Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on L-arabinose and D-xylose

Eric P. Knoshaug (Corresponding Author), Virve Vidgren, Frederico Magalhães, Eric E. Jarvis, Mary Ann Franden, Min Zhang, Arjun Singh

Research output: Contribution to journalArticleScientificpeer-review

20 Citations (Scopus)

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 Km 263mm and Vmax 57nm/mg/min, and PgAxt1p has Km 0.13mm and Vmax 18nm/mg/min. Glucose, galactose and xylose significantly inhibit l-arabinose transport by both transporters. Transport assays using d-xylose showed that KmAxt1p has Km 27mm and Vmax 3.8nm/mg/min, and PgAxt1p has Km 65mm and Vmax 8.7nm/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 Km 371mm and Vmax 341nm/mg/min for l-arabinose, and Km 25mm and Vmax 76nm/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.
Original languageEnglish
Pages (from-to)615-628
JournalYeast
Volume32
Issue number10
DOIs
Publication statusPublished - 2015
MoE publication typeA1 Journal article-refereed

Fingerprint

Kluyveromyces
Arabinose
Xylose
Pichia
Yeast
Saccharomyces cerevisiae
Growth
Galactose
Pentoses
Glucose
Assays
Gene encoding
Monosaccharide Transport Proteins
Sugars
Screening
Biomass
Hexoses
Display devices
Throughput
Libraries

Keywords

  • arabinose
  • xylose
  • transport
  • yeast
  • GAL2

Cite this

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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 = "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 Km 263mm and Vmax 57nm/mg/min, and PgAxt1p has Km 0.13mm and Vmax 18nm/mg/min. Glucose, galactose and xylose significantly inhibit l-arabinose transport by both transporters. Transport assays using d-xylose showed that KmAxt1p has Km 27mm and Vmax 3.8nm/mg/min, and PgAxt1p has Km 65mm and Vmax 8.7nm/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 Km 371mm and Vmax 341nm/mg/min for l-arabinose, and Km 25mm and Vmax 76nm/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.",
keywords = "arabinose, xylose, transport, yeast, GAL2",
author = "Knoshaug, {Eric P.} and Virve Vidgren and Frederico Magalh{\~a}es and Jarvis, {Eric E.} and Franden, {Mary Ann} and Min Zhang and Arjun Singh",
year = "2015",
doi = "10.1002/yea.3084",
language = "English",
volume = "32",
pages = "615--628",
journal = "Yeast",
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}

Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on L-arabinose and D-xylose. / Knoshaug, Eric P. (Corresponding Author); Vidgren, Virve; Magalhães, Frederico; Jarvis, Eric E.; Franden, Mary Ann; Zhang, Min; Singh, Arjun.

In: Yeast, Vol. 32, No. 10, 2015, p. 615-628.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on L-arabinose and D-xylose

AU - Knoshaug, Eric P.

AU - Vidgren, Virve

AU - Magalhães, Frederico

AU - Jarvis, Eric E.

AU - Franden, Mary Ann

AU - Zhang, Min

AU - Singh, Arjun

PY - 2015

Y1 - 2015

N2 - 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 Km 263mm and Vmax 57nm/mg/min, and PgAxt1p has Km 0.13mm and Vmax 18nm/mg/min. Glucose, galactose and xylose significantly inhibit l-arabinose transport by both transporters. Transport assays using d-xylose showed that KmAxt1p has Km 27mm and Vmax 3.8nm/mg/min, and PgAxt1p has Km 65mm and Vmax 8.7nm/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 Km 371mm and Vmax 341nm/mg/min for l-arabinose, and Km 25mm and Vmax 76nm/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.

AB - 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 Km 263mm and Vmax 57nm/mg/min, and PgAxt1p has Km 0.13mm and Vmax 18nm/mg/min. Glucose, galactose and xylose significantly inhibit l-arabinose transport by both transporters. Transport assays using d-xylose showed that KmAxt1p has Km 27mm and Vmax 3.8nm/mg/min, and PgAxt1p has Km 65mm and Vmax 8.7nm/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 Km 371mm and Vmax 341nm/mg/min for l-arabinose, and Km 25mm and Vmax 76nm/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.

KW - arabinose

KW - xylose

KW - transport

KW - yeast

KW - GAL2

U2 - 10.1002/yea.3084

DO - 10.1002/yea.3084

M3 - Article

VL - 32

SP - 615

EP - 628

JO - Yeast

JF - Yeast

SN - 0749-503X

IS - 10

ER -