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<p>Enzymatic synthesis is currently the primary method for preparing 1,3-medium chain- 2-long-chain triacylglycerols (MLM-TAGs), which serve as both a dietary component and clinical nutrient for specific populations. The application of MLM-TAGs is obviously constrained by the high cost of catalysts. Hence, a novel approach was proposed for MLM-TAGs production by engineered yeast. Overexpressing the mutated <i>fas1</i><sup>R1834K</sup> increased the production of medium-chain fatty acids (C8–C12) and resulted in an MLM content of 0.41&#xa0;mol% of the total TAGs. The introduction of <i>Rn</i>ACSM4 enabled the recombinant to produce MLM-TAGs at a level of 4.2&#xa0;mol% when supplemented with 0.2&#xa0;mM sodium laurate. Further deletion of <i>GAT2</i> and <i>LRO1</i> increased the content of MLM-TAGs to 6.7&#xa0;mol%. Iterative optimization involving sodium laurate dosage, culture temperature, and amino acid addition elevated the MLM-TAGs content to 34.4&#xa0;mol%. Under the optimized conditions, the maximum yield of MLM-TAGs reached 18.5&#xa0;mg/g DCW, representing a 135-fold improvement over the original strain. This research presents a promising and sustainable alternative for MLM-TAGs production and demonstrates the feasibility of tailoring the acyl composition of intracellular TAGs.</p> Graphical abstract <p></p>

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Enhanced synthesis of 1, 3-medium chain-2-long chain triacylglycerols by engineered Saccharomyces cerevisiae

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月28日

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  • Zhuangju Peng,
  • Zikun Zhang,
  • Rihan Gao,
  • Li Deng,
  • Fang Wang,
  • Junfeng Liu

摘要

Enzymatic synthesis is currently the primary method for preparing 1,3-medium chain- 2-long-chain triacylglycerols (MLM-TAGs), which serve as both a dietary component and clinical nutrient for specific populations. The application of MLM-TAGs is obviously constrained by the high cost of catalysts. Hence, a novel approach was proposed for MLM-TAGs production by engineered yeast. Overexpressing the mutated fas1R1834K increased the production of medium-chain fatty acids (C8–C12) and resulted in an MLM content of 0.41 mol% of the total TAGs. The introduction of RnACSM4 enabled the recombinant to produce MLM-TAGs at a level of 4.2 mol% when supplemented with 0.2 mM sodium laurate. Further deletion of GAT2 and LRO1 increased the content of MLM-TAGs to 6.7 mol%. Iterative optimization involving sodium laurate dosage, culture temperature, and amino acid addition elevated the MLM-TAGs content to 34.4 mol%. Under the optimized conditions, the maximum yield of MLM-TAGs reached 18.5 mg/g DCW, representing a 135-fold improvement over the original strain. This research presents a promising and sustainable alternative for MLM-TAGs production and demonstrates the feasibility of tailoring the acyl composition of intracellular TAGs.

Graphical abstract