Wang Hao-Nan, Ke Xia, Jia Rui, Huang Liang-Gang, Liu Zhi-Qiang, Zheng Yu-Guo
National and Local Joint Engineering Research Center for Biomanufacturing of Choral Chemicals, Zhejiang University of Technology, Hangzhou 310014, PR China; Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
National and Local Joint Engineering Research Center for Biomanufacturing of Choral Chemicals, Zhejiang University of Technology, Hangzhou 310014, PR China; Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China.
Bioresour Technol. 2022 Nov;364:128033. doi: 10.1016/j.biortech.2022.128033. Epub 2022 Sep 27.
Gibberellic acid (GA) is one of natural phytohormones, widely used in agriculture and downstream fields. Qualified for the nature productivity, Fusarium fujikuroi was currently employed for the industrial biotransformation from agriculture residues into GA. Herein, Multivariate modular metabolic engineering (MMME) was assigned to reconstitute the metabolic balance in F. fujikuroi for enhancing GA production. Three modules including precursor pool, cluster-specific channel and P450-mediated oxidation in GA biosynthetic pathway were defined and optimized separately. The enhancement of both precursor pool and cluster-specific channel pushed metabolic flux transfer into the GA-specific pathway. Moreover, both introduction of Vitreoscilla hemoglobin and reinforcement of NADPH-dependent cytochrome P450 reductase facilitated oxidation cofactor transfer and subsequently boosted mycelium growth and GA biosynthesis. Integration of three modules in the engineered strain accumulated 2.89 g/L GA in shake flask via submerged fermentation, presenting a promising modular metabolic engineering model for efficient microbial transformation in agro-industrial application.
赤霉素（GA）是一种天然植物激素，广泛应用于农业及下游领域。鉴于藤仓镰孢具有天然生产能力，目前它被用于将农业废弃物进行工业生物转化生产GA。在此，多变量模块化代谢工程（MMME）被用于重建藤仓镰孢的代谢平衡以提高GA产量。GA生物合成途径中的三个模块，即前体库、簇特异性通道和P450介导的氧化分别被定义并进行了优化。前体库和簇特异性通道的增强推动代谢流转移至GA特异性途径。此外，透明颤菌血红蛋白的引入和NADPH依赖性细胞色素P450还原酶的强化促进了氧化辅因子的转移，进而促进了菌丝体生长和GA生物合成。工程菌株中三个模块的整合通过深层发酵在摇瓶中积累了2.89 g/L的GA，为农业工业应用中的高效微生物转化提供了一个有前景的模块化代谢工程模型。
