Effects of pre- and pro-sequence of thaumatin on the secretion by Pichia pastoris
Effects of pre- and pro-sequence of thaumatin on the secretion by Pichia pastoris
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DOI：
10.1016/j.bbrc.2007.09.021
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发表时间：
2007-11-23
影响因子：
3.1
通讯作者：
Kitabatake, Naofumi
中科院分区：
文献类型：
--
作者：
Ide, Nobuyuki;Masuda, Tetsuya;Kitabatake, Naofumi
Thaumatin是一种含有8个二硫键的22千瓦时甜味蛋白质。当使用融合了α因子信号序列和酿酒酵母Kex2蛋白酶裂解位点的thaumatin cDNA在毕氏酵母中表达thaumatin时，所分泌的thaumatin分子的n端序列加工不正确。为了研究微波抑制蛋白cdna编码的n端前序列和c端前序列对微波抑制蛋白加工和微波抑制蛋白生产效率的作用，构建了4个具有不同前序列和前序列的表达质粒，并将其转化到巴氏酵母中。含有具有原生植物信号的前突变基因的转化子在培养基中分泌突变分子。对所分泌的thumatin分子的n端氨基酸序列进行了正确处理。该前序列不受c端前序列的影响，且该前序列在P. pastoris中未被加工，说明前序列对合成thaumatin并不是必需的。(C) 2007爱思唯尔公司版权所有。
Thaumatin is a 22-kDa sweet-tasting protein containing eight disulfide bonds. When thaumatin is expressed in Pichia pastoris using the thaumatin cDNA fused with both the alpha-factor signal sequence and the Kex2 protease cleavage site from Saccharomyces cerevisiae, the N-terminal sequence of the secreted thaumatin molecule is not processed correctly. To examine the role of the thaumatin cDNA-encoded N-terminal pre-sequence and C-terminal pro-sequence on the processing of thaumatin and efficiency of thaumatin production in P. pastoris, four expression plasmids with different pre-sequence and pro-sequence were constructed and transformed into P. pastoris. The transformants containing pre-thaumatin gene that has the native plant signal, secreted thaumatin molecules in the medium. The N-terminal amino acid sequence of the secreted thaumatin molecule was processed correctly. The production yield of thaumatin was not affected by the C-terminal pro-sequence, and the pro-sequence was not processed in P. pastoris, indicating that pro-sequence is not necessary for thaumatin synthesis. (C) 2007 Elsevier Inc. All rights reserved.