Cloning and characterization of heterologous transporters in Saccharomyces cerevisiae and identification of important amino acids for xylose utilization
Cloning and characterization of heterologous transporters in Saccharomyces cerevisiae and identification of important amino acids for xylose utilization
复制标题
酿酒酵母异源转运蛋白的克隆和表征以及木糖利用重要氨基酸的鉴定。
DOI：
10.1016/j.ymben.2015.04.007
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发表时间：
2015-07-01
影响因子：
8.4
通讯作者：
Shen, Yu
中科院分区：
文献类型：
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作者：
Wang, Chengqiang;Bao, Xiaoming;Shen, Yu
高效和特异的转运蛋白可以增强酿酒酵母对戊糖的吸收和代谢。在S.啤酒。其中季也蒙Meyerozyrna guilliermondii的转运蛋白Mgt 05196 p的木糖转运活性最高。Mgt 05196 p的几个关键氨基酸残基的结构和序列分析，并通过定点突变的特点。在第七跨膜跨度中的保守的富含芳香残基的基序（YFFYY，位置332-336）在D-木糖转运活性中起重要作用。在位置336处的残基的苯环可以起到防止D-木糖在摄取期间逃逸的作用。F432 A和N360 S突变增强了Mgt 05196 p的D-木糖转运活性。此外，突变株N360 F特异性地转运D-木糖，而没有任何葡萄糖抑制，这突出了其在构建用于生物质精炼的葡萄糖-木糖共发酵菌株中的潜在应用。(C)2015年国际代谢工程学会。爱思唯尔公司出版All rights reserved.
Efficient and specific transporters may enhance pentose uptake and metabolism by Saccharomyces cerevisiae. Eight heterologous sugar transporters were characterized in S. cerevisiae. The transporter Mgt05196p from Meyerozyrna guilliermondii showed the highest xylose transport activity among them. Several key amino acid residues of Mgt05196p were suggested by structural and sequence analysis and characterized by site-directed mutagenesis. A conserved aromatic residue-rich motif (YFFYY, position 332-336) in the seventh trans-membrane span plays an important role in D-xylose transport activity. The phenyl ring of the residue at position 336 may take the function to prevent D-xylose from escaping during uptake. F432A and N360S mutations enhanced the D-xylose transport activities of Mgt05196p. Furthermore, mutant N360F specifically transported D-xylose without any glucose-inhibition, high lighting its potential application in constructing glucose-xylose co-fermentation strains for biomass refining. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.