Method for screening microbe producing polyunsaturated fatty acid

Abstract

The invention discloses a method for rapidly screening produced polyunsaturated fatty acid. In the method, by taking a key enzyme gene as a molecular marker, microbes producing the polyunsaturated fatty acid can be rapidly and directionally screened on the basis of a PCR amplification technology. The method comprises the following steps of: designing a primer according to conserved regions of reported delta 6 fatty acid desaturase (D6) and delta 5 fatty acid desaturase gene (D5), amplifying and screening microbes obtained by separating soil or a water body in sequence to produce GLA by amplifying the gene of D6 and to produce AA and EPA by amplifying the genes of D6 and D5 simultaneously; then fermenting and culturing the microbes capable of producing the gene of D6 and simultaneously producing the genes of D6 and D5 by amplifying and extracting fatty acid; analyzing the fatty acid synthesized by the microbes by using GC and MS; and finally determining the microbe producing the polyunsaturated fatty acid. The invention provides an effective strain or algae seed separating and screening method and a batch of effective starting strains or algae seeds for producing the polyunsaturated fatty acid by using micro to ferment.

Description

technical field [0001] The invention belongs to the technical fields of microbial biotechnology and biopharmaceuticals, and in particular relates to a method for screening microorganisms producing polyunsaturated fatty acids. Background technique [0002] Polyunsaturated fatty acids have a wide range of physiological functions and biological effects in the body, and the more double bonds, the higher the degree of unsaturation and the higher the nutritional value. Polyunsaturated fatty acids have physiological functions such as maintaining the structure and function of biological membranes, treating cardiovascular diseases, anti-inflammation, anti-cancer, promoting brain development, losing weight, and increasing animal litter rate and survival rate. Polyunsaturated fatty acids are usually derived from higher plants, animal offal and fish oil. However, the above-mentioned sources all have the disadvantages that the output cannot meet the market demand and the price is expens...

AI Technical Summary

Problems solved by technology

The carbon starvation detection method has high accuracy, but it is too cumbersome; the fat granule counting method is simple, but lacks accuracy, and the size of the fat granule must be considered when using it; although the Sudan III sludge staining method has high accuracy, However, microscopic observation is required, and the operation is still cumbersome. It will be a very time-consuming task to detect and screen dozens, hundreds, or even thousands of microorganisms

Moreover, a common shortcoming of the above-mentioned methods is that they cannot be targeted for rapid and high-throughput screening of polyunsaturated fatty acid-producing microorganisms. It is not clear whether the oil-containing strains screened by these methods contain the required target polyunsaturated fatty acids.

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