CN108531529A - A method of preparing polypeptide using microbe fermentation method - Google Patents
A method of preparing polypeptide using microbe fermentation method Download PDFInfo
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- CN108531529A CN108531529A CN201810343111.0A CN201810343111A CN108531529A CN 108531529 A CN108531529 A CN 108531529A CN 201810343111 A CN201810343111 A CN 201810343111A CN 108531529 A CN108531529 A CN 108531529A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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Abstract
The present invention discloses a kind of method preparing polypeptide using microbe fermentation method, includes the following steps：(1) culture medium for configuring aspergillus terricola, is inoculated with aspergillus terricola, and culture obtains aspergillus terricola seed liquor；(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, adjust pH value, sterilizing is inoculated with aspergillus terricola seed liquor, fermentation, ultrafiltration, it is dry to get.The method of the present invention is effectively retained bioactive substance and nutritional ingredient in raw material system, and the polypeptide being prepared is more high-quality, and polypeptide yield significantly improves.
Description
Technical field
The present invention relates to deep-processing technical field of agricultural products.It is prepared using microbe fermentation method more particularly, to a kind of
The method of polypeptide.
Background technology
For the mankind by food intake protein, these protein are exactly the source of many biologically active peptides.The how white peptide of egg is
A kind of water-solubility protein, can directly be absorbed rapidly by human body, participate in adjust metabolic processes have to reach extending life
The miraculous effects of delaying senility are imitated, the market demand is extensive.
Microbe fermentation method is by selecting excellent fermentation strain, the protease water generated in growth course using strain
Substrate protein is solved, then tunning is isolated and purified, finally obtains biologically active peptide.Common methods be by fermentation,
Centrifugation, concentration, dry preparation, process is more, is unfavorable for operating in industrialized production, it would be highly desirable to improve.
Corn and soybean, peanut the dregs of rice be the byproduct obtained after extract oil, it is most as livestock and poultry feed, with
The generation of big content of starting materials, the polypeptide prepared by microbe fermentation method is a kind of new type functional food raw material, added value
Higher, market development have a extensive future.
Invention content
The purpose of the present invention is to provide a kind of methods preparing polypeptide using microbe fermentation method.
In order to achieve the above objectives, the present invention uses following technical proposals：
A method of polypeptide being prepared using microbe fermentation method, is included the following steps：
(1) culture medium for configuring aspergillus terricola, is inoculated with aspergillus terricola, and culture obtains aspergillus terricola seed liquor；
(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, pH value, sterilizing is adjusted to be inoculated with aspergillus terricola kind
Sub- liquid, fermentation, ultrafiltration, it is dry to get.
In the preferred embodiment of the present invention, the culture medium of the aspergillus terricola includes each group of following parts by weight
Point：15-19 parts of peptone, 90-110 parts of sodium chloride, 3-4 parts of dipotassium hydrogen phosphate, 2-3 parts of glucose, 900-1000 parts of water.
Further, the preparation method of the culture medium of the aspergillus terricola includes：By peptone, sodium chloride, dipotassium hydrogen phosphate,
Glucose part, water mixing, 6.9-7.2 is adjusted to by pH value, and sterilize 15-20min under the conditions of pressure 103.4kPa, 121 DEG C, will
Temperature is down to 35-45 DEG C.
Further, the inoculum concentration of step (1) described aspergillus terricola is 2-5%.
Further, step (1) culture is small to cultivate 20-24 under the conditions of 220-260 revolutions per minutes, 28-30 DEG C
When.
In the preferred embodiment of the present invention, step (2) the corn germ cake powder, soybean meal powder, peanut meal powder, water
Mass ratio be 1-2:2-3:1-2:3-4.
Further, step (2) described pH value is 7.0-7.2.
Further, step (2) sterilizing is the 18-20min that sterilizes under the conditions of pressure 103.4kPa, 121.3 DEG C of temperature.
Further, the inoculum concentration of step (2) the aspergillus terricola seed liquor is 7-8%.
Further, step (2) fermentation is the 70-72h that ferments under the conditions of 160-180 revolutions per minutes, 28-30 DEG C.
Further, the filter sizes of step (2) described ultrafiltration be 0.05um-1um, pressure 0.1-0.5Mpa.
Further, step (2) described drying is pressure spray dryer, and the pressure spray dryer condition is pressure 5-
10Mpa maintains 10-20s.
Prepared by the method for the present invention so that the yield of polypeptide significantly improves, 95% or more.
Beneficial effects of the present invention are as follows：
Corn germ cake of the present invention contains thick protein, crude fat, crude fibre etc.；Soybean meal contains protein, and oligosaccharide is more
Sugar and cellulose etc.；Peanut meal contains thick protein, arginine etc., nutrition complement after three kinds of raw material mixing, the polypeptide of production
It is more high-quality, while also extending the industrial chain of agricultural product.
The method of the present invention replaces the centrifugation of conventional method and concentration two procedures, process to be not required to increase anyization with ultrafiltration
Reagent is learned, processing procedure without phase-change, to constituent in material without any harmful effect, and detaches, purifies, begins in concentration process
It is in normal temperature state eventually, it is entirely avoided high temperature destroys this drawback to bioactive substance, is effectively retained in raw material system
Bioactive substance and nutritional ingredient, the quality and yield of polypeptide significantly improve.Low energy consumption for device systems, production week
Phase is short, can effectively reduce production cost, is conducive to the popularization of industrialized production, improves Business Economic Benefit.
Specific implementation mode
In order to illustrate more clearly of the present invention, with reference to preferred embodiment, the present invention is described further.Ability
Field technique personnel should be appreciated that following specifically described content is illustrative and be not restrictive, this should not be limited with this
The protection domain of invention.
Aspergillus terricola that the present invention uses, aspergillus awamori is are purchased from Jing Bei Na Chuanlian Bioteknologisk Institut.
A kind of method preparing polypeptide using microbe fermentation method of embodiment 1
A method of polypeptide being prepared using microbe fermentation method, is included the following steps：
(1) peptone 15g, sodium chloride 90g, dipotassium hydrogen phosphate 3g, glucose 2g, water 900g are weighed, mixing adjusts pH value
To 6.9, sterilize 15min under the conditions of pressure 103.4kPa, 121 DEG C, cools the temperature to 35 DEG C, and soil of dwelling is inoculated with 2% inoculum concentration
Aspergillus obtains aspergillus terricola seed liquor to be cultivated 20 hours under the conditions of 220 revolutions per minutes, 28 DEG C；
(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, the corn germ cake powder, soybean meal powder,
Peanut meal powder, water mass ratio be 1:2:1:3, it adjusts pH value to 7.0, sterilizes under the conditions of pressure 103.4kPa, 121.3 DEG C of temperature
18min cools the temperature to 30 DEG C, aspergillus terricola seed liquor is inoculated with 7% inoculum concentration, under the conditions of 160 revolutions per minutes, 28 DEG C
Ferment 70h, and with filter sizes be 0.05um, pressure is 0.1Mpa ultrafiltration, with pressure 5Mpa, maintains 10s progress press atomizations dry
It is dry to get.
After measured, the yield that the above method prepares polypeptide is 95.3%.
A kind of method preparing polypeptide using microbe fermentation method of embodiment 2
A method of polypeptide being prepared using microbe fermentation method, is included the following steps：
(1) peptone 19g, sodium chloride 110g, dipotassium hydrogen phosphate 4g, glucose 3g, water 1000g are weighed, mixing adjusts pH
It is worth 7.2, sterilize 20min under the conditions of pressure 103.4kPa, 121 DEG C, cools the temperature to 45 DEG C, dwells with the inoculation of 5% inoculum concentration
Aspergillus terreus obtains aspergillus terricola seed liquor to be cultivated 24 hours under the conditions of 260 revolutions per minutes, 30 DEG C；
(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, the corn germ cake powder, soybean meal powder,
Peanut meal powder, water mass ratio be 2:3:2:4, it adjusts pH value to 7.2, sterilizes under the conditions of pressure 103.4kPa, 121.3 DEG C of temperature
18-20min cools the temperature to 35 DEG C, aspergillus terricola seed liquor is inoculated with 8% inoculum concentration, in 180 revolutions per minutes, 30 DEG C of conditions
Lower fermentation 72h, with filter sizes be 1um, pressure is 0.5Mpa ultrafiltration, with pressure 10Mpa, maintains 20s progress press atomizations dry
It is dry to get.
After measured, the yield that the above method prepares polypeptide is 95.8%.
A kind of method preparing polypeptide using microbe fermentation method of embodiment 3
A method of polypeptide being prepared using microbe fermentation method, is included the following steps：
(1) peptone 17g, sodium chloride 100g, dipotassium hydrogen phosphate 3.5g, glucose 2.5g, water 950g are weighed, mixing is adjusted
PH value is saved to 7.0, sterilize 17min under the conditions of pressure 103.4kPa, 121 DEG C, cools the temperature to 40 DEG C, is connect with 3% inoculum concentration
Kind aspergillus terricola obtains aspergillus terricola seed liquor to be cultivated 22 hours under the conditions of 240 revolutions per minutes, 29 DEG C；
(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, the corn germ cake powder, soybean meal powder,
Peanut meal powder, water mass ratio be 1:2:2:4, it adjusts pH value to 7.1, sterilizes under the conditions of pressure 103.4kPa, 121.3 DEG C of temperature
19min cools the temperature to 32 DEG C, aspergillus terricola seed liquor is inoculated with 7.5% inoculum concentration, in 170 revolutions per minutes, 29 DEG C of conditions
Lower fermentation 71h, with filter sizes be 0.1um, pressure is 0.4Mpa ultrafiltration, with pressure 7Mpa, maintains 15s progress press atomizations dry
It is dry to get.
After measured, the yield that the above method prepares polypeptide is 95.3%.
A kind of method preparing polypeptide using microbe fermentation method of comparative example 1, includes the following steps：
(1) peptone 17g, sodium chloride 100g, dipotassium hydrogen phosphate 3.5g, glucose 2.5g, water 950g are weighed, mixing is adjusted
PH value is saved to 7.0, sterilize 17min under the conditions of pressure 103.4kPa, 121 DEG C, cools the temperature to 40 DEG C, is connect with 3% inoculum concentration
Kind aspergillus awamori obtains aspergillus awamori seed liquor to be cultivated 22 hours under the conditions of 240 revolutions per minutes, 29 DEG C；
(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, the corn germ cake powder, soybean meal powder,
Peanut meal powder, water mass ratio be 1:2:2:4, it adjusts pH value to 7.1, sterilizes under the conditions of pressure 103.4kPa, 121.3 DEG C of temperature
19min cools the temperature to 32 DEG C, aspergillus awamori seed liquor is inoculated with 7.5% inoculum concentration, in 170 revolutions per minutes, 29 DEG C of conditions
Lower fermentation 71h, with filter sizes be 0.1um, pressure is 0.4Mpa ultrafiltration, with pressure 7Mpa, maintains 15s progress press atomizations dry
It is dry to get.
After measured, the yield that the above method prepares polypeptide is 80.5%.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair
The restriction of embodiments of the present invention may be used also on the basis of the above description for those of ordinary skill in the art
To make other variations or changes in different ways, all embodiments can not be exhaustive here, it is every to belong to this hair
Row of the obvious changes or variations that bright technical solution is extended out still in protection scope of the present invention.
Claims (10)
1. a kind of method preparing polypeptide using microbe fermentation method, which is characterized in that include the following steps：
(1) culture medium for configuring aspergillus terricola, is inoculated with aspergillus terricola, and culture obtains aspergillus terricola seed liquor；
(2) by corn germ cake powder, soybean meal powder, peanut meal powder, water mixing, pH value, sterilizing is adjusted to be inoculated with aspergillus terricola seed
Liquid, fermentation, ultrafiltration, it is dry to get.
2. according to the method described in claim 1, it is characterized in that：The culture medium of the aspergillus terricola, including following weight
The each component of part：15-19 parts of peptone, 90-110 parts of sodium chloride, 3-4 parts of dipotassium hydrogen phosphate, 2-3 parts of glucose, water 900-
1000 parts.
3. according to the method described in claim 2, it is characterized in that：The preparation method of the culture medium of the aspergillus terricola includes：
By peptone, sodium chloride, dipotassium hydrogen phosphate, glucose, water mixing, pH value is adjusted to 6.9-7.2, pressure 103.4kPa,
Sterilize 15-20min under the conditions of 121 DEG C, cools the temperature to 35-45 DEG C.
4. according to the method described in claim 1, it is characterized in that：The inoculum concentration of step (1) described aspergillus terricola is 2-5%.
5. according to the method described in claim 1, it is characterized in that：Step (1) it is described culture be with 220-260 revolutions per minutes,
It is cultivated 20-24 hours under the conditions of 28-30 DEG C.
6. according to the method described in claim 1, it is characterized in that：Step (2) the corn germ cake powder, soybean meal powder, flower
Give birth to dregs of rice powder, the mass ratio of water is 1-2:2-3:1-2:3-4.
7. according to the method described in claim 1, it is characterized in that：Step (2) described pH value is 7.0-7.2；The sterilizing is
Sterilize 18-20min under the conditions of pressure 103.4kPa, 121.3 DEG C of temperature；The fermentation is in 160-180 revolutions per minutes, 28-
Ferment 70-72h under the conditions of 30 DEG C.
8. according to the method described in claim 1, it is characterized in that：The inoculum concentration of step (2) the aspergillus terricola seed liquor is
7-8%.
9. according to the method described in claim 1, it is characterized in that：The filter sizes of step (2) described ultrafiltration are 0.05um-
1um, pressure 0.1-0.5Mpa.
10. according to the method described in claim 1, it is characterized in that：Step (2) described drying is pressure spray dryer, described
Pressure spray dryer condition is pressure 5-10Mpa, maintains 10-20s.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111096368A (en) * | 2019-12-17 | 2020-05-05 | 顾霆 | Method for splitting nutrient substances of soybean milk powder |
| CN112251472A (en) * | 2020-09-21 | 2021-01-22 | 宁夏泰胜生物科技有限公司 | Culture medium and culture method for producing coenzyme Q10 by fermentation of rhodobacter sphaeroides |
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| CN1703968A (en) * | 2004-05-25 | 2005-12-07 | 中国农业大学 | Synbiotic feed addictive of mini-peptide with beneficial microbe and its usage |
| CN104087642A (en) * | 2014-07-29 | 2014-10-08 | 哈尔滨伟平科技开发有限公司 | Method for making protein polypeptide |
| CN104087637A (en) * | 2014-07-29 | 2014-10-08 | 哈尔滨伟平科技开发有限公司 | Method for preparing vegetable protein polypeptide by virtue of microbiological fermentation |
| CN105132509A (en) * | 2015-09-25 | 2015-12-09 | 华南理工大学 | Method for extracting antioxidant protein polypeptide from peanut meal |
-
2018
- 2018-04-17 CN CN201810343111.0A patent/CN108531529A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1703968A (en) * | 2004-05-25 | 2005-12-07 | 中国农业大学 | Synbiotic feed addictive of mini-peptide with beneficial microbe and its usage |
| CN104087642A (en) * | 2014-07-29 | 2014-10-08 | 哈尔滨伟平科技开发有限公司 | Method for making protein polypeptide |
| CN104087637A (en) * | 2014-07-29 | 2014-10-08 | 哈尔滨伟平科技开发有限公司 | Method for preparing vegetable protein polypeptide by virtue of microbiological fermentation |
| CN105132509A (en) * | 2015-09-25 | 2015-12-09 | 华南理工大学 | Method for extracting antioxidant protein polypeptide from peanut meal |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111096368A (en) * | 2019-12-17 | 2020-05-05 | 顾霆 | Method for splitting nutrient substances of soybean milk powder |
| CN112251472A (en) * | 2020-09-21 | 2021-01-22 | 宁夏泰胜生物科技有限公司 | Culture medium and culture method for producing coenzyme Q10 by fermentation of rhodobacter sphaeroides |
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