华东理工大学张立新、谭高翼研究团队报道，通过代谢工程和生物催化生产高产量卟啉。2024年6月5日出版的《自然—生物技术》发表了这项成果。

据介绍，卟啉及其衍生物在医药、食品、能源和材料等领域有着广泛的应用。

在这项研究中，该课题组将高效的细胞工厂——红螺菌（Rhodobacter sphaeroides）与酶催化相结合，生产了卟啉化合物。基于全基因组CRISPRi的筛选，在红螺菌中识别出hemN作为提高共卟啉III（CPIII）生产的目标，并利用PrrA的磷酸化进一步将生物活性CPIII的产量通过补料分批发酵提高到16.5 g L⁻¹。

随后，通过筛选和工程化高活性金属螯合酶和共卟啉脱羧酶，合成了各种金属卟啉，包括血红素和抗肿瘤剂锌卟啉。在进行200升的中试发酵并建立CPIII（纯度>95%）的纯化工艺后，通过酶催化在5升生物反应器中扩大生产血红素和锌卟啉，CPIII分别达到63%和98%的酶转化率，产量分别为10.8 g L⁻¹和21.3 g L⁻¹。

他们的策略为高产量生物生产血红素，及其他具有重要工业和医学应用的卟啉提供了解决方案。

附：英文原文

Title: High-yield porphyrin production through metabolic engineering and biocatalysis

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

Issue&Volume: 2024-06-05

Abstract: 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 16.5gL1 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 (200L) 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 10.8gL1 and 21.3gL1, respectively. Our strategy offers a solution for high-yield bioproduction of heme and other valuable porphyrins with substantial industrial and medical applications.

DOI: 10.1038/s41587-024-02267-3

Source: https://www.nature.com/articles/s41587-024-02267-3