in form material after the fermentation and carry out post processing and make finished product.
Described steps A) includes neutral proteinase, acid protease, alkali protease, non-starch polysaccharide enzyme, pectase, zytase in the complex enzyme in, wherein, neutral proteinase, acid protease and alkali protease are that bacillus or aspergillus niger are secreted, and each ingredients weight parts is respectively: 15 ~ 23 parts of neutral proteinases, 10 ~ 18 parts of acid proteases, 8 ~ 16 parts of alkali proteases, 18 ~ 26 parts of non-starch polysaccharide enzymes, 6 ~ 13 parts of pectases, 3 ~ 12 parts of zytases.
Described steps A) complex enzyme in is that warm water activation 1 ~ 2h of 40 ~ 50oC makes through temperature.
Described steps A) composite bacteria in comprises bacillus lentus, Candida utilis ferment, lactobacillus fermenti, and each ingredients weight parts is respectively: 13 ~ 27 parts of bacillus lentuses, 8 ~ 36 parts of candida utili, 21 ~ 35 parts of lactobacillus fermentis.
Described steps A) each bacterial classification of the composite bacteria in activates through warm water, and wherein, the bacillus lentus activation temperature is 37 ~ 45oC, and the candida utili activation temperature is 35 ~ 37oC, and the lactobacillus fermenti activation temperature is 37 ~ 42oC, and soak time is respectively 1 ~ 2h.
Described step C) in, by weight, the ratio that complex enzyme is added in the dregs of beans is: enzyme 100 ~ 500U/g dregs of beans alive; The ratio that composite bacteria is added in the dregs of beans is: bacterium is counted 1*10
6~ 8*10
6/ g dregs of beans.
Described step C) regulates dregs of beans in to containing moisture 30 ~ 45%; The temperature of used warm water is: 35 ~ 50oC.
Described step D) in stage by stage the temperature controlled fermentation parameter be:
| Stage | Control temperature (oC) | Time (h) |
| 1 | 25-35 | 0-18 |
| 2 | 35-40 | 18-36 |
| 3 | 40-45 | 36-48 |
| 4 | 45-50 | 48-72 |
The temperature control method of the stage by stage temperature controlled fermentation described step D) is: intermittent aeration and intermittently turning.
Described step e) post processing in is: material package is made finished product to the overlay film bag that the polyethylene inner bag is arranged, or be to be dried to moisture in 45 ~ 55oC environment to make finished product less than 12% rear packing with material in temperature.
The present invention is compared with existing technology the advantage that has:
1, substantially removed trypsin inhibitor, urease activity is reduced to 0.086U.Lipoxidase and soybean haemoglutinin are eliminated substantially, and Soybean antigen protein is degraded.
2, the high molecular weight protein in the dregs of beans has obtained effective degraded, little peptide content is brought up to 201mg/g from 18mg/g, molecular weight 140000Da and above protein component account for 6.78%, the molecular weight approximately polypeptide fractions content of 2500Da brings up to 46.34%, and molecular weight 1000Da and the following little peptide composition content of function bring up to 32.51%.Fermented sample all contains the free amino acid about 8%.
3, crude protein content brings up to 51% from 42%, and protein digestibility brings up to 92% from 71%, and lactic acid content is more than or equal to 5% in the dregs of beans, and the beneficial bacterium number reaches 1.87*10
7Cfu/g.
4, temperature-control fermentation method has been realized in the different stages different microorganisms and enzyme being played a role stage by stage, thereby has worked in coordination with to greatest extent the effect of microorganism and enzyme.
The specific embodiment
The following stated only is preferred embodiment of the present invention, does not therefore limit protection scope of the present invention.
In following examples:
1, the selection of dregs of beans.Dregs of beans is taken from oil and foodstuffs processing enterprise, and quality raw materials meet the requirement of GB/T 19541-2004.Also can choose general dregs of beans as raw material, but will carry out strict check to the dregs of beans raw material of different manufacturers, different batches.
2, the selection of water.The fermentation water is chosen sterilization running water or distilled water.As running water selecting, then must not there be harmful bacteria in this running water.
Embodiment 1: enzyme cooperative fermentation dregs of beans prepares the method for active feeding peptide, comprises the steps:
A) the used complex enzyme of preparation cooperative fermentation is total to 10kg.Include neutral proteinase, acid protease, alkali protease, non-starch polysaccharide enzyme, pectase, zytase in this complex enzyme.Wherein, neutral proteinase, acid protease and alkali protease are that bacillus or aspergillus niger are secreted.Simultaneously, each ingredients weight parts is: 15 parts of neutral proteinases, 10 parts of acid proteases, 8 parts of alkali proteases, 18 parts of non-starch polysaccharide enzymes, 6 parts of pectases, 3 parts of zytases.In the preparation, this complex enzyme is the warm water activation 1h of 40oC through temperature.Between pot-life, fully stir, various enzymes are mixed, fully dissolving.
B) the used composite bacteria 10kg of preparation cooperative fermentation.This composite bacteria comprises bacillus lentus, Candida utilis ferment, lactobacillus fermenti.Simultaneously, the weight portion of each component is: the weight portion of each component is respectively: 13 parts of bacillus lentuses, 8 parts of candida utili, 21 parts of lactobacillus fermentis.In the preparation, each bacterial classification of composite bacteria activates through warm water.Wherein, the bacillus lentus activation temperature is that 37oC, soak time are 1h, and the candida utili activation temperature is that 35oC, soak time are 1.5h, and the lactobacillus fermenti activation temperature is 37oC, and soak time is 2h.
C) with the complex enzyme of above preparation be added in the dregs of beans after composite bacteria mixes, regulate dregs of beans with warm water.Wherein, get dregs of beans 1000kg.By weight, complex enzyme is added into the ratio in the dregs of beans and is: the enzyme 100U/g dregs of beans of living.That is, add complex enzyme 1kg.By weight, composite bacteria is added into the ratio in the dregs of beans and is: bacterium is counted 1*10
6/ g dregs of beans.That is, add composite bacteria 2kg.Used warm water temperature is: 35oC.Regulate dregs of beans moisture to 30%.
D) with step C) in contain moisture dregs of beans carry out stage by stage temperature controlled fermentation.That is:
Phase I: the control temperature is that 35oC, fermentation time keep 18h;
Second stage: the control temperature is that 40oC, fermentation time keep 36h;
Phase III: the control temperature is that 45oC, fermentation time keep 48h.
Temperature-controlled process in the above stage is taked intermittent aeration and turning at intermittence mode.
So far, form altogether material 1240kg after the fermentation.
E) with step D) in form material after the fermentation and carry out post processing and make finished product.This post processing mode is: material package is made finished product 1 to the overlay film bag that the polyethylene inner bag is arranged.
Embodiment 2: enzyme cooperative fermentation dregs of beans prepares the method for active feeding peptide, only is with the difference of embodiment 1:
Steps A) in, by weight, the weight portion of each component is respectively in the used complex enzyme of cooperative fermentation: 20 parts of neutral proteinases, 15 parts of acid proteases, 12 parts of alkali proteases, 24 parts of non-starch polysaccharide enzymes, 11 parts of pectases, 8 parts of zytases.In the preparation, this complex enzyme is the warm water activation 1.5h of 45oC through temperature.Between pot-life, fully stir, various enzymes are mixed, fully dissolving.
Step B) in, by weight, the weight portion of used each component of composite bacteria of cooperative fermentation is respectively: 20 parts of bacillus lentuses, 30 parts of candida utili, 30 parts of lactobacillus fermentis.In the preparation, each bacterial classification of composite bacteria activates through warm water.Wherein, the bacillus lentus activation temperature is that 40oC, soak time are 1.5h, and the candida utili activation temperature is that 36oC, soak time are 1h, and the lactobacillus fermenti activation temperature is 40oC, and soak time is 1.5h.
Step C) in, by weight, the ratio that complex enzyme is added in the dregs of beans is: enzyme 300U/g dregs of beans alive.That is, add complex enzyme 3kg.By weight, composite bacteria is added into the ratio in the dregs of beans and is: bacterium is counted the 4*106/g dregs of beans.That is, add composite bacteria 8kg.Used warm water temperature is: 40oC.Regulate dregs of beans moisture to 35%.
Step D) in, the concrete mode of temperature controlled fermentation is stage by stage:
Phase I: the control temperature is that 25oC, fermentation time keep 9h;
Second stage: the control temperature is that 35oC, fermentation time keep 18h;
Phase III: the control temperature is that 40oC, fermentation time keep 36h;
The quadravalence section: the control temperature is that 45oC, fermentation time keep 48h.
Form altogether material 1350kg after the fermentation.
Step e) in, post processing mode is: with material temperature be dried in 45 ~ 55oC environment moisture less than 12% after, finished product 2 is made in packing.
Embodiment 3: enzyme cooperative fermentation dregs of beans prepares the method for active feeding peptide, only is with the difference of embodiment 1:
Steps A) in, by weight, the weight portion of each component is respectively in the used complex enzyme of cooperative fermentation: 23 parts of neutral proteinases, 18 parts of acid proteases, 16 parts of alkali proteases, 26 parts of non-starch polysaccharide enzymes, 13 parts of pectases, 2 parts of zytases.In the preparation, this complex enzyme is the warm water activation 2h of 50oC through temperature.Between pot-life, fully stir, various enzymes are mixed, fully dissolving.
Step B) in, by weight, the weight portion of used each component of composite bacteria of cooperative fermentation is respectively: 27 parts of bacillus lentuses, 36 parts of candida utili, 35 parts of lactobacillus fermentis.In the preparation, each bacterial classification of composite bacteria activates through warm water.Wherein, the bacillus lentus activation temperature is that 45oC, soak time are 2h, and the candida utili activation temperature is that 37oC, soak time are 2h, and the lactobacillus fermenti activation temperature is 42oC, and soak time is 1h.
Step C) in, by weight, the ratio that complex enzyme is added in the dregs of beans is: enzyme 500U/g dregs of beans alive.That is, add complex enzyme 5kg.By weight, composite bacteria is added into the ratio in the dregs of beans and is: bacterium is counted the 8*106/g dregs of beans.That is, add composite bacteria 16kg.Used warm water temperature is: 50oC.Regulate dregs of beans moisture to 45%.
Step D) in, the concrete mode of temperature controlled fermentation is stage by stage:
Phase I: the control temperature is that 30oC, fermentation time keep 9h;
Second stage: the control temperature is that 38oC, fermentation time keep 24h;
Phase III: the control temperature is that 42oC, fermentation time keep 42h;
The quadravalence section: the control temperature is that 48oC, fermentation time keep 60h.
Form altogether material 1450kg after the fermentation.
Step e) in, post processing mode is: with material temperature be dried in 45 ~ 55oC environment moisture less than 12% after, finished product 3 is made in packing.
Embodiment 4: enzyme cooperative fermentation dregs of beans prepares the method for active feeding peptide, and only be with the difference of embodiment 2: step D), the concrete mode of temperature controlled fermentation is stage by stage:
Phase I: the control temperature is that 35oC, fermentation time keep 18h;
Second stage: the control temperature is that 40oC, fermentation time keep 36h;
Phase III: the control temperature is that 45oC, fermentation time keep 48h;
The quadravalence section: the control temperature is that 50oC, fermentation time keep 72h.
Form altogether material 1350kg after the fermentation, form finished product 4.
Effect example: according to the test method of GBT 19541-2004, the content situation of the ANFs of above embodiment 1 ~ 4 gained finished product 1 ~ 4 is detected.The results are shown in Table 1:
Table 1: the ANFs content table of comparisons
* conventional method preparation utilizes dregs of beans to prepare active feeding peptide to come from single bacterium fermentation and protease hydrolyzed dregs of beans sample detection data.
By above data as can be known, prepare for the active feeding peptide with respect to the dregs of beans that utilizes of conventional method preparation, in the prepared active feeding peptide of the method that enzyme cooperative fermentation dregs of beans of the present invention prepares active feeding peptide:
1, urase, the ANFs such as trypsin inhibitor are lower.
2, the high molecular weight protein in the dregs of beans has obtained effective degraded.
3, crude protein content, protein digestibility, beneficial bacterium number average have obtained a larger increase.
Claims (10)
1. bacterium enzyme cooperative fermentation dregs of beans prepares the method for active feeding peptide, comprises the steps:
A) the used complex enzyme of preparation cooperative fermentation;
B) the used composite bacteria of preparation cooperative fermentation;
C) with complex enzyme be added in the dregs of beans after composite bacteria mixes, regulate dregs of beans with warm water;
D) with step C) in contain moisture dregs of beans carry out stage by stage temperature controlled fermentation;
E) with step D) in form material after the fermentation and carry out post processing and make finished product.
2. bacterium enzyme cooperative fermentation dregs of beans according to claim 1 prepares the method for active feeding peptide, it is characterized in that: include neutral proteinase in the complex enzyme described steps A), acid protease, alkali protease, non-starch polysaccharide enzyme, pectase, zytase, wherein, neutral proteinase, acid protease and alkali protease are that bacillus or aspergillus niger are secreted, and each ingredients weight parts is respectively: 15 ~ 23 parts of neutral proteinases, 10 ~ 18 parts of acid proteases, 8 ~ 16 parts of alkali proteases, 18 ~ 26 parts of non-starch polysaccharide enzymes, 6 ~ 13 parts of pectases, 3 ~ 12 parts of zytases.
3. bacterium enzyme cooperative fermentation dregs of beans according to claim 2 prepares the method for active feeding peptide, it is characterized in that: the complex enzyme described steps A) is that warm water activation 1 ~ 2h of 40 ~ 50oC makes through temperature.
4. bacterium enzyme cooperative fermentation dregs of beans according to claim 1 prepares the method for active feeding peptide, it is characterized in that: the composite bacteria described steps A) comprises bacillus lentus, Candida utilis ferment, lactobacillus fermenti, and each ingredients weight parts is respectively: 13 ~ 27 parts of bacillus lentuses, 8 ~ 36 parts of candida utili, 21 ~ 35 parts of lactobacillus fermentis.
5. bacterium enzyme cooperative fermentation dregs of beans according to claim 4 prepares the method for active feeding peptide, it is characterized in that: each bacterial classification of the composite bacteria described steps A) activates through warm water, wherein, the bacillus lentus activation temperature is 37 ~ 45oC, the candida utili activation temperature is 35 ~ 37oC, the lactobacillus fermenti activation temperature is 37 ~ 42oC, and soak time is respectively 1 ~ 2h.
6. bacterium enzyme cooperative fermentation dregs of beans according to claim 1 prepares the method for active feeding peptide, it is characterized in that: described step C), by weight, the ratio that complex enzyme is added in the dregs of beans is: enzyme 100 ~ 500U/g dregs of beans alive; The ratio that composite bacteria is added in the dregs of beans is: bacterium is counted 1*10
6~ 8*10
6/ g dregs of beans.
7. bacterium enzyme cooperative fermentation dregs of beans according to claim 1 prepares the method for active feeding peptide, it is characterized in that: regulate dregs of beans described step C) to containing moisture 30 ~ 45%; The temperature of used warm water is: 35 ~ 50oC.
8. bacterium enzyme cooperative fermentation dregs of beans according to claim 1 prepares the method for active feeding peptide, it is characterized in that: described step D) stage by stage the temperature controlled fermentation parameter be:
9. bacterium enzyme cooperative fermentation dregs of beans according to claim 8 prepares the method for active feeding peptide, and it is characterized in that: the temperature control method of the stage by stage temperature controlled fermentation described step D) is: intermittent aeration and intermittently turning.
According to claim 1 in 9 each described bacterium enzyme cooperative fermentation dregs of beans prepare the method for active feeding peptide, it is characterized in that: the post processing described step e) is: material package is made finished product to the overlay film bag that the polyethylene inner bag is arranged, or be to be dried to moisture in 45 ~ 55oC environment to make finished product less than 12% rear packing with material in temperature.
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