利用混杂的微生物戊烯基转移酶在酵母中合成制备戊烯基化柚皮素

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2021-06-01 DOI:10.1016/j.mec.2021.e00169
Shota Isogai , Nobuyuki Okahashi , Ririka Asama , Tomomi Nakamura , Tomohisa Hasunuma , Fumio Matsuda , Jun Ishii , Akihiko Kondo
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引用次数: 9

摘要

利用类黄酮生物合成途径和微生物中戊烯基转移酶(PTs)重组戊烯基类黄酮是一种有前途的有吸引力的替代植物生产或化学合成的方法。在这里,我们证明了混杂的微生物PTs可以替代区域特异性但大多数未知的植物PTs。为了测试柚皮素的前置化,我们通过基因组整合编码柚皮素生物合成途径成分的6个外源基因,构建了一株能够从l-苯丙氨酸生产柚皮素的酵母菌。利用该平台菌株,对不同微生物PTs进行了产丙烯基柚皮素的试验。体外筛选表明,真菌AnaPT(色氨酸二甲基烯丙基转移酶家族成员)特异性催化柚皮素的C-3 '烯酰化,而植物PT SfN8DT-1特异性催化C-8烯酰化。在体内,表达微生物AnaPT的柚皮素生产菌株表现出异源微生物生产3 ' -烯丙基柚皮素(3 ' -PN),与之前报道的使用植物SfN8DT-1在体内生产8-烯丙基柚皮素(8-PN)形成对比。这些发现为扩大各种烯丙基化化合物的生产提供了策略,包括众所周知的烯丙基柚皮素和新型烯丙基类黄酮。这些结果也表明,有机会取代植物PTs,无论是已知的还是未知的,显示相对严格的区域特异性的戊烯基转移。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Synthetic production of prenylated naringenins in yeast using promiscuous microbial prenyltransferases

Reconstitution of prenylflavonoids using the flavonoid biosynthetic pathway and prenyltransferases (PTs) in microbes can be a promising attractive alternative to plant-based production or chemical synthesis. Here, we demonstrate that promiscuous microbial PTs can be a substitute for regiospecific but mostly unidentified botanical PTs. To test the prenylations of naringenin, we constructed a yeast strain capable of producing naringenin from l-phenylalanine by genomic integration of six exogenous genes encoding components of the naringenin biosynthetic pathway. Using this platform strain, various microbial PTs were tested for prenylnaringenin production. In vitro screening demonstrated that the fungal AnaPT (a member of the tryptophan dimethylallyltransferase family) specifically catalyzed C-3′ prenylation of naringenin, whereas SfN8DT-1, a botanical PT, specifically catalyzed C-8 prenylation. In vivo, the naringenin-producing strain expressing the microbial AnaPT exhibited heterologous microbial production of 3′-prenylnaringenin (3′-PN), in contrast to the previously reported in vivo production of 8-prenylnaringenin (8-PN) using the botanical SfN8DT-1. These findings provide strategies towards expanding the production of a variety of prenylated compounds, including well-known prenylnaringenins and novel prenylflavonoids. These results also suggest the opportunity for substituting botanical PTs, both known and unidentified, that display relatively strict regiospecificity of the prenyl group transfer.

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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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