Purification and biochemical characterization of a novel α-glucosidase from Aspergillus niveus
作者： Tony Marcio Da Silva , Michele Michelin , Andre Ricardo de Lima Damásio , Alexandre Maller , Fausto Bruno Dos Reis Almeida
DOI: 10.1007/S10482-009-9372-1
关键词:
摘要: An extracellular a-glucosidase produced by Aspergillus niveus was purified using DEAE-Fractogel ion-exchange chromatography and Sephacryl S-200 gel filtration. The protein migrated as a single band in 5% PAGE 10% SDS-PAGE. enzyme presented 29% of glycosylation, an isoelectric point 6.8 molecular weight 56 52 kDa estimated SDS-PAGE Bio-Sil-Sec-400 filtration column, respectively. showed typical alpha-glucosidase activity, hydrolyzing p-nitrophenyl alpha-D-glucopyranoside optimum temperature pH 65 degrees C 6.0, In the absence substrate stable for 60 min at C, presenting t(50) 90 C. Hydrolysis polysaccharide substrates decreased order glycogen, amylose, starch amylopectin. Among malto-oligosaccharides preferentially hydrolyzed malto-oligosaccharide (G10), maltopentaose, maltotetraose, maltotriose maltose. Isomaltose, trehalose beta-ciclodextrin were poor substrates, sucrose alpha-ciclodextrin not hydrolyzed. After 2 h incubation, products hydrolysis measured HPLC thin layer only glucose. Mass spectrometry tryptic peptides revealed peptide sequences similar to glucan 1,4-alpha-glucosidases from fumigatus, Hypocrea jecorina. Analysis circular dichroism spectrum predicted a-helical content 31% beta-sheet 16%, which is agreement with values derived analysis crystal structure H. jecorina enzyme.
springer.com
sci-hub.se 参考文章(29)
J.D. Fontana, M. Gebara, M. Blumel, H. Schneider, C.R. MacKenzie, K.G. Johnson, α-4-O-methyl-d-glucuronidase component of xylanolytic complexes☆ Methods in Enzymology. ,vol. 160, pp. 560- 571 ,(1988) , 10.1016/0076-6879(88)60169-8
Torben P. Frandsen, Birte Svensson, Plant α-glucosidases of the glycoside hydrolase family 31. Molecular properties, substrate specificity, reaction mechanism, and comparison with family members of different origin Plant Molecular Biology. ,vol. 37, pp. 1- 13 ,(1998) , 10.1023/A:1005925819741
A. Lewis Farr, Oliver H. Lowry, Rose J. Randall, Nira J. Rosebrough, Protein Measurement with the Folin Phenol Reagent Journal of Biological Chemistry. ,vol. 193, pp. 265- 275 ,(1951)
Michel. DuBois, K. A. Gilles, J. K. Hamilton, P. A. Rebers, Fred. Smith, Colorimetric Method for Determination of Sugars and Related Substances Analytical Chemistry. ,vol. 28, pp. 350- 356 ,(1956) , 10.1021/AC60111A017
Tetsuya Oguma, Hirokazu Matsui, Masatoshi Tanida, Shoichi Takao, Mamoru Honma, Seiya Chiba, Purification and Substrate Specificity of α-Glucosidase from Paecilomyces varioti AHU 9417 Bioscience, Biotechnology, and Biochemistry. ,vol. 56, pp. 1906- 1910 ,(1992) , 10.1271/BBB.56.1906
Isao KOBAYASHI, Masahiro TOKUDA, Hiroyuki HASHIMOTO, Takashi KONDA, Hirofumi NAKANO, Sumio KITAHATA, Purification and Characterization of a New Type of α-Glucosidase from Paecilomyces lilacinus That Has Transglucosylation Activity to Produce α-1,3- and α-1,2-Linked… Bioscience, Biotechnology, and Biochemistry. ,vol. 67, pp. 29- 35 ,(2003) , 10.1271/BBB.67.29
Takeshi Yamamoto, Takehiro Unno, Yoshimi Watanabe, Mikio Yamamoto, Masayuki Okuyama, Haruhide Mori, Seiya Chiba, Atsuo Kimura, Purification and characterization of Acremonium implicatum α-glucosidase having regioselectivity for α-1,3-glucosidic linkage Biochimica et Biophysica Acta. ,vol. 1700, pp. 189- 198 ,(2004) , 10.1016/J.BBAPAP.2004.05.002
Yoshiki YAMASAKI, Toshio MIYAKE, Yukio SUZUKI, Properties of Crystalline α-Glucosidase from Mucor javanicus Agricultural and biological chemistry. ,vol. 37, pp. 251- 259 ,(1973) , 10.1271/BBB1961.37.251
Tomoyuki Nishimoto, Masayuki Nakano, Shoji Ikegami, Hiroto Chaen, Shigeharu Fukuda, Toshiyuki Sugimoto, Masashi Kurimoto, Yoshio Tsujisaka, Existence of a Novel Enzyme Converting Maltose into Trehalose Bioscience, Biotechnology, and Biochemistry. ,vol. 59, pp. 2189- 2190 ,(1995) , 10.1271/BBB.59.2189
Kazuhiko Maruta, Tetsuya Nakada, Michio Kubota, Hiroto Chaen, Toshiyuki Sugimoto, Masashi Kurimoto, Yoshio Tsujisaka, Formation of trehalose from maltooligosaccharides by a novel enzymatic system. Bioscience, Biotechnology, and Biochemistry. ,vol. 59, pp. 1829- 1834 ,(1995) , 10.1271/BBB.59.1829