文献类型：期刊文献

英文题名：High-yield porphyrin production through metabolic engineering and biocatalysis

作者：Chen, Haihong[1];Wang, Yaohong[1];Wang, Weishan[2];Cao, Ting[1];Zhang, Lu[1];Wang, Zhengduo[1];Chi, Xuran[1];Shi, Tong[1];Wang, Huangwei[1];He, Xinwei[1];Liang, Mindong[1];Yang, Mengxue[1];Jiang, Wenyi[1];Lv, Dongyuan[1];Yu, Jiaming[1];Zhu, Guoliang[1];Xie, Yongtao[1];Gao, Bei[1];Wang, Xinye[1];Liu, Xueting[1];Li, Youyuan[1];Ouyang, Limin[1];Zhang, Jingyu[1];Liu, Huimin[1];Li, Zilong[2];Tong, Yaojun[3];Xia, Xuekui[4];Tan, Gao-Yi[1];Zhang, Lixin[1]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]Chinese Acad Sci, Inst Microbiol, State Key Lab Microbial Resources, Beijing, Peoples R China;[3]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai, Peoples R China;[4]Qilu Univ Technol, Shandong Acad Sci, Biol Inst, Key Biosensor Lab Shandong Prov, Jinan, Peoples R China

年份：2025

卷号：43

期号：10

起止页码：1717

外文期刊名：NATURE BIOTECHNOLOGY

收录：;EI(收录号：20242416229650);WOS:【SCI-EXPANDED(收录号:WOS:001243257700001)】；

基金：We thank Q. Chen (School of Biotechnology, East China University of Science and Technology), L. Zhang (School of Chemistry and Molecular Engineering, East China Normal University) and W. Zhang (State Key Laboratory of Microbial Technology, Shandong University) for their technical support in enzyme mining, engineering and characterization. We are also deeply grateful to Z. Zhou (Institute of Plant Physiology and Ecology, Chinese Academy of Sciences) for her generous gift of the ZX-5 strain and to Inner Mongolia Kingdomway Pharmaceutical Co., Ltd. for providing the HY01 strain. We would like to extend our gratitude to D.-Q. Lin from the College of Chemical and Biological Engineering at Zhejiang University for his invaluable guidance in conducting the techno-economic analysis and for providing assistance in using the SuperPro Designer simulation software. During the manuscript preparation, we also greatly appreciate the constructive insights provided by J.-J. Zhong, L. Bai and T. Liu from the School of Life Sciences and Biotechnology, Shanghai Jiao Tong University. This work was financially supported, in part, by grants from the National Key Research and Development Program of China (2020YFA0907800 to Z.L. and 2020YFA0907304 to T.G.-Y.); the National Natural Science Foundation of China (32370064 and 31870040 to T.G.-Y.); 2023 Double World-Class Project-Key Program-Intelligent Biomanufacturing to Z.L.; and the Open Funding Project of the State Key Laboratory of Microbial Metabolism (MMLKF22-03 to T.G.-Y.).

语种：英文

外文关键词：Catalysis - Diagnosis - Fermentation - Medical applications - Metabolic engineering

摘要：Porphyrins and their derivatives find extensive applications in medicine, food, energy and materials. In this study, we produced porphyrin compounds by combining Rhodobacter sphaeroides as an efficient cell factory with enzymatic catalysis. Genome-wide CRISPRi-based screening in R. sphaeroides identifies hemN as a target for improved coproporphyrin III (CPIII) production, and exploiting phosphorylation of PrrA further improves the production of bioactive CPIII to 16.5 g L-1 by fed-batch fermentation. Subsequent screening and engineering high-activity metal chelatases and coproheme decarboxylase results in the synthesis of various metalloporphyrins, including heme and the anti-tumor agent zincphyrin. After pilot-scale fermentation (200 L) and setting up the purification process for CPIII (purity >95%), we scaled up the production of heme and zincphyrin through enzymatic catalysis in a 5-L bioreactor, with CPIII achieving respective enzyme conversion rates of 63% and 98% and yielding 10.8 g L-1 and 21.3 g L-1, respectively. Our strategy offers a solution for high-yield bioproduction of heme and other valuable porphyrins with substantial industrial and medical applications.