新型重组氰酸盐水解酶(rTl-Cyn)在毕赤酵母中的表达、特性及其在氰酸解毒中的应用。
Expression of a novel recombinant cyanate hydratase (rTl-Cyn) in Pichia pastoris, characteristics and applicability in the detoxification of cyanate.
机构信息
Department of Biotechnology and Food Technology, Faculty of Applied Sciences, Durban University of Technology, Durban 4000, South Africa.
Department of Biotechnology and Food Technology, Faculty of Applied Sciences, Durban University of Technology, Durban 4000, South Africa.
出版信息
Bioresour Technol. 2017 Aug;238:582-588. doi: 10.1016/j.biortech.2017.04.091. Epub 2017 Apr 27.
A recombinant Pichia pastoris harbouring the cyanate hydratase gene (rTl-Cyn) from the thermophilic fungus Thermomyces lanuginosus SSBP yielded a high titre of extracellular cyanate hydratase (100±13UmL) which was ∼10-fold higher than the native fungal strain. The purified rTl-Cyn had a molecular mass of ∼20kDa on SDS-PAGE, with K, V, k and k/K values of 0.34mM, 2857.14µmolesmgmin, 2.14×10s and 6.3 ×10Ms, respectively. Its properties of thermostability, pH stability, and heavy metals insensitivity, make it a suitable candidate for bioremediation in extreme environments. The rTl-Cyn was able to degrade toxic cyanate completely with the liberation of ammonia, which was confirmed by FTIR analysis. This is the first report of any known cyanate hydratase that has been expressed in P. pastoris, characterized and effectively evaluated for cyanate detoxification.
一株携带嗜热真菌Thermomyces lanuginosus SSBP 氰酸盐水解酶基因（rTl-Cyn）的重组毕赤酵母产生了高浓度的细胞外氰酸盐水解酶（100±13UmL），比天然真菌菌株高约 10 倍。纯化的 rTl-Cyn 在 SDS-PAGE 上的分子量约为 20kDa，K、V、k 和 k/K 值分别为 0.34mM、2857.14µmolesmgmin、2.14×10s 和 6.3×10Ms。其热稳定性、pH 稳定性和重金属不敏感性等特性使其成为极端环境生物修复的合适候选物。rTl-Cyn 能够完全降解有毒的氰酸盐并释放出氨，这一点通过傅里叶变换红外（FTIR）分析得到了证实。这是首次在毕赤酵母中表达、表征并有效评估氰酸盐解毒功能的已知氰酸盐水解酶。
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