Over-expression of α-bisabolene by metabolic engineering of Yarrowia lipolytica employing a golden gate DNA assembly toolbox

Biotechnology Notes Pub Date : 2023-01-01 DOI:10.1016/j.biotno.2022.12.005

Walid M. El-Sharoud , Samar A. Zalma , Leonardo Rios-Solis , Rodrigo Ledesma-Amaro

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引用次数: 2

Abstract

Yarrowia lipolytica is a modern workhorse for biotechnology that is amenable to genetic manipulations and can produce high levels of various enzymes. The present study was designed to engineer Y. lipolytica for the overexpression of α-bisabolene, a valuable biofuel precursor and pharmaceutical, making use of this yeast's ability to accumulate lipids, and with the use of a golden gate DNA assembly (GG) toolbox. By transforming Y. lipolytica with a GG genetic construct involving truncated 3-hydroxy-3-methyl-glutaryle coenzyme A reductase (tHMG) and α-bisabolene synthase (Bis) genes controlled by the strong TEF promoter and Lip2 terminator, the engineered yeast was able to produce 489 mg l⁻¹ of α-bisabolene. This was increased to 816 mg l⁻¹ by transforming a lipid-over-accumulating Y. lipolytica strain with the same genetic construct. Higher production titers of up to 1243 mg l⁻¹ could be also achieved by varying the culture conditions of the transformed strains.

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利用金门DNA组装工具箱进行脂质体耶氏菌代谢工程对α-双abolene的过表达

解脂Yarrowia是一种现代生物技术的主力，可以进行基因操作，并可以产生高水平的各种酶。本研究旨在利用这种酵母积累脂质的能力，并使用金门DNA组装（GG）工具箱，设计溶脂酵母，使其过表达α-双abolene，这是一种有价值的生物燃料前体和药物。通过用GG基因构建体转化Y.lipolytica，该基因构建体包含由强TEF启动子和Lip2终止子控制的截短的3-羟基-3-甲基戊二酰辅酶a还原酶（tHMG）和α-双abolene合成酶（Bis）基因，工程酵母能够产生489 mg l−1的α-双Abolene。通过转化具有相同基因构建体的脂质过多的溶脂Y。通过改变转化菌株的培养条件，也可以获得高达1243 mg l−1的更高生产滴度。

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Biotechnology Notes

CiteScore

1.70

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0.00%

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