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    题名: Subunit fusion of two yeast D-amino acid oxidases enhances their thermostability and resistance to H2O2
    作者: Wang, Shih-juei;Yu, Chi-yang;Lee, Cheng-kang;Chern, Ming-kai;Kuan, I-ching
    贡献者: 淡江大學生命科學研究所
    关键词: D-Amino acid oxidase;Gene fusion;Rhodosporidium toruloides;Stability;Trigonopsis variabilis
    日期: 2008-08
    上传时间: 2011-10-17 01:01:05 (UTC+8)
    出版者: Dordrecht: Springer Netherlands
    摘要: D-Amino acid oxidases from Rhodosporidium toruloides and Trigonopsis variabilis (RtDAO and TvDAO) are both yeast homodimeric flavoenzymes. Two of their cDNA genes were connected by a hexanucleotide linker and heterologously expressed in E. coli to produce the corresponding double DAOs (dRtDAO and dTvDAO) with two subunits fused into a single polypeptide. The specific activities of double DAOs remained similar to those of native dimeric DAOs, although the catalytic efficiencies (kcat/KM) were decreased due to higher KM values. The Tm value for dRtDAO was shifted 5°C higher while that for dTvDAO was increased only by 2°C, in comparison with the corresponding native counterparts. In the presence of 10 mM H2O2, dRtDAO and dTvDAO exhibited half-lives of about 60 and 40 min, respectively, which were 2- and 1.5-fold, respectively, longer than their native DAOs. These yeast DAOs can therefore be thermally and oxidatively stabilized by linking their subunits together.
    關聯: Biotechnology Letters 30(8), pp.1415-1422
    DOI: 10.1007/s10529-008-9694-5
    显示于类别:[生命科學研究所] 期刊論文

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