CN103243039A - High density culturing method of lactobacillus paracasei - Google Patents
High density culturing method of lactobacillus paracasei Download PDFInfo
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- CN103243039A CN103243039A CN2012100277316A CN201210027731A CN103243039A CN 103243039 A CN103243039 A CN 103243039A CN 2012100277316 A CN2012100277316 A CN 2012100277316A CN 201210027731 A CN201210027731 A CN 201210027731A CN 103243039 A CN103243039 A CN 103243039A
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- fermentation
- high density
- fermentation process
- lactobacillus paraceasi
- lactobacillus paracasei
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Abstract
The invention is suitable for a biological engineering technology field, and provides a method for fermenting lactobacillus paracasei, comprising the following steps of: preparing a liquid medium containing the following components; inoculating active lactobacillus paracasei in a proportion of 4-7% (v/v) onto the liquid medium, stirring in 50-100r/min rotation speed under 37 DEG C, keeping constant PH value of 5.5-6.5, intermittently introducing nitrogen, and fermenting for 15-24h in a pressure non-maintaining condition. In a fermentation process, pH value is monitored, and 12.5% ammonia water is flowedly added as a neutralizer to maintain a pH value at 5.5-6.5. The method for fermenting lactobacillus paracasei in the invention, by optimizing the fermentation, and correcting and improving parameters of an inoculation amount, a throughput, a pH value and other parameters in the fermentation process, implements high density fermentation of lactobacillus paracasei, and substantially reduces production cost.
Description
Technical field
The present invention relates to a kind of high-density culture technology of lactobacillus paraceasi.
Background technology
Lactobacillus paraceasi belongs to the lactobacterium casei in the milk-acid bacteria, extensively is present in the oral cavity, enteron aisle of leavened food and human body.Studies show that lactobacillus paraceasi has the antagonism pathogenic bacterium, regulates functions such as intestinal microflora balance, enhancing body immunizing power.The benefit of lactobacillus paraceasi is given birth to biological function has broad application prospects it in foodstuffs industry, be applied in food fermentation at present, food fresh keeping is anticorrosion and aspect such as health care.
High-density culture does not have definite definition, it is a relative concept, refer to use the fermentation density of certain culture technology and device raising thalline, the more common cultivation of cell density is increased significantly, microorganism is reduced production costs under equal volume equipment, shorten the production cycle, finally improve the specific production rate of specific product.
Conventional fermentation technique is difficult to promote after OD reaches a value again, and the method that this just need find the concentration of thalline in the fermentation density that can improve thalline or the unit volume nutrient solution with comprehensive raising specific production rate, reduces production costs.
Summary of the invention
In view of the above problems, the present invention is intended to provide a kind of lactobacillus paraceasi fermentation process in high density, can avoid characteristics such as the utilization of common fermentation technology substratum is incomplete, the unit viable count is not high, can effectively improve the production efficiency of lactobacillus paraceasi, reduce production costs.
The present invention is achieved through the following technical solutions:
A. the activation of bacterial classification:
The bacterial classification inoculation of-80 ℃ of preservations in 5mL MRS substratum, is cultivated 18h for 37 ℃, go down to posterity and cultivate the bacterial classification that obtains activating for 2 times;
The MRS substratum is composed as follows: glucose 20.0g, yeast powder 4.0g, Tryptones 10.0g, extractum carnis 8.0g, K2HPO42.0g, MgSO47H2O 0.2g, MnSO44H2O 0.05g, ammonium citrate 2.0g, CH3COONa3H2O 5.0g, Tween801.0mL, distilled water 1000mL, pH6.2,121 ℃ of sterilization 15min.
B. fermentation
The 50%-70% modified MRS culture medium of in fermentor tank, packing into, inoculation 4%-7% (V/V) seed liquor, mix rotating speed 50-100rpm/min, 37 ℃, permanent pH5.5-6.5, intermittently logical not pressurize of nitrogen fermentation 15-24h, during the fermentation, monitoring pH value, stream adds 12.5% ammoniacal liquor and keeps pH at 5.5-6.5 as neutralizing agent;
Modified MRS culture medium is composed as follows: glucose 15.0-25.0g, yeast powder 15.0-25.0g, KH
2PO
42.0-3.0g, MgSO
47H
2O 1.5-2.5g, Triammonium citrate 1.7-2.7g, CH
3COONa3H
2O 6.0-7.0g, Tween800.8-1.2mL, distilled water 1000mL, pH 5.5-6.5.
Advantage of the present invention and effect are as follows:
1, the OD value of fermentation back lactobacillus paraceasi is significantly improved;
2, can improve the concentration of thalline in the fermentation density of thalline or the unit volume nutrient solution, and then improve volume productivity;
3, can dwindle the bio-reactor volume, reduce investment of production equipment;
4, strengthen downstream separation and extract, and reduce wastewater flow rate to a certain extent;
5, comprehensively improve specific production rate, the commercialization process of expedite product reduces production costs, and improves the competitive power of product on market.
Embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer, below in conjunction with specific embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explaining the present invention, and be not used in restriction the present invention.
Implement illustration:
Not the improvement the lactobacillus paraceasi fermentation process
The embodiment of the invention is carried out according to following step:
A. the activation of bacterial classification
The bacterial classification inoculation of-80 ℃ of preservations in 5ml MRS substratum, is cultivated 18h for 37 ℃, go down to posterity and cultivate the bacterial classification that obtains activating for 2 times;
The MRS substratum is composed as follows: glucose 20.0g, yeast powder 4.0g, Tryptones 10.0g, extractum carnis 8.0g, K2HPO42.0g, MgSO47H2O 0.2g, MnSO44H2O 0.05g, ammonium citrate 2.0g, CH
3COONa3H
2O 5.0g, Tween801.0ml, distilled water 1000ml, 6.2,121 ℃ of sterilizations of pH 15min;
B. fermentation
The 70%MRS substratum of in fermentor tank, packing into, inoculation 5% (V/V) seed liquor is mixed rotating speed 80r/min, 37 ℃, does not have ventilation spontaneous fermentation 15h;
Modified MRS culture medium is not cultivated composed as follows: glucose 20.0g, yeast powder 4.0g, Tryptones 10.0g, extractum carnis 8.0g, K
2HPO
42.0g, MgSO47H
2O 0.2g, MnSO
44H
2O 0.05g, ammonium citrate 2.0g, CH
3COONa3H
2O 5.0g, Tween801.0ml, distilled water 1000ml, 6.2,121 ℃ of sterilizations of pH 15min;
Fermented liquid viable count with not improved culture medium acquisition is 8.2 * 10
8Cfu/mL.
2. the lactobacillus paraceasi fermentation process in high density of substratum improvement
The embodiment of the invention is carried out according to following step:
A. the activation of bacterial classification
The bacterial classification inoculation of-80 ℃ of preservations in 5ml MRS substratum, is cultivated 18h for 37 ℃, go down to posterity and cultivate the bacterial classification that obtains activating for 2 times;
The MRS substratum is composed as follows: glucose 20.0g, yeast powder 4.0g, Tryptones 10.0g, extractum carnis 8.0g, K
2HPO
42.0g, MgSO
47H
2O 0.2g, MnSO
44H
2O 0.05g, ammonium citrate 2.0g, CH
3COONa3H
2O 5.0g, Tween801.0ml, distilled water 1000ml, 6.2,121 ℃ of sterilizations of pH 15min;
B. fermentation
70% improved culture medium of in fermentor tank, packing into, inoculation 5% (V/V) seed liquor is mixed rotating speed 80r/min, and 37 ℃, permanent pH6.0 does not have ventilation spontaneous fermentation 15h, during the fermentation, and monitoring pH value, stream adds 12.5% ammoniacal liquor and keeps pH 6.0 as neutralizing agent;
Modified MRS culture medium is composed as follows: glucose 20.0g, yeast powder 20.0g, KH
2PO
42.5g, MgSO
47H
2O 2.0g, Triammonium citrate 2.5g, CH
3COONa3H
2O 6.25g, Tween801.0ml, distilled water 1000ml, pH 6.2; 121 ℃ of sterilization 15min;
The fermented liquid viable count that obtains with the substratum after the improvement is 9.6 * 10
9Cfu/mL.
Substratum and fermentation process the improvement the lactobacillus paraceasi fermentation process in high density
The embodiment of the invention is carried out according to following step:
A. the activation of bacterial classification
The bacterial classification inoculation of-80 ℃ of preservations in 5ml MRS substratum, is cultivated 18h for 37 ℃, go down to posterity and cultivate the bacterial classification that obtains activating for 2 times;
The MRS substratum is composed as follows: glucose 20.0g, yeast powder 4.0g, Tryptones 10.0g, extractum carnis 8.0g, K2HPO42.0g, MgSO47H2O 0.2g, MnSO44H
2O 0.05g, ammonium citrate 2.0g, CH
3COONa3H
2O 5.0g, Tween801.0ml, distilled water 1000ml, 6.2,121 ℃ of sterilizations of pH 15min;
B. fermentation
70% improved culture medium of in fermentor tank, packing into, inoculation 5% (V/V) seed liquor, mix rotating speed 80r/min, 37 ℃, permanent pH6.0, intermittently logical not pressurize of nitrogen (every 3h is with the logical nitrogen 5min of the speed of 0.2vvm) fermentation 15h, during the fermentation, monitoring pH value, stream adds 12.5% ammoniacal liquor and keeps pH 6.0 as neutralizing agent;
Modified MRS culture medium is composed as follows: glucose 20.0g, yeast powder 20.0g, KH
2PO
42.5g, MgSO
47H
2O 2.0g, Triammonium citrate 2.5g, CH
3COONa3H
2O 6.25g, Tween801.0ml, distilled water 1000ml, pH 6.2; 121 ℃ of sterilization 15mi n;
The fermented liquid viable count that obtains with the substratum after the improvement is 7.6 * 10
10Cfu/mL.
Above-described only is preferred embodiment of the present invention, not in order to limiting the present invention, all any modifications of doing within the spirit and principles in the present invention, is equal to and replaces and improvement etc., all should be included within protection scope of the present invention.
Claims (6)
1. a lactobacillus paraceasi fermentation process in high density comprises the steps:
A. the activation of bacterial classification
The bacterial classification inoculation of-80 ℃ of preservations in 5ml MRS substratum, is cultivated 18h for 37 ℃, go down to posterity and cultivate the bacterial classification that obtains activating for 2 times;
The MRS substratum is composed as follows: glucose 20.0g, yeast powder 4.0g, Tryptones 10.0g, extractum carnis 8.0g, K2HPO42.0g, MgSO47H
2O 0.2g, MnSO44H
2O 0.05g, ammonium citrate 2.0g, CH
3COONa3H2O 5.0g, Tween801.0ml, distilled water 1000ml, pH6.2,121 ℃ of sterilization 15min;
B. fermentation
The 50%-70% modified MRS culture medium of in fermentor tank, packing into, inoculation 4%-7% (V/V) seed liquor, mix rotating speed 50-100rpm/min, 37 ℃, permanent pH5.5-6.5, intermittently logical not pressurize of nitrogen fermentation 15-24h, during the fermentation, monitoring pH value, stream adds 12.5% ammoniacal liquor and keeps pH at 5.5-6.5 as neutralizing agent;
Modified MRS culture medium is composed as follows: glucose 15.0-25.0g, yeast powder 15.0-25.0g, KH
2PO
42.0-3.0g, MgSO
47H
2O 1.5-2.5g, Triammonium citrate 1.7-2.7g, CH
3COONa3H
2O 6.0-7.0g, Tween800.8-1.2mL, distilled water 1000mL, pH5.5-6.5.
2. lactobacillus paraceasi fermentation process in high density as claimed in claim 1 is characterized in that, the component of described liquid nutrient medium is as follows: glucose 20.0g, yeast powder 20.0g, KH
2PO
42.5g, MgSO
47H
2O2.0g, Triammonium citrate 2.5g, CH
3COONa3H
2O 6.25g, Tween801.0mL, distilled water 1000mL, pH6.2.
3. lactobacillus paraceasi fermentation process in high density as claimed in claim 1 is characterized in that, uses fermentor tank as container in the described fermentation culture step, and the coefficient of described fermentor tank is 0.5-0.7.
4. lactobacillus paraceasi fermentation process in high density as claimed in claim 1 is characterized in that, described fermentation culture is to carry out under the 80rpm/min condition at mixing speed.
5. lactobacillus paraceasi fermentation process in high density as claimed in claim 1 is characterized in that, described fermentation culture is logical not pressurize of nitrogen intermittently, and every 3h is with the logical nitrogen 5min of the speed of 0.2vvm.
As claim 1-5 as described in each fermentation process carry out the lactobacillus paraceasi high density fermentation.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104223305A (en) * | 2014-10-09 | 2014-12-24 | 哈尔滨工业大学 | Coix seed lactobacillus beverage preparation method |
| CN104304450A (en) * | 2014-10-09 | 2015-01-28 | 哈尔滨工业大学 | Semen coicis lactic acid bacteria beverage fermentation process |
| CN106754524A (en) * | 2016-12-28 | 2017-05-31 | 石家庄君乐宝乳业有限公司 | Lactobacillus paracasei N1115 culture mediums and its application |
| CN108048344A (en) * | 2017-11-20 | 2018-05-18 | 广东博沃特生物科技有限公司 | Two plants of deodorization bacterial strains and its application in composite biological deodorant is prepared |
| CN110734884A (en) * | 2019-11-27 | 2020-01-31 | 汉臣氏（沈阳）儿童制品有限公司 | Low-salt culture medium and culture method for lactobacillus paracasei |
| WO2020063515A1 (en) * | 2018-09-30 | 2020-04-02 | 内蒙古伊利实业集团股份有限公司 | Novel application of lactobacillus paracasei k56 capable of regulating balance of gastrointestinal flora |
| CN111286471A (en) * | 2019-12-12 | 2020-06-16 | 湖北华扬科技发展有限公司 | Method for high-density culture of lactobacillus paracasei by using soybean meal hydrolysate |
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| CN101418270A (en) * | 2008-11-06 | 2009-04-29 | 内蒙古农业大学 | The Lactobacillus casei Zhang high-density cultivation method, use them to prepare the method for freeze-dried vaccine powder and resulting freeze-dried vaccine powder and uses thereof |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104223305A (en) * | 2014-10-09 | 2014-12-24 | 哈尔滨工业大学 | Coix seed lactobacillus beverage preparation method |
| CN104304450A (en) * | 2014-10-09 | 2015-01-28 | 哈尔滨工业大学 | Semen coicis lactic acid bacteria beverage fermentation process |
| CN106754524A (en) * | 2016-12-28 | 2017-05-31 | 石家庄君乐宝乳业有限公司 | Lactobacillus paracasei N1115 culture mediums and its application |
| CN106754524B (en) * | 2016-12-28 | 2020-06-16 | 石家庄君乐宝乳业有限公司 | Lactobacillus paracasei N1115 culture medium and application thereof |
| CN108048344A (en) * | 2017-11-20 | 2018-05-18 | 广东博沃特生物科技有限公司 | Two plants of deodorization bacterial strains and its application in composite biological deodorant is prepared |
| CN108048344B (en) * | 2017-11-20 | 2019-05-31 | 广东博沃特生物科技有限公司 | Two plants of deodorization bacterial strains and its application in preparation composite biological deodorant |
| WO2020063515A1 (en) * | 2018-09-30 | 2020-04-02 | 内蒙古伊利实业集团股份有限公司 | Novel application of lactobacillus paracasei k56 capable of regulating balance of gastrointestinal flora |
| US11666613B2 (en) | 2018-09-30 | 2023-06-06 | Inner Mongolia Yili Industrial Group Co., Ltd. | Use of Lactobacillus paracasei subsp. paracasei K56 capable of regulating gastrointestinal flora balance |
| CN110734884A (en) * | 2019-11-27 | 2020-01-31 | 汉臣氏（沈阳）儿童制品有限公司 | Low-salt culture medium and culture method for lactobacillus paracasei |
| CN110734884B (en) * | 2019-11-27 | 2024-04-09 | 江西仁仁健康微生态科技有限公司 | Lactobacillus paracasei low-salt culture medium and culture method |
| CN111286471A (en) * | 2019-12-12 | 2020-06-16 | 湖北华扬科技发展有限公司 | Method for high-density culture of lactobacillus paracasei by using soybean meal hydrolysate |
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Application publication date: 20130814 |