Purification and Characterization of a New Type of α-Glucosidase from<i>Paecilomyces lilacinus</i>That Has Transglucosylation Activity to Produce α-1,3- and α-1,2-Linked…

Kobayashi, Isao; Tokuda, Masahiro; Hashimoto, Hiroyuki; Konda, Takashi; Nakano, Hirofumi; Kitahata, Sumio

doi:10.1271/bbb.67.29

Cited by 37 publications

(27 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…1, lanes c and d). The molecular weight estimated by SDS-PAGE and gel filtration column was 56 and 52 kDa, respectively, which is similar to the molecular weight of 54 kDa presented by the monomeric a-glucosidase from Paecilomyces lilacinus (Kobayashi et al 2003). It is noteworthy that gel filtration analysis of dimeric fungal a-glucosidases from Mortierella alliacea, Chaetomium thermophilum var.…”

Section: Resultssupporting

confidence: 72%

“…coprophilum and Aspergillus nidulans presented molecular weights of 92, 107 and 129 kDa (Tanaka et al 2002;Giannesi et al 2006;Kato et al 2002). Electrofocusing of the purified aglucosidase of A. niveus revealed a pI of 6.8, which is similar to value of the a-glucosidase from Mortierella alliceae (pI 6.3) (Tanaka et al 2002), in contrast, an a-glucosidase from Paecilomyces lilacinus presented a pI of 9.1 (Kobayashi et al 2003). The A. niveus a-glucosidase is a glycoprotein, and contains 29% carbohydrate, which is in agreement with other fungal a-glucosidases (Giannesi et al 2006;Yamamoto et al 2004).…”

Section: Resultsmentioning

confidence: 82%

“…1 Electrophoresis analysis of purified a-glucosidase: a molecular weight ladder; b purified a-glucosidase in 10% SDS-PAGE; c developed for glucoamylase activity using iodine; d 5% PAGE of the purified a-glucosidase revealed with comassie blue lower than that reported to Chaetomium thermophilum var. coprophilum (Giannesi et al 2006) and Paecilomyces lilacinus (Kobayashi et al 2003). The classification of the A. niveus enzyme as an a-glucosidase was based both on the hydrolysis of synthetic substrate (a-PNPG), a specific substrate for a-glucosidase (Yamamoto et al 2004;Mackenzie et al 2000), and the products of starch (see Fig.…”

Section: Hydrolysis Of Different Substrates and Kinetic Parametersmentioning

confidence: 99%

See 2 more Smart Citations

Purification and biochemical characterization of a novel α-glucosidase from Aspergillus niveus

Silva

Michelin

Damásio

et al. 2009

Antonie van Leeuwenhoek

View full text Add to dashboard Cite

An extracellular a-glucosidase produced by Aspergillus niveus was purified using DEAE-Fractogel ion-exchange chromatography and Sephacryl S-200 gel filtration. The purified protein migrated as a single band in 5% PAGE and 10% SDS-PAGE. The enzyme presented 29% of glycosylation, an isoelectric point of 6.8 and a molecular weight of 56 and 52 kDa as estimated by SDS-PAGE and Bio-Sil-Sec-400 gel filtration column, respectively. The enzyme showed typical alpha-glucosidase activity, hydrolyzing p-nitrophenyl alpha-D-glucopyranoside and presented an optimum temperature and pH of 65 degrees C and 6.0, respectively. In the absence of substrate the purified alpha-glucosidase was stable for 60 min at 60 degrees C, presenting t(50) of 90 min at 65 degrees C. Hydrolysis of polysaccharide substrates by alpha-glucosidase decreased in the order of glycogen, amylose, starch and amylopectin. Among malto-oligosaccharides the enzyme preferentially hydrolyzed malto-oligosaccharide (G10), maltopentaose, maltotetraose, maltotriose and maltose. Isomaltose, trehalose and beta-ciclodextrin were poor substrates, and sucrose and alpha-ciclodextrin were not hydrolyzed. After 2 h incubation, the products of starch hydrolysis measured by HPLC and thin layer chromatography showed only glucose. Mass spectrometry of tryptic peptides revealed peptide sequences similar to glucan 1,4-alpha-glucosidases from Aspergillus fumigatus, and Hypocrea jecorina. Analysis of the circular dichroism spectrum predicted an a-helical content of 31% and a beta-sheet content of 16%, which is in agreement with values derived from analysis of the crystal structure of the H. jecorina enzyme.

show abstract

Section: Resultssupporting

confidence: 72%

Section: Resultsmentioning

confidence: 82%

Section: Hydrolysis Of Different Substrates and Kinetic Parametersmentioning

confidence: 99%

See 1 more Smart Citation

Purification and biochemical characterization of a novel α-glucosidase from Aspergillus niveus

Silva

Michelin

Damásio

et al. 2009

Antonie van Leeuwenhoek

View full text Add to dashboard Cite

show abstract

“…As described previously (Chiba 1997), the glucosyl residue is replaced by a proton from H 2 O or an acceptor, namely an exchange reaction between the glucosyl residue and the proton in both hydrolysis and transglycosylation. Various α-glucosidases can transfer glucosyl groups to 6-OH of the acceptor to synthesize isomaltose and panose, or to other hydroxyl groups (2-OH, 3-OH, 4-OH), such as the α-glucosidase from Paecilomyces lilacinus (Kobayashi et al 2003). In this study, with 400 g/L maltose and 0.05 mg/mL recombinant α-glucosidase presented at the reaction solution, 42% maltose was transferred to isomaltose, panose and isomaltriose (about 168 g/L) after 10 h. The results strongly suggested that the recombinant α-glucosidase from T. ethanolicus JW200 was able to transfer glucosyl groups to the 4-OH and 6-OH of a glucose residue.…”

Section: Biochemical Characterization Of the Recombinant α-Glucosidasementioning

confidence: 99%

Expression and characterization of an α-glucosidase from Thermoanaerobacter ethanolicus JW200 with potential for industrial application

et al. 2009

View full text Add to dashboard Cite

In this study, a new α-glucosidase gene from Thermoanaerobacter ethanolicus JW200 was cloned and expressed in Escherichia coli by a novel heat-shock vector pHsh. The recombinant α-glucosidase exhibited its maximum hydrolytic activity at 70• C and pH 5.0∼5.5. With p-nitrophenyl-α-D-glucoside as a substrate and under the optimal condition (70 • C, pH 5.5), Km and Vmax of the enzyme was 1.72 mM and 39 U/mg, respectively. The purified α-glucosidase could hydrolyze oligosaccharides with both α-1,4 and α-1,6 linkages. The enzyme also had strong transglycosylation activity when maltose was used as sugar donor. The transglucosylation products towards maltose are isomaltose, maltotriose, panose, isomaltotriose and tetrasaccharides. The enzyme could convert 400 g/L maltose to oligosaccharides with a conversion rate of 52%, and 83% of the oligosaccharides formed were prebiotic isomaltooligosaccharides (containing isomaltose, panose and isomaltotriose).

show abstract

“…12) In bacterial -glucosidases, a small number of enzymes have been found to catalyze transglucosylation reactions 13,14) that are utilized in biotechnology to produce food oligosaccharides 15,16) or to conjugate sugars with biologically useful materials but these enzyme properties cannot explain the formation of the highly branched -glucan.…”

mentioning

confidence: 99%

Purification and Characterization of Highly Branched α-Glucan–Producing Enzymes fromPaenibacillussp. PP710

Tsusaki

Watanabe

Yamamoto

et al. 2012

Bioscience, Biotechnology, and Biochemistry

View full text Add to dashboard Cite

Purification and Characterization of a New Type of α-Glucosidase fromPaecilomyces lilacinusThat Has Transglucosylation Activity to Produce α-1,3- and α-1,2-Linked…

Cited by 37 publications

References 15 publications

Purification and biochemical characterization of a novel α-glucosidase from Aspergillus niveus

Purification and biochemical characterization of a novel α-glucosidase from Aspergillus niveus

Expression and characterization of an α-glucosidase from Thermoanaerobacter ethanolicus JW200 with potential for industrial application

Purification and Characterization of Highly Branched α-Glucan–Producing Enzymes fromPaenibacillussp. PP710

Contact Info

Product

Resources

About