Jump to main content
Jump to site search

Issue 13, 2010
Previous Article Next Article

Simultaneous C7- and N1-prenylation of cyclo-L-Trp-L-Trp catalyzed by a prenyltransferase from Aspergillus oryzae

Author affiliations

Abstract

A putative prenyltransferase gene cTrpPT was amplified from Aspergillus oryzae DSM1147, cloned into pQE70 and overexpressed in Escherichia coli. The overproduced His6-CTrpPT was purified to near homogeneity and incubated with L-tryptophan or tryptophan-containing cyclic dipeptides in the presence of dimethylallyl diphosphate. The formation of the enzyme products was monitored with HPLC. It was shown that CTrpPT differed clearly from other known indole prenyltransferases in several aspects. This enzyme showed higher substrate specificity towards aromatic substrates, but lower regioselectivity regarding the prenylation position than other indole prenyltransferases. Cyclo-L-Trp-L-Trp was much better accepted than other cyclic dipeptides tested in this study. In comparison to other indole prenyltransferases with one dominant enzyme product, at least two product peaks were detected in the reaction mixtures of CTrpPT. 1H- and 13C-NMR analyses, including long-range 1H–13C connectivities in Heteronuclear Multiple-Bond Correlation (HMBC) and Nuclear Overhauser Effect Spectroscopy (NOESY), proved the structures of the enzyme products as C7- and N1-prenylated derivatives with a ratio of 1 : 1.2 using cyclo-L-Trp-L-Trp as substrate. The KM values were determined at about 2.5 mM for dimethylallyl diphosphate and 0.3 mM for cyclo-L-Trp-L-Trp with a turnover number of 0.33 s−1.

Graphical abstract: Simultaneous C7- and N1-prenylation of cyclo-l-Trp-l-Trp catalyzed by a prenyltransferase from Aspergillus oryzae

Back to tab navigation
Please wait while Download options loads

Supplementary files

Publication details

The article was received on 12 Feb 2010, accepted on 16 Apr 2010 and first published on 14 May 2010


Article type: Paper
DOI: 10.1039/C002850A
Citation: Org. Biomol. Chem., 2010,8, 3037-3044
  •   Request permissions

    Simultaneous C7- and N1-prenylation of cyclo-L-Trp-L-Trp catalyzed by a prenyltransferase from Aspergillus oryzae

    H. Zou, X. Xie, U. Linne, X. Zheng and S. Li, Org. Biomol. Chem., 2010, 8, 3037
    DOI: 10.1039/C002850A

Search articles by author