Polyketide Synthases; non-ribosomal peptide synthase; naringenin; Flavonoids; Peptide Synthases; Malonyl Coenzyme A; Phylogeny; Tandem Mass Spectrometry; Peptide Synthases/metabolism; Catalysis; Polyketide Synthases/genetics; Polyketide Synthases/metabolism; Chemistry (all); Biochemistry, Genetics and Molecular Biology (all); Physics and Astronomy (all)
Abstract :
[en] Biosynthesis of the flavonoid naringenin in plants and bacteria is commonly catalysed by a type III polyketide synthase (PKS) using one p-coumaroyl-CoA and three malonyl-CoA molecules as substrates. Here, we report a fungal non-ribosomal peptide synthetase -polyketide synthase (NRPS-PKS) hybrid FnsA for the naringenin formation. Feeding experiments with isotope-labelled precursors demonstrate that FnsA accepts not only p-coumaric acid (p-CA), but also p-hydroxybenzoic acid (p-HBA) as starter units, with three or four malonyl-CoA molecules for elongation, respectively. In vitro assays and MS/MS analysis prove that both p-CA and p-HBA are firstly activated by the adenylation domain of FnsA. Phylogenetic analysis reveals that the PKS portion of FnsA shares high sequence homology with type I PKSs. Refactoring the biosynthetic pathway in yeast with the involvement of fnsA provides an alternative approach for the production of flavonoids such as isorhamnetin and acacetin.
Disciplines :
Agriculture & agronomy
Author, co-author :
Zhang, Hongjiao ✱; State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China ; Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China
Li, Zixin ✱; Université de Liège - ULiège > TERRA Research Centre ; State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China ; Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China
Zhou, Shuang; State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China ; Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China
Li, Shu-Ming ; Institut für Pharmazeutische Biologie und Biotechnologie, Fachbereich Pharmazie, Philipps-Universität Marburg, Marburg, 35037, Germany
Ran, Huomiao; State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China
Song, Zili; State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China ; Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China
Yu, Tao ; Center for Synthetic Biochemistry, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes for Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China
Yin, Wen-Bing ; State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. yinwb@im.ac.cn ; Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China. yinwb@im.ac.cn
✱ These authors have contributed equally to this work.
Language :
English
Title :
A fungal NRPS-PKS enzyme catalyses the formation of the flavonoid naringenin.
We thank Professor Jens Nielsen (Chalmers University of Technology, Sweden) for kindly sharing the QL35 strain. We thank Professors Yihua Chen, Zhaosheng Kong and Yu Fu from Institute of Microbiology, Chinese Academy of Sciences for kindly sharing BAP1 E. coli strain, plant Arabidopsis thaliana , and the Liquid nitrogen freezing homogenizer, respectively. We thank Professor Yi Zou (Southwest University, China) for helpful discussion and suggestion of the manuscript. We thank Drs. Jinwei Ren and Wenzhao Wang (Institute of Microbiology, CAS) for NMR and MS data collection. This work was supported by National Key Research and Development Program of China [grant no. 2020YFA0907801]; National Natural Science Foundation of China [grant no. 31861133004]; Deutsche Forschungsgemeinschaft (DFG, German Re-search Foundation) − Li844/11-1; Key Research Program of Frontier Sciences, CAS [grant no. ZDBS-LY-SM016 to W.-B.Y.]; Construction of the Registry and Database of Bioparts for Synthetic Biology, CAS [grant no. ZSYS-016 to W.-B.Y.], and China Postdoctoral Science Foundation [YJ20200309 and 2021M693362].We thank Professor Jens Nielsen (Chalmers University of Technology, Sweden) for kindly sharing the QL35 strain. We thank Professors Yihua Chen, Zhaosheng Kong and Yu Fu from Institute of Microbiology, Chinese Academy of Sciences for kindly sharing BAP1 E. coli strain, plant Arabidopsis thaliana, and the Liquid nitrogen freezing homogenizer, respectively. We thank Professor Yi Zou (Southwest University, China) for helpful discussion and suggestion of the manuscript. We thank Drs. Jinwei Ren and Wenzhao Wang (Institute of Microbiology, CAS) for NMR and MS data collection. This work was supported by National Key Research and Development Program of China [grant no. 2020YFA0907801]; National Natural Science Foundation of China [grant no. 31861133004]; Deutsche Forschungsgemeinschaft (DFG, German Re-search Foundation) − Li844/11-1; Key Research Program of Frontier Sciences, CAS [grant no. ZDBS-LY-SM016 to W.-B.Y.]; Construction of the Registry and Database of Bioparts for Synthetic Biology, CAS [grant no. ZSYS-016 to W.-B.Y.], and China Postdoctoral Science Foundation [YJ20200309 and 2021M693362].
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