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Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices.

ABSTRACT: This study was conducted to evaluate the efficacy of purified free and immobilized xylanase in enrichment of fruit juices. Extracellular xylanase produced from Bacillus pumilus VLK-1 was purified to apparent homogeneity by 15.4-fold with 88.3 % recovery in a single step using CM-Sephadex C-50. Purified xylanase showed a single band on SDS-polyacrylamide gel with a molecular mass of 22.0 kDa. The purified enzyme was immobilized on glutaraldehyde-activated aluminum oxide pellets and the immobilization process parameters were optimized statistically through response surface methodology. The bound enzyme displayed an increase in optimum temperature from 60 to 65?ºC and pH from 8.0 to 9.0. The pH and temperature stability of the enzyme was also enhanced after immobilization. It could be reused for 10 consecutive cycles with 58 % residual enzyme activity. The potential of purified xylanase (free and immobilized) in juice enrichment from grape (Vitis amurensis) and orange (Citrus sinensis) pulps has been investigated. The optimization of this process using free xylanase revealed maximum juice yield, clarity and reducing sugar on treatment with 20 IU/g fruit pulp for 30 min at 50?ºC. Treatment of both the fruit pulps with xylanase under optimized conditions resulted in an increase in juice yield, clarity, reducing sugars, titratable acidity, and filterability but a decline in turbidity and viscosity. Immobilized enzyme was more effective in improving juice quality as compared to its soluble counterpart. The results showed B. pumilus VLK-1 xylanase, in both free and immobilized form, as a potential candidate for use in fruit juice enrichment.

SUBMITTER: Kumar L

PROVIDER: S-EPMC4152508 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices.

Kumar Lalit L Nagar Sushil S Mittal Anuradha A Garg Neelam N Gupta Vijay Kumar VK

Journal of food science and technology 20140204 9

This study was conducted to evaluate the efficacy of purified free and immobilized xylanase in enrichment of fruit juices. Extracellular xylanase produced from Bacillus pumilus VLK-1 was purified to apparent homogeneity by 15.4-fold with 88.3 % recovery in a single step using CM-Sephadex C-50. Purified xylanase showed a single band on SDS-polyacrylamide gel with a molecular mass of 22.0 kDa. The purified enzyme was immobilized on glutaraldehyde-activated aluminum oxide pellets and the immobiliza ...[more]

PMID: 25190829

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Project description:BackgroundThe 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.ObjectivesIn 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.Materials and methodsFollowing 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.ResultsThe 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.ConclusionCellulase 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.

Dataset Information

Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices.

Publications

Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices.

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