Oda K, Atomi H, Ueda M, Kondo J, Teranishi Y, Tanaka A
Department of Industrial Chemistry, Faculty of Engineering, Kyoto University, Japan.
Arch Microbiol. 1991;156(6):439-43. doi: 10.1007/BF00245389.
The genomic DNA of peroxisomal isocitrate lyase (ICL) isolated from an n-alkane-assimilating yeast, Candida tropicalis, was truncated to utilize the original open reading frame under the control of the GAL7 promoter and was expressed in Saccharomyces cerevisiae. The recombinant ICL was synthesized as a functionally active enzyme with a specific activity similar to the enzyme purified from C. tropicalis, and was accounted for approximately 30% of the total extractable proteins in the yeast cells. This recombinant enzyme was easily purified to homogeneity. N-Terminal amino acid sequence, molecular masses of native form and subunit, amino acid composition, peptide maps, and kinetic parameters of the recombinant ICL were essentially the same as those of ICL purified from C. tropicalis. From these facts, S. cerevisiae was suggested to be an excellent micro-organism to highly express the genes encoding peroxisomal proteins of C. tropicalis.
从能同化正构烷烃的热带假丝酵母中分离出的过氧化物酶体异柠檬酸裂解酶（ICL）的基因组DNA，在GAL7启动子的控制下被截短以利用原始开放阅读框，并在酿酒酵母中表达。重组ICL作为一种功能活性酶被合成，其比活性与从热带假丝酵母中纯化的酶相似，并且占酵母细胞中总可提取蛋白的约30%。这种重组酶很容易纯化至均一。重组ICL的N端氨基酸序列、天然形式和亚基的分子量、氨基酸组成、肽图以及动力学参数与从热带假丝酵母中纯化的ICL基本相同。基于这些事实，酿酒酵母被认为是高效表达热带假丝酵母过氧化物酶体蛋白编码基因的优良微生物。
