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An automatic feedback feeding method for high-density culture of lactic acid bacteria

A technology for high-density culture and lactic acid bacteria, applied in the field of automatic feedback feeding of high-density cultured lactic acid bacteria, can solve problems such as the inapplicability of exponential flow addition, to ensure stability, short culture time, and avoid insufficient or excessive feeding. Effect

Active Publication Date: 2019-11-26
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in order to obtain a higher concentration of lactic acid bacteria cells, it is still necessary to carry out fed alkali supplementation culture in the late logarithmic phase of batch culture. At this time, the cell concentration will continue to increase, but not in an exponential manner. method does not apply
At present, in terms of high-density culture of lactic acid bacteria, there is no effective batch culture fed-batch method

Method used

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  • An automatic feedback feeding method for high-density culture of lactic acid bacteria
  • An automatic feedback feeding method for high-density culture of lactic acid bacteria
  • An automatic feedback feeding method for high-density culture of lactic acid bacteria

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Embodiment 1 automatic feedback feeding high-density culture Lactobacillus casei

[0036] (1) Preparation of seed solution

[0037] Draw 200μL of Lactobacillus casei from the glycerol storage tube into 10mL MRS liquid medium, incubate at 42°C for 18 hours for activation, and then transfer it to 100mL MRS liquid medium with a 5% (v / v) inoculation amount for the second time After activating, cultivate at 42°C for 12 hours to obtain the seed solution.

[0038] (2) Preparation before vaccination and vaccination process

[0039] Prepare 600 mL of MRS liquid medium with half the concentration of carbon source and nitrogen source and pour it into a 1L fermenter. After the fermenter body is sterilized at 121°C, it is cooled to 42°C in a water bath, and the temperature is kept constant. When starting the inoculation, pour the seed solution into the fermenter with an inoculation amount of 5% (v / v) above the alcohol flame circle, stir at a speed of 150-200 r / min, and cultivate a...

Embodiment 2

[0048] Example 2 Automatic Feedback Feeding High Density Cultivation of Bifidobacterium adolescentis

[0049] (1) Preparation of seed solution

[0050] Draw 200 μL of Bifidobacterium adolescentis from the glycerol storage tube into 10mL MRS liquid medium (adding 1g / L cysteine ​​hydrochloride), put it into the anaerobic workstation for 24 hours at 37°C, and then add 5% (v The inoculum amount of / v) was transferred to 100 mL of liquid medium for secondary activation, and the seed liquid was obtained by anaerobic cultivation at a constant temperature of 37° C. for 15 hours.

[0051] (2) Preparation before vaccination and vaccination process

[0052] Prepare 600 mL of MRS liquid medium (adding 1 g / L cysteine ​​hydrochloride) with half the concentration of carbon source and nitrogen source and pour it into a 1L fermenter. °C and keep the temperature constant. When starting the inoculation, pour the seed liquid into the fermenter with an inoculum amount of 5% (v / v) above the alco...

Embodiment 3

[0061] Example 3 Automatic Feedback Feeding High Density Cultivation of Plant Bacteria

[0062] The automatic feedback feeding culture of Lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus rhamnosus, Lactobacillus acidophilus and other homofermentative lactobacilli is the same as in Example 1, and the culture temperature is the optimum growth temperature of each strain. Such as figure 2 As shown, Lactobacillus plantarum reached a final viable concentration of 1.3±0.1×10 after 16 hours of culture 10 cfu / mL.

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Abstract

The invention discloses an automatic feedback supplementing method for high-density culture of lactic acid bacteria, and belongs to the field of microbial fermentation engineering. According to the automatic feedback supplementing method disclosed by the invention, in the culturing process of the lactic acid bacteria, alkali liquor is automatically added according to a pH value so as to keep a neutral environment of a culture system; a supplementing pump is associated with an alkali supplementing pump, so that a supplementing culture medium and the alkali liquor are automatically added into the culture system according to a certain ratio, thereby ensuring stability of concentration of a substrate in the culture system. According to the automatic feedback supplementing method disclosed by the invention, not only is the requirement of bacteria for the substrate ensured, but also the substrate is automatically added according to consumption of the bacteria without exceeding the standard. The method disclosed by the invention is applicable to all homofermentative lactobacillus fermentium, such as lactobacillus casei, lactobacillus plantarum, lactobacillus rhamnosus, lactobacillus acidophilus, lactobacillus helveticus, lactobacillus bulgaricus and the like, and all bifidobacterium, such as bifidobacterium bifidum, bifidobacterium adolescentis, bifidobacterium animalis, bifidobacterium longum, bifidobacterium breve and the like.

Description

technical field [0001] The invention relates to an automatic feedback feeding method for high-density culture of lactic acid bacteria, which belongs to the field of microbial fermentation engineering. Background technique [0002] Lactic acid bacteria can not only improve the quality and flavor of fermented food through their own metabolism, but also improve human health. Lactic acid bacteria are widely used in fermented dairy products, fermented sausages, fermented fruits and vegetables, direct-throwing kimchi and other fermented foods. In recent decades, the functional research of lactic acid bacteria has also received extensive attention. Functional probiotics that improve the stomach, treat constipation, antagonize enteritis, and improve immunity are deeply loved by consumers. To improve the production efficiency of lactic acid bacteria, high-density culture of lactic acid bacteria is very important for the promotion of lactic acid bacteria products. [0003] A variety...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/245C12R1/25C12R1/225C12R1/23C12R1/01
CPCC12N1/20
Inventor 张灏崔树茂赵建新田丰伟王刚陈卫
Owner JIANGNAN UNIV
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