Purification and characterization of a new type of alpha-glucosidase from Paecilomyces lilacinus that has transglucosylation activity to produce alpha-1,3- and alpha-1,2-linked oligosaccharides.
A fungus producing an alpha-glucosidase that synthesizes alpha-1,3- and alpha-1,2-linked glucooligosaccharides by transglucosylation was isolated and identified as Paecilomyces lilacinus. The cell-bound enzyme responsible for the synthesis was extracted by suspension of mycelia with 0.1 M phosphate buffer (pH 8.0), and the extract was purified. The molecular weight and the isoelectric point were estimated to be 54,000 and 9.1, respectively. The enzyme was most active at pH 5.0 and 65 degres C. The enzyme hydrolyzed maltose, nigerose, and kojibiose. The enzyme also hydrolyzed soluble starch and amylose with the rate toward maltose. p-Nitro-phenyl alpha-glucoside and isomaltose were not good substrates. The enzyme had high transglucosylation activity to synthesize oligosaccharides containing alpha-1,3- and alpha-1,2-linkages. At an early stage of the reaction, considerable maltotriose, 4-O-alpha-nigerosyl-D-glucose, and 4-O-alpha-kojibiosyl-D-glucose were synthesized. Afterwards, nigerose and kojibiose were accumulated gradually with glucose as an acceptor.
Kobayashi
Isao
I
Honen Corporation, 1-2-3 Ohtemachi, Chiyoda-ku, Tokyo 100-8150, Japan. ikobayashi@honen.co.jp
Tokuda
Masahiro
M
Hashimoto
Hiroyuki
H
Konda
Takashi
T
Nakano
Hirofumi
H
Kitahata
Sumio
S
D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D003470:Culture Media; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D010143:Paecilomyces / Q000254:growth & development / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D000520:alpha-Glucosidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D003470:Culture Media; D008670:Metals; D009844:Oligosaccharides; D000520:alpha-Glucosidases