Characterization of a New (R)-Hydroxynitrile Lyase from the Japanese Apricot Prunus mume and cDNA Cloning and Secretory Expression of One of the Isozymes in Pichia pastoris

• FUKUTA Yasuhisa

  Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University

• NANDA Samik

  Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University

• KATO Yasuo

  Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University

• YURIMOTO Hiroya

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• SAKAI Yasuyoshi

  Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University

• KOMEDA Hidenobu

  Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University

• ASANO Yasuhisa

  Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University

Bibliographic Information

Other Title

• Characterization of a New (<i>R</i>)-Hydroxynitrile Lyase from the Japanese Apricot<i>Prunus mume</i>and cDNA Cloning and Secretory Expression of One of the Isozymes in<i>Pichia pastoris</i>
• Characterization of a new (R)-hydroxynitrile lyase from the Japanese apricot Prunus mume and cDNA cloning and secretory expression of one of the isozymes in Pichia pastoris, Biosci

Abstract

PmHNL, a hydroxynitrile lyase from Japanese apricot ume (Prunus mume) seed was purified to homogeneity by ammonium sulfate fractionation and chromatographic steps. The purified enzyme was a monomer with molecular mass of 58 kDa. It was a flavoprotein similar to other hydroxynitrile lyases of the Rosaceae family. It was active over a broad temperature, and pH range. The N-terminal amino acid sequence (20 amino acids) was identical with that of the enzyme from almond (Prunus dulcis). Based on the N-terminal sequence of the purified enzyme and the conserved amino acid sequences of the enzymes from Pr. dulcis, inverse PCR method was used for cloning of a putative PmHNL (PmHNL2) gene from a Pr. mume seedling. Then the cDNA for the enzyme was cloned. The deduced amino acid sequence was found to be highly similar (95%) to that of an enzyme from Pr. serotina, isozyme 2. The recombinant Pichia pastoris transformed with the PmHNL2 gene secreted an active enzyme in glycosylated form.

Journal

• Bioscience, Biotechnology, and Biochemistry

  Bioscience, Biotechnology, and Biochemistry 75 (2), 214-220, 2011

  Japan Society for Bioscience, Biotechnology, and Agrochemistry

Keywords

• (<I>R</I>)-hydroxynitrile lyase
• <I>Prunus mume</I>
• mandelonitrile
• cyanogenic plant
• <I>Pichia pastoris</I>

Details

• CRID: 1390282681455161088
• NII Article ID: 10027897395
• NII Book ID: AA10824164
• DOI: 10.1271/bbb.100187
• ISSN: 13476947
• ISSN: 09168451
• NDL BIB ID: 10993100
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I10993100
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.100187
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed