Synthetic polyunsaturated fatty acid conjugated phospholipid yeast engineering bacteria, construction method and application thereof

By constructing a phospholipid synthesis pathway in yeast cells and using the CRISPR/Cas9 system to knock out and integrate related genes, the problems of low production efficiency and high cost of polyunsaturated fatty acid-bound phospholipids in existing technologies have been solved, enabling efficient and economical large-scale production.

CN121699770BActive Publication Date: 2026-06-26YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
Filing Date
2026-02-12
Publication Date
2026-06-26

AI Technical Summary

Technical Problem

Existing technologies make it difficult to achieve large-scale production of polyunsaturated fatty acid-bound phospholipids (PUFAs-PL) efficiently and economically. Chemical synthesis methods carry the risk of double bond destruction, while enzyme-catalyzed synthesis methods are costly and have low reaction efficiency.

Method used

Phospholipid synthesis pathways were constructed in yeast cells using genetic engineering techniques. Related genes were knocked out and genes related to phospholipid synthesis, pyruvate synthesis, and polyunsaturated fatty acid synthesis were integrated to form an expression module. Gene knockout and homologous recombination were performed using the CRISPR/Cas9 system to construct the yeast engineered strain PL15.

Benefits of technology

The efficient synthesis of PUFAs-PL was achieved, increasing phospholipid yield by 7 times to 151.19 mg/g, with an EPA content of 13.2%, thus solving the problems of production efficiency and cost in existing technologies.

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Abstract

The present application relates to a kind of synthetic polyunsaturated fatty acid binding phospholipid yeast engineering bacteria and construction method and application, belong to microbial engineering technical field, the engineering bacteria is with Yarrowia lipolytica as bottom cell, knock out the Mhy1, Iro1, IDH1, CEX1 gene in its genome for generating mycelium, synthesis of triglyceride, citric acid metabolism, citric acid efflux;Phospholipid synthesis, pyruvate synthesis, polyunsaturated fatty acid synthesis and acyltransferase CDS1, OPI3, Mce2, LplA, △9Elo, △8Des, △5Des, △17Des, △5Elo, △15Des, LPCAT3 gene are sequentially connected together, as an expression module is integrated in the URA site of genome.The yeast engineering bacteria of the present application obtains polyunsaturated fatty acid binding phospholipid in one step efficiently, and phospholipid yield is 151.19 mg / g, and in binding fatty acid, EPA content is 13.2%.
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