Zhao Wei, Fan Aili, Tarcz Sylwia, Zhou Kang, Yin Wen-Bing, Liu Xiao-Qing, Li Shu-Ming
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.
Appl Microbiol Biotechnol. 2017 Mar;101(5):1989-1998. doi: 10.1007/s00253-016-7966-x. Epub 2016 Nov 10.
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.
烟曲霉的真菌环二肽异戊烯基转移酶FtmPT1催化短杆菌肽F（环-L-色氨酸-L-脯氨酸）的常规C2-异戊烯基化反应，并参与多种生物活性天然产物的生物合成，包括色曲菌素、螺色曲菌素、疣孢菌素和烟曲霉震颤素。与二甲基烯丙基色氨酸合酶超家族的其他成员一样，FtmPT1对含色氨酸的环二肽和其他一些芳香族底物具有较高的底物选择性。先前的一项研究表明FtmPT1可以接受1-萘酚，但产物收率非常低。在本研究中，我们报道了FtmPT1_G115A、六个FtmPT1_Y205X单突变体以及FtmPT1_G115A_Y205C对1-萘酚和其他羟基萘的接受度显著提高。这些结果为通过定点诱变创建具有更高催化活性的生物催化剂提供了一个实例。
