CN102321710A - Method for preparing soy peptide with immobilized cell - Google Patents
Method for preparing soy peptide with immobilized cell Download PDFInfo
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- CN102321710A CN102321710A CN201110195717A CN201110195717A CN102321710A CN 102321710 A CN102321710 A CN 102321710A CN 201110195717 A CN201110195717 A CN 201110195717A CN 201110195717 A CN201110195717 A CN 201110195717A CN 102321710 A CN102321710 A CN 102321710A
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- soybean peptides
- immobilized cell
- soy peptide
- nutrient solution
- somatic cells
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Abstract
The invention discloses a method for preparing soy peptide with an immobilized cell, which is characterized in that: fermented and transformed culture fluid is adopted as a substrate; and the immobilized cell which contains bacillusli cheniformis is prepared into the soy peptide through bacterium activation, the expanded culture of bacteria, bacterium cell collection, bacterium cell immobilization, biological transformation, concentration and spray drying. The method for preparing soy peptide with the immobilized cell prepares the soy peptide with the immobilized cell which contains bacillusli cheniformis, not only can conveniently collect the transformation fluid to prepare the soy peptide but also can be used repeatedly, so that a process is simplified, the utilization rate of the substrate is improved, the soy peptide transformation rate of the substrate can reach above 50 percents, the product yield can reach above 45 percents, and benefits the automatic control in an industrialized production process.
Description
Technical field
The present invention relates to utilize fixation of microbial cell to prepare the method for soybean peptides, belong to technical field of enzyme engineering.
Background technology
The low-peptide mixer that soybean peptides is made up of 2-10 amino-acid residue; It has and be prone to absorb, is prone to digestion, promotes many-sided physicochemical properties such as mineral substance absorbs, anti-oxidant, strengthening immunity; Except being rich in nutrition, easy digestion, also have low antigenicity, suppress SUV, promote biological functions such as metabolism of fat and promotion microbial fermentation.Plain as the super nutrient protein of a new generation, soybean peptides has been widely used in health care of food and feedstuff industry, has wide application and market outlook.
At present, the working method of soybean peptides mainly contains enzymolysis process and fermentation method.Enzymolysis process has been owing to the enzyme that uses high price has improved the production cost of soybean peptides, thereby limited its application; And fermentation method can make and contains a large amount of tropinas and other impurity in the fermented liquid, is unfavorable for the separation and purification of product, also causes production cost to increase.
Summary of the invention
The object of the present invention is to provide a kind of immobilized cell to prepare the method for soybean peptides; Collection conversion fluid that can be easy carries out the preparation of soybean peptides, and uses repeatedly, simplifies technological process; Improve substrate utilization ratio, help the automatic control in the commercial process.
The technical scheme that the present invention adopted is following:
In the present invention, be used for bacterial classification that soybean peptides produces be Bacillus licheniformis (
Bacillus licheniformis).
Substratum and culture condition
⑴ seed culture medium
Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g adds water to 1000mL, pH7.2-7.4,30min is subsequent use through the 0.1MPa autoclaving;
⑵ shake-flask culture base
Soybean protein isolate 30g-45g, NaCl 5g-6g adds water to 1000mL, pH value nature, 30min is subsequent use through the 0.1MPa autoclaving;
⑶ fermentation transforms nutrient solution
Soybean protein isolate 80g-120g adds water to 1000mL, pH value nature, and 30min is subsequent use through the 0.1MPa autoclaving.
This method utilizes immobilized cell to prepare soybean peptides, and its step is following:
⑴ actication of culture
Get slant preservation Bacillus licheniformis (
Bacillus licheniformis) bacterial classification inserts in the seed culture medium 37 ℃ and cultivate 24h;
⑵ the collection of strain expanded culture and somatic cells
Above-mentioned bacterial classification is received in the shake-flask culture base, and liquid amount is the 50-100mL/250mL triangular flask, and inoculum size is 2%-5%, in 35 ℃-37 ℃, 150r/min-200r/min shaking culture 18h-24h, then medium centrifugal is collected thalline;
⑶ somatic cells immobilization
Collected somatic cells and a certain amount of embedding medium are mixed, splash into curing molding in the forming agent, promptly get the immobilized bacterium somatocyte;
⑷ the conversion of soybean peptides
With said fixing cell colloid dress post, will ferment transforms the nutrient solution gel column of flowing through and carries out bio-transformation, collects effusive conversion fluid, and conversion fluid can repeat upper prop, and the substrate of 80%-90% only is converted in nutrient solution.
⑸ the preparation of soy peptide powder
Above-mentioned conversion fluid is concentrated into suitable concn, can obtains the soybean peptides liquid concentrator, the soybean peptides liquid concentrator promptly gets soy peptide powder through spraying drying.
Embedding medium described in the above-mentioned steps ⑶ is a sodium-alginate; Forming agent is a calcium chloride solution.
The concentration of the embedding medium sodium-alginate described in the above-mentioned steps ⑶ is 1-5%, and the amount of somatic cells is l0-20%; The concentration of forming agent calcium chloride is 1-5%, and be 2-3h set time, and solidification value is 30-40 ℃.
It is 200-500mL/h that fermentation described in the above-mentioned steps ⑷ transforms the nutrient solution flow velocity, and invert point is 35-40 ℃.
The present invention utilizes the method production soybean peptides of Bacillus licheniformis immobilized cell; Collection conversion fluid that not only can be easy carries out the preparation of soybean peptides, and can use repeatedly, has simplified technological process; Improved the utilization ratio of substrate; The soybean peptides transformation efficiency of substrate can be reached more than 50%, and the product yield can reach more than 45%, and helps the automatic control in the commercial process.
Embodiment
Embodiment 1
1. substratum
⑴ seed culture medium
Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g adds water to 1000mL, pH7.2-7.4,30min is subsequent use through the 0.1MPa autoclaving;
⑵ shake-flask culture base
Soybean protein isolate 30g, NaCl 5g adds water to 1000mL, pH value nature, 30min is subsequent use through the 0.1MPa autoclaving;
⑶ fermentation transforms nutrient solution
Soybean protein isolate 80g adds water to 1000mL, pH value nature, and 30min is subsequent use through the 0.1MPa autoclaving.
2. the collection of the enlarged culturing of bacterial classification and somatic cells
Get slant preservation Bacillus licheniformis (
Bacillus licheniformis) bacterial classification inserts in the seed culture medium 37 ℃ and cultivate 24h.Above-mentioned bacterial classification is received in the shake-flask culture base, and liquid amount is the 50-100mL/250mL triangular flask, and inoculum size is 5%, in 37 ℃, 180r/min shaking culture 20h, then medium centrifugal is collected thalline.
3. somatic cells immobilization
Is that 5% sodium alginate to embed agent mixes with collected somatic cells in the ratio of 1:5 and concentration, splashes into concentration and be curing molding 2h in 5% calcium chloride, the forming agent that temperature is 40 ℃, promptly gets and fixes
Change somatic cells.
4. the conversion of soybean peptides
With said fixing cell dress post; To ferment transforms the nutrient solution gel column of flowing through and carries out bio-transformation, and fermentation transforms the nutrient solution Flow Control and is made as 300mL/h, collects effusive conversion fluid; Conversion fluid can repeat upper prop, and the substrate of the 80%-90% in nutrient solution is converted into and ends.
5. the preparation of soy peptide powder
Above-mentioned conversion fluid is concentrated into suitable concn, can obtains the soybean peptides liquid concentrator, the soybean peptides liquid concentrator promptly gets soy peptide powder through spraying drying.
Embodiment 2
1. substratum
⑴ seed culture medium
Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g adds water to 1000mL, pH7.2-7.4,30min is subsequent use through the 0.1MPa autoclaving;
⑵ shake-flask culture base
Soybean protein isolate 35g, NaCl 5g adds water to 1000mL, pH value nature, 30min is subsequent use through the 0.1MPa autoclaving;
⑶ fermentation transforms nutrient solution
Soybean protein isolate 100g adds water to 1000mL, pH value nature, and 30min is subsequent use through the 0.1MPa autoclaving.
2. the collection of the enlarged culturing of bacterial classification and somatic cells
Get slant preservation Bacillus licheniformis (
Bacillus licheniformis) bacterial classification inserts in the seed culture medium 37 ℃ and cultivate 24h.Above-mentioned bacterial classification is received in the shake-flask culture base, and liquid amount is the 50mL/250mL triangular flask, and inoculum size is 3%, in 35 ℃, 200r/min shaking culture 24h, then medium centrifugal is collected thalline.
3. somatic cells immobilization
Is that 5% sodium alginate to embed agent mixes with collected somatic cells in the ratio of 1:5 and concentration, splashes into concentration and be curing molding 3h in 1% calcium chloride, the forming agent that temperature is 35 ℃, promptly gets and fixes
Change somatic cells.
4. the conversion of soybean peptides
With said fixing cell dress post; To ferment transforms the nutrient solution gel column of flowing through and carries out bio-transformation, and fermentation transforms the nutrient solution Flow Control and is made as 250mL/h, collects effusive conversion fluid; Conversion fluid can repeat upper prop, and the substrate of the substrate 80%-90% in nutrient solution is converted into and ends.
5. the preparation of soy peptide powder
Above-mentioned conversion fluid is concentrated into suitable concn, can obtains the soybean peptides liquid concentrator, the soybean peptides liquid concentrator promptly gets soy peptide powder through spraying drying.
Embodiment 3
1. substratum
⑴ seed culture medium
Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g adds water to 1000mL, pH7.2-7.4,30min is subsequent use through the 0.1MPa autoclaving;
⑵ shake-flask culture base
Soybean protein isolate 45g, NaCl 6g adds water to 1000mL, pH value nature, 30min is subsequent use through the 0.1MPa autoclaving;
⑶ fermentation transforms nutrient solution
Soybean protein isolate 120g adds water to 1000mL, pH value nature, and 30min is subsequent use through the 0.1MPa autoclaving.
2. the collection of the enlarged culturing of bacterial classification and somatic cells
Get slant preservation Bacillus licheniformis (
Bacillus licheniformis) bacterial classification inserts in the seed culture medium 37 ℃ and cultivate 24h.Above-mentioned bacterial classification is received in the shake-flask culture base, and liquid amount is the 80mL/250mL triangular flask, and inoculum size is 3%, in 37 ℃, 180r/min shaking culture 18h, then medium centrifugal is collected thalline.
3. somatic cells immobilization
Is that 1% sodium alginate to embed agent mixes with collected somatic cells in the ratio of 1:5 and concentration, splashes into concentration and be curing molding 2.5h in 5% calcium chloride, the forming agent that temperature is 38 ℃, promptly gets and consolidates
Surely change somatic cells.
4. the conversion of soybean peptides
With said fixing cell dress post; To ferment transforms the nutrient solution gel column of flowing through and carries out bio-transformation, and fermentation transforms the nutrient solution Flow Control and is made as 200mL/h, collects effusive conversion fluid; Conversion fluid can repeat upper prop, and the substrate of the substrate 80%-90% in nutrient solution is converted into and ends.
5. the preparation of soy peptide powder
Above-mentioned conversion fluid is concentrated into suitable concn, can obtains the soybean peptides liquid concentrator, the soybean peptides liquid concentrator promptly gets soy peptide powder through spraying drying.
Embodiment 4
1. substratum
⑴ seed culture medium
Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g adds water to 1000mL, pH7.2-7.4,30min is subsequent use through the 0.1MPa autoclaving;
⑵ shake-flask culture base
Soybean protein isolate 35g, NaCl 5g adds water to 1000mL, pH value nature, 30min is subsequent use through the 0.1MPa autoclaving;
⑶ fermentation transforms nutrient solution
Soybean protein isolate 100g adds water to 1000mL, pH value nature, and 30min is subsequent use through the 0.1MPa autoclaving.
2. the collection of the enlarged culturing of bacterial classification and somatic cells
Get slant preservation Bacillus licheniformis (
Bacillus licheniformis) bacterial classification inserts in the seed culture medium 37 ℃ and cultivate 24h.Above-mentioned bacterial classification is received in the shake-flask culture base, and liquid amount is the 50mL/250mL triangular flask, and inoculum size is 3%, in 35 ℃, 200r/min shaking culture 24h, then medium centrifugal is collected thalline.
3. somatic cells immobilization
Is that 3% sodium alginate to embed agent mixes with collected somatic cells in the ratio of 1:5 and concentration, splashes into concentration and be curing molding 3h in 2% calcium chloride, the forming agent that temperature is 30 ℃, promptly gets and fixes
Change somatic cells.
4. the conversion of soybean peptides
With said fixing cell dress post; To ferment transforms the nutrient solution gel column of flowing through and carries out bio-transformation, and fermentation transforms the nutrient solution Flow Control and is made as 250mL/h, collects effusive conversion fluid; Conversion fluid can repeat upper prop, and the substrate of the substrate 80%-90% in nutrient solution is converted into and ends.
5. the preparation of soy peptide powder
Above-mentioned conversion fluid is concentrated into suitable concn, can obtains the soybean peptides liquid concentrator, the soybean peptides liquid concentrator promptly gets soy peptide powder through spraying drying.
Claims (6)
1. method of utilizing immobilized cell to prepare soybean peptides, its step is following:
⑴ the preparation of substratum
1. seed culture medium
Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g adds water to 1000mL, pH7.2-7.4,30min is subsequent use through the 0.1MPa autoclaving;
2. shake-flask culture base
Soybean protein isolate 30g-45g, NaCl 5g-6g adds water to 1000mL, pH value nature, 30min is subsequent use through the 0.1MPa autoclaving;
3. fermentation transforms nutrient solution
Soybean protein isolate 80g-120g adds water to 1000mL, pH value nature, and 30min is subsequent use through the 0.1MPa autoclaving;
⑵ actication of culture
Get slant preservation Bacillus licheniformis (
Bacillus licheniformis) bacterial classification inserts in the seed culture medium 37 ℃ and cultivate 24h;
⑶ the collection of strain expanded culture and somatic cells
Above-mentioned bacterial classification is received in the shake-flask culture base, and liquid amount is the 50-100mL/250mL triangular flask, and inoculum size is 3%-5%, in 35 ℃-37 ℃, 150r/min-200r/min shaking culture 18h-24h, then with the centrifugal collection thalline of fermented liquid;
⑷ somatic cells immobilization
Collected somatic cells and a certain amount of embedding medium are mixed, splash into curing molding in the forming agent, promptly get the immobilized bacterium somatocyte;
⑸ the conversion of soybean peptides
With said fixing cell colloid dress post, will ferment transforms the nutrient solution gel column of flowing through and carries out bio-transformation, collects effusive conversion fluid, and conversion fluid can repeat upper prop, and the substrate of 80%-90% only is converted in nutrient solution;
⑹ the preparation of soy peptide powder
Above-mentioned conversion fluid is concentrated into suitable concn, can obtains the soybean peptides liquid concentrator, the soybean peptides liquid concentrator promptly gets soy peptide powder through spraying drying.
2. the method for utilizing immobilized cell to prepare soybean peptides according to claim 1 is characterized in that: the bacterial classification described in the step ⑵ be Bacillus licheniformis (
Bacillus licheniformis).
3. the method for utilizing immobilized cell to prepare soybean peptides according to claim 1 is characterized in that: the embedding medium described in the step ⑷ is a sodium-alginate.
4. the method for utilizing immobilized cell to prepare soybean peptides according to claim 1 is characterized in that: the forming agent described in the step ⑷ is a calcium chloride solution.
5. the method for utilizing immobilized cell to prepare soybean peptides according to claim 1; It is characterized in that: the concentration of the embedding medium sodium-alginate described in the step ⑷ is 1-5%; The amount of somatic cells is l0-20%; The concentration of forming agent calcium chloride is 1-5%, and be 2-3h set time, and solidification value is 30-40 ℃.
6. the method for utilizing immobilized cell to prepare soybean peptides according to claim 1 is characterized in that: it is 200-500mL/h that the fermentation described in the step ⑸ transforms the nutrient solution flow velocity, and invert point is 35-40 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110195717A CN102321710A (en) | 2011-07-13 | 2011-07-13 | Method for preparing soy peptide with immobilized cell |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110195717A CN102321710A (en) | 2011-07-13 | 2011-07-13 | Method for preparing soy peptide with immobilized cell |
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| CN102321710A true CN102321710A (en) | 2012-01-18 |
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| CN201110195717A Pending CN102321710A (en) | 2011-07-13 | 2011-07-13 | Method for preparing soy peptide with immobilized cell |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676482A (en) * | 2012-05-11 | 2012-09-19 | 河南省南街村（集团）有限公司 | Method for producing alkali protease from highland bacillus |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1546648A (en) * | 2003-12-15 | 2004-11-17 | 宜昌市微生物研究所 | A bacterial strain and carotenoid production method using said strain |
| CN101779721A (en) * | 2010-01-25 | 2010-07-21 | 哈尔滨商业大学 | Method for preparing soybean protein |
| CN101979646A (en) * | 2010-11-05 | 2011-02-23 | 河南省南街村（集团）有限公司 | Method for preparing guanosine by using immobilized cell |
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2011
- 2011-07-13 CN CN201110195717A patent/CN102321710A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1546648A (en) * | 2003-12-15 | 2004-11-17 | 宜昌市微生物研究所 | A bacterial strain and carotenoid production method using said strain |
| CN101779721A (en) * | 2010-01-25 | 2010-07-21 | 哈尔滨商业大学 | Method for preparing soybean protein |
| CN101979646A (en) * | 2010-11-05 | 2011-02-23 | 河南省南街村（集团）有限公司 | Method for preparing guanosine by using immobilized cell |
Non-Patent Citations (2)
| Title |
|---|
| 乐超银等: "一株地衣芽孢杆菌的分离及在大豆肽发酵中的应用", 《中国酿造》 * |
| 王顺新等: "酶工程技术在开发活性多肽方面的研究进展", 《食品工程》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676482A (en) * | 2012-05-11 | 2012-09-19 | 河南省南街村（集团）有限公司 | Method for producing alkali protease from highland bacillus |
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Application publication date: 20120118 |