Xuefeng Mei, Deping Hua, Na Liu, Lilin Zhang, Xiaowen Zhao, Yujing Tian, Baiping Zhao, Jinhai Huang, Lei Zhang
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引用次数: 0
Abstract
Background: Anthocyanins are water-soluble flavonoids in plants, which give plants bright colors and are widely used as food coloring agents, nutrients, and cosmetic additives. There are several limitations for traditional techniques of collecting anthocyanins from plant tissues, including species, origin, season, and technology. The benefits of using engineering microbial production of natural products include ease of use, controllability, and high efficiency.
Results: In this study, ten genes encoding enzymes involved in the anthocyanin biosynthetic pathway were successfully cloned from anthocyanin-rich plant materials blueberry fruit and purple round eggplant rind. The Yeast Fab Assembly technology was utilized to construct the transcriptional units of these genes under different promoters. The transcriptional units of PAL and C4H, 4CL and CHS were fused and inserted into Chr. XVI and IV of yeast strain JDY52 respectively using homologous recombination to gain Strain A. The fragments containing the transcriptional units of CHI and F3H, F3'H and DFR were inserted into Chr. III and XVI to gain Strain B1. Strain B2 has the transcriptional units of ANS and 3GT in Chr. IV. Several anthocyanidins, including cyanidin, peonidin, pelargonidin, petunidin, and malvidin, were detected by LC-MS/MS following the predicted outcomes of the de novo biosynthesis of anthocyanins in S. cerevisiae using a multi-strain co-culture technique.
Conclusions: We propose a novel concept for advancing the heterologous de novo anthocyanin biosynthetic pathway, as well as fundamental information and a theoretical framework for the ensuing optimization of the microbial synthesis of anthocyanins.
背景:花青素是植物中的水溶性类黄酮,它赋予植物鲜艳的色彩,被广泛用作食品着色剂、营养素和化妆品添加剂。从植物组织中采集花青素的传统技术存在一些局限性,包括物种、产地、季节和技术等。利用工程微生物生产天然产品的好处包括使用方便、可控性强、效率高:本研究从富含花青素的植物材料蓝莓果实和紫圆茄外皮中成功克隆了十个编码花青素生物合成途径中酶的基因。利用酵母Fab组装技术,在不同启动子下构建了这些基因的转录单元。利用同源重组技术,将 PAL 和 C4H、4CL 和 CHS 的转录单元分别融合并插入到酵母菌株 JDY52 的 XVI 和 IV 链中,得到菌株 A;将含有 CHI 和 F3H、F3'H 和 DFR 转录单元的片段插入到 III 和 XVI 链中,得到菌株 B1。菌株 B2 的染色体 IV 中含有 ANS 和 3GT 的转录单元。通过 LC-MS/MS,利用多菌株共培养技术,按照花青素在 S. cerevisiae 中从头开始生物合成的预测结果,检测到了几种花青素,包括青花素、芍药苷、芍药苷、佩兰苷和麦冬苷:我们提出了一种推进异源从头开始花青素生物合成途径的新概念,并为随后优化花青素的微生物合成提供了基础信息和理论框架。
期刊介绍:
Microbial Cell Factories is an open access peer-reviewed journal that covers any topic related to the development, use and investigation of microbial cells as producers of recombinant proteins and natural products, or as catalyzers of biological transformations of industrial interest. Microbial Cell Factories is the world leading, primary research journal fully focusing on Applied Microbiology.
The journal is divided into the following editorial sections:
-Metabolic engineering
-Synthetic biology
-Whole-cell biocatalysis
-Microbial regulations
-Recombinant protein production/bioprocessing
-Production of natural compounds
-Systems biology of cell factories
-Microbial production processes
-Cell-free systems