L-Arabinose/D-galactose 1-dehydrogenase of Rhizobium leguminosarum bv. trifolii characterised and applied for bioconversion of L-arabinose to L-arabonate with Saccharomyces cerevisiae

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L-Arabinose/D-galactose 1-dehydrogenase of Rhizobium leguminosarum bv. trifolii characterised and applied for bioconversion of L-arabinose to L-arabonate with Saccharomyces cerevisiae. / Aro-Karkkainen, Niina; Toivari, Mervi; Maaheimo, Hannu; Ylilauri, Mikko; Pentikainen, Olli T.; Andberg, Martina; Oja, Merja; Penttilä, Merja; Wiebe, Marilyn G.; Ruohonen, Laura; Koivula, Anu.

In: Applied Microbiology and Biotechnology, Vol. 98, No. 23, 12.2014, p. 9653-9665.

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Aro-Karkkainen, Niina ; Toivari, Mervi ; Maaheimo, Hannu ; Ylilauri, Mikko ; Pentikainen, Olli T. ; Andberg, Martina ; Oja, Merja ; Penttilä, Merja ; Wiebe, Marilyn G. ; Ruohonen, Laura ; Koivula, Anu. / L-Arabinose/D-galactose 1-dehydrogenase of Rhizobium leguminosarum bv. trifolii characterised and applied for bioconversion of L-arabinose to L-arabonate with Saccharomyces cerevisiae. In: Applied Microbiology and Biotechnology. 2014 ; Vol. 98, No. 23. pp. 9653-9665.

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@article{9201ca7dd5e341b696582a328a1e1f01,
title = "L-Arabinose/D-galactose 1-dehydrogenase of Rhizobium leguminosarum bv. trifolii characterised and applied for bioconversion of L-arabinose to L-arabonate with Saccharomyces cerevisiae",
abstract = "Four potential dehydrogenases identified through literature and bioinformatic searches were tested for l-arabonate production from l-arabinose in the yeast Saccharomyces cerevisiae. The most efficient enzyme, annotated as a d-galactose 1-dehydrogenase from the pea root nodule bacterium Rhizobium leguminosarum bv. trifolii, was purified from S. cerevisiae as a homodimeric protein and characterised. We named the enzyme as a l-arabinose/d-galactose 1-dehydrogenase (EC 1.1.1.-), Rl AraDH. It belongs to the Gfo/Idh/MocA protein family, prefers NADP(+) but uses also NAD(+) as a cofactor, and showed highest catalytic efficiency (k (cat)/K (m)) towards l-arabinose, d-galactose and d-fucose. Based on nuclear magnetic resonance (NMR) and modelling studies, the enzyme prefers the alpha-pyranose form of l-arabinose, and the stable oxidation product detected is l-arabino-1,4-lactone which can, however, open slowly at neutral pH to a linear l-arabonate form. The pH optimum for the enzyme was pH 9, but use of a yeast-in-vivo-like buffer at pH 6.8 indicated that good catalytic efficiency could still be expected in vivo. Expression of the Rl AraDH dehydrogenase in S. cerevisiae, together with the galactose permease Gal2 for l-arabinose uptake, resulted in production of 18 g of l-arabonate per litre, at a rate of 248 mg of l-arabonate per litre per hour, with 86 {\%} of the provided l-arabinose converted to l-arabonate. Expression of a lactonase-encoding gene from Caulobacter crescentus was not necessary for l-arabonate production in yeast.",
keywords = "L-Arabonic acid, L-Arabinose, L-Arabinose transport, L-Arabinose dehydrogenase, Platform chemical, Galactose permease, Saccharomyces cerevisiae, DIMERIC DIHYDRODIOL DEHYDROGENASE, ENGINEERED ESCHERICHIA-COLI, D-XYLONATE, D-XYLOSE, PSEUDOMONAS-SACCHAROPHILA, ALTERNATIVE PATHWAY, METABOLISM, YEAST, IDENTIFICATION, EXPRESSION",
author = "Niina Aro-Karkkainen and Mervi Toivari and Hannu Maaheimo and Mikko Ylilauri and Pentikainen, {Olli T.} and Martina Andberg and Merja Oja and Merja Penttil{\"a} and Wiebe, {Marilyn G.} and Laura Ruohonen and Anu Koivula",
year = "2014",
month = "12",
doi = "10.1007/s00253-014-6039-2",
language = "English",
volume = "98",
pages = "9653--9665",
journal = "Applied Microbiology & Biotechnology",
issn = "0175-7598",
number = "23",

}

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TY - JOUR

T1 - L-Arabinose/D-galactose 1-dehydrogenase of Rhizobium leguminosarum bv. trifolii characterised and applied for bioconversion of L-arabinose to L-arabonate with Saccharomyces cerevisiae

AU - Aro-Karkkainen, Niina

AU - Toivari, Mervi

AU - Maaheimo, Hannu

AU - Ylilauri, Mikko

AU - Pentikainen, Olli T.

AU - Andberg, Martina

AU - Oja, Merja

AU - Penttilä, Merja

AU - Wiebe, Marilyn G.

AU - Ruohonen, Laura

AU - Koivula, Anu

PY - 2014/12

Y1 - 2014/12

N2 - Four potential dehydrogenases identified through literature and bioinformatic searches were tested for l-arabonate production from l-arabinose in the yeast Saccharomyces cerevisiae. The most efficient enzyme, annotated as a d-galactose 1-dehydrogenase from the pea root nodule bacterium Rhizobium leguminosarum bv. trifolii, was purified from S. cerevisiae as a homodimeric protein and characterised. We named the enzyme as a l-arabinose/d-galactose 1-dehydrogenase (EC 1.1.1.-), Rl AraDH. It belongs to the Gfo/Idh/MocA protein family, prefers NADP(+) but uses also NAD(+) as a cofactor, and showed highest catalytic efficiency (k (cat)/K (m)) towards l-arabinose, d-galactose and d-fucose. Based on nuclear magnetic resonance (NMR) and modelling studies, the enzyme prefers the alpha-pyranose form of l-arabinose, and the stable oxidation product detected is l-arabino-1,4-lactone which can, however, open slowly at neutral pH to a linear l-arabonate form. The pH optimum for the enzyme was pH 9, but use of a yeast-in-vivo-like buffer at pH 6.8 indicated that good catalytic efficiency could still be expected in vivo. Expression of the Rl AraDH dehydrogenase in S. cerevisiae, together with the galactose permease Gal2 for l-arabinose uptake, resulted in production of 18 g of l-arabonate per litre, at a rate of 248 mg of l-arabonate per litre per hour, with 86 % of the provided l-arabinose converted to l-arabonate. Expression of a lactonase-encoding gene from Caulobacter crescentus was not necessary for l-arabonate production in yeast.

AB - Four potential dehydrogenases identified through literature and bioinformatic searches were tested for l-arabonate production from l-arabinose in the yeast Saccharomyces cerevisiae. The most efficient enzyme, annotated as a d-galactose 1-dehydrogenase from the pea root nodule bacterium Rhizobium leguminosarum bv. trifolii, was purified from S. cerevisiae as a homodimeric protein and characterised. We named the enzyme as a l-arabinose/d-galactose 1-dehydrogenase (EC 1.1.1.-), Rl AraDH. It belongs to the Gfo/Idh/MocA protein family, prefers NADP(+) but uses also NAD(+) as a cofactor, and showed highest catalytic efficiency (k (cat)/K (m)) towards l-arabinose, d-galactose and d-fucose. Based on nuclear magnetic resonance (NMR) and modelling studies, the enzyme prefers the alpha-pyranose form of l-arabinose, and the stable oxidation product detected is l-arabino-1,4-lactone which can, however, open slowly at neutral pH to a linear l-arabonate form. The pH optimum for the enzyme was pH 9, but use of a yeast-in-vivo-like buffer at pH 6.8 indicated that good catalytic efficiency could still be expected in vivo. Expression of the Rl AraDH dehydrogenase in S. cerevisiae, together with the galactose permease Gal2 for l-arabinose uptake, resulted in production of 18 g of l-arabonate per litre, at a rate of 248 mg of l-arabonate per litre per hour, with 86 % of the provided l-arabinose converted to l-arabonate. Expression of a lactonase-encoding gene from Caulobacter crescentus was not necessary for l-arabonate production in yeast.

KW - L-Arabonic acid

KW - L-Arabinose

KW - L-Arabinose transport

KW - L-Arabinose dehydrogenase

KW - Platform chemical

KW - Galactose permease

KW - Saccharomyces cerevisiae

KW - DIMERIC DIHYDRODIOL DEHYDROGENASE

KW - ENGINEERED ESCHERICHIA-COLI

KW - D-XYLONATE

KW - D-XYLOSE

KW - PSEUDOMONAS-SACCHAROPHILA

KW - ALTERNATIVE PATHWAY

KW - METABOLISM

KW - YEAST

KW - IDENTIFICATION

KW - EXPRESSION

U2 - 10.1007/s00253-014-6039-2

DO - 10.1007/s00253-014-6039-2

M3 - Article

VL - 98

SP - 9653

EP - 9665

JO - Applied Microbiology & Biotechnology

JF - Applied Microbiology & Biotechnology

SN - 0175-7598

IS - 23

ER -

ID: 9225848