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首页> 外文期刊>Applied Microbiology and Biotechnology >Function of the prosequence for in vivo folding and secretion of active Rhizopus oryzae lipase in Saccharomyces cerevisiae
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Function of the prosequence for in vivo folding and secretion of active Rhizopus oryzae lipase in Saccharomyces cerevisiae

机译：前序对于酿酒酵母体内折叠和分泌活性米根霉脂肪酶的功能

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摘要

The role of the prosequence of Rhizopus oryzae lipase (ROL) with a preprosequence was analyzed by an expression system using Saccharomyces cerevisiae. When the mature portion of ROL (mROL) fused to the pre-alpha -factor leader sequence was expressed, secretion of active mROL was not observed. However, when mROL was synthesized together with the prosequence in trans (individually and coincidentally), secretion of active mROL was observed. The results indicate that the prosequence of ROL helped correct folding of mROL and its subsequent secretion from the yeast cells, and that physical linkage (cis) of the prosequence to the mature region was not prerequisite. From the expression of the ROL mutants with deletions at the N-terminal end of the prosequence together with mROL in trans, the residues from 20 to 37 in the prosequence were essential for the secretion, and those from 38 to 57 were essential for the formation of the active ROL and might play a role as an intramolecular chaperone. The results using the fragment of the prosequence confirmed that these residues (20-57) were significant for in vivo folding and secretion of active mROL. [References: 42]
机译：通过使用酿酒酵母的表达系统分析了米根霉脂肪酶（ROL）与前序列的关系。当表达与前α因子前导序列融合的ROL的成熟部分（mROL）时，未观察到活性mROL的分泌。然而，当mROL与反式序列一起合成时（个别地和同时地），观察到活性mROL的分泌。结果表明，ROL的序列有助于纠正mROL的折叠及其随后从酵母细胞中的分泌，并且序列与成熟区域的物理连接（顺式）不是必要条件。从前序列N末端缺失的ROL突变体的表达以及反式mROL的表达来看，前序列中20到37的残基对于分泌是必不可少的，而38到57的残基对于形成是必不可少的ROL的活性，并可能充当分子内伴侣。使用序列片段的结果证实，这些残基（20-57）对于活体mROL的体内折叠和分泌是重要的。 [参考：42]

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