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Title: Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on l ‐arabinose and d ‐xylose

Abstract

Abstract Genes encoding l ‐arabinose transporters in Kluyveromyces marxianus and Pichia guilliermondii were identified by functional complementation of Saccharomyces cerevisiae whose growth on l ‐arabinose was dependent on a functioning l ‐arabinose transporter, or by screening a differential display library, respectively. These transporters also transport d ‐xylose and were designated KmAXT1 (arabinose–xylose transporter) and PgAXT1 , respectively. Transport assays using l ‐arabinose showed that KmAxt1p has K m 263 m m and V max 57 n m /mg/min, and PgAxt1p has K m 0.13 m m and V max 18 n m /mg/min. Glucose, galactose and xylose significantly inhibit l ‐arabinose transport by both transporters. Transport assays using d ‐xylose showed that KmAxt1p has K m 27 m m and V max 3.8 n m /mg/min, and PgAxt1p has K m 65 m m and V max 8.7 n m /mg/min. Neither transporter is capable of recovering growth on glucose or galactose in a S. cerevisiae strain deleted for hexose and galactose transporters. Transport kinetics of S. cerevisiae Gal2p showed K m 371 m m and V max 341 n m /mg/min for l ‐arabinose, and K m 25 m m and V max 76 n m /mg/min for galactose. Due to the ability of Gal2p and these two newly characterized transportersmore » to transport both l ‐arabinose and d ‐xylose, one scenario for the complete usage of biomass‐derived pentose sugars would require only the low‐affinity, high‐throughput transporter Gal2p and one additional high‐affinity general pentose transporter, rather than dedicated d ‐xylose or l ‐arabinose transporters. Additionally, alignment of these transporters with other characterized pentose transporters provides potential targets for substrate recognition engineering. Accession Nos: KmAXT1 : GZ791039; PgAXT1 : GZ791040 Copyright © 2015 John Wiley & Sons, Ltd.« less

Authors:
 [1];  [2];  [2];  [1];  [1];  [1];  [1]
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  1. National Renewable Energy Laboratory National Bioenergy Centre Golden CO USA
  2. VTT Technical Research Centre of Finland PO Box 1000 FI‐02044 VTT Finland
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1400718
Resource Type:
Publisher's Accepted Manuscript
Journal Name:
Yeast
Additional Journal Information:
Journal Name: Yeast Journal Volume: 32 Journal Issue: 10; Journal ID: ISSN 0749-503X
Publisher:
Wiley Blackwell (John Wiley & Sons)
Country of Publication:
United Kingdom
Language:
English

Citation Formats

Knoshaug, Eric P., Vidgren, Virve, Magalhães, Frederico, Jarvis, Eric E., Franden, Mary Ann, Zhang, Min, and Singh, Arjun. Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on l ‐arabinose and d ‐xylose. United Kingdom: N. p., 2015. Web. doi:10.1002/yea.3084.
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Knoshaug, Eric P., Vidgren, Virve, Magalhães, Frederico, Jarvis, Eric E., Franden, Mary Ann, Zhang, Min, & Singh, Arjun. Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on l ‐arabinose and d ‐xylose. United Kingdom. https://doi.org/10.1002/yea.3084
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Knoshaug, Eric P., Vidgren, Virve, Magalhães, Frederico, Jarvis, Eric E., Franden, Mary Ann, Zhang, Min, and Singh, Arjun. Mon . "Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on l ‐arabinose and d ‐xylose". United Kingdom. https://doi.org/10.1002/yea.3084.
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@article{osti_1400718,
title = {Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on l ‐arabinose and d ‐xylose},
author = {Knoshaug, Eric P. and Vidgren, Virve and Magalhães, Frederico and Jarvis, Eric E. and Franden, Mary Ann and Zhang, Min and Singh, Arjun},
abstractNote = {Abstract Genes encoding l ‐arabinose transporters in Kluyveromyces marxianus and Pichia guilliermondii were identified by functional complementation of Saccharomyces cerevisiae whose growth on l ‐arabinose was dependent on a functioning l ‐arabinose transporter, or by screening a differential display library, respectively. These transporters also transport d ‐xylose and were designated KmAXT1 (arabinose–xylose transporter) and PgAXT1 , respectively. Transport assays using l ‐arabinose showed that KmAxt1p has K m 263 m m and V max 57 n m /mg/min, and PgAxt1p has K m 0.13 m m and V max 18 n m /mg/min. Glucose, galactose and xylose significantly inhibit l ‐arabinose transport by both transporters. Transport assays using d ‐xylose showed that KmAxt1p has K m 27 m m and V max 3.8 n m /mg/min, and PgAxt1p has K m 65 m m and V max 8.7 n m /mg/min. Neither transporter is capable of recovering growth on glucose or galactose in a S. cerevisiae strain deleted for hexose and galactose transporters. Transport kinetics of S. cerevisiae Gal2p showed K m 371 m m and V max 341 n m /mg/min for l ‐arabinose, and K m 25 m m and V max 76 n m /mg/min for galactose. Due to the ability of Gal2p and these two newly characterized transporters to transport both l ‐arabinose and d ‐xylose, one scenario for the complete usage of biomass‐derived pentose sugars would require only the low‐affinity, high‐throughput transporter Gal2p and one additional high‐affinity general pentose transporter, rather than dedicated d ‐xylose or l ‐arabinose transporters. Additionally, alignment of these transporters with other characterized pentose transporters provides potential targets for substrate recognition engineering. Accession Nos: KmAXT1 : GZ791039; PgAXT1 : GZ791040 Copyright © 2015 John Wiley & Sons, Ltd.},
doi = {10.1002/yea.3084},
journal = {Yeast},
number = 10,
volume = 32,
place = {United Kingdom},
year = {Mon Aug 17 00:00:00 EDT 2015},
month = {Mon Aug 17 00:00:00 EDT 2015}
}
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https://doi.org/10.1002/yea.3084
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