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Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum

A technology of Lactobacillus plantarum and conjugated linoleic acid, which is applied in the field of microbial fermentation, can solve the problems of cumbersome purification process and poor separation efficiency of active CLA, and achieve the effects of convenient product separation, avoiding coenzyme regeneration, and high yield

Inactive Publication Date: 2011-10-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the chemical synthesis method inevitably produces a series of CLA isomer mixtures, saturated or unsaturated fatty acids; although the operating cost is low, the separation efficiency is poor, requiring repeated operations, and the active CLA purification process is cumbersome

Method used

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  • Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum
  • Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum
  • Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum

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Experimental program
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Effect test

Embodiment 1

[0023] After the Lactobacillus plantarum (Lactobacillus plantarum) lp15-2-1 is activated twice, it is inoculated into 300ml MRS liquid medium with 1% inoculum size, and the emulsified linoleic acid solution is added so that the LA weight percent concentration reaches 0.5%, After anaerobic shaking culture (180r / min) at 30°C for 24h, high-speed refrigerated centrifugation for 5-20min (10000r / min, 4°C), the bacteria were collected and washed three times with normal saline to obtain resting cells.

[0024] Resuspend the resting cells in citric acid-sodium citrate buffer (0.15mol / L, pH5.5) to a concentration of 0.3mg / mL, and add the emulsified substrate LA to a concentration of 3mg / ml , 30°C shaking culture (180r / min) for 84h, and the output of CLA was detected and calculated by the following method, which reached 0.32mg / ml, and the conversion rate was 10%.

[0025] UV spectrophotometry was used for detection. Conjugated linoleic acid had an obvious absorption peak at 233nm. Using ...

Embodiment 2

[0029] The present invention also investigates the effects of different pre-cultivation methods, different storage conditions of resting cells, different shaking culture temperatures and different LA concentrations on the final CLA production.

[0030] (1) Effect of pre-culture method on CLA production

[0031] Such as figure 1 As shown, a total of 6 pre-culture conditions were set up, which were paraffin sealing + no addition of LA, paraffin sealing + addition of 0.5% LA, no paraffin sealing + no addition of LA, no paraffin sealing + addition of 0.5% LA, anaerobic bottle culture + No addition of LA, anaerobic bottle culture + addition of 0.5% LA, cell concentration of resting cells 0.3mg / mL, citric acid-sodium citrate buffer (0.1mol / L, pH6.0), reaction temperature 30°C, rotation speed 180r / min, reaction time 24h, LA concentration in citric acid-sodium citrate buffer solution is 0.5mg / ml.

[0032] from figure 1 It can be concluded that under the pre-culture condition of ana...

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Abstract

The invention discloses a method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum, which comprises the following steps: inoculating Lactobacillus plantarum CGMCC NO.3785 to an MRS (Man, Rogosa and Sharpe) culture medium containing linoleic acid, preculturing, and carrying out refrigerated centrifugation to obtain resting cells; and inoculating the resting cells to a citric acid-sodium citrate buffer solution containing 3.0-25 mg / ml of linoleic acid, and carrying out shake culture for 84-120 hours. The method for converting LA (linoleic acid) into CLA (conjugated linoleic acid) does not need a high-temperature high-pressure environment, and the product can be separated conveniently; and the resting cells obtained by centrifugation are used for catalyzing the synthesis of CLA, thereby preventing the overhigh-concentration LA from inhibiting the growth of bacteria and further achieving the goal of high CLA yield.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a method for biotransformation of conjugated linoleic acid by Lactobacillus plantarum. Background technique [0002] Conjugated linoleic acid (CLA) is a general term for a series of octadecadienoic acids containing conjugated double bonds and having positional and conformational isomerism. It is an essential fatty acid linoleic acid (LA for short). ) is a group of isomers. The most important isomer in humans and animals is cis-9, trans-11-octadecadienoic acid (c9, t11-CLA), which is named rumenic acid. [0003] The synthesis of CLA mainly has two kinds of chemical method and microbial method. According to different reaction mechanisms, chemical methods can be divided into: carbanion process, carbocation process, free radical process, addition and elimination process, conjugation catalyzed by Group VIII metals or their compounds, etc. However, the chemical synthe...

Claims

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Application Information

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IPC IPC(8): C12P7/64C12R1/25
Inventor 何国庆刘佩阮晖周倩
Owner ZHEJIANG UNIV
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