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A digestive beta-glucosidase from the silkworm, Bombyx mori: cDNA cloning, expression and enzymatic characterization.
Comp Biochem Physiol B Biochem Mol Biol. 2005 Aug; 141(4):418-27.CB

Abstract

A digestive beta-glucosidase cDNA was cloned from the silkworm, Bombyx mori. The B. mori beta-glucosidase cDNA contains an open reading frame of 1473 bp encoding 491 amino acid residues. The B. mori beta-glucosidase possesses the amino acid residues involved in catalysis and substrate binding conserved in glycosyl hydrolase family 1. Southern blot analysis of genomic DNA suggested the B. mori beta-glucosidase to be a single gene. Northern blot analysis of B. mori beta-glucosidase gene confirmed larval midgut-specific expression. The B. mori beta-glucosidase mRNA expression in larval midgut was detectable only during feeding period, whereas its expression was downregulated during starvation. The B. mori beta-glucosidase cDNA was expressed as a 57-kDa polypeptide in baculovirus-infected insect Sf9 cells, and the recombinant beta-glucosidase was active on cellobiose and lactose, but not active on salicin, indicating that the B. mori beta-glucosidase possesses the characteristics of the Class 2 enzyme. The enzyme activity of the purified recombinant beta-glucosidase expressed in baculovirus-infected insect cells was approximately 665 U per microg of recombinant B. mori beta-glucosidase. The purified recombinant B. mori beta-glucosidase showed the highest activity at 35 degrees C and pH 6.0, and were stable at 50 degrees C at least for 10 min. Treatment of recombinant virus-infected Sf9 cells with tunicamycin, a specific inhibitor of N-glycosylation, revealed that the recombinant B. mori beta-glucosidase is N-glycosylated, but the carbohydrate moieties are not essential for enzyme activity.

Authors+Show Affiliations

College of Natural Resources and Life Science, Dong-A University, Busan 604-714, Korea.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15970451

Citation

Byeon, Gyeong Min, et al. "A Digestive Beta-glucosidase From the Silkworm, Bombyx Mori: cDNA Cloning, Expression and Enzymatic Characterization." Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology, vol. 141, no. 4, 2005, pp. 418-27.
Byeon GM, Lee KS, Gui ZZ, et al. A digestive beta-glucosidase from the silkworm, Bombyx mori: cDNA cloning, expression and enzymatic characterization. Comp Biochem Physiol B, Biochem Mol Biol. 2005;141(4):418-27.
Byeon, G. M., Lee, K. S., Gui, Z. Z., Kim, I., Kang, P. D., Lee, S. M., Sohn, H. D., & Jin, B. R. (2005). A digestive beta-glucosidase from the silkworm, Bombyx mori: cDNA cloning, expression and enzymatic characterization. Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology, 141(4), 418-27.
Byeon GM, et al. A Digestive Beta-glucosidase From the Silkworm, Bombyx Mori: cDNA Cloning, Expression and Enzymatic Characterization. Comp Biochem Physiol B, Biochem Mol Biol. 2005;141(4):418-27. PubMed PMID: 15970451.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A digestive beta-glucosidase from the silkworm, Bombyx mori: cDNA cloning, expression and enzymatic characterization. AU - Byeon,Gyeong Min, AU - Lee,Kwang Sik, AU - Gui,Zhong Zheng, AU - Kim,Iksoo, AU - Kang,Pil Don, AU - Lee,Sang Mong, AU - Sohn,Hung Dae, AU - Jin,Byung Rae, PY - 2005/01/13/received PY - 2005/05/02/revised PY - 2005/05/03/accepted PY - 2005/6/23/pubmed PY - 2005/10/19/medline PY - 2005/6/23/entrez SP - 418 EP - 27 JF - Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology JO - Comp. Biochem. Physiol. B, Biochem. Mol. Biol. VL - 141 IS - 4 N2 - A digestive beta-glucosidase cDNA was cloned from the silkworm, Bombyx mori. The B. mori beta-glucosidase cDNA contains an open reading frame of 1473 bp encoding 491 amino acid residues. The B. mori beta-glucosidase possesses the amino acid residues involved in catalysis and substrate binding conserved in glycosyl hydrolase family 1. Southern blot analysis of genomic DNA suggested the B. mori beta-glucosidase to be a single gene. Northern blot analysis of B. mori beta-glucosidase gene confirmed larval midgut-specific expression. The B. mori beta-glucosidase mRNA expression in larval midgut was detectable only during feeding period, whereas its expression was downregulated during starvation. The B. mori beta-glucosidase cDNA was expressed as a 57-kDa polypeptide in baculovirus-infected insect Sf9 cells, and the recombinant beta-glucosidase was active on cellobiose and lactose, but not active on salicin, indicating that the B. mori beta-glucosidase possesses the characteristics of the Class 2 enzyme. The enzyme activity of the purified recombinant beta-glucosidase expressed in baculovirus-infected insect cells was approximately 665 U per microg of recombinant B. mori beta-glucosidase. The purified recombinant B. mori beta-glucosidase showed the highest activity at 35 degrees C and pH 6.0, and were stable at 50 degrees C at least for 10 min. Treatment of recombinant virus-infected Sf9 cells with tunicamycin, a specific inhibitor of N-glycosylation, revealed that the recombinant B. mori beta-glucosidase is N-glycosylated, but the carbohydrate moieties are not essential for enzyme activity. SN - 1096-4959 UR - https://www.unboundmedicine.com/medline/citation/15970451/A_digestive_beta_glucosidase_from_the_silkworm_Bombyx_mori:_cDNA_cloning_expression_and_enzymatic_characterization_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1096-4959(05)00089-8 DB - PRIME DP - Unbound Medicine ER -