Characterization of a unique Caulobacter crescentus aldose-aldose oxidoreductase having dual activities

Martina Andberg, Hannu Maaheimo, Esa-Pekka Kumpula, Harry Boer, Mervi Toivari, Merja Penttilä, Anu Koivula

Research output: Contribution to journalArticleScientificpeer-review

2 Citations (Scopus)

Abstract

We describe here the characterization of a novel enzyme called aldose-aldose oxidoreductase (Cc AAOR; EC 1.1.99) from Caulobacter crescentus. The Cc AAOR exists in solution as a dimer, belongs to the Gfo/Idh/MocA family and shows homology with the glucose-fructose oxidoreductase from Zymomonas mobilis. However, unlike other known members of this protein family, Cc AAOR is specific for aldose sugars and can be in the same catalytic cycle both oxidise and reduce a panel of monosaccharides at the C1 position, producing in each case the corresponding aldonolactone and alditol, respectively. Cc AAOR contains a tightly-bound nicotinamide cofactor, which is regenerated in this oxidation-reduction cycle. The highest oxidation activity was detected on d-glucose but significant activity was also observed on d-xylose, l-arabinose and d-galactose, revealing that both hexose and pentose sugars are accepted as substrates by Cc AAOR. The configuration at the C2 and C3 positions of the saccharides was shown to be especially important for the substrate binding. Interestingly, besides monosaccharides, Cc AAOR can also oxidise a range of 1,4-linked oligosaccharides having aldose unit at the reducing end, such as lactose, malto- and cello-oligosaccharides as well as xylotetraose. 1H NMR used to monitor the oxidation and reduction reaction simultaneously, demonstrated that although d-glucose has the highest affinity and is also oxidised most efficiently by Cc AAOR, the reduction of d-glucose is clearly not as efficient. For the overall reaction catalysed by Cc AAOR, the l-arabinose, d-xylose and d-galactose were the most potent substrates.
Original languageEnglish
Pages (from-to)673-685
JournalApplied Microbiology and Biotechnology
Volume100
Issue number2
DOIs
Publication statusPublished - 2016
MoE publication typeA1 Journal article-refereed

Fingerprint

Caulobacter crescentus
Oxidoreductases
Arabinose
Monosaccharides
Xylose
Oligosaccharides
Galactose
Glucose
Oxidation-Reduction
Zymomonas
Sugar Alcohols
Pentoses
Hexoses
Niacinamide
Lactose
Enzymes
Proteins

Keywords

  • carbohydrate
  • enzyme catalysis
  • glucose-fructose oxidoreductase
  • nuclear magnetic resonance
  • tightly-bound cofactor

Cite this

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title = "Characterization of a unique Caulobacter crescentus aldose-aldose oxidoreductase having dual activities",
abstract = "We describe here the characterization of a novel enzyme called aldose-aldose oxidoreductase (Cc AAOR; EC 1.1.99) from Caulobacter crescentus. The Cc AAOR exists in solution as a dimer, belongs to the Gfo/Idh/MocA family and shows homology with the glucose-fructose oxidoreductase from Zymomonas mobilis. However, unlike other known members of this protein family, Cc AAOR is specific for aldose sugars and can be in the same catalytic cycle both oxidise and reduce a panel of monosaccharides at the C1 position, producing in each case the corresponding aldonolactone and alditol, respectively. Cc AAOR contains a tightly-bound nicotinamide cofactor, which is regenerated in this oxidation-reduction cycle. The highest oxidation activity was detected on d-glucose but significant activity was also observed on d-xylose, l-arabinose and d-galactose, revealing that both hexose and pentose sugars are accepted as substrates by Cc AAOR. The configuration at the C2 and C3 positions of the saccharides was shown to be especially important for the substrate binding. Interestingly, besides monosaccharides, Cc AAOR can also oxidise a range of 1,4-linked oligosaccharides having aldose unit at the reducing end, such as lactose, malto- and cello-oligosaccharides as well as xylotetraose. 1H NMR used to monitor the oxidation and reduction reaction simultaneously, demonstrated that although d-glucose has the highest affinity and is also oxidised most efficiently by Cc AAOR, the reduction of d-glucose is clearly not as efficient. For the overall reaction catalysed by Cc AAOR, the l-arabinose, d-xylose and d-galactose were the most potent substrates.",
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Characterization of a unique Caulobacter crescentus aldose-aldose oxidoreductase having dual activities. / Andberg, Martina; Maaheimo, Hannu; Kumpula, Esa-Pekka; Boer, Harry; Toivari, Mervi; Penttilä, Merja; Koivula, Anu.

In: Applied Microbiology and Biotechnology, Vol. 100, No. 2, 2016, p. 673-685.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Characterization of a unique Caulobacter crescentus aldose-aldose oxidoreductase having dual activities

AU - Andberg, Martina

AU - Maaheimo, Hannu

AU - Kumpula, Esa-Pekka

AU - Boer, Harry

AU - Toivari, Mervi

AU - Penttilä, Merja

AU - Koivula, Anu

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N2 - We describe here the characterization of a novel enzyme called aldose-aldose oxidoreductase (Cc AAOR; EC 1.1.99) from Caulobacter crescentus. The Cc AAOR exists in solution as a dimer, belongs to the Gfo/Idh/MocA family and shows homology with the glucose-fructose oxidoreductase from Zymomonas mobilis. However, unlike other known members of this protein family, Cc AAOR is specific for aldose sugars and can be in the same catalytic cycle both oxidise and reduce a panel of monosaccharides at the C1 position, producing in each case the corresponding aldonolactone and alditol, respectively. Cc AAOR contains a tightly-bound nicotinamide cofactor, which is regenerated in this oxidation-reduction cycle. The highest oxidation activity was detected on d-glucose but significant activity was also observed on d-xylose, l-arabinose and d-galactose, revealing that both hexose and pentose sugars are accepted as substrates by Cc AAOR. The configuration at the C2 and C3 positions of the saccharides was shown to be especially important for the substrate binding. Interestingly, besides monosaccharides, Cc AAOR can also oxidise a range of 1,4-linked oligosaccharides having aldose unit at the reducing end, such as lactose, malto- and cello-oligosaccharides as well as xylotetraose. 1H NMR used to monitor the oxidation and reduction reaction simultaneously, demonstrated that although d-glucose has the highest affinity and is also oxidised most efficiently by Cc AAOR, the reduction of d-glucose is clearly not as efficient. For the overall reaction catalysed by Cc AAOR, the l-arabinose, d-xylose and d-galactose were the most potent substrates.

AB - We describe here the characterization of a novel enzyme called aldose-aldose oxidoreductase (Cc AAOR; EC 1.1.99) from Caulobacter crescentus. The Cc AAOR exists in solution as a dimer, belongs to the Gfo/Idh/MocA family and shows homology with the glucose-fructose oxidoreductase from Zymomonas mobilis. However, unlike other known members of this protein family, Cc AAOR is specific for aldose sugars and can be in the same catalytic cycle both oxidise and reduce a panel of monosaccharides at the C1 position, producing in each case the corresponding aldonolactone and alditol, respectively. Cc AAOR contains a tightly-bound nicotinamide cofactor, which is regenerated in this oxidation-reduction cycle. The highest oxidation activity was detected on d-glucose but significant activity was also observed on d-xylose, l-arabinose and d-galactose, revealing that both hexose and pentose sugars are accepted as substrates by Cc AAOR. The configuration at the C2 and C3 positions of the saccharides was shown to be especially important for the substrate binding. Interestingly, besides monosaccharides, Cc AAOR can also oxidise a range of 1,4-linked oligosaccharides having aldose unit at the reducing end, such as lactose, malto- and cello-oligosaccharides as well as xylotetraose. 1H NMR used to monitor the oxidation and reduction reaction simultaneously, demonstrated that although d-glucose has the highest affinity and is also oxidised most efficiently by Cc AAOR, the reduction of d-glucose is clearly not as efficient. For the overall reaction catalysed by Cc AAOR, the l-arabinose, d-xylose and d-galactose were the most potent substrates.

KW - carbohydrate

KW - enzyme catalysis

KW - glucose-fructose oxidoreductase

KW - nuclear magnetic resonance

KW - tightly-bound cofactor

U2 - 10.1007/s00253-015-7011-5

DO - 10.1007/s00253-015-7011-5

M3 - Article

VL - 100

SP - 673

EP - 685

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 2

ER -