Immobilization kinetics of CMCase and β-glucosidase from Aspergillus niger in calcium alginate beads

NEELIMA GARG; DEVENDRA KUMAR; KAUSHLESH K YADAV; MUTHUKUMAR M

doi:10.56093/ijas.v85i1.45937

Authors

NEELIMA GARG Principal Scientist, Division of PHM, Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow 226 101
DEVENDRA KUMAR SRF, Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow 226 101
KAUSHLESH K YADAV SRF, Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow 226 101
MUTHUKUMAR M Scientist, Division of Crop Improvement, Central Institute for Subtropical Horticulture, Rehmankhera, Lucknow 226 101

https://doi.org/10.56093/ijas.v85i1.45937

Keywords:

Aspergillus niger, Î²-glucosidase, CMCase, Immobilization, Mango peel

Abstract

Carboxymethyl cellulase (CMCase) and Î²-glucosidase were produced by Aspergillus niger using mango peel residue as substrate and immobilized on calcium alginate beads through entrapment technique. The catalytic properties of the immobilized CMCase and β-glucosidase were compared to free enzyme. The enzyme efficiency of the immobilized CMCase and β-glucosidase were found to be 89.52 and 86.61%, respectively. Maximum immobilization efficiency was achieved on beads formed by 5% (m/v) of sodium alginate and bead size of 2 mm and 1.5 mm, for CMCase and β-glucosidase, respectively. The immobilized enzyme showed higher stability over a wider pH and temperature range. Metal ions, viz. Cu²+, Mo²⁺, Mn²⁺, Ca²⁺, B³⁺, Zn²⁺, Mg²⁺ and Fe²⁺ showed positive effect for immobilized β-glucosidase activity, while Mg²⁺ on immobilized CMCase activity. The immobilized enzymes could express 20-25% relative enzyme activity even after 4^th repeated use. Immobilization improved the thermal stability over the free enzyme and affected the enzyme kinetic constants, viz. K_M, V_maxand activation energy.

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References

Abdel-Naby M A. 1993. Immobilization of Aspergillus niger NRC107 xylanase and b-xylosidase, and properties of the immobilized enzymes. Applied Biochemistry and Biotechnology 38: 69–81. DOI: https://doi.org/10.1007/BF02916413

Andriani D, Sunwoo C, Ryu H W, Prasetya B and Park D H. 2012. Immobilization of cellulase from newly isolated strain Bacillus subtilis TD6 using calcium alginate as a support material. Bioprocess and Biosystems Engineering 35: 29–33. DOI: https://doi.org/10.1007/s00449-011-0630-z

Anwar A, Qader S A, Raiz A, Iqbal S and Azhar A. 2009. Calcium alginate: A support material for immobilization of proteases from newly isolated strain of Bacillus subtilis KIBGE-HAS. World Applied Science Journal 7(10): 1 281–6.

Attitalla I H and Salleh B. 2010. Improvement of carboxymethyl cellulase and xylase production by alginate immobilized Trichoderma harzianum. Biotechnology 9(4): 529–32. DOI: https://doi.org/10.3923/biotech.2010.529.532

Busto M D, Ortega N and Perez-Mateos M. 1998. Characterization of microbial endo-b-glucanase immobilized in alginate beads. Acta Biotechnology 18(3): 189–200. DOI: https://doi.org/10.1002/abio.370180303

Chakrabarti A C and Storey K B. 1988. Immobilization of cellulase using polyurethane foam. Applied Biochemistry Biotechnology 19: 189–207. DOI: https://doi.org/10.1007/BF02921483

Chen P J, Wei T C, Chang Y T and Lin L P. 2004. Purification and characterization of carboxymethyl cellulase from Sinorhizobium fredii. Botanical Bulletin of Academia Sinica 45: 111–8.

Dey G, Mitra A, Banerjee R and Maiti B R. 2001. Enhanced production of amylase by optimization of nutritional constituents using response surface methodology. Biochemical Engineering Journal 7(3): 227–31. DOI: https://doi.org/10.1016/S1369-703X(00)00139-X

Doaa A, Mahmoud R and Helmy W A. 2009. Potential application of immobilization technology in enzyme and biomass production. Journal Applied Science Resource 5(12): 2 466– 76.

Figueira J A, Dias F F G, Sato H H and Fernandes P. 2011. Screening of supports for the immobilization -glucosidase. Enzyme Resource 8: 1–8. DOI: https://doi.org/10.4061/2011/642460

Drevon G F. 2002. Enzyme immobilization into polymers and coatings. Doctoral thesis, University of Pittsburgh, USA.

Garg N, Kumar D and Ashfaque M. 2008. Production and characterization of extracellular cellulases from a cellulolytic Bacillus sp using mango peel as substrate. Proceeing of National Conference on Eco-friendly approaches in sustainable agriculture and horticulture production, 28-30 Nov, Lucknow, India, pp 184.

Garg N and Ashfaque M. 2010. Mango peel as substrate for production of extra cellular polygalacturonase from Aspergillus fumigatus. Indian Journal of Horticulture 67: 140–3.

Garg N, Tandon D K and Kalra S K. 1994. Utilization of fruit industry waste. Beverage and Food World 21: 16–20.

Garg N, Tandon D K and Kalra S K. 1998. Biodegradation of mango peel fiber. Indian Food Packer 52: 32–5.

Garg N, Tandon D K and Kalra S K. 2000. Protein enrichment of mango peel through solid state fermentation using Aspergillus niger for utilization as feed. Indian Food Packer 54: 62–5.

Iqbal H M N, Kamal S, Ahmed I and Naveed M T. 2012. Enhanced bio-catalytic and tolerance properties of an indigenous cellulase through xerogel immobilization. Advance in Bioscience and Biotechnology 3(4): 308–13. DOI: https://doi.org/10.4236/abb.2012.34044

Jabasingh S A and Nachiyar C V. 2011. Optimization and immobilization kinetics of Aspergillus nidulans cellulase onto modified chitin by response surface methodology. DOI: https://doi.org/10.1260/0263-6174.29.9.897

Biotechnology Bioinformatics and Bioengineering 1: 201–20.

Kumar D, Ashfaque M, Muthukumar M, Singh M and Garg N. 2012. Production and characterization of carboxymethyl cellulase from Paenibacillus polymyxa using mango peel as substrate. Journal of Environmental Biology 33: 81–4.

Kumar D, Muthukumar M and Garg N. 2012. Kinetics of fungal extracellular -amylase from Fusarium solani immobilized in calcium alginate beads. Journal of Environmental Biology 33: 1 021–5.

Kumar D, Yadav K K, Muthukumar M and Garg N. 2013. Production and characterization of a amylase from mango kernel by Fusarium solani NAMICC-F-02956 using submerge fermentation. Journal of Environmental Biology 34: 1 053–8.

Larrauri J A, Ruperez P, Borroto B and Saura-Calixto F. 1996. Mango peels as a new tropical fiber: preparation and characterization. Lebensmittel-Wissenschaft Technology 29(8): 729–33. DOI: https://doi.org/10.1006/fstl.1996.0113

Liang W and Cao X. 2012. Preparation of a pH-sensitive polyacrylate amphiphilic copolymer and its application in cellulase immobilization. Bioresource Technology 16: 140–6. DOI: https://doi.org/10.1016/j.biortech.2012.03.082

Lowry O H, Rosenbrough N J, Far A L and Randoll R J. 1951. Protein measurement with the folin phenol reagent. Journal of Biology and Chemistry 193: 265–75. DOI: https://doi.org/10.1016/S0021-9258(19)52451-6

Narasimha G, Sridevi A, Viswanath B, Chandra S M and Reddy B R. 2005. Nutrient affects on production of cellulolytic enzymes by Aspergillus niger. African Journal of Biotechnology 5: 472–6.

Plahuta P and Raspor P. 1996. Cellulase immobilization on Caalginate beads. Food Technology and Biotechnology 34(2-3): 77–80.

Tandon D K and Garg N. 1999. Mango waste: A potential source of pectin fiber and starch. Indian Journal of Environmental Protectection 19: 924–7.

Wongkhalaung C, Kashiwagi Y, Mahae Y, Ohta T and Sasaki T. 1985. Cellulase immobilized on a soluble polymer. Applied Microbiology and Biotechnology 21: 37–41. DOI: https://doi.org/10.1007/BF00252359

Xin F and Geng A. 2010. Horticultural waste as the substrate for cellulase and hemicellulase production by Trichoderma reesei under solid-state fermentation. Applied Biochemistry Biotechnology 162: 295–306. DOI: https://doi.org/10.1007/s12010-009-8745-2

Xu J, Huo S, Yuan Z, Zhang Y, Xu H, Guo Y, Liang C and Zhuang X. 2011. Characterization of direct cellulase immobilization with superparamagnetic nanoparticles. Biocatalyst Biotransformatics 29: 71–6. DOI: https://doi.org/10.3109/10242422.2011.566326

Yuan X Y, Shen N X, Sheng J and Wei X. 1999. Immobilization of cellulase using acrylamide grafted acrylonitrile copolymer membranes. Journal of Membrane Science 155(1): 101–6. DOI: https://doi.org/10.1016/S0376-7388(98)00313-5

Zeng Y, Hung Y, Chen M, Peng C, Tzen J T and Liu J. 2009. Simultaneous refolding, purification, and immobilization of recombinant Fibrobacter succinogenes 1,3-1,4-b-D-glucanase on artificial oil bodies. Journal of Chemical Technology and Biotechnology 84: 1 480–5. DOI: https://doi.org/10.1002/jctb.2205

Zhang M, Tu X, Kurabi A, Gilkes N, Mabee W and Saddler J. 2006. Immobilization of -glucosidase on eupergit C for lignocelluloses hydrolysis, Biotechnology Letter 28: 151–6. DOI: https://doi.org/10.1007/s10529-005-5328-3