Expression and characterization of Pichia etchellsii beta-glucosidase in Escherichia coli

Gene
M Pandey, S Mishra

Abstract

The beta-glucosidase enzyme is important as the terminal enzyme involved in hydrolysis of cellobiose and short-chain cellodextrins generated during enzymatic cellulose degradation. Under controlled reaction conditions the enzyme also displays cello-oligosaccharide synthesizing ability (based on either the thermodynamic or kinetic approach). We present here the purification of the enzyme beta-glucosidase (BGL) of Pichia etchellsii from recombinant pBG55 Escherichia coli clone. The kinetic parameters, substrate specificity and oligosaccharide synthesizing ability of the purified enzyme are also reported. The purified 200-kDa protein (tetramer of 50 kDa) was identified as a broad-substrate-specificity enzyme exhibiting increased temperature and glucose tolerance compared to the native yeast enzyme. Temperature directed substrate specificity for aryl beta,1-4 linkage, and beta (1-2), beta(1-4), beta(1-6) and beta(2-1) linkages in various natural disaccharides was observed. Glycosylation of the enzyme was found to be unimportant for enzyme activity. With both cellobiose and glucose, oligosaccharide synthesis was detected. The implications of this information with regard to cellulose hydrolysis and oligosaccharide synthesis are discu...Continue Reading

References

Jan 1, 1989·Critical Reviews in Biotechnology·V S Bisaria, S Mishra
Apr 27, 1989·Nature·W R Brown
Jan 1, 1967·Journal of Bacteriology·L W Fleming, J D Duerksen
Feb 1, 1993·Enzyme and Microbial Technology·C P KubicekE M Kubicek-Pranz
Nov 1, 1994·The Journal of Applied Bacteriology·I RosiP Domizio

❮ Previous
Next ❯

Citations

Nov 3, 2010·Applied Biochemistry and Biotechnology·Roopa K GaonkarM A Vijayalakshmi
Jun 24, 2003·Biochimica Et Biophysica Acta·Anu Wallecha, Saroj Mishra
Jan 9, 2013·Journal of Agricultural and Food Chemistry·Joelise A FigueiraPedro Fernandes
Oct 26, 2001·Bioscience, Biotechnology, and Biochemistry·S I YunH S Choi
May 3, 2014·Archives of Insect Biochemistry and Physiology·Hatibe Ertürk KaraSelma Sinan
Sep 3, 2005·Biochemical and Biophysical Research Communications·Pranita RoyTapan K Chaudhuri
Dec 19, 2002·Critical Reviews in Biotechnology·Yukti BhatiaV S Bisaria
Jul 6, 2020·Applied Biochemistry and Biotechnology·Jingjing SunJianhua Hao
Nov 16, 2017·International Journal of Biological Macromolecules·Mohammad Asif ShahTapan Kumar Chaudhuri
Apr 16, 2011·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hatibe Ertürk KaraYusuf Turan

❮ Previous
Next ❯

Related Concepts

Related Feeds

Barrett Esophagus

Barrett’s esophagus if a serious complication of gastroesophageal reflux disease during which the normal esophageal lining changes to tissue that resembles intestinal lining. Here is the latest research.