CN102550874A - Method for producing active polypeptide rich product by solid state fermentation and application - Google Patents
Method for producing active polypeptide rich product by solid state fermentation and application Download PDFInfo
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- CN102550874A CN102550874A CN2012100156523A CN201210015652A CN102550874A CN 102550874 A CN102550874 A CN 102550874A CN 2012100156523 A CN2012100156523 A CN 2012100156523A CN 201210015652 A CN201210015652 A CN 201210015652A CN 102550874 A CN102550874 A CN 102550874A
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Abstract
The invention relates to a method for producing an active polypeptide rich product by solid state fermentation and application. The method comprises the following steps of: preparing a solid state fermentation culture medium by using bean pulp and bran as raw materials, sterilizing, cooling, inoculating a seed solution of activated aspergillus niger or bacillus subtilis, adding KH2PO4, performing fermentation culture at the temperature of between 28 and 32 DEG C for 60 to 70 hours, heating to the temperature of between 50 and 52 DEG C after fermentation is finished, hydrolyzing, drying the fermentation product till the water content is less than 8 percent, crushing, sieving with a sieve of 80 meshes, and thus obtaining the active polypeptide rich feed product. The soybean active polypeptide product prepared by using the method basically does not contain anti-nutritional factors, has small molecular weight, is easily absorbed, and has a remarkable promoting effect on the production performance of weaned pigs.
Description
Technical field
The invention belongs to food processing technology field, relate to method and application that the active peptides goods are rich in a kind of solid state fermentation production.
Background technology
The dregs of beans crude protein content is abundant; Amino acid is formed rationally, in addition, contains other nutriments and many active materials that can produce biological effect such as fat, carbohydrate, vitamin; Like soyabean oligosaccharides, isoflavones, soybean active peptide etc., be a kind of good plant-derived albumen feed.
Contain multiple ANFs (anti-nutritional factors in the dregs of beans; ANF), like trypsin ihhibitor, urase etc., discover; These ANFs have influenced digestion, the absorption of livestock and poultry to nutritional labeling, influence health and the production performance of animal.This just need handle ANFs, thereby improves the digestibility of dregs of beans, improves nutritive value, and processing method commonly used so far is summarised as: physical method, chemical method and biological method.
Method through microbial fermentation prepares protease, and with this hydrolysis process dregs of beans, the high molecular weight protein in the dregs of beans is hydrolyzed into micromolecular peptide by protease etc., and the utilization rate of protein is improved.Soybean Peptide has good physicochemical property, and boundless application prospect is arranged aspect food and medicine, and fermented bean dregs is at animal health, and is especially respond well aspect the enteric microorganism of regulating young animal, is widely used in herding and feed industry circle.In addition, the microbial fermentation dregs of beans need not heat or add chemical reagent, and the ruined possibility of feed nutrient is little, and application safety is practical.
Soybean Peptide has unique physical and chemical properties and multiple biologically active.Method through microbial fermentation can be removed the multiple ANFs in the dregs of beans effectively, reduces cost, and improves the utilization rate of product, has wide significance in actual production with in using.Yet the preparation of protease receives the influence of envirment factor.As: the 1. influence of pH.The protease kind that culture medium produced of different pH values is different, the optimum pH of the generally close and participation enzymatic reaction of the pH value of the protease that produces.2. the influence of ventilation.The process that deep layer is cultivated; Need to ventilate stir how many influences that how much receive ventilation of the growth of fungal hyphae and yield of enzyme, how much in actual production, will the controlling well of ventilation; To be beneficial on the one hand the growth of thalline, effect that on the other hand can the CKIs enzyme.3. Influence of Temperature.It is different that different microbial fermentation produces the required optimum temperature of enzyme, and it is too high or cross low influence that the growth of thalline, the cycle of producing enzyme all can receive temperature, is unfavorable for the generation of highly active protein enzyme like this.In addition; The composition of fermentation medium and the generation of other metabolites also can have influence on the fermenting and producing of protease, therefore, in the actual production should according to screen fermentation, the hydrolysising condition that the microorganism that obtains gropes to suit; Thereby obtain sufficient protease, further improve the Soybean Peptide conversion ratio.
Summary of the invention
Order of the present invention is to provide a kind of solid state fermentation production to be rich in the method and the application of active peptides goods.
In order to realize the object of the invention, the inventor has finally obtained following technical scheme through a large amount of experimental studies:
The method of active peptides goods is rich in a kind of solid state fermentation production, comprises the steps:
(1) dregs of beans, wheat bran and water are mixed with solid-state fermentation culture medium by the weight ratio of 1:0.15-0.2:1.4-1.6; Sterilization; Cooling; Account for solid fermentation culture medium dry weight mass percent in seed liquor, aspergillus niger (Aspergillus niger, ACCC30787) seed liquor 8.5%, bacillus subtilis (the Bacillus subtilis of inoculation activation; ACCC01746) seed liquor 1.4%, and cultivates at following condition bottom fermentation: adding and accounting for solid fermentation culture medium dry weight mass percent is 1.2% KH
2PO
4, fermentation temperature 28-32 ℃, fermentation time 60-70h;
(2) after the fermentation ends, be warming up to 50-52 ℃, hydrolysis 4-5h is dried to water content less than 8% with tunning then, pulverizes, and crosses 80 mesh sieves, must be rich in the feed goods of active peptides.
Preferably; The method of active peptides goods is rich in described a kind of solid state fermentation production; Wherein said aspergillus niger (Aspergillus niger; ACCC30787) seed liquor prepares as follows: slant strains is transferred in the liquid activation medium, places shaking table, shaken cultivation 32-40h under 28-32 ℃, 200r/min condition; In weight portion, the prescription of described liquid activation medium is: NaNO
30.2 part, K
2HPO
40.1 part, 0.05 part of KCl, MgSO
40.05 part, FeSO
40.001 part, 3.0 parts of sucrose, 100 parts of distilled water, natural pH value.
Preferably; The method of active peptides goods is rich in described a kind of solid state fermentation production; Wherein said bacillus subtilis (Bacillus subtilis; ACCC01746) seed liquor prepares as follows: slant strains is transferred in the liquid activation medium, places shaking table, shaken cultivation 28-30h under 33-35 ℃, 200r/min condition; In weight portion, the prescription of described liquid activation medium is: 0.2 part of glucose, 0.5 part of NaCl, 0.5 part of yeast extract, 1 part of peptone, 100 parts of distilled water, pH7.0.
Discover through animal experiment, utilize the soybean active polypeptide product of method for preparing having significant effect aspect the raising piglet production performance.Therefore, second purpose of the present invention is to provide a kind of new use of product, promptly utilizes the application of active peptides goods in the preparation animal feed of method for preparing.Preferably, the application of the active peptides goods of method for preparing in the preparation pig starter feed.
The present invention adopts the method for substep fermentation; Produce the character of protease and the fundamental property of protease hydrolytic according to growth of microorganism metabolism in the sweat; Production technology to Soybean Peptide is optimized; Make each stage of solid state fermentation reach optimal culture conditions separately, such solid ferment process had both helped the generation of highly active protein enzyme, helped protease again and acted on substrate protein generation Soybean Peptide; Thereby reach the purpose that shortens fermentation time, improves the Soybean Peptide conversion ratio, for the Application and Development of Soybean Peptide provides the production route.In addition, utilize the soybean active polypeptide product of method preparation of the present invention not contain ANFs basically, molecular weight is little, is prone to absorb, and the weanling pig production performance is had comparatively significant facilitation.
The specific embodiment
Below be concrete preparation example and the application examples that is rich in the active peptides goods of the present invention, technical scheme of the present invention is done further the description, but protection scope of the present invention be not limited to these embodiment.Every do not deviate from the change of the present invention design or be equal to substitute include within protection scope of the present invention.
The preparation of embodiment 1 active peptides goods
(1) dregs of beans, wheat bran and water are mixed with solid-state fermentation culture medium by the weight ratio of 1:0.18:1.5; Sterilization, cooling accounts for solid fermentation culture medium dry weight mass percent in seed liquor; Aspergillus niger (the Aspergillus niger of inoculation activation; ACCC30787) (Bacillus subtilis, ACCC01746) seed liquor 1.4%, and cultivates at following condition bottom fermentation: adding and accounting for solid fermentation culture medium dry weight mass percent is 1.2% KH for seed liquor 8.5%, bacillus subtilis
2PO
4, 30 ℃ of fermentation temperatures, fermentation time 65h;
Wherein, Aspergillus niger (Chinese agriculture microorganism fungus kind preservation administrative center provides for Aspergillus niger, ACCC30787) seed liquor prepares as follows: slant strains is transferred in the liquid activation medium; Place shaking table, shaken cultivation 32-40h under 30 ℃, 200r/min condition; Slant medium prescription: NaNO3 0.2g, K2HPO4 0.1g, KCl 0.05g, MgSO4 0.05g, FeSO4 0.001g, sucrose 3.0g, distilled water 100mL, agar 1.5～2.0g, 121 ℃, sterilization 20min; The prescription of liquid activation medium is: NaNO
30.2g, K
2HPO
40.1g, KCl 0.05g, MgSO
40.05g, FeSO
40.001g, sucrose 3.0g, distilled water 100ml, natural pH value, 121 ℃, sterilization 20min.
Bacillus subtilis (Bacillus subtilis; ACCC01746; Chinese agriculture microorganism fungus kind preservation administrative center provides) seed liquor prepares as follows: slant strains is transferred in the liquid activation medium; Place shaking table, shaken cultivation 28-30h under 33 ℃, 200r/min condition; Slant medium prescription: peptone 1g, beef extract 0.3g, NaCl 0.5g, agar 2.0g, distilled water 100ml, pH7.0,121 ℃, sterilization 20min; The prescription of liquid activation medium is: glucose 0.2g, NaCl 0.5g, yeast extract 0.5g, peptone 1g, distilled water 100ml, pH7.0,121 ℃, sterilization 20min.
(2) after the fermentation ends, be warming up to 50 ℃, hydrolysis 4h is dried to water content less than 8% with tunning then, pulverizes, and crosses 80 mesh sieves, must be rich in the feed goods of active peptides.
Embodiment 2 active peptides goods are to the influence test of piglet production performance
48 of the Landraces (♂) of experiment selected 28 ages in days wean, average weight 7 scholar 0.1kg * Large White (♀) two-way cross piglet; Press the single factor experiment design principle; If 3 processed group (referring to table 1), each handles 4 repetitions, and each repeats 4 pigs (public female half and half); After raising 3 days in advance, carry out the test in 5 weeks by a definite date.
Table 1 test is divided into groups and the diet preparation
| Handle | Diet is formed |
| Control group | The basis diet |
| Test I group | The active peptides goods of embodiment 1 preparation with 5% substitute the dregs of beans of basal diet moderate |
| Test II group | The active peptides goods of embodiment 1 preparation with 10% substitute the dregs of beans of basal diet moderate |
Experiment selected corn-dregs of beans-expanded soybean type basis diet with reference to the nutritional need of NRC (1998) pig, divides behind the weaned piglet two stages of 1-3 week and 4-5 week to carry out diet and prepares, feed formula be (by weight): corn 58%; Dregs of beans 17%, expanded full-fat bean 17%, fish meal 4.5%, whey powder 4.5%; Inferior powder 5.0%, calcium monohydrogen phosphate 1.5%, salt 0.25%; Stone flour 1.1%, B B-complex-microelement pre-mix material 1.0%, lysine 0.15%.Wherein, contain in the per kilogram feed: vitamin A 1500IU, vitamin D 200IU, vitamin E 10IU, vitamin K 0.5mg, biotin 0.05mg, folic acid 0.3mg, pantothenic acid 10mg, nicotinic acid 10mg, Cobastab
23.6mg, Cobastab
11.0mg, Cobastab
61.5mg, Cu 6mg, Fe 100mg, Zn 110mg, Mn 3mg, Se 0.30mg, I 0.14mg.
Test is carried out at piglet child care house, and the child care house adopts slatted floor child care equipment, and the house temperature control is at 25 ~ 28 ℃, and relative humidity is controlled at 50 ~ 70%.Piglet is regularly fed, and feeding time is respectively at 8:00,12:00,16:00 and the 20:00 of every day.Feed adopts powder to mix wet mode and feeds, and each feeding capacity is advisable so that a small amount of clout to be arranged in the hopper.Piglet restricted scale of rationing in initial 3 days after wean, free choice feeding after 3 days, duration of test is freely drunk water.Clean the child care house every day once, whenever the child care house is carried out spraying disinfection once at a distance from four days.Piglet immunological program and other feedings and managements are undertaken by the pig farm routine.
When on-test and end, respectively each piglet is carried out empty stomach and weigh.Write down the inventory and the surplus material amount (clout amount+waste) of every circle, actual feed intake=inventory-surplus material amount.Calculate the material anharmonic ratio based on average daily gain and average daily ingestion amount.
The weanling pig production performance is seen table 2.Can be found out by table 2: in the whole test phase, test I group is compared with control group, can improve average daily gain 11.1%, reduces material anharmonic ratio 12.6% (P＜0.05); Test II group piglet is compared with control group, can improve average daily gain 18.5% (P＜0.05), reduces material anharmonic ratio 12.0% (P＜0.05); But in the whole test phase, average daily ingestion amount difference all not significantly (P＞0.05) between each processed group piglet.This shows that the soybean active polypeptide product of the present invention's preparation has comparatively significant facilitation to the weanling pig production performance.
Table 2 green feed additive is to the influence of weanling pig production performance
| Index | Control group | Test I group | Test II group |
| Starting weight | 7.12±0.14 | 7.07±0.09 | 7.11±0.10 |
| The end is heavy | 16.68±0.86 | 17.52±0.95 * | 18.23±0.87 ** |
| 1-5 week average daily gain | 0.27±0.05 | 0.30±0.10 | 0.32±0.04 * |
| 1-5 week is daily ingestion amount on average | 0.45±0.06 | 0.44±0.11 | 0.47±0.04 |
| 1-5 week material anharmonic ratio | 1.67±0.09 | 1.46±0.08 * | 1.47±0.12 * |
Compare * with control group
P＜0.05, * *
P＜0.01
Claims (5)
1. the method for active peptides goods is rich in a solid state fermentation production, comprises the steps:
(1) dregs of beans, wheat bran and water are mixed with solid-state fermentation culture medium by the weight ratio of 1:0.15-0.2:1.4-1.6; Sterilization; Cooling; Account for solid fermentation culture medium dry weight mass percent in seed liquor, aspergillus niger (Aspergillus niger, ACCC30787) seed liquor 8.5%, bacillus subtilis (the Bacillus subtilis of inoculation activation; ACCC01746) seed liquor 1.4%, and cultivates at following condition bottom fermentation: adding and accounting for solid fermentation culture medium dry weight mass percent is 1.2% KH
2PO
4, fermentation temperature 28-32 ℃, fermentation time 60-70h;
(2) after the fermentation ends, be warming up to 50-52 ℃, hydrolysis 4-5h is dried to water content less than 8% with tunning then, pulverizes, and crosses 80 mesh sieves, must be rich in the feed goods of active peptides.
2. the method for active peptides goods is rich in a kind of solid state fermentation production according to claim 1; It is characterized in that: described aspergillus niger (Aspergillus niger; ACCC30787) seed liquor prepares as follows: slant strains is transferred in the liquid activation medium; Place shaking table, shaken cultivation 32-40h under 28-32 ℃, 200r/min condition; In weight portion, the prescription of described liquid activation medium is: NaNO
30.2 part, K
2HPO
40.1 part, 0.05 part of KCl, MgSO
40.05 part, FeSO
40.001 part, 3.0 parts of sucrose, 100 parts of distilled water, natural pH value.
3. the method for active peptides goods is rich in a kind of solid state fermentation production according to claim 1; It is characterized in that: described bacillus subtilis (Bacillus subtilis; ACCC01746) seed liquor prepares as follows: slant strains is transferred in the liquid activation medium; Place shaking table, shaken cultivation 28-30h under 33-35 ℃, 200r/min condition; In weight portion, the prescription of described liquid activation medium is: 0.2 part of glucose, 0.5 part of NaCl, 0.5 part of yeast extract, 1 part of peptone, 100 parts of distilled water, pH7.0.
4. the application of active peptides goods in the preparation animal feed for preparing according to each said method of claim 1-3.
5. the application of active peptides goods in the preparation pig starter feed for preparing according to each said method of claim 1-3.
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| CN102660622A (en) * | 2012-04-28 | 2012-09-12 | 浙江大飞龙动物保健品有限公司 | Method for preparing bioactive soybean peptide by mixed culture solid-state fermentation |
| CN103829040A (en) * | 2014-03-18 | 2014-06-04 | 广东轻工职业技术学院 | Method for compounding and fermenting soybean meal by Chenxiang tea and bacillus subtilis and application |
| CN103918875A (en) * | 2014-04-11 | 2014-07-16 | 沈阳市康普利德生物科技有限公司 | Preparation method of cured fermented feed |
| CN104372060A (en) * | 2014-08-11 | 2015-02-25 | 江苏大学 | Method for preparation of oligopeptides by solid state fermentation cake |
| CN106720935A (en) * | 2016-12-30 | 2017-05-31 | 四川省旺达饲料有限公司 | A kind of fermentation wheat bran rich in bioactive micro peptide and preparation method thereof |
| CN106977292A (en) * | 2017-04-18 | 2017-07-25 | 烟台泓源生物肥料有限公司 | A kind of calcic water-soluble compound fertilizer and its preparation technology |
| CN107410771A (en) * | 2017-08-15 | 2017-12-01 | 兰溪市酉泽饲料技术服务有限公司 | Crab and prawn mixed breeding special feed |
| CN109362942A (en) * | 2018-12-29 | 2019-02-22 | 广东旺大集团股份有限公司 | A kind of wean sucking pig fermented feed and preparation method thereof |
| CN113699201A (en) * | 2021-08-25 | 2021-11-26 | 吉林省畜牧兽医科学研究院 | Method for preparing bioactive peptide |
| CN115944022A (en) * | 2022-12-27 | 2023-04-11 | 成都大帝汉克生物科技有限公司 | Preparation method of animal feeding promoting agent |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660622A (en) * | 2012-04-28 | 2012-09-12 | 浙江大飞龙动物保健品有限公司 | Method for preparing bioactive soybean peptide by mixed culture solid-state fermentation |
| CN103829040B (en) * | 2014-03-18 | 2015-10-07 | 广东轻工职业技术学院 | A kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans and application |
| CN103829040A (en) * | 2014-03-18 | 2014-06-04 | 广东轻工职业技术学院 | Method for compounding and fermenting soybean meal by Chenxiang tea and bacillus subtilis and application |
| CN103918875A (en) * | 2014-04-11 | 2014-07-16 | 沈阳市康普利德生物科技有限公司 | Preparation method of cured fermented feed |
| CN104372060B (en) * | 2014-08-11 | 2018-04-03 | 江苏大学 | A kind of method that solid state fermentation grouts prepare oligopeptide |
| CN104372060A (en) * | 2014-08-11 | 2015-02-25 | 江苏大学 | Method for preparation of oligopeptides by solid state fermentation cake |
| CN106720935A (en) * | 2016-12-30 | 2017-05-31 | 四川省旺达饲料有限公司 | A kind of fermentation wheat bran rich in bioactive micro peptide and preparation method thereof |
| CN106977292A (en) * | 2017-04-18 | 2017-07-25 | 烟台泓源生物肥料有限公司 | A kind of calcic water-soluble compound fertilizer and its preparation technology |
| CN106977292B (en) * | 2017-04-18 | 2018-03-30 | 烟台泓源生物肥料有限公司 | A kind of calcic water-soluble compound fertilizer and its preparation technology |
| CN107410771A (en) * | 2017-08-15 | 2017-12-01 | 兰溪市酉泽饲料技术服务有限公司 | Crab and prawn mixed breeding special feed |
| CN109362942A (en) * | 2018-12-29 | 2019-02-22 | 广东旺大集团股份有限公司 | A kind of wean sucking pig fermented feed and preparation method thereof |
| CN113699201A (en) * | 2021-08-25 | 2021-11-26 | 吉林省畜牧兽医科学研究院 | Method for preparing bioactive peptide |
| CN115944022A (en) * | 2022-12-27 | 2023-04-11 | 成都大帝汉克生物科技有限公司 | Preparation method of animal feeding promoting agent |
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Application publication date: 20120711 |