| 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 |
2008-11-1
|
| 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 |
3.53
|
| Times Cited |
15
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| Resource |
| Arabidopsis / Cultured plant cells, genes |
pda08860 |
