CN102382772A - Industrialized production method of Aspergillus oryzae strain for fermentation - Google Patents
Industrialized production method of Aspergillus oryzae strain for fermentation Download PDFInfo
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- CN102382772A CN102382772A CN2011103083303A CN201110308330A CN102382772A CN 102382772 A CN102382772 A CN 102382772A CN 2011103083303 A CN2011103083303 A CN 2011103083303A CN 201110308330 A CN201110308330 A CN 201110308330A CN 102382772 A CN102382772 A CN 102382772A
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- bacterial classification
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Abstract
An industrialized production method of Aspergillus oryzae strain for fermentation. Aspergillus oryzae Shanghai brewing 3.042 strains are activated, treated with erlenmeyer flask enlargement culture and cultured in a culture room, namely the erlenmeyer flask strains are treated with further enlargement culture in the culture room. A medium formula is as below: 100 jin of a mixture of wheat bran and soybean meal in a ratio of 4:6 is added with 48-52 jin of water; The medium material is steamed in a charging steam basket for 30 min, cooled to 35-40 DEG C and inoculated with an inoculation proportion of 0.3%. The inoculated yeast material is placed in a yeast fan in a culture room for culture, and the yeast material has a thickness of 2.5-3 cm, and a culture time is 7d. The method of the invention can be utilized to produce Aspergillus oryzae strain with high enzyme production vitality and is capable of fermenting soybean paste, fermented soya bean and soy rapidly.
Description
Technical field
The invention belongs to the working method of fermentation with the aspergillus oryzae bacterial classification, be specifically related to the method for large-scale production aspergillus oryzae bacterial classification, the bacterial classification of employing is that 3.042 aspergillus oryzae bacterial classifications are made in Shanghai.
Background technology
Zymotechnique comprises two kinds of traditional zymotic technology and manual fermentation technologies; Growth cycle is long, manageability, product health do not have the characteristics of guarantee because traditional zymotic technology has; More and more do not catch up with the needs of modern production; In order to enhance productivity, ensure market supply, the manual fermentation process quilt generally adopts at present.Shanghai is made 3.042 aspergillus oryzae bacterial classifications and is had that growth is fast, manageability, characteristics that the product enzyme activity is high, often is used in the inoculation fermentation of traditional soya sauce, fermented soya bean and soy sauce.Along with the expansion of suitability for industrialized production scale, aspergillus oryzae bacterial classification demand will increase, and large-scale production aspergillus oryzae bacterial classification will have great importance.Many reports about induction mutation of bacterium, bacterial screening and different sorts synergistic bacterium fermentation aspect are arranged at present, the report of rare large-scale production bacterial classification, the present invention has solved the technical problem of large-scale production bacterial classification from the enlarged culturing aspect of aspergillus oryzae bacterial classification.
Two key factors in the spawn culture process are the prescription of substratum and the control of culturing process.Wheat bran and dregs of beans often are used to do the substratum that mould is cultivated, and in the process of cultivating, wheat bran plays loose logical oxygen, and dregs of beans provides nutritive substance, optimizes the process and the quality product of spawn culture through the ratio of confirming the two; The addition of water also has material impact to the fermentation of bacterial classification in the substratum, and the ratio snapshot of oneself that adds water rings the utilization of bacterial classification nutritive ingredient, in large-scale production process, adds the growth that water then influences inner mycelia too much, and " burning bent " phenomenon takes place in culturing process easily.The control of culturing process is most important to the cultivation of aspergillus oryzae bacterial classification; Especially in culturing room, produce then more difficult control in enormous quantities; Thereby the temperature and humidity through control culturing room makes the vigorous pollution that suppresses assorted bacterium of growth, the design of culturing room will help the circulation of heat and stable.
Summary of the invention
The object of the invention provides a kind of large-scale method for producing of aspergillus oryzae bacterial classification.
The objective of the invention is to realize like this: a kind of fermentation with the industrialized preparing process of aspergillus oryzae bacterial classification may further comprise the steps:
1), promptly carries out activation culture to the female test tube slant that is inoculated in the making of employing fermented bean drink substratum of planting of aspergillus oryzae with aspergillus oryzae bacterial classification activation culture;
2) will pass through activatory aspergillus oryzae test tube strains and be inoculated in the triangular flask enlarged culturing 4d under 30 ℃ of temperature;
3) triangular flask enlarged culturing bacterial classification further enlarged culturing in culturing room is carried out suitability for industrialized production, adopt wheat bran: dregs of beans=4:6, add the culture medium prescription of 48-52 jin water in the compound of per 100 jin of wheat brans and dregs of beans; Culture base-material is steamed 30min with steam in special steaming charging basket; The cold of drying in the air after taking out is promptly inoculated after temperature is reduced to about 35 ℃, and inoculative proportion is 0.3%; Postvaccinal song is expected to cultivate during being placed on song winnows with a dustpan; Bent material thickness is 2.5-3cm, and incubation time is 7d, obtains fermentation and uses the aspergillus oryzae bacterial classification.
The aspergillus oryzae bacterial classification that adopts in the step 1) is that 3.042 aspergillus oryzae bacterial classifications are made in Shanghai.
The triangular flask culture medium prescription is: in weight part: wheat bran: dregs of beans=6:4, wheat bran and dregs of beans mix, and add the water of 48-55ml in the compound of every 100g wheat bran and dregs of beans, obtain after 30 minutes through autoclaving.
Fermented bean drink in the step 1) is trained basic preparation technology: fermented bean drink: 1000mL (4-5 ° of Be), Zulkovsky starch 20g, MgSO
4.7H
2O 0.5g, (NH
4)
2SO
40.5g, regulate pH value 6.5-7.0 after the agar powder 20-25g mixing of materials, 0.1Mpa down sterilization 30min to obtain the fermented bean drink training basic.
The preparation process of fermented bean drink is: select fresh soyabean 100g to be soaked in water and pull out after the imbibition, add the clear water of 5-6 times of soybean amount then, boiled 2-5 hour, cross with absorbent cotton and filter the 100mL fermented bean drink, this fermented bean drink is the fermented bean drink of 4-5 ° of Be.
Through fermentation provided by the invention is taken a sample with the resulting aspergillus oryzae bacterial classification of the industrialized preparing process of aspergillus oryzae bacterial classification finished product; According to SB/T10317-1999 " proteinase activity measuring method " the enzyme work of aspergillus oryzae bacterial classification is measured; The enzyme slip-knot of 8 tests is 7039 ± 449U/g (butt) for the aspergillus oryzae bacterial classification enzyme work that mass-producing is cultivated really, and the enzyme work of the aspergillus oryzae bacterial classification of inoculation triangular flask enlarged culturing is 7120 ± 322U/g (butt).
The present invention optimizes fermentative medium formula, and through test of many times the enlarged culturing process is optimized control, has realized the large-scale production of aspergillus oryzae bacterial classification, and the enzyme work of the bacterial classification of large-scale production reaches 7039 ± 449U/g (butt).Improve the output of aspergillus oryzae bacterial classification, can reduce the cost of aspergillus oryzae bacterial classification significantly.
For the safety of the aspergillus oryzae bacterial classification method that fully proves large-scale production, the contriver has done simultaneous test to the bacterial classification of large-scale production and the bacterial classification of triangular flask enlarged culturing to the fermentative action of fermented soya bean, and concrete steps are:
1) get 100 jin of soybean and in soaking cylinder, soak, soak time is that winter and spring are 22-24 hour, and summer and autumn are 16-18 hour, and it is no sandwich to be dipped to inside, center full of water branch, and the no wrinkle of beans expansion soaks back soybean moisture and should reach 55-60%.
2) soaked soybean is taken out to be placed on steam material on the food steamer, steaming the material time is about 2.5 hours, and all evenly well-done with beans, it is filbert that look is; The distinctive fragrance of ripe beans is arranged, and particle is kept perfectly, and extruding can become mud to be as the criterion.
3) soybean to be cooked is dried in the air and coolly after about 35 ℃-40 ℃, promptly inoculates; In the seeded process soybean that cooks is divided into two parts; 50 jin every part, the ratio inoculation aspergillus oryzae bacterial classification in 0.6%, the aspergillus oryzae bacterial classification of a copy of it inoculation triangular flask enlarged culturing; The aspergillus oryzae bacterial classification of another part inoculation large-scale production needs bacterial classification and the abundant mixing of soybean in the seeded process.
4) postvaccinal soybean is placed on makes song in the dustpan then, two parts of trial targets are placed on same system deep closet system song, the dustpans of placing two parts of inoculation different strains are carried out mark.The temperature of control system deep closet is about 30 ℃, and humidity is 75%, and the bent time of system is 7d, treats that mycelia penetrates soybean inside, and spore is promptly made bent the end after being covered with the soybean surface.
After washing in a pan the spore elutriation totally of enzyme chamber with the soybean surface, transfer to proving room and ferment, the temperature of control proving room is the finished product fermented soya bean after stacking 15 days under the airy condition of interval between 30-40 ℃.The total acid and the amino acid nitrogen content of sampling and measuring finished product fermented soya bean; Adopt the method for measuring total acid and amino-nitrogen among GB/T 5009.39-2003 " analytical procedure of sauce sanitary standard " and the GB/T 5009.40-2003 " analytical procedure of sauce hygienic standard " to measure; Through collecting the total acid and the amino acid nitrogen content of the finished product fermented soya bean of testing for 8 times; The total acid content that obtains the fermented soya bean of two kinds of inoculation different strains is distributed between the 2.6-3.2g/100g, and amino acid nitrogen content is distributed between the 0.7-1.1g/100g.The finished product fermented soya bean of inoculating different bacterial classifications do not have significant difference on the physical and chemical index of fermentation, finished product fermented soya bean local flavor does not have difference yet, and the two is as broad as long to the fermentative action of fermented soya bean.
Description of drawings
Below in conjunction with accompanying drawing and embodiment the present invention is described further.
Fig. 1, Fig. 2 are the total acid content and the amino acid nitrogen content distribution plan of the fermented soya bean of two kinds of inoculation different strains.
Fig. 3 is the comparison diagram that triangular flask enlarged culturing bacterial classification and suitability for industrialized production bacterial classification enzyme are lived.
Embodiment
Embodiment one:
1) 3.042 aspergillus oryzae bacterial classifications is made in Shanghai and carried out activation culture; On fermented bean drink substratum test tube slant; 3.042 aspergillus oryzae bacterial classifications are made in Shanghai be inoculated under the aseptic condition on this substratum, fermented bean drink culture medium preparation method is: fermented bean drink: 1000mL (4-5 ° of Be), Zulkovsky starch 20g, MgSO
4.7H
2O 0.5g, (NH
4)
2SO
40.5g, agar powder 20-25g, pH value 6.5-7.0; 0.1Mpa following sterilization 30min, wherein the preparation process of fermented bean drink is pulled out after the imbibition for selecting fresh soyabean 100g to be soaked in water, and adds the clear water of 5-6 times of soybean amount then; Boiled 2-5 hour; To note moisturizing when boiling beans, make to cross with absorbent cotton at last to filter the 100mL fermented bean drink that this fermented bean drink is the fermented bean drink of 5 ° of Be.The control culture temperature is 30 ℃ in the culturing process, and incubation time is 4d, treats that the yellow-green colour spore is covered with the inclined-plane and gets final product.
2) then in the triangular flask with the above-mentioned bacterial classification 250ml that the 10g substratum is housed that access has been sterilized under aseptic condition, the control culture temperature is 30 ℃, and incubation time is 4d, adopts the mode of button flask culture, cultivates to shake the bottle back behind the 2d and then cultivate.The concrete prescription of substratum is wheat bran: dregs of beans=6:4, adds the water (final moisture of substratum is between 45%-48%) of 48-55ml in the compound of every 100g wheat bran and dregs of beans, sterilization 30min under 0.1Mpa.
3) with the aspergillus oryzae bacterial classification of above-mentioned triangular flask enlarged culturing further enlarged culturing in culturing room.Adopt wheat bran: dregs of beans=4:6, add the culture medium prescription of 48-52 jin water in the compound of per 100 jin of wheat brans and dregs of beans, culture base-material is steamed 30min with steam in steaming charging basket; The cold of drying in the air after taking out; After temperature is reduced between 35 ℃-40 ℃, promptly inoculate, inoculative proportion is 0.3%, will guarantee in the seeded process that the bacterial classification of triangular flask enlarged culturing is evenly distributed in the substratum; Mix to fumigate culturing room with formaldehyde and potassium permanganate before cultivating, close the stifling 12h of culturing room's door.Then postvaccinal bacterium culture medium is placed in the beanstalk of winnowing with a dustpan and cultivates, substratum thickness is 2.5-3cm, and the height of the every layer of beanstalk of winnowing with a dustpan differs 30cm.When temperature during less than 20 ℃; Bacterial classification is at room temperature through promptly beginning to adopt the electricradiator heat temperature raising in 20-24 hour, and when temperature during greater than 20 ℃, the heat that then leans on growth to produce comes nature to heat up; Need be by external thermal source heating; When the article temperature of substratum reaches 30 ℃, stop to heat up, temperature-rise period needs wet gauze is covered on the substratum, prevents moisture evaporation.Then whenever promptly carried out once aeration-cooling at a distance from two hours; Make the substratum temperature maintenance between 28-35 ℃, when mycelia penetrates substratum, promptly whenever led to a wind at a distance from one hour; The substratum temperature still maintains between 28-35 ℃, is the period of mycelia to the spore conversion at this moment.Treat that spore all transforms the rear venting cooling, temperature is stirred bacterial classification up and down after reducing to room temperature, feasible wherein moisture rapid evaporation, and the final moisture content of finished product bacterial classification is 13%, incubation time needs 7 days.
Claims (5)
1. one kind ferments with the industrialized preparing process of aspergillus oryzae bacterial classification, it is characterized in that: may further comprise the steps:
1), promptly carries out activation culture to the female test tube slant that is inoculated in the making of employing fermented bean drink substratum of planting of aspergillus oryzae with aspergillus oryzae bacterial classification activation culture;
2) will pass through activatory aspergillus oryzae test tube strains and be inoculated in the triangular flask enlarged culturing 4d under 30 ℃ of temperature;
3) triangular flask enlarged culturing bacterial classification further enlarged culturing in culturing room is carried out suitability for industrialized production, adopt wheat bran: dregs of beans=4:6, add the culture medium prescription of 48-52 jin water in the compound of per 100 jin of wheat brans and dregs of beans; Culture base-material is steamed 30min with steam in special steaming charging basket; The cold of drying in the air after taking out is promptly inoculated after temperature is reduced to about 35 ℃, and inoculative proportion is 0.3%; Postvaccinal song is expected to cultivate during being placed on song winnows with a dustpan; Bent material thickness is 2.5-3cm, and incubation time is 7d, obtains fermentation and uses the aspergillus oryzae bacterial classification.
2. fermentation according to claim 1 is characterized in that with the industrialized preparing process of aspergillus oryzae bacterial classification: the aspergillus oryzae starter kind that adopts in the step 1) is that 3.042 aspergillus oryzae bacterial classifications are made in Shanghai.
3. fermentation according to claim 1 is with the industrialized preparing process of aspergillus oryzae bacterial classification; It is characterized in that: the triangular flask culture medium prescription is: in weight part: wheat bran: dregs of beans=6:4; Wheat bran and dregs of beans mix; Add the water of 48-55ml in the compound of every 100g wheat bran and dregs of beans, obtain after 30 minutes through autoclaving.
4. fermentation according to claim 1 is characterized in that with the industrialized preparing process of aspergillus oryzae bacterial classification: the fermented bean drink in the step 1) is trained basic preparation technology and is: fermented bean drink: 1000mL (4-5 ° of Be), Zulkovsky starch 20g, MgSO
4.7H
2O 0.5g, (NH
4)
2SO
40.5g after agar powder 20-25g mixing, regulate pH value 6.5-7.0, it is basic that sterilization 30min obtains the fermented bean drink training under the 0.1Mpa.
5. fermentation according to claim 4 is with the industrialized preparing process of aspergillus oryzae bacterial classification; It is characterized in that: the preparation process of fermented bean drink is: select fresh soyabean 100g to be soaked in water and pull out after the imbibition; The clear water that adds 5-6 times of soybean amount then; Boiled 2-5 hour, and crossed with absorbent cotton and filter the 100mL fermented bean drink, this fermented bean drink is the fermented bean drink of 4-5 ° of Be.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103087876A (en) * | 2013-01-29 | 2013-05-08 | 会稽山绍兴酒股份有限公司 | Preparation method of rice-koji |
| CN104172053A (en) * | 2014-07-18 | 2014-12-03 | 江苏省宿迁市三园调味品有限公司 | Process for preparing sweet oil |
| CN105192593A (en) * | 2015-08-18 | 2015-12-30 | 广东美味鲜调味食品有限公司 | Production technology for aspergillus oryzae type fermented soybeans |
| CN105558512A (en) * | 2014-11-05 | 2016-05-11 | 上海鼎丰酿造食品有限公司 | Strain culturing method |
| CN107080006A (en) * | 2017-06-21 | 2017-08-22 | 云南农业大学 | A kind of preparation method of high theophylline Pu'er cooked tea |
| CN109463639A (en) * | 2018-12-24 | 2019-03-15 | 左汶骏 | A kind of oil thick broad-bean sauce and preparation method thereof |
| CN110484453A (en) * | 2019-09-02 | 2019-11-22 | 千禾味业食品股份有限公司 | It is a kind of for koji preparation culture medium and base with this soy sauce koji preparation method |
| CN112790340A (en) * | 2021-01-11 | 2021-05-14 | 浙江敬存仁生物科技有限公司 | Preparation method for chili sauce fermented soya beans |
| CN114410486A (en) * | 2022-03-22 | 2022-04-29 | 中国科学院天津工业生物技术研究所 | Aspergillus oryzae strain and application thereof in development of feed protein |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087876A (en) * | 2013-01-29 | 2013-05-08 | 会稽山绍兴酒股份有限公司 | Preparation method of rice-koji |
| CN104172053A (en) * | 2014-07-18 | 2014-12-03 | 江苏省宿迁市三园调味品有限公司 | Process for preparing sweet oil |
| CN104172053B (en) * | 2014-07-18 | 2016-08-24 | 江苏省宿迁市三园调味品有限公司 | A kind of preparation technology of sweet oil |
| CN105558512A (en) * | 2014-11-05 | 2016-05-11 | 上海鼎丰酿造食品有限公司 | Strain culturing method |
| CN105192593A (en) * | 2015-08-18 | 2015-12-30 | 广东美味鲜调味食品有限公司 | Production technology for aspergillus oryzae type fermented soybeans |
| CN105192593B (en) * | 2015-08-18 | 2019-07-23 | 广东美味鲜调味食品有限公司 | A kind of aspergillus oryzae type fermented soya bean production technology |
| CN107080006A (en) * | 2017-06-21 | 2017-08-22 | 云南农业大学 | A kind of preparation method of high theophylline Pu'er cooked tea |
| CN109463639A (en) * | 2018-12-24 | 2019-03-15 | 左汶骏 | A kind of oil thick broad-bean sauce and preparation method thereof |
| CN110484453A (en) * | 2019-09-02 | 2019-11-22 | 千禾味业食品股份有限公司 | It is a kind of for koji preparation culture medium and base with this soy sauce koji preparation method |
| CN112790340A (en) * | 2021-01-11 | 2021-05-14 | 浙江敬存仁生物科技有限公司 | Preparation method for chili sauce fermented soya beans |
| CN114410486A (en) * | 2022-03-22 | 2022-04-29 | 中国科学院天津工业生物技术研究所 | Aspergillus oryzae strain and application thereof in development of feed protein |
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Application publication date: 20120321 |