Acid-resistance lactococcus lactis and application thereof

A technology of Lactococcus lactis and yogurt, applied in the direction of streptococcus/lactococcus, application, bacteria, etc., can solve the problems affecting the production efficiency of cell metabolism activity, and achieve simple and easy operation, obvious effect, good growth performance and acid resistance receptive effect

Active Publication Date: 2017-01-04
JIANGNAN UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

On the other hand, with the accumulation of intracellular acid production, the pH in the microbial cells continues to drop, and the activities of related enzymes in the cells that maintain the normal physiological functions of the cells are inhibited, which in turn affects the metabolic activity and production efficiency of the cells. And bury many hidden dangers for production cost, downstream processing and subsequent environmental treatment of industrial emissions

Method used

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  • Acid-resistance lactococcus lactis and application thereof

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Experimental program
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Effect test

Embodiment 1

[0010] The breeding method of embodiment 1 Lactococcus lactis (CCTCC NO:M 2016234)

[0011] Lactococcus lactis NZ9000 was used as the starting strain, cultured in GM17 medium to the logarithmic growth phase, and the concentration of the bacterial solution was adjusted to 1*10 7 Unit / CFU, the supernatant was removed after the sample was centrifuged (5000rpm; 10min), washed and resuspended with 0.85% normal saline, and repeated twice. Add an equal volume of GM17 medium containing 0.5% v / v diethyl sulfate (DES) to resuspend, 100rpm, treat at 30°C for 30 minutes, immediately wash and resuspend with 0.85% normal saline, repeat 5 times, add an equal volume of GM17 After the culture medium was resuspended, it was cultured statically at 30°C for 1.5h.

[0012] Add 1 ml of GM17 (pH 5.0) medium to 2.2 ml of 96 deep-well plates, transfer the above-mentioned post-cultivation culture solution to 96 deep-well plates with a 2% inoculation amount, and culture at 30°C for 48 hours. Investiga...

Embodiment 2

[0013] The growth performance of embodiment 2 Lactococcus lactis (CCTCC NO:M 2016234) acid stress condition

[0014] The starting strain Lactococcus lactis NZ9000 and the selected Lactococcus lactis WH102 stored in a glycerol tube at -80°C were inserted into GM17 medium at an inoculation amount of 2%, and cultured statically at 30°C for 12 hours.

[0015] Transfer the activated Lactococcus lactis NZ9000 and Lactococcus lactis WH102 to GM17 (pH 4.5) medium with an inoculum amount of 2%, and culture them statically at 30°C for 48 hours. After the fermentation, measure the bacterial concentration of the fermentation broth . Under the condition of pH 4.5, the mutant strain OD 600 The value reached 0.527, which was 4.5 times higher than before breeding. The results are shown in Table 1.

[0016] Thalline growth performance under acid stress conditions in table 1

[0017] strain Lactococcus lactis NZ9000 Lactococcus lactis WH102 OD 600 0.081 0.371

Embodiment 3

[0018] Embodiment 3 acid tolerance experiment

[0019] The starting strain Lactococcus lactis NZ9000 and the selected Lactococcus lactis WH102 stored in a glycerol tube at -80°C were inserted into GM17 medium at an inoculation amount of 2%, and cultured statically at 30°C for 12 hours.

[0020] The activated Lactococcus lactis NZ9000 and Lactococcus lactis WH102 were put into the GM17 medium with an inoculation amount of 2%, and cultured statically at 30°C until the logarithmic growth phase.

[0021] Take the cells in the logarithmic growth phase, collect the cells by centrifugation at 5000rpm for 10 minutes, wash and centrifuge the cells twice with 0.85% normal saline, resuspend in an equal volume in GM17 (pH 4.0) medium, and take samples at different times of stress, and re- Centrifuge and wash the cells twice with the same normal saline, resuspend in an equal volume of normal saline, take 100 μl of the bacterial solution and spread it on a plate after appropriate dilution, ...

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Abstract

The invention discloses an acid-resistance lactococcus lactis and application thereof, and belongs to the field of food biotechnology. A separated bacterial strain is named lactococcus lactis WH102; under a condition that a pH value is 4.5, an OD600 value reached 0.371, which is increased by 2.6 times compared with the OD600 value of the strain before mutation; if the strain is intimidated for 2 hours in lactic acid with a pH value of 4.0, the survival rate is 6.09%; and if the strain after mutation is intimidated for 2 hours under lactic acid with the pH value of 4.0, the survival rate of the strain after mutation is 6.1 times of the strain before mutation under the same treatment condition. The acid-resistance lactococcus lactis has better acid resistance and a huge application value in the fields of foods, fermentation and the like.

Description

technical field [0001] The invention relates to a strain of acid-resistant Lactococcus lactis and application thereof, belonging to the field of food biotechnology. Background technique [0002] As an industrialized microbial cell factory, Lactococcus lactis has been widely used in food, fermentation and other fields. Acidogenic properties become an essential component of microorganisms in carrying out the fermentative production of industrial products required in the aforementioned fields. On the one hand, metabolic acid production can help promote cell energy conversion, maintain the balance of osmotic pressure inside and outside the cell, and enhance the environmental competitiveness of the body cells. On the other hand, with the accumulation of intracellular acid production, the pH in the microbial cells continues to drop, and the activities of related enzymes in the cells that maintain the normal physiological functions of the cells are inhibited, which in turn affects...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L29/00A23L19/20A23C9/123C12R1/46
CPCA23C9/1236C12R2001/46C12N1/205A23V2400/231
Inventor 张娟朱政明杨佩珊陈坚堵国成
Owner JIANGNAN UNIV
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