TY - JOUR
T1 - Identification and Analysis of Sugar Transporters Capable of Co-transporting Glucose and Xylose Simultaneously
AU - Kuanyshev, Nurzhan
AU - Deewan, Anshu
AU - Jagtap, Sujit Sadashiv
AU - Liu, Jingjing
AU - Selvam, Balaji
AU - Chen, Li-Qing
AU - Shukla, Diwakar
AU - Rao, Christopher V
AU - Jin, Yong-Su
N1 - Funding Information:
This material is based on the work supported by the US Department of Energy, Office of Science, Office of Biological and Environmental Research under Award Number(s) DE‐SC0018420. The authors thank Blue Waters Supercomputing Facility funded by National Science Foundation (OCI‐0725070 and ACI‐1238993) and the state of Illinois for the computer time. D.S acknowledges New Innovator Award from the Foundation for Food and Agricultural Research (FFAR) and NSF Early CAREER Award, NSF MCB 18–45606 for the research support.
Publisher Copyright:
© 2021 Wiley-VCH GmbH
PY - 2021/11
Y1 - 2021/11
N2 - Simultaneous co-fermentation of glucose and xylose is a key desired trait of engineered Saccharomyces cerevisiae for efficient and rapid production of biofuels and chemicals. However, glucose strongly inhibits xylose transport by endogenous hexose transporters of S. cerevisiae. We identified structurally distant sugar transporters (Lipomyces starkeyi LST1_205437 and Arabidopsis thaliana AtSWEET7) capable of co-transporting glucose and xylose from previously unexplored oleaginous yeasts and plants. Kinetic analysis showed that LST1_205437 had lenient glucose inhibition on xylose transport and AtSWEET7 transported glucose and xylose simultaneously with no inhibition. Modelling studies of LST1_205437 revealed that Ala335 residue at sugar binding site can accommodates both glucose and xylose. Docking studies with AtSWEET7 revealed that Trp59, Trp183, Asn145, and Asn179 residues stabilized the interactions with sugars, allowing both xylose and glucose to be co-transported. In addition, we altered sugar preference of LST1_205437 by single amino acid mutation at Asn365. Our findings provide a new mechanistic insight on glucose and xylose transport mechanism of sugar transporters and the identified sugar transporters can be employed to develop engineered yeast strains for producing cellulosic biofuels and chemicals.
AB - Simultaneous co-fermentation of glucose and xylose is a key desired trait of engineered Saccharomyces cerevisiae for efficient and rapid production of biofuels and chemicals. However, glucose strongly inhibits xylose transport by endogenous hexose transporters of S. cerevisiae. We identified structurally distant sugar transporters (Lipomyces starkeyi LST1_205437 and Arabidopsis thaliana AtSWEET7) capable of co-transporting glucose and xylose from previously unexplored oleaginous yeasts and plants. Kinetic analysis showed that LST1_205437 had lenient glucose inhibition on xylose transport and AtSWEET7 transported glucose and xylose simultaneously with no inhibition. Modelling studies of LST1_205437 revealed that Ala335 residue at sugar binding site can accommodates both glucose and xylose. Docking studies with AtSWEET7 revealed that Trp59, Trp183, Asn145, and Asn179 residues stabilized the interactions with sugars, allowing both xylose and glucose to be co-transported. In addition, we altered sugar preference of LST1_205437 by single amino acid mutation at Asn365. Our findings provide a new mechanistic insight on glucose and xylose transport mechanism of sugar transporters and the identified sugar transporters can be employed to develop engineered yeast strains for producing cellulosic biofuels and chemicals.
KW - co-fermentation
KW - rational evolution
KW - substrate specificity
KW - sugar membrane transporter
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U2 - 10.1002/biot.202100238
DO - 10.1002/biot.202100238
M3 - Article
C2 - 34418308
SN - 1860-6768
VL - 16
SP - e2100238
JO - Biotechnology Journal
JF - Biotechnology Journal
IS - 11
M1 - 2100238
ER -