CN100560731C - A kind of is the technology of raw material production nisin with the plant cake - Google Patents
A kind of is the technology of raw material production nisin with the plant cake Download PDFInfo
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- CN100560731C CN100560731C CNB200610019565XA CN200610019565A CN100560731C CN 100560731 C CN100560731 C CN 100560731C CN B200610019565X A CNB200610019565X A CN B200610019565XA CN 200610019565 A CN200610019565 A CN 200610019565A CN 100560731 C CN100560731 C CN 100560731C
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Abstract
The present invention relates to the production technology of the plain based food additive of a kind of biological antibiotic.A kind of is the technology of raw material production nisin with the plant cake, it is characterized in that it is raw material with the plant cake, with plant cake through enzymolysis, fermentation, solid-liquid separation, concentrate, saltout, dry and be packaged to be finished product.Adopt this technology, the output of nisin increases substantially, the fermentation unit of fermented liquid is about 3000IU/ml in the prior art suitability for industrialized production, and can reach 4500IU/ml with the highest fermentation unit of fermented liquid that this zymotechnique makes, improved 50% than existing technology, thereby also greatly reduce production cost of products, and improved the output of product.
Description
Technical field
The present invention relates to the production technology of the plain based food additive of a kind of biological antibiotic, be specifically related to a kind of technology of utilizing plant cake to produce nisin.
Background technology
Nisin is a kind of fermentating metabolism product of Lactococcus lactis, can suppress the growth and breeding of gram positive bacterium and gemma thereof effectively.It is a kind of micromolecule polypeptide class material that is made of 34 amino-acid residues, can be decomposed rapidly by the proteolytic enzyme in the human body alimentary canal after entering human body alimentary canal, thereby can be absorbed by the body utilization, thereby be a kind of safe, nontoxic food preservatives, and can not influence the profitable strain in the enteron aisle, more can not develop immunity to drugs.Nisin is natural existence in milk preparation, is one of putative natural, nontoxic biological antibiotic based food sanitas of minority, and China stipulates that in GB GB2760-1996 it can be used as food preservatives.
Chinese invention patent CN0015935.6 discloses a kind of " streptococcus acidi lactici preparation technology ".This invention is to make raw material with mung bean, pea etc., scalds through purifying that bubble, the defibrination dregs of rice remove slag, sterilize, add that the fermentation of female slurry, aggegation fecula purifying bacterial classification, serum sterilization breed, solid-liquid separation, liquid acid concentrate, accent PH salt precipitation, dry packing form.But it is relatively more expensive that this invention exists cost of material, the shortcoming that nisin output is lower.
Summary of the invention
For overcoming deficiency of the prior art, the invention provides a kind of output height, what production cost was low is the technology of raw material production nisin with the plant cake.
A kind of is the technology of raw material production nisin with the plant cake, it is characterized in that it is raw material with the plant cake, with plant cake through enzymolysis, fermentation, solid-liquid separation, concentrate, saltout, dry and be packaged to be finished product.
Described plant cake is dregs of rapeseed cake, peanut dregs, cotton cake dregs or soybean cake dregs.
Described enzymolysis is: the aqueous suspensions that plant cake is mixed with concentration 8-12g/100ml, boil and keep 30min, regulating pH after being cooled to temperature 30-60 ℃ is 2-10, be incorporated as the enzyme of plant cake weight 1-8% then, enzymolysis solution with gained behind the process 8-20h carries out filtering and impurity removing, gets vegetable-protein enzymolysis liquid.
The adding of described enzyme divides secondary to add, and the enzyme that adds is inequality with the enzyme that adds for the second time for the first time; The amount of the enzyme that adds is the 1-7% of plant cake weight for the first time.
Described enzyme is one or both the mixing wherein of aspartic protease, neutral protease, Sumizyme MP, trypsinase, papoid, and the proportioning when wherein any two kinds of enzymes mix is 1-5: 1-7.
Described fermentation is: the vegetable-protein enzymolysis liquid of gained is added water with sugar, yeast extract and phosphoric acid salt be mixed with the mixed solution that nitrogen content is 0.5-4g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the 5-15ml/100ml of mixed solution, the add-on of sugar are that the 1-5g/100ml of mixed solution, the add-on of yeast extract are that the 1-5g/100ml of mixed solution, phosphatic add-on are the 1-3g/100ml of mixed solution;
Mixed solution inserts the good Lactococcus lactis breast subspecies of activation after the sterilization cooling, the access amount of Lactococcus lactis breast subspecies is the 1-10% of mixeding liquid volume, and inoculation fermentation 12-30h gets fermented liquid.
Described sugar is one or more the mixing in sucrose, glucose, lactose, fructose, the maltose, and more than one are any proportioning when mixing.
Described phosphoric acid salt is one or more the mixing in potassium primary phosphate, SODIUM PHOSPHATE, MONOBASIC, dipotassium hydrogen phosphate, Sodium phosphate dibasic, the ammonium phosphate, and more than one are any proportioning when mixing.
Described solid-liquid separation is: the fermented liquid of above-mentioned steps gained is regulated the pH value to 2-4, adopt solid-liquid isolation method removal thalline and impurity to get filtered liquid.
Described simmer down to: because the filtered liquid volume that obtains behind solid-liquid separation process is bigger, therefore by rising the 1/2-1/20 that concentration technologies such as film, falling liquid film, multiple-effect or ultrafiltration are concentrated into filtered liquid original volume.So that nisin can dissociate from thalline fully, production cost also decreases simultaneously.
Described saltouing in concentrated solution, adding salt, through leaving standstill the centrifugal throw out that gets 6-10 hour to saturated; Selected salt is wherein one or both such as sodium-chlor, ammonium sulfate, sodium sulfate, sal epsom, sodium phosphate in the technology of saltouing, the proportioning when selecting for use two kinds to mix for arbitrarily than.
Described drying is: the salt precipitation thing through 40-80 ℃ low-temperature vacuum drying, makes finished product more than micronizing to 200 order.
The present invention compares with existing preparation technology, has following outstanding advantage:
1) choosing of raw material: utilized the by product plant cake to be raw material, reduced the cost of raw material significantly.
2) adopt the enzymolysis operation among the present invention, and selected for use aspartic protease, neutral protease, Sumizyme MP, trypsinase, papoid to carry out enzymolysis in the operation, the yield of product is higher.
3) adopt the Lactococcus lactis subsp.lactis inoculation can significantly improve the yield of product.
4) after fermented liquid carried out solid-liquid separation, the filtered liquid that obtains is carried out ultrafiltration and concentration can reduce production costs.
In sum, adopt this technology, the output of nisin increases substantially, the fermentation unit of fermented liquid is about 3000IU/ml in the prior art suitability for industrialized production, and can reach 4500IU/ml with the highest fermentation unit of fermented liquid that this zymotechnique makes, improved 50% than existing technology, thereby also greatly reduced production cost of products, and improved the output of product.
Embodiment
Further illustrate content of the present invention below in conjunction with embodiment, but content of the present invention not only is confined to the following examples.
Embodiment 1:
Dregs of rapeseed cake is mixed with the aqueous suspensions of 8g/100ml, stirring and evenly mixing, boil 30min, 50 ℃ of postcooling holding temperatures, sodium hydroxide with 5mol/l is regulated pH9.00, be incorporated as dregs of rapeseed cake weight 4% trypsinase while stirring, keeping enzymolysis solution pH with 5mol/l sodium hydroxide in the enzymolysis process is 9.00, enzymolysis 8h; Regulate enzymolysis liquid temp to 35 ℃, pH is 7.00, is incorporated as the neutral protease of dregs of rapeseed cake weight 4% while stirring, and keeping enzymolysis solution pH with the sodium hydroxide of 5mol/l is 7.00, and enzymolysis 4h gets enzymolysis solution.Enzymolysis solution adds the diatomite of 1g/100ml through the Plate Filtration removal of impurities, and filtered liquid is vegetable-protein enzymolysis liquid.The nitrogen content recovery rate of the vegetable-protein enzymolysis liquid that makes with this technology is 32%.
The vegetable-protein enzymolysis liquid that makes is added water with sugar, yeast extract and phosphoric acid salt be mixed with the mixed solution 3L that nitrogen content is 2g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the add-on of 8ml/100ml, the sucrose of mixed solution is that the add-on of the 1.5g/100ml of mixed solution, yeast extract is that the 1g/100ml of mixed solution, the add-on of Sodium phosphate dibasic are the 2g/100ml of mixed solution.Mixed solution mixes the back at 112.3 ℃ of 30min that sterilize down, be cooled to 35 ℃, with Lactococcus lactis breast subspecies ATCC11454 (Lactococcus lactis, as follows) be fermented bacterium, insert in the mixed solution and be the fermented bacterium that has activated of mixeding liquid volume 7% (activation of Lactococcus lactis breast subspecies is a known technology), fermentation culture 14h gets fermented liquid; Gained fermented liquid nisin is tired and is 4200IU/ml.
Regulate the pH value to 2 of fermented liquid, make nisin from thalline, fully dissociate, the diatomite that adds 2g/100ml in the fermented liquid carries out that solid-liquid separation is removed thalline and impurity gets filtered liquid, filtered liquid is risen membrane concentration to 1/8 of original fermented solution volume through vacuum, add ammonium sulfate to saturated saltouing, must precipitate through centrifugal then, throw out through 60 ℃ low-temperature vacuum drying, more than micronizing to 200 order after with the salt stdn finished product 10.1g (1000IU/mg).
Embodiment 2:
Soybean meal is mixed with the aqueous suspensions of 12g/100ml, stirring and evenly mixing, boil 30min, 50 ℃ of cooling and holding temperatures, regulating pH with the sodium hydroxide of 5mol/l is 8.50, the papoid that adds grouts weight 4% concentration of substrate while stirring, keeping enzymolysis solution pH with 5mol/l sodium hydroxide in the enzymolysis process is 8.50, enzymolysis 8h; Regulate enzymolysis liquid temp to 40 ℃, pH is 8.00, adds the trypsinase of grouts weight 4% concentration of substrate while stirring, and keeping enzymolysis solution pH with the sodium hydroxide of 5mol/l is 8.00, and enzymolysis 4h must enzymolysis solution.Enzymolysis solution adds 1% diatomite through the Plate Filtration removal of impurities, and filtered liquid is vegetable-protein enzymolysis liquid.The nitrogen content recovery rate of the vegetable-protein enzymolysis liquid that makes with this technology is 33%.
The vegetable-protein enzymolysis liquid that makes and glucose, Secondary ammonium phosphate and yeast extract are added water be mixed with the mixed solution 3L that nitrogen content is 3g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the add-on of add-on 2g/100ml, the yeast extract of 8.5ml/100ml, the glucose of mixed solution is that the add-on of mixed solution 1g/100ml, ammonium phosphate is mixed solution 2g/100ml; Mixed solution is mixed the back at 121 ℃ of 30min that sterilize down, be cooled to 35 ℃, mixed solution inserts the Lactococcus lactis breast subspecies ATCC11454 bacterial classification that has activated for mixeding liquid volume 10%, and fermentation culture 22h, gained fermented liquid nisin tire and be 4320IU/ml.
Regulate the pH value to 4 of fermented liquid, the diatomite of adding 0.8% carries out solid-liquid separation removal thalline and impurity in the fermented liquid, with the filtered liquid of gained through membrane concentration (damming more than the molecular weight 1Kd) to 1/8 of original fermented solution volume, add ammonium sulfate to saturated saltouing, must precipitate through centrifugal then, the throw out drying, pulverize and with after the salt stdn finished product 10.7g (1000IU/mg).
Embodiment 3:
With soybean meal through purifying pulverization process, be mixed with the aqueous suspensions of 8g/100ml, stirring and evenly mixing, boil 30min, 50 ℃ of postcooling holding temperatures, regulating pH with the sodium hydroxide of 5mol/L is 9.00, (prozyme A is Sumizyme MP and papoid to add the prozyme A of soybean meal weight 4% while stirring, by Sumizyme MP: papoid=proportioning formed in 2: 1), keeping enzymolysis solution pH with 5mol/L sodium hydroxide in the enzymolysis process is 9.00, enzymolysis 8h; Regulate enzymolysis liquid temp to 35 ℃, pH is 7.00, (prozyme B is neutral protease and trypsinase to add the prozyme B of soybean meal weight 4% while stirring, by neutral protease: trypsinase=proportioning formed in 1: 1), keeping enzymolysis solution pH with the sodium hydroxide of 5mol/l is 7.00, and enzymolysis 4h gets enzymolysis solution.The diatomite that enzymolysis solution is added 1g/100ml is through the Plate Filtration removal of impurities, and filtered liquid is vegetable-protein enzymolysis liquid.The nitrogen content recovery rate of the vegetable-protein enzymolysis liquid that makes with this technology is 40%.
The vegetable-protein enzymolysis liquid that makes and sucrose, Secondary ammonium phosphate and yeast extract are added water be mixed with the mixed solution 3L that nitrogen content is 2g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the add-on of add-on 4g/100ml, the yeast extract of 6ml/100ml, the glucose of mixed solution is that the add-on of mixed solution 1g/100ml, Secondary ammonium phosphate is mixed solution 3g/100ml; Mixed solution is mixed the back at 112.3 ℃ of 30min that sterilize down, be cooled to 35 ℃, the access amount is the bacterial classification Lactococcus lactis breast subspecies ATCC11454 that 7% of mixeding liquid volume has activated, and fermentation culture 14h, gained fermented liquid nisin tire and be 4450IU/ml.
Regulate fermented liquid pH value to 4, the treated nisin that makes of fermented liquid fully dissociates from thalline, the diatomite of adding 2% carries out solid-liquid separation removal thalline and impurity in the fermented liquid, filtered liquid is risen membrane concentration to 1/8 of original fermented solution volume through vacuum, add ammonium sulfate to saturated saltouing, must precipitate through centrifugal then, the throw out drying, pulverize and with after the salt stdn finished product 11.6g (1000IU/mg).
Embodiment 4:
Peanut dregs is mixed with the aqueous suspensions of 10g/ml through the purification pulverization process, stirring and evenly mixing, boil 30min, 40 ℃ of postcooling holding temperatures, regulating pH with the sodium hydroxide of 5mol/l is 8.00, adds the prozyme C (prozyme C is neutral protease and papoid, and by neutral protease: papoid=proportioning formed in 2: 3) of 8% concentration of substrate while stirring, keeping enzymolysis solution pH with 5mol/l sodium hydroxide in the enzymolysis process is 8.00, enzymolysis 12h.Enzymolysis solution adds the diatomite of 1g/100ml through the Plate Filtration removal of impurities, and filtered liquid is vegetable-protein enzymolysis liquid.The nitrogen content recovery rate of the vegetable-protein enzymolysis liquid that makes with this technology is 32%.
The vegetable-protein enzymolysis liquid that makes and sucrose, Secondary ammonium phosphate and yeast extract are added water be mixed with the mixed solution 3L that nitrogen content is 1.5g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the add-on of add-on 4g/100ml, the yeast extract of 5ml/100ml, the glucose of mixed solution is that the add-on of mixed solution 1g/100ml, Secondary ammonium phosphate is mixed solution 3g/100ml; Mixed solution is mixed the back at 115 ℃ of 30min that sterilize down, be cooled to 35 ℃, the access amount is the bacterial classification Lactococcus lactis breast subspecies ATCC11454 that 7% of mixeding liquid volume has activated, and fermentation culture 14h, gained fermented liquid nisin tire and be 4000IU/ml.
Regulate fermented liquid pH value to 4, the treated nisin that makes of fermented liquid fully dissociates from thalline, the diatomite of adding 2% carries out solid-liquid separation removal thalline and impurity in the fermented liquid, filtered liquid is risen membrane concentration to 1/8 of original fermented solution volume through vacuum, add ammonium sulfate to saturated saltouing, must precipitate through centrifugal then, the throw out drying, pulverize and with after the salt stdn finished product 10g (1000IU/mg).
Embodiment 5:
Soybean cake dregs is mixed with the aqueous suspensions of 10g/ml through the purification pulverization process, stirring and evenly mixing, boil 30min, 50 ℃ of postcooling holding temperatures, regulating pH with the sodium hydroxide of 5mol/l is 9.00, add the Sumizyme MP of 8% concentration of substrate (be soybean cake dregs weight 8%) while stirring, keeping enzymolysis solution pH with 5mol/l sodium hydroxide in the enzymolysis process is 9.00, enzymolysis 12h.Enzymolysis solution adds the diatomite of 1g/100ml through the Plate Filtration removal of impurities, and filtered liquid is vegetable-protein enzymolysis liquid.The nitrogen content recovery rate of the vegetable-protein enzymolysis liquid that makes with this technology is 28%.
The vegetable-protein enzymolysis liquid that makes and sucrose, Secondary ammonium phosphate and yeast extract are added water be mixed with the mixed solution 3L that nitrogen content is 1.5g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the add-on of add-on 4g/100ml, the yeast extract of 5 ml/100ml, the glucose of mixed solution is that the add-on of mixed solution 1g/100ml, Secondary ammonium phosphate is mixed solution 3g/100ml; Mixed solution is mixed the back at 115 ℃ of 30min that sterilize down, be cooled to 35 ℃, the access amount is the bacterial classification Lactococcus lactis breast subspecies ATCC11454 that 7% of mixeding liquid volume has activated, and fermentation culture 14h, gained fermented liquid nisin tire and be 3800IU/ml.
Regulate fermented liquid pH value to 4, the treated nisin that makes of fermented liquid fully dissociates from thalline, the diatomite of adding 2% carries out solid-liquid separation removal thalline and impurity in the fermented liquid, filtered liquid is risen membrane concentration to 1/8 of original fermented solution volume through vacuum, add ammonium sulfate to saturated saltouing, must precipitate through centrifugal then, the throw out drying, pulverize and with after the salt stdn finished product 9.6g (1000IU/mg).
The bound value and the interval value of each raw material of the present invention can both be realized the present invention, just do not enumerate embodiment one by one at this.
Claims (5)
1. one kind is the technology of raw material production nisin with the plant cake, it is characterized in that it is raw material with the plant cake, with plant cake through enzymolysis, fermentation, solid-liquid separation, concentrate, saltout, dry and be packaged to be finished product;
Described enzymolysis is: the aqueous suspensions that plant cake is mixed with concentration 8-12g/100ml, boil 30min, regulating pH after being cooled to temperature 30-60 ℃ is 2-10, be incorporated as the enzyme of plant cake weight 1-8% then, enzymolysis solution with gained behind the process 8-20h carries out filtering and impurity removing, gets vegetable-protein enzymolysis liquid;
Described enzyme is one or both the mixing wherein of aspartic protease, neutral protease, Sumizyme MP, trypsinase, papoid, and the proportioning when wherein any two kinds of enzymes mix is 1-5: 1-7;
Described fermentation is: the vegetable-protein enzymolysis liquid of gained is added water with sugar, yeast extract and phosphoric acid salt be mixed with the mixed solution that nitrogen content is 0.5-4g/100ml, wherein the add-on of vegetable-protein enzymolysis liquid is that the 5-15ml/100ml of mixed solution, the add-on of sugar are that the 1-5g/100ml of mixed solution, the add-on of yeast extract are that mixed solution 1-5g/100ml, phosphatic add-on are mixed solution 1-3g/100ml;
Mixed solution inserts the good Lactococcus lactis breast subspecies of activation after the sterilization cooling, the access amount of Lactococcus lactis breast subspecies is the 1-10% of mixeding liquid volume, and inoculation fermentation 12-30h gets fermented liquid;
Described solid-liquid separation is: the fermented liquid of gained is regulated the pH value to 2-4, adopt solid-liquid isolation method removal thalline and impurity to get filtered liquid;
Described simmer down to: by rising the 1/2-1/20 that film, falling liquid film, multiple-effect or ultrafiltration concentration process are concentrated into filtered liquid original volume.
2. according to claim 1 a kind of be the technology of raw material production nisin with the plant cake, it is characterized in that: described plant cake is dregs of rapeseed cake, peanut dregs, cotton cake dregs or soybean cake dregs.
3. according to claim 1 a kind of be the technology of raw material production nisin with the plant cake, it is characterized in that: the adding of described enzyme divides secondary to add, the enzyme that adds is inequality with the enzyme that adds for the second time for the first time; The amount of the enzyme that adds is the 1-7% of plant cake weight for the first time.
4. according to claim 1 a kind of be the technology of raw material production nisin with the plant cake, it is characterized in that: described sugar is one or more the mixing in sucrose, glucose, lactose, fructose, the maltose, and more than one are any proportioning when mixing.
5. according to claim 1 a kind of be the technology of raw material production nisin with the plant cake, it is characterized in that: described phosphoric acid salt is one or more the mixing in potassium primary phosphate, SODIUM PHOSPHATE, MONOBASIC, dipotassium hydrogen phosphate, Sodium phosphate dibasic, the ammonium phosphate, and more than one are any proportioning when mixing.
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| CNB200610019565XA CN100560731C (en) | 2006-07-07 | 2006-07-07 | A kind of is the technology of raw material production nisin with the plant cake |
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| CNB200610019565XA CN100560731C (en) | 2006-07-07 | 2006-07-07 | A kind of is the technology of raw material production nisin with the plant cake |
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| CN100560731C true CN100560731C (en) | 2009-11-18 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105567770A (en) * | 2016-02-03 | 2016-05-11 | 常州达奥新材料科技有限公司 | Preparation method of nisin |
| CN106480140A (en) * | 2016-10-12 | 2017-03-08 | 天津大学 | Lactococcus lactis bacteria fermentation culture medium and preparation method based on dregs of beans protein enzymatic hydrolyzate |
| CN107513548B (en) * | 2017-10-31 | 2021-01-22 | 荣成市日鑫水产有限公司 | Method for preparing peptone by using fish meal squeezed liquid |
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2006
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Non-Patent Citations (4)
| Title |
|---|
| 乳酸链球菌固态发酵豆粕的研究. 季伟，徐学明.粮食与饲料工业，第5期. 2006 |
| 乳酸链球菌固态发酵豆粕的研究. 季伟，徐学明.粮食与饲料工业，第5期. 2006 * |
| 微生物防腐剂Nisin的研究与应用. 汤凤霞，蔡慧农.中国食品报第B02版. 2003 |
| 微生物防腐剂Nisin的研究与应用. 汤凤霞，蔡慧农.中国食品报第B02版. 2003 * |
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