Cloning and functional analysis of novel short-chain cis-prenyltransferases

Biochemical and Biophysical Research Communications
Takanori AmboTanetoshi Koyama

Abstract

cis-Prenyltransferase catalyzes the synthesis of Z,E-mixed prenyl diphosphates by sequential condensation of isopentenyl diphosphate with allylic diphosphate. cis-Prenyltransferases can be classified into three subgroups: short-, medium-, and long-chain cis-prenyltransferase, according to their product chain lengths. cis-Farnesyl diphosphate synthase from Mycobacterium tuberculosis has been the only example as short-chain cis-prenyltransferase so far characterized. In this study, we cloned the novel short-chain cis-prenyltransferases from three different bacteria, and characterized their enzymatic activities to compare and elucidate a common feature of the short-chain cis-prenyltransferases. Furthermore, we identified a specific isoleucine that is conserved in short-chain cis-prenyltransferases and located in close proximity of the omega-end of the geranyl diphosphate. Several site-directed mutants with respect to the isoleucine residue synthesized longer prenyl chain products and showed broader allylic substrate specificity. These results suggested that the isoleucine plays an important role in the substrate specificity and chain length determination mechanism of cis-prenyltransferase.

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Citations

Apr 24, 2013·Metabolic Engineering·Chonglong WangSeon-Won Kim
Jul 28, 2009·Cellular and Molecular Life Sciences : CMLS·Sophie VandermotenMichel Cusson
Feb 14, 2012·Plant Science : an International Journal of Experimental Plant Biology·Martin LohrJürgen E W Polle
May 19, 2009·Journal of Bioscience and Bioengineering·Takanori AmboTanetoshi Koyama
Nov 9, 2012·The Plant Journal : for Cell and Molecular Biology·Tariq A AkhtarEran Pichersky
Jan 26, 2010·Medicinal Research Reviews·Francis E UmesiriSteven J Sucheck
Jun 4, 2013·Biochemical and Biophysical Research Communications·Takeshi MoriHisashi Hemmi
Apr 2, 2020·Annual Review of Biochemistry·Satoshi Yamashita, Seiji Takahashi
Jul 13, 2016·The Journal of Biological Chemistry·Kariona A GrabińskaWilliam C Sessa
Jun 25, 2015·Microbiology Spectrum·Mamadou DafféMary Jackson

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