CN102008006B - Preparation method of polypeptide for fermented feed with molecular weight of less than 15KDa - Google Patents
Preparation method of polypeptide for fermented feed with molecular weight of less than 15KDa Download PDFInfo
- Publication number
- CN102008006B CN102008006B CN2009101695011A CN200910169501A CN102008006B CN 102008006 B CN102008006 B CN 102008006B CN 2009101695011 A CN2009101695011 A CN 2009101695011A CN 200910169501 A CN200910169501 A CN 200910169501A CN 102008006 B CN102008006 B CN 102008006B
- Authority
- CN
- China
- Prior art keywords
- dregs
- fermentation
- molecular weight
- grouts
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to the technical field of biological fermentation and in particular to a solid-state fermentation process for preparing a plant polypeptide for a feed having a molecular weight lower than 15KDa. In the preparation method, bean dregs, cottonseed dregs and bran are used as raw materials, a microbial fermentation technology is used to effectively remove soybean anti-nutritional factors in the bean pulp and gossypol in the cottonseed pulp, and simultaneously, ahigh-quality fermented plant dreg cake which is rich in bacillus coagulans is obtained. The preparation method comprises the following steps of: activating one original bacillus coagulans strain, amplifying the bacillus coagulans strain, inoculating the bacillus coagulans strain to a plant dreg cake fermenting base material (wherein, the bean dregs and the cottonseed dregs are crushed to be 20 mesh , and the ratio of the bean dregs to the cottonseed dregs to the bran is 4:4:2, and the ratio of the materials to water is 1:0.9) by using a culture solution of which the OD600nm reaches 4 as a dreg cake fermenting inoculation solution according to an inoculation size of 10%, uniformly mixing, and fermenting at 30 DEG C for 48 hours; and drying at 60 DEG C, and analyzing nutritional ingredients of the bean dregs which are not fermented and the nutritional ingredients of the bean dregs which are fermented.
Description
(1) technical field
The present invention relates to technical field of biological fermentation, take dregs of beans, cotton dregs and wheat bran as raw material, utilize microbial fermentation technology effectively to remove Main Antinutritional Factors in dregs of beans and the cotton dregs, obtain simultaneously being rich in the high-quality fermented bean dregs of bacillus coagulans, especially a kind of preparation molecular weight of solid state fermentation is lower than the technique of 15 dalton's (KD) forage plant property polypeptide.
(2) background technology
In recent years, import high-quality fish meal, whey powder price are unprecedented soaring, bring unprecedented pressure for the production of feed processing enterprise, bring unprecedented opportunity to develop also for biofermentation grouts product.Develop-and-import scheme high-quality animal protein (such as fish meal, plasma protein, goldbeater's skin albumen etc.) substitute products are the sustainable developments that realize feed industry and aquaculture, keep the Important Action of newborn piglet raiser profitability
The fermentation grouts refer to it is by modern biofermentation technique raw material dregs of beans, cotton dregs, the vegetable seeds dregs of rice etc. to be carried out product after the fermentation process.Can produce the plurality of enzymes such as protease, non-starch polysaccharide enzyme in the sweat lives, eliminate ANFs, clear up Soybean antigen protein and realize the soybean biological desensitization, the poisonous free gossypol of Degradation and Transformation, the cake protein matter of macromolecule is decomposed into polypeptide, oligopeptides even little peptide, thereby increase water-solublely, improve digestibility, be beneficial to animal digestion and absorb.Fiber substance is decomposed into sugar, and part sugar is converted into lactic acid, and produces a large amount of beneficial microbes, makes grouts change into the functional feed of high nutritive value.
The technology of the present invention is mainly for the above-mentioned key issue that dregs of beans and cotton dregs nutritive value are provided, and by the technological parameter of research bacillus solid fermentation, finally obtains a kind of little molecule (＜15KD) preparation method of the feeding polypeptide of fermentation that is rich in.The high-quality feed protein resource is in short supply to provide good protein raw materials and probio that technical support is provided in particular for young animal in order to solve.
(3) summary of the invention
The object of the invention is to by the solid-state biofermentation of bacillus coagulans, provide a kind of little molecule (＜15KD) fermentation feeding polypeptide the preparation method, promote dregs of beans and the effective application of cotton dregs in young animal is fed, solve the problem in short supply of high-quality protein feed resource.
Purpose of the present invention is achieved by the following technical programs.
A kind of little molecule provided by the invention (＜15KD) fermentation feeding polypeptide the preparation method, its step is as follows:
A. original starting strain bacillus coagulans line 30 ℃, is left standstill cultivation 48h and is activated on the beef extract-peptone agar slant culture-medium.
B. picking one ring thalline from the inclined-plane is inoculated in the beef extract-peptone fluid nutrient medium, and 30 ℃, 200-250rpm cultivates 12-16h, as seed liquor.Enlarge in the beef extract-peptone fluid nutrient medium at liquid, with 5% inoculum concentration inoculation seed liquor, 30 ℃, 200-250rpm cultivates OD
600nm=4 o'clock, as the fermentation inoculation liquid.
C. the fermentation inoculation liquid among the b is inoculated into 10% inoculum concentration that (dregs of beans and cotton dregs were pulverized 20 orders, dregs of beans: cotton dregs: wheat bran=4: 4: 2, material-water ratio=1: 0.9 in the grouts fermentation base-material.), mix, in 30 ℃ of fermentation 48h.In 60 ℃ of oven dry, the grouts before and after the fermentation are carried out Analysis of Nutritive Composition.
The beef extract-peptone agar medium of the activation thalline described in the step a of the present invention contains 1.0% peptone by mass percentage, 0.5% sodium chloride, 0.3% beef extract, 2.0% agar.
Thalline seed liquor described in the step b of the present invention and expansion culture medium contain 1.0% peptone, 0.5% sodium chloride, 0.3% beef extract by mass percentage.
Bacterial classification described in the step a of the present invention is bacillus coagulans (Bacillus Coagulans, CGMCC 1.2009) 1 strain.
The present invention has following beneficial effect:
Compare with physical method with other chemistry, the used microbe fermentation method of the present invention has the characteristics less with destruction on the impact of feed nutrient.Reduced simultaneously the content of dregs of beans and cotton dregs ANFs, albumen solubility increases, and macro-molecular protein is by obvious degradation, thereby has greatly improved the nutritional quality of dregs of beans and cotton dregs.By phosphate buffer extract (PBS, the pH7.4:KH of special-purpose software (GlyKo BandScanVersion 5.0) for the albumen before and after the grouts fermentation
2PO
40.24g/L, Na
2HPO
41.44g/L, NaCl 8.0g/L, KCl 0.2g/L) trihydroxy methyl glycine--lauryl sodium sulfate--polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) band concentration analysis shows, molecular weight on average accounts for 40% greater than the protein of 45KDa in the grouts that do not ferment, average out to 2% in the fermentation grouts; Molecular weight does not on average account for 55% in the protein between 15KDa and the 45KDa is fermenting grouts, average out to 35% in the fermentation grouts; Molecular weight on average accounts for 5% less than the protein of 15KDa in the grouts that do not ferment, improve average out to 63% (table 2) in the grouts that ferment. and trypsin inhibitor content is down to the rear 0.082mg/g of fermentation by the front 1.749mg/g of fermentation; Degree of hydrolysis is 22.72% after the fermentation; Can find out from table 2 and Fig. 1: poisonous free gossypol is all degraded effectively in 5 subunits (molecular weight is all greater than 58KDa) of fermentation main antigen protein such as β-companion's globulin α subunit (molecular weight is 57-76KDa) and glycinin the grouts and the cotton dregs, thereby can reduce its antigenicity and toxicity, the probio bacillus coagulans reaches 5.0 * 10
8CFU/g.8 large key indexs of beneficial microbe fermenting plant grouts (dregs of beans and cotton dregs) base-material comprise: less than the small-molecular peptides mass ratio of 15kD＞60%, degree of hydrolysis＞20%, the apparent content of true protein improves 2.55%, main allergic protein degradation rate＞90%, trypsin inhibitor content＜0.20mg/g, proteinase activity＞1000U/g, gossypol content＜80mg/Kg, probio coagulated bacillus living quantity＞1.0 * 10
8CFU/g is a kind of high value forage protein that the important application future is arranged in Modern Animal Husbandry and feed industry.
Description of drawings
The Tricine-SDS-PAGE protein electrophorese collection of illustrative plates of Fig. 1 grouts before and after fermentation
The present invention will be further described below in conjunction with accompanying drawing.
Fig. 1 is the SDS-PAGE protein electrophorese collection of illustrative plates before and after the grouts fermentation, and wherein swimming lane 1 is protein molecular weight standard, is followed successively by from top to bottom: 94.0kDa, 66.2kDa, 45.0kDa, 35.0kDa, 28.5kDa, 20.0kDa, 14.4kDa; Swimming lane 2 is the base-material sample that do not ferment; Swimming lane 3 is fermentation grouts product.
(4) specific embodiment
A kind of little molecule provided by the invention (＜15KD) preparation method's of fermentation forage plant property polypeptide the specific embodiment is as follows:
The used bacterial strain of the present embodiment is bacillus coagulans (Bacillus Coagulans, CGMCC 1.2009).
A.10g peptone, 5g sodium chloride, the 3g beef extract is settled to 1L, adds 20g agar, after the heating for dissolving, minute install in the test tube, 121 ℃ of steam sterilizings, 20min makes the beef extract-peptone agar slant culture-medium of activation thalline.The 10g peptone, 5g sodium chloride, the 3g beef extract is settled to 1L, divides to install to triangular flask, 121 ℃ of steam sterilizings, 20min makes thalline seed liquor and scale-up medium body culture medium.
B. original starting strain bacillus coagulans line is on the beef extract-peptone agar slant culture-medium, and 30 ℃, static cultivation 48h activates.
C. picking one ring thalline from the inclined-plane is inoculated in the beef extract-peptone fluid nutrient medium, and 30 ℃, 200-250rpm cultivates 12-16h, as seed liquor.Enlarge in the beef extract-peptone fluid nutrient medium at liquid, with 5% inoculum concentration inoculation seed liquor, 30 ℃, 200-250rpm cultivates OD
600nm=4 o'clock, as the fermentation inoculation liquid.
D. the inoculation liquid of fermenting was inoculated in the vegetalitas grouts fermentation base-material (dregs of beans and cotton dregs were pulverized 20 orders, dregs of beans: cotton dregs: wheat bran=4: 4: 2, material-water ratio=1: 0.9) with 10% inoculum concentration, mixed, in 30 ℃ of fermentation 48h.In 60 ℃ of oven dry.
E. the dregs of beans before and after the fermentation is carried out Analysis of Nutritive Composition.
The plant-derived micromolecule polypeptide of microbe fermentation method preparation is compared with fermenting plant grouts not in above-mentioned example, nutritive peculiarity be improved significantly.Behind the bean pulp fermentation, albumen solubility increases, and macro-molecular protein is by obvious degradation, by PBS extract (the PBS pH7.4:KH of special-purpose software (GlyKo BandScan Version 5.0) for the albumen before and after dregs of beans, the cotton dregs fermentation
2PO
40.24g/L, Na
2HPO
41.44g/L, NaCl 8.0g/L, KCl 0.2g/L) Tricine-SDS-PAGE electrophoretic band (Fig. 1) concentration analysis show, molecular weight on average accounts for 40% greater than the protein of 45KDa in the grouts that do not ferment, the fermentation grouts in average out to 2%; Molecular weight does not on average account for 55% in the protein between 15KDa and the 45KDa is fermenting grouts, average out to 35% in the fermentation grouts; Molecular weight on average accounts for 5% less than the protein of 15KDa in the grouts that do not ferment, improve average out to 63% (table 2) in the grouts that ferment. and trypsin inhibitor content is down to the rear 0.082mg/g of fermentation by the front 1.749mg/g of fermentation; Neutral proteinase enzyme work＞1331U/g, degree of hydrolysis is 22.72% after the fermentation; Main antigen protein such as 5 subunits (molecular weight is all greater than 58KDa) of β-companion's globulin α subunit (molecular weight is 57-76KDa) and glycinin are all degraded (table 2 and Fig. 1) effectively in the dregs of beans, thereby can reduce its antigenicity; The contained poisonous free gossypol of cotton dregs is reduced to 77mg/Kg by 330mg/Kg in the fermentation medium simultaneously, reduces by 76.67%; The probio bacillus coagulans reaches 5.0 * 10
8CFU/g.Take dregs of beans and cotton dregs as main forage protein culture medium, nutritive value is obviously improved by the bacillus coagulans solid state fermentation, and its effect sees Table 1 and table 2.8 large key indexs of beneficial microbe fermenting plant grouts (dregs of beans and cotton dregs) base-material products obtained therefrom are respectively: less than the small-molecular peptides mass ratio of 15kD＞60%, degree of hydrolysis＞20%, the apparent content of true protein improves 2.55%, main allergic protein degradation rate＞90%, trypsin inhibitor content＜0.20mg/g, proteinase activity＞1000U/g, gossypol content＜80mg/Kg, probio coagulated bacillus living quantity＞1.0 * 10
8CFU/g is a kind of high value forage protein that the important application future is arranged in Modern Animal Husbandry and feed industry.
The comparison of composition before and after the fermentation of table 1 culture medium
Molecular weight of albumen allocation proportion before and after the fermentation of table 2 culture medium
*,+/ _ CK (%)=(processing costs-control value) * 100/ control value
Claims (5)
1. a molecular weight the steps include: less than the ferment preparation method of feeding polypeptide of the little molecule of 15KD
A. original starting strain bacillus coagulans (Bacillus Coagulans) line 30 ℃, is left standstill cultivation 48h and is activated on the beef extract-peptone agar slant culture-medium;
B. picking one ring thalline from the inclined-plane is inoculated in the beef extract-peptone fluid nutrient medium, and 30 ℃, 200-250rpm, cultivate 12-16h, as seed liquor, enlarge in the beef extract-peptone fluid nutrient medium at liquid, the inoculum concentration inoculation seed liquor with 5%, 30 ℃, 200-250rpm cultivates OD
600nm=4 o'clock, as the fermentation inoculation liquid;
C. the fermentation inoculation liquid among the b is inoculated in the vegetalitas grouts fermentation base-material with 10% inoculum concentration, the preparation of vegetalitas grouts fermentation base-material: dregs of beans and cotton dregs were pulverized 20 orders, dregs of beans: cotton dregs: wheat bran=4: 4: 2, material-water ratio=1: 0.9, mix, in 30 ℃ of fermentation 48h, in 60 ℃ of oven dry, the grouts before and after the fermentation are carried out Analysis of Nutritive Composition.
2. molecular weight according to claim 1 is less than the ferment preparation method of feeding polypeptide of the little molecule of 15KD, the beef extract-peptone agar slant culture-medium of the described activation thalline of its step a, by the 10g peptone, 5g sodium chloride, 3g beef extract, be settled to 1L, add 20g agar, after the heating for dissolving, divide to install in the test tube, 121 ℃ of steam sterilizings, 20min makes.
3. molecular weight according to claim 1 is less than the ferment preparation method of feeding polypeptide of the little molecule of 15KD, the described beef extract-peptone fluid nutrient medium of step b and liquid enlarge the beef extract-peptone fluid nutrient medium, by the 10g peptone, 5g sodium chloride, the 3g beef extract is settled to 1L, divides to install to triangular flask, 121 ℃ of steam sterilizings, 20min makes.
4. molecular weight according to claim 1 is less than the ferment preparation method of feeding polypeptide of the little molecule of 15KD, and the described bacterial classification of its step a is bacillus coagulans (CGMCC 1.2009) 1 strain.
5. each described molecular weight is less than the little molecule of the 15KD fermenting plant grouts that the preparation method of feeding polypeptide makes that ferment according to claim 1-4, the 8 large key indexs that it is characterized in that grouts are as follows: less than the small-molecular peptides mass ratio of 15KD＞60%, degree of hydrolysis＞20%, the apparent content of true protein improves 2.55%, proteinase activity＞1000U/g, main allergic protein degradation rate＞90%, trypsin inhibitor content＜0.20mg/g, gossypol content＜80mg/Kg, probio coagulated bacillus living quantity＞1.0 * 10
8CFU/g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101695011A CN102008006B (en) | 2009-09-08 | 2009-09-08 | Preparation method of polypeptide for fermented feed with molecular weight of less than 15KDa |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101695011A CN102008006B (en) | 2009-09-08 | 2009-09-08 | Preparation method of polypeptide for fermented feed with molecular weight of less than 15KDa |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102008006A CN102008006A (en) | 2011-04-13 |
| CN102008006B true CN102008006B (en) | 2013-01-09 |
Family
ID=43838509
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101695011A Active CN102008006B (en) | 2009-09-08 | 2009-09-08 | Preparation method of polypeptide for fermented feed with molecular weight of less than 15KDa |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102008006B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104531575A (en) * | 2014-12-18 | 2015-04-22 | 湖北华扬科技发展有限公司 | Bacillus coagulans solid-state fermentation production method |
| CN107467347A (en) * | 2017-07-31 | 2017-12-15 | 兰溪市酉泽饲料技术服务有限公司 | The preparation method of Penaeus Vannmei growth-promoting feed |
| CN107467413A (en) * | 2017-07-31 | 2017-12-15 | 兰溪市酉泽饲料技术服务有限公司 | The preparation method of loach scrod feed |
| CN107683954B (en) * | 2017-09-17 | 2018-07-31 | 浙江大学 | An a kind of step adds the method that bacterium step fermentation prepares low anti-nutritional factors acidifying feed |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101390571A (en) * | 2007-09-21 | 2009-03-25 | 北京大北农科技集团有限责任公司 | Coagulation bacillus feedstuff supplement and preparation method thereof |
| CN101455267A (en) * | 2008-11-18 | 2009-06-17 | 广东省农业科学院农业生物技术研究所 | Preparation method of fermented bean pulp rich in function peptide for feeding |
-
2009
- 2009-09-08 CN CN2009101695011A patent/CN102008006B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101390571A (en) * | 2007-09-21 | 2009-03-25 | 北京大北农科技集团有限责任公司 | Coagulation bacillus feedstuff supplement and preparation method thereof |
| CN101455267A (en) * | 2008-11-18 | 2009-06-17 | 广东省农业科学院农业生物技术研究所 | Preparation method of fermented bean pulp rich in function peptide for feeding |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102008006A (en) | 2011-04-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102948614B (en) | Method for preparing peptide used for active feed by bacteria and enzyme synergistic fermentation bean pulp | |
| CN104054902B (en) | A kind of mixed bacteria solid state fermentation produces the technique of fermented bean dregs | |
| CN103609862B (en) | Method for improving feeding nutritive value of sesame seed meal by combining enzymolysis method with fermentation method | |
| CN103271221B (en) | Method for producing hydrolyzed wheat protein | |
| CN105661011A (en) | Functional bio-protein feed ferment and fermented protein feed | |
| CN104920806A (en) | Bran protein feed preparing method by using mixed bacteria multi-step fermentation | |
| CN102823725B (en) | Method for producing biologic protein feed by using solid-state fermentation fiber dreg | |
| CN1559261A (en) | Two-effects microbiological additives of forage specially used for ruminants | |
| CN1207873A (en) | Straw feed fermenting process | |
| CN105494890A (en) | Enzymolysis and fermentation combined method for improving feeding nutritional value of rapeseed meal | |
| TW201915165A (en) | Bacillus subtilis strain for preparing fermented feather meal and use thereof for producing feather powder and proteolytic enzyme and keratinolytic enzyme strains | |
| CN104000021B (en) | The working method of a kind of vegetalitas fermented feed | |
| CN103211084A (en) | Method for preparing biological enzyme-enriched camellia seed meal fermented feed | |
| CN103859150A (en) | Production technology of aspergillus oryzae fermented soybean meal | |
| CN102008006B (en) | Preparation method of polypeptide for fermented feed with molecular weight of less than 15KDa | |
| CN106173204A (en) | A kind of method preparing high protein feed for base material fermentation with citric acid corn starch residue and mycelia slag | |
| CN111748598A (en) | Small molecule peptide protein powder and preparation method thereof | |
| CN109757603A (en) | A kind of method that fermentation soybean grouts prepare pannage | |
| CN107279464B (en) | Process for engineering fermentation of biomass feed | |
| CN106260589A (en) | A kind of method that recycling wheat bran prepares albumen feedstuff | |
| CN101990984A (en) | Method for preparing high-antioxidation low-sensitization forage fermented soybean meal | |
| CN111296723A (en) | Probiotic agent for multi-strain combined solid state fermentation of rapeseed meal and fermentation method thereof | |
| CN110301526A (en) | Complex micro organism fungicide and its method for preparing bioactive feed | |
| CN108203729B (en) | Preparation method of kelp antioxidant peptide | |
| CN102907568A (en) | Cold-region fermented soybean meal industrialized production method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |