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Immobilization of thermostable, bacterial cellulase from Stenotrophomonas maltophilia in agar-agarose matrices and its characterization

By: Rohini Tamilanban, Sam Ebenezer Rajadas, Vignesh Sounderrajan, Shakila Harshavardhan

Key Words: Thermal stability, Agarose, Agar, Immobilization, Entrapment.

Int. J. Biosci. 13(3), 198-208, September 2018.

DOI: http://dx.doi.org/10.12692/ijb/13.3.198-208

Certification: ijb 2018 0002 [Generate Certificate]

Abstract

Cellulase the biocatalyst for conversion of cellulose into simple sugars is gaining global economic importance owing to its wide application in various industrial and clinical domains. Commercial cellulase is preferred to be stable in adverse conditions and to be recovered from the reactor once the process is done. Hence, a search for such a cellulase still exists even though it was discovered six decades ago. In this study, purified thermostable cellulase from Stenotrophomonas maltophilia was immobilized on an agar-agarose matrix, and the properties of immobilized cellulase were studied. The optimum temperature and pH for immobilized enzyme activity were found to be 50oC and 8.0 respectively. The immobilized enzyme exhibited its stability at much wider alkaline pH ranges and higher temperatures even after 24 hours incubation. Km and Vmax values of the immobilized enzyme were 6.618 mg/ml and 131.578 µmol/min/mg of protein respectively. Both free and immobilized forms of enzymes were inhibited significantly by Hg2+ metal ion and the activity of the latter was affected in the presence of detergents and additives at higher concentration. The agar-agarose immobilized enzyme could be reused up to 5 repeated cycles and it is stable for at least 1 month when stored at 4oC. Hence this immobilized cellulase with good storage stability than the soluble one can be considered for commercial applications.

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Immobilization of thermostable, bacterial cellulase from Stenotrophomonas maltophilia in agar-agarose matrices and its characterization

Abraham RE, Verma ML, Barrow CJ, Puri M. 2014. Suitability of magnetic nanoparticle immobilised cellulases in enhancing enzymatic saccharification of pretreated hemp biomass. Biotechnology for biofuels 7, 90.

http://dx.doi.org/10.1186/1754-6834-7-90.

Alahakoon T, Koh JW, Chong XWC,  Lim WTL. 2012. Immobilization of cellulases on amine and aldehyde functionalized Fe2O3 magnetic nanoparticles. Preparative Biochemistry and Biotechnology 42, 234–248.

http://dx.doi.org/10.1080/10826068.2011.602800.

Barbara K, Maciej L, Wiesawa Z. 1990. Urease immobilized on chitosan membrane: Preparation and properties. Journal of Chemical Technology & Biotechnology 48, 337–350.

http://dx.doi.org/10.1002/jctb.280480309.

Bhat MK. 2000. Cellulases and related enzymes in biotechnology. Biotechnology advances 18, 355–383.

Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.

http://dx.doi.org/10.1016/0003-2697(76)90527-3.

Cherry JR, Fidantsef AL. 2003. Directed evolution of industrial enzymes: an update. Current opinion in biotechnology 14, 438–443.

http://dx.doi.org/10.1016/S0958-1669(03)00099-5

Dey G, Nagpal V, Banerjee R. 2002. Immobilization of alpha-amylase from Bacillus circulans GRS 313 on coconut fiber. Applied

biochemistry and biotechnology 102–103, 303–313.

http://dx.doi.org/10.1385/ABAB:102-103:1-6:303.

Dinçer A, Telefoncu A. 2007. Improving the stability of cellulase by immobilization on modified polyvinyl alcohol coated chitosan beads. Journal of Molecular Catalysis B: Enzymatic 45, 10–14.

http://dx.doi.org/10.1016./j.molcatb.2006.10.005.

El-hadi AA, Kamel Z, Hammad A, El-nour SA, Anwar M. 2014. Optimization of Aspergillus hortai Cellulase Immobilized on Poly ( Acrylamide-Co-Acrylic Acid ) for Hydroxylation of Cellulose Rice Straw Wastes. Global Journal of Pharmacology  8, 681–687.

http://dx.doi.org/10.5829/idosi.gjp.2014.8.4.86175.

Guisan JM. 2006. Immobilization of Enzymes and Cells. Humana Press, Totowa, NJ.

Gulay S, Sanli-Mohamed G. 2012. Immobilization of thermoalkalophilic recombinant esterase enzyme by entrapment in silicate coated Ca-alginate beads and its hydrolytic properties. International journal of biological macromolecules 50(3), 545–551.

http://dx.doi.org/10.1016/j.ijbiomac.2012.01.017.

Huang PJ, Chang KL, Hsieh JF, Chen ST. 2015. Catalysis of rice straw hydrolysis by the combination of immobilized cellulase from Aspergillus niger on β -Cyclodextrin-Fe3O4 nanoparticles and ionic liquid. BioMed Research International 2015.

http://dx.doi.org/10.1155/2015.409103.

Iqbal H, Ahmed I, Zia MA, Irfan M. 2011. Purification and characterization of the kinetic parameters of cellulase produced from wheat straw by Trichoderma viride under SSF and its detergent compatibility. Advances in Bioscience and Biotechnology 2, 149–156.

http://dx.doi.org/10.4236/abb.2011.23024.

Jordan J, Kumar CSSR, Theegala C. 2011. Preparation and characterization of cellulase-bound magnetite nanoparticles. Journal of Molecular Catalysis B: Enzymatic 68, 139–146.

Khoshnevisan K, Bordbar AK, Zare D, Davoodi D, Noruzi M, Barkhi M, Tabatabaei M. 2011. Immobilization of cellulase enzyme on superparamagnetic nanoparticles and determination of its activity and stability. Chemical Engineering Journal 171, 669–673.

Kim JY, Hur SH, Hong JH. 2005. Purification and characterization of an alkaline cellulase from a newly isolated  alkalophilic Bacillus sp. HSH-810. Biotechnology letters 27, 313–316.

http://dx.doi.org/10.1007/s10529-005-0685-5.

Kim KC, Yoo SS, Oh YA, Kim SJ. 2003. Isolation and characteristics of Trichoderma harzianum FJ1 producing cellulases and xylanase. Journal of Microbiology and Biotechnology 13, 1–8.

Krajewska B. 2004. Application of chitin- and chitosan-based materials for enzyme immobilizations: a review. Enzyme and Microbial Technology 35,  126–139.

http://dx.doi.org/10.1016/j.enzmictec.2003.12.013.

Kumar A, Singh S, Nain L. 2018. Magnetic Nanoparticle Immobilized Cellulase Enzyme for Saccharification of Paddy Straw. International journal of Current Microbiology and Applied Sciences 7, 881–893.

http://dx.doi.org/10.20546/ ijcmas.2018.704.095.

Lalonde J, Margolin A. 2008. Immobilization of Enzymes. Enzyme Catalysis in Organic Synthesis. http://dx.doi.org/10.1002/ 9783527618262.ch6.

Lupoi JS, Smith EA. 2011. Evaluation of nanoparticle-immobilized cellulase for improved ethanol yield in simultaneous saccharification and fermentation reactions. Biotechnology and Bioengineering 108, 2835–2843.

http://dx.doi.org/10.1002/bit.23246.

Miller GL. 1959. Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Analytical Chemistry 31, 426–428.

http://dx.doi.org/10.1021/ac60147a030.

 Perez J, Munoz-Dorado J, de la Rubia T, Martinez J. 2002. Biodegradation and biological treatments of cellulose, hemicellulose and lignin:  an overview. International microbioligy: the official journal of the Spanish Society for Microbiology 5, 53–63.

http://dx.doi.org/10.1007/s10123-002-0062-3.

Prakash O, Jaiswal N. 2011. Immobilization of a thermostable α-amylase on agarose and agar matrices and its application in starch stain removal. World Applied Sciences Journal 13, 572–577.

Prasad M, Palanivelu P. 2013. A novel method for the immobilization of a thermostable fungal chitinase and the properties of the immobilized enzyme. Biotechnology and Applied Biochemistry 61, 441–445.

http://dx.doi.org/10.1002.bab.1179.

Roger AS. 2007. Enzyme Immobilization: The Quest for Optimum Performance. Advanced Synthesis & Catalysis 349, 1289–1307.

http://dx.doi.org/10.1002.adsc.200700082.

Rohini T, Santhi VS, Saranya S,  Sam ebenezer R, Shakila H. 2017. Optimization of AFEX Pretreated Agrowaste Media for Endoglucanase and Xylanase Production by Stenotrophomonas maltophilia. International Journal For Research In Applied Science & Enginnering Technology 5, 2361–2374.

http://dx.doi.org/10.4103.ijmr.

Tamilanban R, Velayudhan SS, Rajadas SE, Harshavardhan S. 2017. Purification and characterization of an extracellular cellulase produced using alkali pretreated rice straw by Stenotrophomonas maltophilia. International Journal of Biology Research 2, 2455–6548.

Tao QL, Li Y, Shi Y, Liu RJ, Zhang YW, Guo J. 2016. Application of Molecular Imprinted Magnetic Fe3O4@SiO2 Nanoparticles for Selective Immobilization of Cellulase. Journal of nanoscience and nanotechnology 16, 6055–6060.

Teather RM, Wood PJ. 1982. Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Applied and Environmental Microbiology 43, 777–780.

Tumturk H, demirel G, Altinok H, Aksoy S, Hasirci N. 2008. Immobilization Of Glucose Isomerase In Surface-Modified Alginate Gel Beads. Journal of Food Biochemistry 32, 234–246.

http://dx.doi.org/10.1111/j.1745-4514.2008.00171.X.

Tümtürk H, Karaca N, Demirel G, Şahin F. 2007. Preparation and application of poly(N,N-dimethylacrylamide-co-acrylamide) and poly(N-isopropylacrylamide-co-acrylamide)/κ-Carrageenan hydrogels for immobilization of lipase. International Journal of Biological Macromolecules 40, 281–285.

Wen Z, Liao W, Chen S. 2005. Production of cellulase by Trichoderma reesei from dairy manure. Bioresource technology 96, 491—499.

http://dx.doi.org/10.1016/j.biortech.2004.05.021.

Xuepu M, Gangjun G, Jinfeng H, Zhiyun D, Zhishu H, Lin M, Pei L, Lianquan G. 2005. A novel method to prepare chitosan powder and its application in cellulase immobilization. Journal of Chemical Technology & Biotechnology 81, 189–195. http://dx.doi.org/10.1002/bit.260370912.

Zhongliang S, Xingyu Y, Luli, Hongbo S, Shitao Y. 2012. Cellulase immobilization properties and their catalytic effect on cellulose hydrolysis in ionic liquid. African Journal of Microbiology Research 6, 64–70.

http://dx.doi.org/10.5897/AJMR11.922.

Rohini Tamilanban, Sam Ebenezer Rajadas, Vignesh Sounderrajan, Shakila Harshavardhan.
Immobilization of thermostable, bacterial cellulase from Stenotrophomonas maltophilia in agar-agarose matrices and its characterization.
Int. J. Biosci. 13(3), 198-208, September 2018.
https://innspub.net/ijb/immobilization-thermostable-bacterial-cellulase-stenotrophomonas-maltophilia-agar-agarose-matrices-characterization/
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