Cloning and Functional Expression of an α-Galactosidase fromYersinia pestis biovar Microtusstr. 91001

Cao, Yanan; Huang, Huoqing; Meng, Kun; Yang, Peilong; Shi, Pengjun; Wang, Yaru; Luo, Huiying; Zhang, Zhifang; Wu, Ningfeng; Yao, Bin

doi:10.1271/bbb.80145

Cited by 10 publications

(5 citation statements)

References 12 publications

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“…3.2.1.22) (α-Gals) catalyze the hydrolysis of galactosyl residues from the nonreducing ends of a variety of oligosaccharides and polysaccharides, as well as galactolipids and glycoproteins. This substrate diversity is matched by a wide taxonomic distribution of α-Gal-producing organisms including archaea, 2 bacteria, [3][4][5] fungi, 6,7 plants 8 and animals. 9 GHs are classified into the sequence-based CAZy (Carbohydrate-Active EnZymes †) database 10 currently comprising 125 GH families organized in mechanistically and structurally related clans (A through N).…”

Section: Introductionmentioning

confidence: 99%

Crystal Structure of α-Galactosidase from Lactobacillus acidophilus NCFM: Insight into Tetramer Formation and Substrate Binding

Fredslund

Hachem

Larsen

et al. 2011

Journal of Molecular Biology

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Section: Introductionmentioning

confidence: 99%

Crystal Structure of α-Galactosidase from Lactobacillus acidophilus NCFM: Insight into Tetramer Formation and Substrate Binding

Fredslund

Hachem

Larsen

et al. 2011

Journal of Molecular Biology

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“…α-D-Galactosidase (EC 3.2.1.22), which catalyzes terminal α-galactose hydrolysis, has been classified into six glycosyl hydrolase (GH) families based on amino acid sequence similarity: GH4, GH27, GH36, GH57, GH97, and GH110. In particular, GH36 α-galactosidases are widely distributed in archaea, 1) bacteria, 2) 3) 4) fungi, 5) 6) 7) 8) and plant. 9) 10) 11) To date, several bifidobacterial α-galactosidases belonging to GH36 were cloned and characterized from Bifidobacterium longum subsp.…”

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confidence: 99%

Characterization of a GH36 α-D-Galactosidase Associated with Assimilation of Gum Arabic in Bifidobacterium longum subsp. longum JCM7052

et al. 2021

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We recently characterized a 3-O-α-D-galactosyl-α-L-arabinofuranosidase (GAfase) for the release of α-D-Gal-(1→3)-L-Ara from gum arabic arabinogalactan protein (AGP) in Bifidobacterium longum subsp. longum JCM7052. In the present study, we cloned and characterized a neighboring αgalactosidase gene (BLGA_00330; blAga3). It contained an Open Reading Frame of 2151-bp nucleotides encoding 716 amino acids with an estimated molecular mass of 79,587 Da. Recombinant BlAga3 released galactose from α-D-Gal-(1→3)-L-Ara, but not from intact gum arabic AGP, and a little from the related oligosaccharides. The enzyme also showed the activity toward blood group B liner trisaccharide. The specific activity for α-D-Gal-(1→3)-L-Ara was 4.27-and 2.10-fold higher than those for melibiose and raffinose, respectively. The optimal pH and temperature were 6.0 and 50 °C, respectively. BlAga3 is an intracellular α-galactosidase that cleaves α-D-Gal-(1→3)-L-Ara produced by GAfase; it is also responsible for a series of gum arabic AGP degradation in B. longum JCM7052.

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“…Moreover, galV was more stable than most GH36 α-galactosidases, such as the α-galactosidase from Bacillus megaterium [ 18 ], Yersinia pestisbiovar Microtus str . 91,001 [ 30 ], Aspergillus oryzae YZ1 [ 22 ] and Paceilomyces thermophila [ 19 ]. Thus, the enzyme showed activity and stability over a broad range of temperature, which made it a potential candidate in various industrial processes.…”

Section: Resultsmentioning

confidence: 99%

Identification and Characterization of a Thermostable GH36 α-Galactosidase from Anoxybacillus vitaminiphilus WMF1 and Its Application in Synthesizing Isofloridoside by Reverse Hydrolysis

Wang

Cao

Chen

et al. 2021

IJMS

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An α-galactosidase-producing strain named Anoxybacillus vitaminiphilus WMF1, which catalyzed the reverse hydrolysis of d-galactose and glycerol to produce isofloridoside, was isolated from soil. The α-galactosidase (galV) gene was cloned and expressed in Escherichia coli. The galV was classified into the GH36 family with a molecular mass of 80 kDa. The optimum pH and temperature of galV was pH 7.5 and 60 °C, respectively, and it was highly stable at alkaline pH (6.0–9.0) and temperature below 65 °C. The specificity for p-nitrophenyl α-d-galactopyranoside was 70 U/mg, much higher than that for raffinose and stachyose. Among the metals and reagents tested, galV showed tolerance in the presence of various organic solvents. The kinetic parameters of the enzyme towards p-nitrophenyl α-d-galactopyranoside were obtained as Km (0.12 mM), Vmax (1.10 × 10−3 mM s−1), and Kcat/Km (763.92 mM−1 s−1). During the reaction of reverse hydrolysis, the enzyme exhibited high specificity towards the glycosyl donor galactose and acceptors glycerol, ethanol and ethylene glycol. Finally, the isofloridoside was synthesized using galactose as the donor and glycerol as the acceptor with a 26.6% conversion rate of galactose. This study indicated that galV might provide a potential enzyme source in producing isofloridoside because of its high thermal stability and activity.

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Cloning and Functional Expression of an α-Galactosidase fromYersinia pestis biovar Microtusstr. 91001

Cited by 10 publications

References 12 publications

Crystal Structure of α-Galactosidase from Lactobacillus acidophilus NCFM: Insight into Tetramer Formation and Substrate Binding

Crystal Structure of α-Galactosidase from Lactobacillus acidophilus NCFM: Insight into Tetramer Formation and Substrate Binding

Characterization of a GH36 α-D-Galactosidase Associated with Assimilation of Gum Arabic in <i>Bifidobacterium longum </i>subsp.<i> longum </i>JCM7052

Identification and Characterization of a Thermostable GH36 α-Galactosidase from Anoxybacillus vitaminiphilus WMF1 and Its Application in Synthesizing Isofloridoside by Reverse Hydrolysis

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