Optimization of the Cellulase Free Xylanase Production by Immobilized Bacillus Pumilus

The extracellular xylanase secreted by microorganisms is a hydrolytic enzyme, which arbitrarily cleaves the β-1, 4 backbone of the polysaccharide xylan; an enzyme used in the food processing, bio-pulping and bio-bleaching. The commercial production of the xylanase is limited because of a higher cost involvement, which can be overcome by the cost-effective production of the xylanase through immobilization of the microbial cell by the non-toxic substances. In this work, the optimization of the extra-cellular cellulase free xylanase production by the immobilized cell of the Bacillus pumilus IMAU80221 strain using Ca-alginate beads along with standardization of the various parameters for a higher xylanase production were studied. Following to sterilization, the Na-alginate solution was mixed with the bacterial suspension of the Bacillus pumilus IMAU80221 and was added drop by drop into the 1 M calcium chloride solution for 1 h for obtaining a uniform sized polymeric bead of the Ca-alginate. For xylanase production, the Ca-alginate beads were then transferred into 100 mL Erlenmeyer flasks with 20 mL of the culture medium containing (w/v) 0.02% NaCl, 0.02% MgSO4, 0.04% (KH4)2PO4, 0.1% peptone, and 0.5% xylan and incubated at 34 °C in an incubator shaker (150 rpm) for 24 h. The resultant supernatant (crude enzyme) was used for enzyme assay. The maximum xylanase production by the free cell (1.9 U.mL-1.min-1) was recorded at 48 h which was 40.5% lower than the xylanase production by the immobilized cell (2.67 U.mL-1.min-1) at the same time. The beads containing the immobilized cells could be reused up to eight fermentation cycles for xylanase production and retained 83.5% of the productivity at the fourth cycle. The entrapped cells were stable after six months of storage at 4 °C and retained 68% of the xylanase productivity. Cellulase free xylanase production from the immobilized Bacillus pumilus IMAU80221 was optimized. The xylanase production by the immobilized cells of Bacillus pumilus was higher by 40.5 and 132.6 % over the free cells respectively after 48 and 72 h of the incubation.