Mutation on Gly115 and Tyr205 of the cyclic dipeptide C2-prenyltransferase FtmPT1 increases its catalytic activity toward hydroxynaphthalenes.
The fungal cyclic dipeptide prenyltransferase FtmPT1 from Aspergillus fumigatus catalyzes a regular C2-prenylation of brevianamide F (cyclo-L-Trp-L-Pro) and is involved in the biosynthesis of a number of biologically active natural products including tryprostatins, spirotryprostatins, verruculogen, and fumitremorgins. FtmPT1, like other members of the dimethylallyltryptophan synthase superfamily, was shown to have high substrate promiscuity for tryptophan-containing cyclic dipeptides and a few other aromatic substrates. A previous study demonstrated the acceptance of 1-naphthol by FtmPT1, but with very low product yield. In this study, we report the significantly increased acceptance of 1-naphthol and other hydroxynaphthalenes by FtmPT1_G115A and six FtmPT1_Y205X single mutants as well as FtmPT1_G115A_Y205C. These results provided an example for creation of biocatalysts with improved catalytic activity by site-directed mutagenesis.
College of Life Sciences, Capital Normal University, No.105 Xisanhuan Beilu, Beijing, 100048, China.|Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, 35037, Marburg, Germany.
Wei
W
Zhao
State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
Aili
A
Fan
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, 35037, Marburg, Germany.
Sylwia
S
Tarcz
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, 35037, Marburg, Germany.
Kang
K
Zhou
State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
Wen-Bing
WB
Yin
College of Life Sciences, Capital Normal University, No.105 Xisanhuan Beilu, Beijing, 100048, China. liuxq@mail.cnu.edu.cn.
Xiao-Qing
XQ
Liu
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, 35037, Marburg, Germany. shuming.li@staff.uni-marburg.de.
Shu-Ming
SM
Li
D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000235:genetics*; D055162:Biocatalysis; D004122:Dimethylallyltranstransferase / Q000235:genetics* / Q000378:metabolism*; D026121:Indole Alkaloids / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009284:Naphthols / Q000378:metabolism*; D054876:Prenylation
D026121:Indole Alkaloids; D009284:Naphthols; C000590722:brevianamide F; C029350:1-naphthol; D004122:Dimethylallyltranstransferase; C534854:FtmPT1 protein, Aspergillus fumigatus
Dimethylallyltryptophan synthase; Enzyme catalysis; Friedel–Crafts alkylation; Hydroxynaphthalenes; Prenyltransferase