CN103194403A - Simple and practical lactobacillus liquid fermentation method - Google Patents

Simple and practical lactobacillus liquid fermentation method Download PDF

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Publication number
CN103194403A
CN103194403A CN2012100051432A CN201210005143A CN103194403A CN 103194403 A CN103194403 A CN 103194403A CN 2012100051432 A CN2012100051432 A CN 2012100051432A CN 201210005143 A CN201210005143 A CN 201210005143A CN 103194403 A CN103194403 A CN 103194403A
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China
Prior art keywords
lactobacillus
fermentation
fermentation method
nutrient medium
animal
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CN2012100051432A
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Chinese (zh)
Inventor
张捷
郑文官
潘韵
彭永鹤
李永新
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SHENZHEN SUNSMILE BIOTECHNOLOGY CO Ltd
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SHENZHEN SUNSMILE BIOTECHNOLOGY CO Ltd
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Priority to CN2012100051432A priority Critical patent/CN103194403A/en
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Abstract

The invention is applicable to the technical field of fermentation engineering, and provides a simple and practical lactobacillus liquid fermentation method. The method comprises the following steps: preparing a liquid culture medium comprising the components specified as in the specification; inoculating lactobacillus with inoculum age of 250000000-500000000 / mL by an inoculum size of 2.5-7% (V / V) into the liquid culture medium; and fermenting and culturing under conditions of pH of 6-6.5, ventilation of 0.06-0.2vvm and temperature of 30-37 DEG C for 22-24 h. The lactobacillus fermentation method provided by the invention through the optimization of fermentation culture medium realizes a same fermentation bacterial count as a traditional fermentation culture medium, has simple preparation method and obviously reduced cost, and is more suitable for industrial need.

Description

A kind of simple and practical lactobacillus liquid fermentation process
Technical field
The invention belongs to the fermentation engineering field, relate in particular to a kind of lactobacillus fermentation methods and applications.
Background technology
Lactobacillus is that lactic acid bacteria class is a kind of, produces the bacterium of a large amount of lactic acid after the common general reference of milk-acid bacteria can utilize carbohydrate to ferment.Milk-acid bacteria extensively exists in nature and people and animals' Digestive tract, and wherein the overwhelming majority all is present in the enteron aisle of human body and animal, is the flora that body is essential and have important physiological function.Probiotic lactobacillus commonly used mostly is the lactobacillus class.Also have no lack of animal lactobacillus goods on the market.
At present, for the cultivation of lactobacillus, utilize solid-state substrate or semi-solid state substrate cultivation base (for example milk-product, juice product etc.) more, leavened prod is directly used, but the product category that these class methods are produced is single, and cost is higher, range of application is narrow, is not suitable for the animal healthcare products.Also the someone utilize solid substances fermentative production animal food prods (number of patent application is: 200910040347.8), its prescription is feedstuff raw material really, but its zymotechnique complexity, complex operation, the material physical condition is required height, and behind the fermentation 48h, bacterial content does not reach still very high.
For the liquid fermentation and culture of lactobacillus, method is limited, and common is exactly to utilize the MRS liquid nutrient medium to cultivate, and obtains the liquid fermenting lactobacillus.
The MRS culture medium prescription is as follows:
Figure BSA00000654722100021
Cost height such as this peptone used in filling a prescription, beef powder, yeast powder are unsuitable for industrial production.
Summary of the invention
In view of the above problems, the present invention aims to provide a kind of lactobacillus liquid fermentation process, can simplify the liquid fermenting of lactobacillus, and guarantees that fermentation back viable count is consistent with the traditional zymotic substratum, makes its better application in producing animal lactobacillus goods.
The present invention is achieved in that
A kind of lactobacillus fermentation method comprises the steps:
Preparation comprises the liquid nutrient medium of following component;
Being age that the lactobacillus of hundred million/mL of 2.5-5 is that 2.5-7.0% (v/v) is inoculated in described liquid nutrient medium by inoculum size with the bacterial classification kind, is that 6-6.5, air flow are 0.06-0.2vvm at pH, and temperature is under the 30-37 ℃ of condition, fermentation culture 22-24 hour.
This nutrient media components is as follows:
Figure BSA00000654722100031
The embodiment of the invention further provides the application of the prepared lactobacillus of above-mentioned lactobacillus liquid fermentation process in animal probiotic bacterium healthcare products.
Advantage of the present invention and effect are as follows:
1, the culture medium prescription raw materials cost is cheap, is fit to the industrialization needs;
2, the liquid fermenting effect can be consistent with traditional substratum;
3, technology is simple, is fit to suitability for industrialized production;
4, have wide range of applications, be fit to produce various animals lactobacillus goods.
Embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer, below in conjunction with implementing illustration, 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.
The embodiment of the invention provides a kind of lactobacillus fermentation method: comprise the steps:
S01, preparation comprise the liquid nutrient medium of following component;
S02, being age that the lactobacillus of hundred million/mL of 2.5-5 is that 2.5-7.0% (v/v) is inoculated in described liquid nutrient medium by inoculum size with the bacterial classification kind, is that 6-6.5, air flow are 0.06-0.2vvm at pH, and temperature is under the 30-37 ℃ of condition, fermentation culture 22-24 hour.
This nutrient media components is as follows:
Figure BSA00000654722100041
Particularly, used lactobacillus among the step S02 can be plant lactobacillus, also can be in lactobacterium casei, the lactobacillus acidophilus any one.These 3 kinds of lactobacilluss are all available from microbial strains preservation center, Guangdong Province.Its numbering is respectively: plant lactobacillus: GIM1.140=AS1.3; Lactobacterium acidophilum: GIM1.208=AS1.2686; Lactobacterium casei: GIM1.325=ATCC7469.
Particularly, among the step S02, use alkaline agent to regulate pH value in the fermenting process, this alkaline agent without limits, for example, sodium hydroxide, ammoniacal liquor etc.
Particularly, among the step S02, bacterial classification kind age is hundred million/mL of 2.5-5, and its culturing process is: picking colony from MRS solid slant culture base, insert in the 30mL MRS liquid nutrient medium, and to cultivate in 37 ℃ of constant temperature shaking tables, rotating speed is 200rpm, cultivates 16-18 hour.
Implement illustration
1, picking colony from MRS solid slant culture base inserts in the 30mL MRS liquid nutrient medium, cultivates in 37 ℃ of constant temperature shaking tables, and rotating speed is 200rpm, cultivates 16-18 hour.Be hundred million/mL of 4-5 bacterial classification kind age of plant lactobacillus.
The MRS nutrient media components is as follows:
Figure BSA00000654722100051
Solid medium need add agar powder 15.0g.Mentioned component is added in people's distilled water, and heating for dissolving is proofreaied and correct pH 6.2,121 ℃ of autoclaving 15min-20min after the packing.
2, in the 20L fermentor tank, insert the described liquid nutrient medium of 15L, after 121 ℃ of sterilization 30min are cooled to 32 ℃, be the plant lactobacillus of hundred million/mL of 4-5 with bacterial classification kind age, be 3.3% (v/v) by inoculum size, be inoculated in the fermentor tank, be 6.4 at pH, air flow is 0.1vvm, temperature is under 37 ℃ of conditions, fermentation culture 24 hours.
Described liquid nutrient medium component is as follows:
Figure BSA00000654722100061
3, the fermentation result measures: by the method for blood counting chamber counting, measured fermentation termination, plant lactobacillus viable bacteria content: reach 1.1 * 10 10Cfu/mL.The culture effect of effect and MRS substratum is suitable.
4, above-mentioned tunning can directly add in the animal daily ration, also can be made into solid product, more can be added in the daily tap water of animal etc., and is respond well, promotes breeding performonce fo animals.
The above is preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; can also make some improvements and modifications, these improvements and modifications also are considered as protection scope of the present invention.

Claims (7)

1. a lactobacillus fermentation method comprises the steps:
Preparation comprises the liquid nutrient medium of following component;
Being age that the lactobacillus of hundred million/mL of 2.5-5 is that 2.5-7.0% (v/v) is inoculated in described liquid nutrient medium by inoculum size with the bacterial classification kind, is that 6-6.5, air flow are 0.06-0.2vvm at pH, and temperature is under the 30-37 ℃ of condition, fermentation culture 22-24 hour,
Described nutrient media components is as follows:
Figure FSA00000654722000011
Lactobacillus as claimed in claim 1 can in plant lactobacillus, lactobacterium casei, the lactobacillus acidophilus any one.
3. lactobacillus fermentation method as claimed in claim 1 is characterized in that, the component of described liquid nutrient medium is as follows:
4. lactobacillus fermentation method as claimed in claim 1 is characterized in that, uses the encloses container that can sterilize as fermenting container in the described fermentation culture step, also can use fermentor tank as fermenting container, and the coefficient of described fermenting container is 0.5-0.75.
5. lactobacillus fermentation method as claimed in claim 1 is characterized in that, described fermentation culture is under quiescent conditions or near carrying out under the quiescent conditions (mixing speed is about 25-50rpm).
6. lactobacillus fermentation method as claimed in claim 1 is characterized in that, described fermentation culture is to carry out under the 0.03-0.08Mpa condition at pressure.
As claim 1-6 as described in each prepared lactobacillus fermentation liquid of fermentation process at animal lactobacillus goods, as the application in animal health product, animal-feed.
CN2012100051432A 2012-01-09 2012-01-09 Simple and practical lactobacillus liquid fermentation method Pending CN103194403A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105400713A (en) * 2015-10-09 2016-03-16 中国科学院烟台海岸带研究所 Method for fermenting mixed lactic acid bacteria

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7300787B2 (en) * 2002-07-05 2007-11-27 Archer-Daniels-Midland Company Lactobacillus strains and use thereof in fermentation for L-lactic acid production

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7300787B2 (en) * 2002-07-05 2007-11-27 Archer-Daniels-Midland Company Lactobacillus strains and use thereof in fermentation for L-lactic acid production

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
L. F. COELHO 等: ""LACTIC ACID PRODUCTION BY NEW Lactobacillus plantarum LMISM6 GROWN IN MOLASSES: OPTIMIZATION OF MEDIUM COMPOSITION"", 《BRAZILIAN JOURNAL OF CHEMICAL ENGINEERING》, vol. 28, no. 1, 31 March 2011 (2011-03-31) *
梁晓华 等: ""水产动物益生菌乳酸杆菌LH发酵工艺的研究"", 《中国微生态学杂志》, vol. 21, no. 8, 31 December 2009 (2009-12-31), pages 679 - 2 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105400713A (en) * 2015-10-09 2016-03-16 中国科学院烟台海岸带研究所 Method for fermenting mixed lactic acid bacteria
CN105400713B (en) * 2015-10-09 2019-04-26 中国科学院烟台海岸带研究所 A kind of mixing lactic acid bacteria fermentation process

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Application publication date: 20130710