PJB-2018-662
PURIFICATION AND CHARACTERIZATION OF CLONED Β-XYLOSIDASE FROM BACILLUS LICHENIFORMIS ATCC 14580 INTO E. COLI BL21
Asma Zafar
Abstract
Present research work aimed to purify and characterize a recombinant β-xylosidase enzyme which was previously cloned from Bacillus licheniformis ATCC 14580 in to Escherichia coli BL21. Purification of recombinant enzyme was carried out by using ammonium sulphate precipitation method followed by single step immobilized metal ion affinity chromatography. Specific activity of purified recombinant β-xylosidase enzyme was 20.78 Umg-1 with 2.58 purification fold and 33.75% recovery. Molecular weight of purified β-xylosidase was calculated by sodium dodecyl sulphate polyacrylamide gel electrophoresis as 52 kDa. Purified enzyme showed stability up to 90°C within a pH range of 3-8 with and optimal temperature and pH, 55ºC and 7.0, respectively. The enzyme activity was not considerably affected in the presence of EDTA. An increase in the enzyme activity was found in the manifestation of Mg+2. Enzyme activity was also increased by 6%, 18% and 22% in the presence of 1% Tween 80, β-mercaptoethanol and DTT, respectively. Higher concentrations (10 – 40%) of organic solvents did not showed any effect upon activity of enzyme. The recombinant β-xylosidase enzyme is capable to hydrolyze the natural plant biomass like sugarcane bagass, wheat straw and rice straw into simple sugars. So it could be a potential candidate for biofuel industry.
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