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2017 Fiscal Year Final Research Report

Enzymological characterization of novel L-amino acid dehydrogenase for development of high specific agrochemicals

Research Project

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Project/Area Number 16K18689
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Applied biochemistry
Research InstitutionTokai University

Principal Investigator

YONEDA Kazunari  東海大学, 農学部, 准教授 (00469397)

Project Period (FY) 2016-04-01 – 2018-03-31
KeywordsL-スレオニン酸脱水素酵素 / 酵素化学 / 結晶構造解析 / 植物疫病菌
Outline of Final Research Achievements

A gene encoding an L-Threonine dehydrogenase (ThrDH; PITG_05140) homologue was identified in the Phytophthora infestans. After transforming E. coli with pET-PITG_05140, an expression vector harboring the gene, the transformant cells exhibited a high level of L-ThrDH activity, and the enzyme was readily purified from the cells’ crude extract in one simple step: Talon metal affinity chromatography. The purified enzyme showed a single protein band on SDS-PAGE, and its N-terminal sequence was determined to be SDKIIHLTDD. SDS-PAGE showed the subunit molecular mass of P. infestans ThrDH to be about 55 kDa, which is consistent with the molecular weight calculated from the amino acid sequence. Non-tagged P. infestans ThrDH acted on L-threonine and DL-3-hydroxynorvaline, and the enzyme was partially inhibited by L-cysteine and N-acetylglycine.

Free Research Field

応用生物化学

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Published: 2019-03-29  

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