工程解脂耶氏菌生物合成香叶醇

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2023-11-02 DOI:10.1016/j.mec.2023.e00228
Ayushi Agrawal , Zhiliang Yang , Mark Blenner
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引用次数: 1

摘要

香叶醇是一种单萜类化合物,在食品、化妆品、制药等行业有着广泛的应用。微生物生产在很大程度上使用了缺乏单萜烯生产有利特性的模式生物。在这项工作中,我们在代谢工程的解脂耶氏菌中生产香叶醇。首先,在前人报道的基础上,对两种植物源性香叶醇合成酶(GES)进行了活性检测。通过与MVA途径酶tHMG1和IDI1共表达,检测了GES野生型和截断突变体(不靶向叶绿体的信号肽)。截断的CrGES (tCrGES)产生的香叶醇最多,因此用于进一步的实验。通过过表达截断的HMG1、IDI和tCrGES获得初始菌株。乙酰辅酶a前体库通过过表达甲羟戊酸途径基因如ERG10、HMGS或MVK、PMK而增强。最终菌株在摇瓶发酵中过表达3份tCrGES和单份ERG10、HMGS、tHMG1、IDI,产量约为1 g/L。这是首次证明在解脂耶氏菌中产生香叶醇,也是迄今为止在酵母中报道的最高的新生滴度。
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Engineering Yarrowia lipolytica for the biosynthesis of geraniol

Geraniol is a monoterpene with wide applications in the food, cosmetics, and pharmaceutical industries. Microbial production has largely used model organisms lacking favorable properties for monoterpene production. In this work, we produced geraniol in metabolically engineered Yarrowia lipolytica. First, two plant-derived geraniol synthases (GES) from Catharanthus roseus (Cr) and Valeriana officinalis (Vo) were tested based on previous reports of activity. Both wild type and truncated mutants of GES (without signal peptide targeting chloroplast) were examined by co-expressing with MVA pathway enzymes tHMG1 and IDI1. Truncated CrGES (tCrGES) produced the most geraniol and thus was used for further experimentation. The initial strain was obtained by overexpression of the truncated HMG1, IDI and tCrGES. The acetyl-CoA precursor pool was enhanced by overexpressing mevalonate pathway genes such as ERG10, HMGS or MVK, PMK. The final strain overexpressing 3 copies of tCrGES and single copies of ERG10, HMGS, tHMG1, IDI produced approximately 1 g/L in shake-flask fermentation. This is the first demonstration of geraniol production in Yarrowia lipolytica and the highest de novo titer reported to date in yeast.

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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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