酿酒酵母中柚皮素异源合成途径构建与优化
Construction and optimization of synthetic pathway of naringin from Saccharomyces cerevisiae
任枫 1周一征 1杨利文 1王舜 1何如怡 1杨彪 1田宇 1刘志国1
作者信息
- 1. 武汉轻工大学生命科学与技术学院,武汉 430023
- 折叠
摘要
本研究构建酿酒酵母柚皮素细胞合成工厂,优化关键合成步骤,以提升柚皮素产量.采用同源重组技术,在酿酒酵母的rDNA位点构建从对香豆酸到柚皮素的合成途径,包括4-香豆酸辅酶A连接酶(4CL)、查尔酮合成酶(CHS)、查尔酮异构酶(CHI)三个酶基因的完整表达阅读框.同时,单因素法优化YPD发酵培养基中酵母提取物,葡萄糖和胰蛋白胨浓度.结果表明:得到了5株产柚皮素的酿酒酵母改造菌株S1、S2、S3、S4、S5,在4％的酵母提取物,10％的胰蛋白胨和2％的葡萄糖的最优配比下,5 L发酵罐扩大培养,通过分批补料发酵,最高柚皮素产量为438 mg/L,成功实现了在酿酒酵母中异源合成柚皮素途径的构建与优化,为工业化微生物合成柚皮素提供了新的途径和参考价值.
Abstract
In this study,a naringenin cell synthesis plant of Saccharomyces cerevisiae was constructed to op-timize the key synthetic steps and increase naringenin production.A synthetic pathway from para-coumaric acid to naringin was constructed at the rDNA site of Saccharomyces cerevisiae using homologous recombi-nation technology,including a complete expression reading frame of 4-coumaric acid CoA ligase(4CL),Chalcone synthetase(CHS)and Chalcone isomerase(CHI).At the same time,the single factor method was used to optimize the yeast extract in YPD fermentation medium.Glucose and tryptone concentrations.The results showed that 5 strains S1,S2,S3,S4 and S5 of S.cerevisiae producing naringin were obtained.The maximum naringin yield was 438mg/L through batch feeding fermentation under the optimal ratio of 4％yeast extract,10％tryptonate and 2％glucose.The construction and optimization of heterologous syn-thesis pathway of naringenin in Saccharomyces cerevisiae were successfully realized,which provided a new way and reference value for industrial microbial synthesis of naringenin.
关键词
柚皮素/酿酒酵母/异源合成/同源整合/发酵罐Key words
naringenin/saccharomyces cerevisiae/allosynthesis/homologous integration/fermenter引用本文复制引用
出版年
2024
NETL
NSTL
万方数据