米根霉脂肪酶前导序列的克隆、表达、特性及作用
Cloning, expression, characterization and role of the leader sequence of a lipase from Rhizopus oryzae.
作者信息
Beer H D, McCarthy J E, Bornscheuer U T, Schmid R D
机构信息
AG Werner, MPI-Biochemie, Martinsried, Germany.
出版信息
Biochim Biophys Acta. 1998 Aug 20;1399(2-3):173-80. doi: 10.1016/s0167-4781(98)00104-3.
A lipase from Rhizopus oryzae DSM 853 (ROL) was cloned from a chromosomal gene bank, sequenced and overexpressed in Escherichia coli. ROL and its precursors ProROL and PreProROL were purified and their pH and temperature profile was determined. In contrast to ROL, ProROL and PreProROL had considerably higher thermostability and a slightly higher pH optimum. Moreover, it could be demonstrated by in vitro experiments that the natural leader sequence of ROL is able to inhibit the folding supporting properties of the prosequence, resulting in a retardation of folding. In addition, there is strong evidence that all different lipase forms derived from Rhizopus sp. described in the literature are a result of different proteolytic processing and originate from the same gene.
从米根霉DSM 853中克隆出一种脂肪酶（ROL），该酶来自染色体基因文库，进行了测序并在大肠杆菌中过表达。对ROL及其前体ProROL和PreProROL进行了纯化，并测定了它们的pH和温度曲线。与ROL相比，ProROL和PreProROL具有更高的热稳定性和略高的最适pH值。此外，体外实验表明，ROL的天然前导序列能够抑制前序列的折叠支持特性，导致折叠延迟。此外，有强有力的证据表明，文献中描述的源自根霉属的所有不同脂肪酶形式都是不同蛋白水解加工的结果，并且起源于同一个基因。
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