[Purification and physico-chemical properties of glycosidase of Aspergillus niger 185sh].
A scheme has been developed for isolation and purification of the enzyme with alpha-N-acetylgalactosaminidase and alpha-galactosidase activities which included fractionation by ammonium sulphate and chromatography on TSK-gels Toyopearl HW-60 and Fractogel DEAE-650-s and Sepharose 6B. The enzyme was purified 600 times with the yield of 28%. The enzyme preparation did not contain fucosidase, invertase and proteolytic activities. Molecular mass of the enzyme from the data of gel-filtration on Sepharose 6B was 430 kDa, according to the data of electrophoresis in DS-PAAG--70 kDa. It is shown that acidic and hydrophobic aminoacids prevail in the enzyme molecule, the carbohydrate component containing galactose, mannose, glucosamine and two nonidentified hexosamines is also present there. The enzyme preparation is stable during 48 hours at 20 degrees C; its pH-optimum is at pH 3.5-4.1. Michaelis constants concerning n-nitrophenyl-alpha-N-acetylgalactopyranoside and n-nitrophenyl-alpha-D-galactopyranoside were 1.18 and 1.25 mM, respectively.
Borzova
N V
NV
Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, 154 Zabolotny St., Kyiv, 03143, Ukraine.
Varbanets'
L D
LD
D000645:Ammonium Sulfate; D001234:Aspergillus niger / Q000201:enzymology*; D002850:Chromatography, Gel; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006596:Hexosaminidases / Q000737:chemistry / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D011108:Polymers; D012685:Sepharose; D013696:Temperature; D000519:alpha-Galactosidase / Q000737:chemistry / Q000302:isolation & purification; D048809:alpha-N-Acetylgalactosaminidase
D011108:Polymers; D012685:Sepharose; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D000519:alpha-Galactosidase; D048809:alpha-N-Acetylgalactosaminidase; D000645:Ammonium Sulfate