Cloning and characterization of two a-glucosidases from Bifidobacterium adolescentis DSM20083.

L.A.M. van den Broek, K. Struijs, J.C. Verdoes, G. Beldman, A.G.J. Voragen

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Two alpha-glucosidase encoding genes (aglA and aglB) from Bifidobacterium adolescentis DSM 20083 were isolated and characterized. Both alpha-glucosidases belong to family 13 of the glycosyl hydrolases. Recombinant AglA (EC 3.2.1.10) and AglB (EC 3.2.1.20), expressed in Escherichia coli, showed high hydrolytic activity towards isomaltose and pnp-alpha-glucoside. The K,, for pnp-alpha-glucoside was 1.05 and 0.47 mM and the V-max was 228 and 113 U mg(-1) for AglA and AglB, respectively. Using pnp-a-glucoside as substrate, the pH optimum for AglA was 6.6 and the temperature optimum was 37degreesC. For AglB, values of pH 6.8 and 47degreesC were found. AglA also showed high hydrolytic activity towards isomaltotriose and, to a lesser extent, towards trehalose. AglB has a high preference for maltose and less activity towards sucrose; minor activity was observed towards melizitose, low molecular weight dextrin, maltitol, and maltotriose. The recombinant a-glucosidases were tested for their transglucosylation activity. AglA was able to synthesize oligosaccharides from trehalose and sucrose. AglB formed oligosaccharides from sucrose, maltose, and melizitose.
Original languageEnglish
Pages (from-to)55-60
JournalApplied Microbiology and Biotechnology
Volume61
DOIs
Publication statusPublished - 2003

Fingerprint

Glucosidases
Glucosides
Sucrose
Organism Cloning
Trehalose
alpha-Glucosidases
Maltose
Oligosaccharides
Isomaltose
Oligo-1,6-Glucosidase
Hydrolases
Molecular Weight
Escherichia coli
Temperature
Genes
Bifidobacterium adolescentis

Keywords

  • intestinal microflora
  • beta-galactosidase
  • enzymatic methods
  • oligosaccharides
  • purification
  • bacteria
  • growth
  • longum
  • rats

Cite this

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title = "Cloning and characterization of two a-glucosidases from Bifidobacterium adolescentis DSM20083.",
abstract = "Two alpha-glucosidase encoding genes (aglA and aglB) from Bifidobacterium adolescentis DSM 20083 were isolated and characterized. Both alpha-glucosidases belong to family 13 of the glycosyl hydrolases. Recombinant AglA (EC 3.2.1.10) and AglB (EC 3.2.1.20), expressed in Escherichia coli, showed high hydrolytic activity towards isomaltose and pnp-alpha-glucoside. The K,, for pnp-alpha-glucoside was 1.05 and 0.47 mM and the V-max was 228 and 113 U mg(-1) for AglA and AglB, respectively. Using pnp-a-glucoside as substrate, the pH optimum for AglA was 6.6 and the temperature optimum was 37degreesC. For AglB, values of pH 6.8 and 47degreesC were found. AglA also showed high hydrolytic activity towards isomaltotriose and, to a lesser extent, towards trehalose. AglB has a high preference for maltose and less activity towards sucrose; minor activity was observed towards melizitose, low molecular weight dextrin, maltitol, and maltotriose. The recombinant a-glucosidases were tested for their transglucosylation activity. AglA was able to synthesize oligosaccharides from trehalose and sucrose. AglB formed oligosaccharides from sucrose, maltose, and melizitose.",
keywords = "intestinal microflora, beta-galactosidase, enzymatic methods, oligosaccharides, purification, bacteria, growth, longum, rats",
author = "{van den Broek}, L.A.M. and K. Struijs and J.C. Verdoes and G. Beldman and A.G.J. Voragen",
year = "2003",
doi = "10.1007/s00253-002-1179-1",
language = "English",
volume = "61",
pages = "55--60",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",

}

Cloning and characterization of two a-glucosidases from Bifidobacterium adolescentis DSM20083. / van den Broek, L.A.M.; Struijs, K.; Verdoes, J.C.; Beldman, G.; Voragen, A.G.J.

In: Applied Microbiology and Biotechnology, Vol. 61, 2003, p. 55-60.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Cloning and characterization of two a-glucosidases from Bifidobacterium adolescentis DSM20083.

AU - van den Broek, L.A.M.

AU - Struijs, K.

AU - Verdoes, J.C.

AU - Beldman, G.

AU - Voragen, A.G.J.

PY - 2003

Y1 - 2003

N2 - Two alpha-glucosidase encoding genes (aglA and aglB) from Bifidobacterium adolescentis DSM 20083 were isolated and characterized. Both alpha-glucosidases belong to family 13 of the glycosyl hydrolases. Recombinant AglA (EC 3.2.1.10) and AglB (EC 3.2.1.20), expressed in Escherichia coli, showed high hydrolytic activity towards isomaltose and pnp-alpha-glucoside. The K,, for pnp-alpha-glucoside was 1.05 and 0.47 mM and the V-max was 228 and 113 U mg(-1) for AglA and AglB, respectively. Using pnp-a-glucoside as substrate, the pH optimum for AglA was 6.6 and the temperature optimum was 37degreesC. For AglB, values of pH 6.8 and 47degreesC were found. AglA also showed high hydrolytic activity towards isomaltotriose and, to a lesser extent, towards trehalose. AglB has a high preference for maltose and less activity towards sucrose; minor activity was observed towards melizitose, low molecular weight dextrin, maltitol, and maltotriose. The recombinant a-glucosidases were tested for their transglucosylation activity. AglA was able to synthesize oligosaccharides from trehalose and sucrose. AglB formed oligosaccharides from sucrose, maltose, and melizitose.

AB - Two alpha-glucosidase encoding genes (aglA and aglB) from Bifidobacterium adolescentis DSM 20083 were isolated and characterized. Both alpha-glucosidases belong to family 13 of the glycosyl hydrolases. Recombinant AglA (EC 3.2.1.10) and AglB (EC 3.2.1.20), expressed in Escherichia coli, showed high hydrolytic activity towards isomaltose and pnp-alpha-glucoside. The K,, for pnp-alpha-glucoside was 1.05 and 0.47 mM and the V-max was 228 and 113 U mg(-1) for AglA and AglB, respectively. Using pnp-a-glucoside as substrate, the pH optimum for AglA was 6.6 and the temperature optimum was 37degreesC. For AglB, values of pH 6.8 and 47degreesC were found. AglA also showed high hydrolytic activity towards isomaltotriose and, to a lesser extent, towards trehalose. AglB has a high preference for maltose and less activity towards sucrose; minor activity was observed towards melizitose, low molecular weight dextrin, maltitol, and maltotriose. The recombinant a-glucosidases were tested for their transglucosylation activity. AglA was able to synthesize oligosaccharides from trehalose and sucrose. AglB formed oligosaccharides from sucrose, maltose, and melizitose.

KW - intestinal microflora

KW - beta-galactosidase

KW - enzymatic methods

KW - oligosaccharides

KW - purification

KW - bacteria

KW - growth

KW - longum

KW - rats

U2 - 10.1007/s00253-002-1179-1

DO - 10.1007/s00253-002-1179-1

M3 - Article

VL - 61

SP - 55

EP - 60

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

ER -