Method for biosynthesizing conjugated α-linolenic acid isomers in organic medium by coated bacteria

Abstract

The method for the biosynthesis of conjugated α-linolenic acid isomers in an organic medium by the coated thalline comprises the following steps: (1) each gram of Lactobacillus casei CGMCC 1.574 bacterium is resuspended in 5 mL of the permeabilization treatment solution, 37 Treat at ℃ for 20 minutes, and collect the permeabilized bacteria by centrifugation; (2) After washing with 50mM, pH5.8 phosphate buffer, collect the bacteria by centrifugation, and resuspend each gram of permeabilized bacteria in 20mL of 50mM, pH5.8 Add 0.5-2.0% Tween-20 to the phosphate buffer solution of the bacterial suspension, mix evenly, treat at 4-40°C for 2-6h, collect the bacterial cells by centrifugation, and obtain the coated bacterial cells after freeze-drying; (3) Add each gram of coated bacteria into 600mL of n-hexane, stir evenly, add α-linolenic acid with a volume of 0.1-0.3% of n-hexane, and react at 4-40°C for 1-12h; (4) Centrifuge to separate the coated bacteria , recovery of n-hexane, the product is c9, t11, c15-conjugated α-linolenic acid isomers; the reaction time of the present invention is short, the coated bacteria can be reused many times, the yield has been significantly improved, no environmental pollution, significant reduce manufacturing cost.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. Background technique [0002] Conjugated α-linolenic acid is a conjugated isomer of α-linolenic acid, which is a general term for a group of octadecyl-conjugated trienoic acids. There are various positional isomers and geometric isomers, such as: c 9, t 11, c 15-conjugated alpha-linolenic acid and t 10, c 12, c 15-conjugated α-linolenic acid isomers, etc. Studies have shown that conjugated α-linolenic acid has physiological functions such as anti-cancer, prevention of atherosclerosis, and weight loss, and its physiological activities are isomer-specific, such as: c 9, t 11, c 15-conjugated α-linolenic acid isomers have anticancer effects. In nature, conjugated α-linolenic acid mainly exists in the milk and meat fat of ruminants, mainly composed of c 9, t 11, c 15-conjugated α-linolenic acid and other isomers, but its content is usually very low, which cannot meet the development a...

AI Technical Summary

Problems solved by technology

At present, microbial fermentation is carried out in aqueous solution. Since the added fermentation substrate - α-linolenic acid is a fat-soluble substance, α-linolenic acid needs to be emulsified before being added to the fermentation broth, and the amount of addition is limited. From the fermentation broth The product of conjugated α-linolenic acid needs to be extracted by organic solvent, and the whole production process has the disadvantages of long fermentation period, high production cost and low yield.

Invention patent CN200410060670.9 reports specific biosynthesis by Lactobacillus casei CGMCC 1.574 c 9, t 11, c 15-conjugated α-linolenic acid isomer method, the substrate addition amount is 1mg / mL, the fermentation time is as long as 42h, the conversion rate of the substrate is less than 50%, and the bacteria cannot be reused