Characterization of chimeric enzymes constructed between two distinct α-Amylase cDNAs from cultured rice cells

Rei Abe, Kensuke Yoshida, Masanobu Aoyagi, Shin Kasahara, Tasuku Nakajima

    Research output: Contribution to journalArticlepeer-review

    3 Citations (Scopus)

    Abstract

    Cultured cells of rice (Oryza sativa cv Sasanishiki) produce two α-amylase isozymes, AMY-I and AMY-III. Using a bacterial expression system, eight chimeric genes constructed with various combination of AMY-I and AMY-III cDNA fragments were expressed, and each recombinant chimeric protein was characterized. Four of the eight recombinant enzymes having region c (one of the four regions having unconserved base sequences between AMY-I and AMY-III cDNAs) of AMY-I showed the same enzyme characteristics as that of native AMY-I, which had high temperature optimum at 50°C. The other four chimeric proteins carrying region c of AMY-III showed the AMY-III type characteristics, which were a low temperature optimum at 25°C and susceptibility to a higher maltooligosaccharide (G17) substrate. The unconserved region c is involved in the decision of the characteristic of AMY-I or AMY-III.

    Original languageEnglish
    Pages (from-to)1329-1335
    Number of pages7
    JournalBioscience, Biotechnology and Biochemistry
    Volume63
    Issue number8
    DOIs
    Publication statusPublished - 1999 Jan 1

    Keywords

    • Chimeric protein
    • Gene expression
    • Suspension-cultured rice cells
    • α-amylase

    ASJC Scopus subject areas

    • Biotechnology
    • Analytical Chemistry
    • Biochemistry
    • Applied Microbiology and Biotechnology
    • Molecular Biology
    • Organic Chemistry

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