Withania coagulans tryptophan decarboxylase gene cloning, heterologous expression, and catalytic characteristics of the recombinant enzyme.
Tryptophan decarboxylase (EC 4.1.1.28) catalyzes pyridoxal 5′-phosphate (PLP)-dependent decarboxylation of tryptophan to produce tryptamine for recruitment in a myriad of biosynthetic pathways of metabolites possessing indolyl moiety. A recent report of certain indolyl metabolites in Withania specie...
| Publicado en: | Protoplasma Vol. 254; no. 1; pp. 181 - 193 |
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| Autores principales: | , , , , |
| Formato: | Journal Article |
| Publicado: |
Springer Nature
Jan2017
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=120569779&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 120569779 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 0033183X 2CF jtl: Protoplasma issn: 0033183X maglogo: N pubinfo: dt: Jan2017 vid: 254 iid: 1 pid: 237 pub: Springer Nature place: New York, New York artinfo: ui: 120569779 10.1007/s00709-015-0929-8 120569779 ppf: 181 ppct: 12 formats: fmt: @attributes: type: P tig: atl: Withania coagulans tryptophan decarboxylase gene cloning, heterologous expression, and catalytic characteristics of the recombinant enzyme. aug: au: Jadaun, Jyoti Sangwan, Neelam Narnoliya, Lokesh Tripathi, Sandhya Sangwan, Rajender affil: CSIR-Central Institute of Medicinal and Aromatic Plants , Lucknow 226015 India sug: ab: Tryptophan decarboxylase (EC 4.1.1.28) catalyzes pyridoxal 5′-phosphate (PLP)-dependent decarboxylation of tryptophan to produce tryptamine for recruitment in a myriad of biosynthetic pathways of metabolites possessing indolyl moiety. A recent report of certain indolyl metabolites in Withania species calls for a possible predominant functional role of tryptophan decarboxylase (TDC) in the genome of Withania species to facilitate production of the indolyl progenitor molecule, tryptamine. Therefore, with this metabolic prospection, we have identified and cloned a full-length cDNA sequence of TDC from aerial tissues of Withania coagulans. The functional WcTDC gene comprises of 1506 bp open reading frame (ORF) encoding a 502 amino acid protein with calculated molecular mass and pI value of 56.38 kDa and 8.35, respectively. The gene was expressed in Escherichia coli, and the recombinant enzyme was affinity-purified to homogeneity to discern its kinetics of catalysis. The enzyme (WcTDC) exhibited much higher K value for tryptophan than for pyridoxal 5′-phosphate and was dedicated to catalyze decarboxylation of only tryptophan or, to a limited extent, of its analogue (like 5-hydroxy tryptophan). The observed optimal catalytic functionality of the enzyme on the slightly basic side of the pH scale and at slightly higher temperatures reflected adaptability of the plant to hot and arid regions, the predominant natural habitat of the herb. This pertains to be the first report on cloning and characterization of heterologously expressed recombinant enzyme from W. coagulans and forms a starting point to further understanding of withanamide biosynthesis. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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