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Aspergillus oryzae β galactosidase

Manufactured by Merck Group
Sourced in United States, Germany

Aspergillus oryzae β-galactosidase is an enzyme derived from the fungus Aspergillus oryzae. It catalyzes the hydrolysis of lactose into glucose and galactose.

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4 protocols using aspergillus oryzae β galactosidase

1

Neem Leaf β-Galactosidase Assay

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Aspergillus oryzae β-galactosidase, ONPG and zinc acetate dihydrate was obtained from Sigma Aldrich, USA. Azadirachta indica (neem) leaves were collected from nearby garden. All reagents were used without further purifications.
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2

Synthesis and Characterization of Glycans

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Lacto-N-fucopentaose I (LNFP I: Fucα1-2Galβ1-3GlcNAcβ1-3Galβ1-4Glc), p-nitrophenyl 6-O-sulfo-β-GlcNAc (GlcNAc6Sβ-pNP), and GlcNAcβ-O-Ser were purchased from Carbosynth. GlcNAcβ-pNP was obtained from Nacalai Tesque, and core 2-pNP [Galβ1-3(GlcNAcβ1-6)GalNAcα-pNP] was obtained from Toronto Research Chemicals. N,N′-Diacetylchitobiose (GlcNAcβ1-4GlcNAc), core 2-Thr [Galβ1-3(GlcNAcβ1-6)GalNAcα-O-Thr], core 3-Thr [GlcNAcβ1-3GalNAcα-O-Thr], and core 4-Thr [GlcNAcβ1-3(GlcNAcβ1-6)GalNAcα-O-Thr] were purchased from Tokyo Chemical Industry. GalNAcβ-pNP, GlcNAcβ1-2Man, and porcine gastric mucin (PGM, type III) were from Sigma-Aldrich. PGM was dialyzed against water and lyophilized prior to use. Lacto-N-triose II (GlcNAcβ1-3Galβ1-4Glc) was prepared by digesting lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc, LNnT) by Aspergillus oryzae β-galactosidase (Sigma-Aldrich) followed by separation with Toyopearl HW-40C gel chromatography (20 mm × 600 mm) (Tosoh). LNnT was a gift from Glycom A/S. The purity was confirmed by high-performance anion exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) as described later.
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3

Enzymatic Conversion of Lactose

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Aspergillus oryzae β-galactosidase (E. C. 3.2.1.23), lactulose, d-fructose, 6-galactobiose, and 50% glutaraldehyde were purchased from Sigma-Aldrich (Steinheim, Germany). Sepharose 4B was supplied by Pharmacia Biotechnology (Uppsala, Sweden). d-galactose, raffinose and o-nitrophenyl-β-d-galactopyranoside (ONPG) from Fluka (Steimheim, Germany). Fructooligosaccharides (Orafti P95) and inulin (Orafti HP) were kindly gifted by Orafti (Tienen, Belgium).
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4

Recombinant Sialidase Expression and Purification

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Recombinant sialidases were expressed and purified as reported previously for human cytosolic sialidase hNEU2,54 (link) as well as bacterial sialidases from Streptococcus pneumoniae (SpNanA,52 SpNanB,52 and SpNanC18 (link)), Pasteurella multocida sialyltransferase 1 with α2–3-sialidase activity (PmST1),41 (link) and Bifidobacterium infantis sialidase BiNanH2.53 (link) Commercially available bacterial sialidases including those from Arthrobacter ureafaciens (Prozyme), Clostridium perfringens CpNanI (Sigma-Aldrich), and Vibrio cholerae were from Sigma-Aldrich. Aspergillus oryzae β-galactosidase was purchased from Sigma-Aldrich. Pasteurella multocida sialic acid aldolase (PmNanA),39 (link)Neisseria meningitidis CMP-sialic acid synthetase (NmCSS),40 (link) and Pasteurella multocida sialyltransferase 1 M144D mutant (PmST1_M144D)41 (link),42 (link) were expressed and purified as described previously. Siaα2–3GalβpNP44 (link) used for substrate specificity studies were synthesized as described previously.
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