期刊
JOURNAL OF BIOTECHNOLOGY
卷 231, 期 -, 页码 183-192出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2016.06.007
关键词
Plant natural product; Phenylpropanoids; Metabolic engineering; mRNA structure
资金
- National Natural Science Foundation of China [31201422, 31371807]
- Fundamental Research Funds for the Central Universities [KYZ201419]
- Key Projects in the National Science & Technology Pillar Program of China [2013BAD18B01-4]
- High-Tech Research and Development Program of China [2011AA100903]
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
Developing efficient microbial processes to produce flavonoids has been a metabolic engineering goal over the past decade due to their important functions. Previously, the de novo production of the main flavonoid precursor (2S)-pinocembrin was achieved. However, low productivity and two separate fermentation steps made it inappropriate for industrial scale low-cost production. Here, a stepwise modular engineering approach was introduced to systematically identify and eliminate metabolic pathway bottlenecks. The overall pathway was firstly divided into four modules and analysis then revealed that efficient conversion of L-phenylalanine to (2S)-pinocembrin is the major limiting factor. Therefore, the pathway from L-phenylalanine to (2S)-pinocembrin was re-cast into three modules to alleviate this bottleneck by modifying both the gene copy number and promoter strength. Furthermore, the expression of the rate limiting enzyme PAL was found to be correlated with 5' region of mRNA structure. The efficiency of the synthetic pathway was then improved by customizing the PAL expression level based on modification of the mRNA secondary structure. Fed-batch cultivation of engineered strains in a 3-L fermentor resulted in a final (2S)-pinocembrin production of 432.4 mg/L. The results presented here pave the way for the development of an economical and simple process for microbial production of flavonoids. (C) 2016 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据