RRC ID 4844
Author Park M, Kang K, Park S, Kim YS, Ha SH, Lee SW, Ahn MJ, Bae JM, Back K.
Title Expression of serotonin derivative synthetic genes on a single self-processing polypeptide and the production of serotonin derivatives in microbes.
Journal Appl. Microbiol. Biotechnol.
Abstract The plant-specific serotonin derivatives feruloylserotonin (FS) and 4-coumaroylserotonin (CS) are synthesized by the enzymes 4-coumarate:coenzyme A ligase (4CL) and serotonin N-hydroxycinnamoyltransferase (SHT). To express these genes coordinately, SHT was fused in-frame with the self-processing FDMV 2A sequence followed by 4CL in a single open reading frame and introduced into Escherichia coli or Saccharomyces cerevisiae. The transgenes were abundantly expressed in both recombinant microbes, but functional expression was achieved only in yeast, with cleavage at the 2A sequence yielding monomeric SHT-2A and 4CL as judged by immunoblot and product analyses. In the presence of an exogenous supply of precursors such as serotonin and ferulic acid, the recombinant yeast synthesized 4.5 mg l(-1) FS in the medium while 0.02 mg l(-1) FS was produced in the cells. Time-course analysis indicated peak accumulation of FS at 24 h after induction, and this level was maintained until 96 h. The optimum precursor concentration was 2 mM. A series of serotonin derivatives was produced by adding various cinnamate derivative precursors with serotonin; 2.5 mg l(-1) caffeoylserotonin (CaS) and 1.4 mg l(-1) CS were produced, whereas no sinapoylserotonin or cinnamoylserotonin was yielded.
Volume 81(1)
Pages 43-9
Published 2008-11
DOI 10.1007/s00253-008-1634-8
PMID 18709368
MeSH Biomass Capsicum / enzymology Coenzyme A Ligases / genetics* Coenzyme A Ligases / metabolism Escherichia coli / genetics Escherichia coli / metabolism* Gene Expression* Genetic Engineering Plant Proteins / genetics* Plant Proteins / metabolism Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / metabolism* Serotonin / analogs & derivatives Serotonin / chemistry Serotonin / metabolism* Transferases / genetics* Transferases / metabolism
IF 3.67
Times Cited 12
Arabidopsis / Cultured plant cells, genes pda08860