title: Functional Analysis of Two l-Arabinose Transporters from Filamentous Fungi Reveals Promising Characteristics for Improved Pentose Utilization in Saccharomyces cerevisiae.
abstract: Limited uptake is one of the bottlenecks for l-arabinose fermentation from lignocellulosic hydrolysates in engineered Saccharomyces cerevisiae. This study characterized two novel l-arabinose transporters, LAT-1 from Neurospora crassa and MtLAT-1 from Myceliophthora thermophila. Although the two proteins share high identity (about 83%), they display different substrate specificities. Sugar transport assays using the S. cerevisiae strain EBY.VW4000 indicated that LAT-1 accepts a broad substrate spectrum. In contrast, MtLAT-1 appeared much more specific for l-arabinose. Determination of the kinetic properties of both transporters revealed that the Km values of LAT-1 and MtLAT-1 for l-arabinose were 58.12 ± 4.06 mM and 29.39 ± 3.60 mM, respectively, with corresponding Vmax values of 116.7 ± 3.0 mmol/h/g dry cell weight (DCW) and 10.29 ± 0.35 mmol/h/g DCW, respectively. In addition, both transporters were found to use a proton-coupled symport mechanism and showed only partial inhibition by d-glucose during l-arabinose uptake. Moreover, LAT-1 and MtLAT-1 were expressed in the S. cerevisiae strain BSW2AP containing an l-arabinose metabolic pathway. Both recombinant strains exhibited much faster l-arabinose utilization, greater biomass accumulation, and higher ethanol production than the control strain. In conclusion, because of higher maximum velocities and reduced inhibition by d-glucose, the genes for the two characterized transporters are promising targets for improved l-arabinose utilization and fermentation in S. cerevisiae.
authors.affiliation: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
authors.forename: Jingen
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China College of Life Sciences, Hubei University, Wuhan, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.affiliation: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
authors.forename: Pengli
authors.initials: P
authors.lastname: Cai
authors.affiliation: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
authors.forename: Bang
authors.initials: B
authors.lastname: Wang
authors.affiliation: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China.
authors.forename: Yanhe
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Holzforschung München, TUM School of Life Sciences Weihenstephan, Technische Universität München, Freising, Germany benz@hfm.tum.de tian_cg@tib.cas.cn.
authors.forename: J Philipp
authors.initials: JP
authors.lastname: Benz
authors.affiliation: Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China benz@hfm.tum.de tian_cg@tib.cas.cn.
authors.forename: Chaoguang
authors.initials: C
authors.lastname: Tian
mesh_terms: D001089:Arabinose / Q000378:metabolism*; D001692:Biological Transport; D018533:Biomass; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation / Q000235:genetics; D005818:Genetic Engineering; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D053858:Metabolic Networks and Pathways; D009004:Monosaccharide Transport Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D009492:Neurospora crassa / Q000235:genetics*; D010429:Pentoses / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D020037:Sordariales / Q000235:genetics* / Q000378:metabolism; D014994:Xylose / Q000378:metabolism
chemical_list: D009004:Monosaccharide Transport Proteins; D010429:Pentoses; D000431:Ethanol; D014994:Xylose; D001089:Arabinose; D005947:Glucose
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