title: Biosynthesis of Neocarazostatin A Reveals the Sequential Carbazole Prenylation and Hydroxylation in the Tailoring Steps.
abstract: Neocarazostatin A (NZS) is a bacterial alkaloid with promising bioactivities against free radicals, featuring a tricyclic carbazole nucleus with a prenyl moiety at C-6 of the carbazole ring. Here, we report the discovery and characterization of the biosynthetic pathway of NZS through genome mining and gene inactivation. The in vitro assays characterized two enzymes: NzsA is a P450 hydroxylase and NzsG is a new phytoene-synthase-like prenyltransferase (PTase). This is the first reported native PTase that specifically acts on the carbazole nucleus. Finally, our in vitro reconstituted experiment demonstrated a coupled reaction catalyzed by NzsG and NzsA tailoring the NZS biosynthesis.
authors.affiliation: Key Laboratory of Combinatory Biosynthesis and Drug Discovery (Ministry of Education), School of Pharmaceutical Sciences, Wuhan University, 185 East Lake Road, Wuhan 430071, P.R. China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Huang
authors.affiliation: Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, UK.
authors.forename: Somayah Sameer
authors.initials: SS
authors.lastname: Elsayed
authors.affiliation: Key Laboratory of Combinatory Biosynthesis and Drug Discovery (Ministry of Education), School of Pharmaceutical Sciences, Wuhan University, 185 East Lake Road, Wuhan 430071, P.R. China.
authors.forename: Meinan
authors.initials: M
authors.lastname: Lv
authors.affiliation: Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, UK.
authors.forename: Jioji
authors.initials: J
authors.lastname: Tabudravu
authors.affiliation: Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, UK; Pharmacognosy Department, School of Pharmacy, Beni-Suef University, Beni-Suef 32514, Egypt.
authors.forename: Mostafa E
authors.initials: ME
authors.lastname: Rateb
authors.affiliation: Department of Chemistry, University of Ghana, P.O. Box LG56, Legon-Accra, Ghana.
authors.forename: Roland
authors.initials: R
authors.lastname: Gyampoh
authors.affiliation: Department of Chemistry, University of Ghana, P.O. Box LG56, Legon-Accra, Ghana.
authors.forename: Kwaku
authors.initials: K
authors.lastname: Kyeremeh
authors.affiliation: Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, UK.
authors.forename: Rainer
authors.initials: R
authors.lastname: Ebel
authors.affiliation: Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, UK.
authors.forename: Marcel
authors.initials: M
authors.lastname: Jaspars
authors.affiliation: Key Laboratory of Combinatory Biosynthesis and Drug Discovery (Ministry of Education), School of Pharmaceutical Sciences, Wuhan University, 185 East Lake Road, Wuhan 430071, P.R. China.
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
authors.affiliation: Key Laboratory of Combinatory Biosynthesis and Drug Discovery (Ministry of Education), School of Pharmaceutical Sciences, Wuhan University, 185 East Lake Road, Wuhan 430071, P.R. China. Electronic address: yu_yi@whu.edu.cn.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, UK. Electronic address: h.deng@abdn.ac.uk.
authors.forename: Hai
authors.initials: H
authors.lastname: Deng
mesh_terms: D053898:Biosynthetic Pathways; D002227:Carbazoles / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006900:Hydroxylation; D008969:Molecular Sequence Data; D015394:Molecular Structure; D054876:Prenylation; D053719:Tandem Mass Spectrometry
chemical_list: D002227:Carbazoles; C041514:carbazole; C071015:neocarazostatin A
keywords: P450 enzyme; biosynthesis; carbazole alkaloids; neocarazostatin A; prenyltransferase; reconstitution