非特异性过氧合酶GmaUPO的重组表达及生化表征  

Recombinant Expression and Biochemical Characterization of Unspecific Peroxygenase GmaUPO

作　　者：蓝东明[1] 乜晓爽 马云建 王永华[1] LAN Dongming;NIE Xiaoshuang;MA Yunjian;WANG Yonghua(School of Food Science and Engineering,South China University of Technology,Guangzhou 510640,Guangdong,China)

机构地区：[1]华南理工大学食品科学与工程学院,广东广州510640

出　　处：《华南理工大学学报（自然科学版）》2021年第9期22-28,共7页Journal of South China University of Technology(Natural Science Edition)

基　　金：国家自然科学基金资助项目(31930084,31725022,32001633)。

摘　　要：非特异性过氧合酶(Unspecific Peroxygenase,UPO)因在催化有机底物时具有独特的立体选择性和对映选择性而在工业生产中具有广阔的应用前景。本研究从纹缘盔孢伞(Galerina marginata)的基因组中发现了一个假定的非特异性过氧合酶基因(以下简称GmaUPO),通过全基因合成技术获得了GmaUPO的基因序列,并构建了GmaUPO诱导型表达的毕赤酵母X-33基因工程菌。在最佳的发酵条件下,发酵液酶活力达到6 U/L,重组GmaUPO分子质量约为40 ku,其最适反应条件为35℃、pH 9.0。重组GmaUPO可在温和条件下利用H_(2)O_(2)高效催化芳环类物质中的惰性C—H键羟化,合成芳环类含氧化合物,在香精香料等领域具有巨大的应用潜力。Unspecific peroxygenase(UPO)has broad application prospects due to its unique stereoselectivity and enantioselectivity in catalyzing organic substrate.In this study,a putative unspecific peroxygenase gene(GmaUPO)was found from the genome of Galerina marginata,and the corresponding gene sequence was obtained by means of the whole gene synthesis technology.Moreover,Pichia pastoris X-33 gene engineering bacteria of GmaUPO inducible expression was constructed.Under the optimal fermentation condition,the enzyme activity of the fermentation broth can reach 6 U/L,the relative molecular mass of GmaUPO is about 40 ku,the optimal reaction temperature is 35℃and the optimal pH is 9.0.The recombinant GmaUPO can synthesize aromatic oxygen-containing compounds by using H_(2)O_(2) to catalyze the hydroxylation of the inert C—H bond in aromatic substances under mild condition,which means that it possesses great application potential in the fields of flavors and fragrances.

关 键 词：非特异性过氧合酶 纹缘盔孢伞 克隆 表达 毕赤酵母 

分 类 号：Q786[生物学—分子生物学] Q814