Metabolic engineering of Saccharomyces cerevisiae for bioconversion of D-xylose to D-xylonate

Mervi Toivari; Yvonne Nygard; Esa-Pekka Kumpula; Maija-Leena Vehkomaki; Mojca Bencina; Mari Valkonen; Hannu Maaheimo; Martina Andberg; Anu Koivula; Laura Ruohonen; Merja Penttila; Marilyn G. Wiebe

doi:10.1016/j.ymben.2012.03.002

Metabolic engineering of Saccharomyces cerevisiae for bioconversion of D-xylose to D-xylonate

Mervi Toivari^*, Yvonne Nygard, Esa-Pekka Kumpula, Maija-Leena Vehkomaki, Mojca Bencina, Mari Valkonen, Hannu Maaheimo, Martina Andberg, Anu Koivula, Laura Ruohonen, Merja Penttila, Marilyn G. Wiebe

^*Corresponding author for this work

Not published at Aalto University

VTT Technical Research Centre of Finland

Research output: Contribution to journal › Article › Scientific › peer-review

Abstract

An NAD(+)-dependent D-xylose dehydrogenase, XylB, from Caulobacter crescentus was expressed in Saccharomyces cerevisiae, resulting in production of 17 +/- 2 g D-xylonate l(-1) at 0.23 g l(-1) h(-1) from 23 g D-xylose l(-1) (with glucose and ethanol as co-substrates). D-Xylonate titre and production rate were increased and xylitol production decreased, compared to strains expressing genes encoding T. reesei or pig liver NADP(+)-dependent D-xylose dehydrogenases. D-Xylonate accumulated intracellularly to similar to 70 mg g(-1); xylitol to similar to 18 mg g(-1). The aldose reductase encoding gene GRE3 was deleted to reduce xylitol production. Cells expressing D-xylonolactone lactonase xylC from C crescentus with xylB initially produced more extracellular o-xylonate than cells lacking xylC at both pH 5.5 and pH 3, and sustained higher production at pH 3. Cell vitality and viability decreased during o-xylonate production at pH 3.0. An industrial S. cerevisiae strain expressing xylB efficiently produced 43 g D-xylonate l(-1) from 49 g D-xylose l(-1). (C) 2012 Elsevier Inc. All rights reserved.

Original language	English
Pages (from-to)	427-436
Number of pages	10
Journal	Metabolic Engineering
Volume	14
Issue number	4
DOIs	https://doi.org/10.1016/j.ymben.2012.03.002
Publication status	Published - Jul 2012
MoE publication type	A1 Journal article-refereed

Funding

Technical assistance of Jenni Kaija, Tarja Laakso, Outi Kononen and Tuuli Teikari is gratefully acknowledged. This study was financially supported by the Academy of Finland through the Centre of Excellence in White Biotechnology - Green Chemistry (grant 118573). The pH studies were funded with Academy of Finland researcher mobility grant (132169) and Slovenian Research Agency (grant BI-FI/11-12-019). Financial support from the VU Graduate School is acknowledged (Yvonne Nygard). The financial support of the European Commission through the Sixth Framework Programme Integrated Project BioSynergy (038994-5E56) and the Seventh Framework Programme (FP7/2007-2013) under grant agreement No. FP7-241566 BIOCORE are also gratefully acknowledged.

Keywords

D-xylose dehydrogenase
D-xylonic acid
D-xylose
Saccharomyces cerevisiae
Bioconversion
PSEUDOMONAS-FRAGI
GLUCONOBACTER-OXYDANS
INTRACELLULAR PH
STEAMED HEMICELLULOSE
ACID PRODUCTION
LACTIC-ACID
PIG-LIVER
DEHYDROGENASE
OXIDATION
REDUCTASE

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10.1016/j.ymben.2012.03.002

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@article{9a75fd98ae944e4e9ae05e2d9d057f48,

title = "Metabolic engineering of Saccharomyces cerevisiae for bioconversion of D-xylose to D-xylonate",

abstract = "An NAD(+)-dependent D-xylose dehydrogenase, XylB, from Caulobacter crescentus was expressed in Saccharomyces cerevisiae, resulting in production of 17 +/- 2 g D-xylonate l(-1) at 0.23 g l(-1) h(-1) from 23 g D-xylose l(-1) (with glucose and ethanol as co-substrates). D-Xylonate titre and production rate were increased and xylitol production decreased, compared to strains expressing genes encoding T. reesei or pig liver NADP(+)-dependent D-xylose dehydrogenases. D-Xylonate accumulated intracellularly to similar to 70 mg g(-1); xylitol to similar to 18 mg g(-1). The aldose reductase encoding gene GRE3 was deleted to reduce xylitol production. Cells expressing D-xylonolactone lactonase xylC from C crescentus with xylB initially produced more extracellular o-xylonate than cells lacking xylC at both pH 5.5 and pH 3, and sustained higher production at pH 3. Cell vitality and viability decreased during o-xylonate production at pH 3.0. An industrial S. cerevisiae strain expressing xylB efficiently produced 43 g D-xylonate l(-1) from 49 g D-xylose l(-1). (C) 2012 Elsevier Inc. All rights reserved.",

keywords = "D-xylose dehydrogenase, D-xylonic acid, D-xylose, Saccharomyces cerevisiae, Bioconversion, PSEUDOMONAS-FRAGI, GLUCONOBACTER-OXYDANS, INTRACELLULAR PH, STEAMED HEMICELLULOSE, ACID PRODUCTION, LACTIC-ACID, PIG-LIVER, DEHYDROGENASE, OXIDATION, REDUCTASE",

author = "Mervi Toivari and Yvonne Nygard and Esa-Pekka Kumpula and Maija-Leena Vehkomaki and Mojca Bencina and Mari Valkonen and Hannu Maaheimo and Martina Andberg and Anu Koivula and Laura Ruohonen and Merja Penttila and Wiebe, \{Marilyn G.\}",

year = "2012",

month = jul,

doi = "10.1016/j.ymben.2012.03.002",

language = "English",

volume = "14",

pages = "427--436",

journal = "Metabolic Engineering",

issn = "1096-7176",

publisher = "Elsevier",

number = "4",

}

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T1 - Metabolic engineering of Saccharomyces cerevisiae for bioconversion of D-xylose to D-xylonate

AU - Toivari, Mervi

AU - Nygard, Yvonne

AU - Kumpula, Esa-Pekka

AU - Vehkomaki, Maija-Leena

AU - Bencina, Mojca

AU - Valkonen, Mari

AU - Maaheimo, Hannu

AU - Andberg, Martina

AU - Koivula, Anu

AU - Ruohonen, Laura

AU - Penttila, Merja

AU - Wiebe, Marilyn G.

PY - 2012/7

Y1 - 2012/7

N2 - An NAD(+)-dependent D-xylose dehydrogenase, XylB, from Caulobacter crescentus was expressed in Saccharomyces cerevisiae, resulting in production of 17 +/- 2 g D-xylonate l(-1) at 0.23 g l(-1) h(-1) from 23 g D-xylose l(-1) (with glucose and ethanol as co-substrates). D-Xylonate titre and production rate were increased and xylitol production decreased, compared to strains expressing genes encoding T. reesei or pig liver NADP(+)-dependent D-xylose dehydrogenases. D-Xylonate accumulated intracellularly to similar to 70 mg g(-1); xylitol to similar to 18 mg g(-1). The aldose reductase encoding gene GRE3 was deleted to reduce xylitol production. Cells expressing D-xylonolactone lactonase xylC from C crescentus with xylB initially produced more extracellular o-xylonate than cells lacking xylC at both pH 5.5 and pH 3, and sustained higher production at pH 3. Cell vitality and viability decreased during o-xylonate production at pH 3.0. An industrial S. cerevisiae strain expressing xylB efficiently produced 43 g D-xylonate l(-1) from 49 g D-xylose l(-1). (C) 2012 Elsevier Inc. All rights reserved.

AB - An NAD(+)-dependent D-xylose dehydrogenase, XylB, from Caulobacter crescentus was expressed in Saccharomyces cerevisiae, resulting in production of 17 +/- 2 g D-xylonate l(-1) at 0.23 g l(-1) h(-1) from 23 g D-xylose l(-1) (with glucose and ethanol as co-substrates). D-Xylonate titre and production rate were increased and xylitol production decreased, compared to strains expressing genes encoding T. reesei or pig liver NADP(+)-dependent D-xylose dehydrogenases. D-Xylonate accumulated intracellularly to similar to 70 mg g(-1); xylitol to similar to 18 mg g(-1). The aldose reductase encoding gene GRE3 was deleted to reduce xylitol production. Cells expressing D-xylonolactone lactonase xylC from C crescentus with xylB initially produced more extracellular o-xylonate than cells lacking xylC at both pH 5.5 and pH 3, and sustained higher production at pH 3. Cell vitality and viability decreased during o-xylonate production at pH 3.0. An industrial S. cerevisiae strain expressing xylB efficiently produced 43 g D-xylonate l(-1) from 49 g D-xylose l(-1). (C) 2012 Elsevier Inc. All rights reserved.

KW - D-xylose dehydrogenase

KW - D-xylonic acid

KW - D-xylose

KW - Saccharomyces cerevisiae

KW - Bioconversion

KW - PSEUDOMONAS-FRAGI

KW - GLUCONOBACTER-OXYDANS

KW - INTRACELLULAR PH

KW - STEAMED HEMICELLULOSE

KW - ACID PRODUCTION

KW - LACTIC-ACID

KW - PIG-LIVER

KW - DEHYDROGENASE

KW - OXIDATION

KW - REDUCTASE

U2 - 10.1016/j.ymben.2012.03.002

DO - 10.1016/j.ymben.2012.03.002

M3 - Article

SN - 1096-7176

VL - 14

SP - 427

EP - 436

JO - Metabolic Engineering

JF - Metabolic Engineering

IS - 4

ER -

Metabolic engineering of Saccharomyces cerevisiae for bioconversion of D-xylose to D-xylonate

Abstract

Funding

Keywords

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