title: Cloning and characterization of heterologous transporters in Saccharomyces cerevisiae and identification of important amino acids for xylose utilization.
abstract: 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 Meyerozyma 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.
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Chengqiang
authors.initials: C
authors.lastname: Wang
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Bao
authors.affiliation: Environment Research Institute, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Li
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Chunlei
authors.initials: C
authors.lastname: Jiao
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Jin
authors.initials: J
authors.lastname: Hou
authors.affiliation: Environment Research Institute, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Qingzhu
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Weixin
authors.initials: W
authors.lastname: Zhang
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Liu
authors.affiliation: The State Key Laboratory of Microbial Technology, Shandong University, Shan Da Nan Road 27, Jinan, 250100, PR China. Electronic address: shenyu@sdu.edu.cn.
authors.forename: Yu
authors.initials: Y
authors.lastname: Shen
mesh_terms: D019943:Amino Acid Substitution; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis / Q000235:genetics; D020125:Mutation, Missense; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014994:Xylose / Q000235:genetics / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D009004:Monosaccharide Transport Proteins; D014994:Xylose
keywords: Budding yeast; Heterologous; Pentose; Protein engineering; Transporter; d-xylose