Research Article | Volume: 3, Issue: 2, March-April, 2015

Comparative analysis of two catalytically distinct endoglucanases from Aspergillus nidulans

Baljit Kaur H.S. Oberoi B.S. Chadha   

Open Access   

Published:  Apr 27, 2015

DOI: 10.7324/JABB.2015.3205
Abstract

This study reports purification and characterization of two catalytically distinct endoglucanases (EGI and EGII) from a thermotolerant fungus Aspergillus nidulans. The endoglucanases (EGI and EGII) exhibited molecular masses of 56 and 31 kDa and pIs of 3.6 and 3.8, respectively. EGI was putatively classified as GH7 family member catalyzed carboxymethyl cellulose, xyloglucan, barley β-glucan as well as pNP-β-D-lactopyranoside and pNP-cellobioside, and was optimally active at 50°C and pH 4.0. Whereas, EGII lacking CBD preferentially recognized barley β-glucan when compared substrate CMC, xyloglucan and lichenan and was putatively classified as GH12 member. Interestingly, EGII was characterized to be thermoacidophilic exhibiting 96% its activity at pH 2.0 and at 60 °C. Hydrolysis of barley β-glucan and CMC by EGI and EGII liberated cellobiose as a major product. HPLC analysis showed that barley β-glucan hydrolysate obtained by action of EGI showed high levels of glucose in addition to cellobiose indicating towards an exo type action of this enzyme.


Keyword:     Thermoacidophilic endoglucanasesGH7GH12barley β-glucan.


Citation:

Baljit Kaur, H.S. Oberoi, B.S. Chadha. Comparative analysis of two catalytically distinct endoglucanases from Aspergillus nidulans. J App Biol Biotech. 2015; 3 (02): 022-029.

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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Reference

1. Saha, S., Roy, R., Sen, S.K., Ray, A.K. Characterization of cellulase producing bacteria from the digestive tract of tilapia, Oreochromis mossambica (Peters) and grass carp. (tenopharyngodon idella (Valenciennes). Aquaculture Research. 2006; 37(4):380-388. http://dx.doi.org/10.1111/j.1365-2109.2006.01442.x

2. Lynd, L.R., Weimer, P.J., Van Zyl, W.H., Pretorius, I.S. Microbial cellulose utilization: fundamental and biotechnology. Microbiology and Molecular Biology Reviews. 2002; 66(3): 506-507. http://dx.doi.org/10.1128/MMBR.66.3.506-577.2002

3. Ragothaman, M.Y., Andrew, J.R., Jeffrey, D.W., Tazez, Z.S. Thermostability in endoglucanase is fold specific. BMC Structural Biology. 2011; 11: 10. http://dx.doi.org/10.1186/1472-6807-11-10

4. Horn, S.J., Vaaje-Kolstad, G., Westereng, B., Eijsink, V.G. Novel enzymes for the degradation of cellulose. Biotechnology for Biofuels. 2012; 5: 45. http://dx.doi.org/10.1186/1754-6834-5-45

5. Narra, M., Dixit, G., Divecha, J., Kumar, K., Madamwar, D. and Shah, A.R. Production, purification and characterization of a novel GH12 family endoglucanase from Aspergillus terreus and its application in enzymatic degradation of delignified rice straw. 2014. International journal of Biodeterioration and Biodegradation. 2014; 88: 150-161. http://dx.doi.org/10.1016/j.ibiod.2013.12.016

6. Soni, R., Nazir, A., Chadha, B.S. and Saini, H.S. Novel sources of fungal cellulases for efficient deinking of composite paper waste. Bioresources. 2008; 3: 234-246.

7. Kaur, B., Oberoi, H. and Chadha, B.S. Enhanced cellulase producing mutants developed from heterokaryotic Aspergillus strain. Bioresource Technology. 2014; 156: 100-107. http://dx.doi.org/10.1016/j.biortech.2014.01.016

8. Wang, K., Luo, H., Shi, P., Huang, H., Bai, Y. and Yao, Bin. A highly-active endo-1,3-1,4-β-glucanase from thermophilic Talaromyces emersonii CBS394.64 with application potential in the brewing and feed industries. Process Biochemistry. 2014; 49(9): 1448-1456. http://dx.doi.org/10.1016/j.procbio.2014.06.003

9. Sujit, S.J., Saurabh, S.D., Tae-SU, K., In-Won, K. and Jung-Kul,L. Characterization of a novel endo-β-1,4-glucanase from Armillaria gemina and its application in biomass hydrolysis. Applied Microbiology and Biotechnology. 2013; 98:661-669.

10. Nakamura, A., Watanabe, H., Ishida, T., Uchihashi, T., Wada, M., Ando, T., Igarashi, K. and Samejima, M. Trade-off between processivity and hydrolytic velocity of cellobiohydrolases at the surface of crystalline cellulose. Journal of the American Chemical Society. 2014; 136(12):5484-4592. http://dx.doi.org/10.1021/ja4119994

11. Sharma, M., Chadha, B.S, Kaur, M., Ghatora, S.K. and Saini, H.S. Molecular characterization of multiple xylanase producing thermophilic/thermotolerant fungi isolated from composting materials. Letters in Applied Microbiology. 2008; 46 (5):526-535. http://dx.doi.org/10.1111/j.1472-765X.2008.02357.x

12. Bradford, M.M. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 1976; 72 (1-2):248-254. http://dx.doi.org/10.1016/0003-2697(76)90527-3

13. Miller, G.L. Use of dinitrosalicylic acid reagent for determination of reducing sugars. Analytical Chemistry. 1959; 31 (3):426-428. http://dx.doi.org/10.1021/ac60147a030

14. Kenneth, M.B., Alejandrro, P.R., Xin-Liang, L., Siqing, L. and Stephen, R.H. Purification and characterization of a family 5 endoglucanase from a moderately thermophilic strain of Bacillus licheniformis. Biotechnology Letters. 2006; 28: 1761-1765. http://dx.doi.org/10.1007/s10529-006-9153-0

15. Kaur,B., Sharma, M., Soni, R., Oberoi, H. and Chadha, B.S. Proteome based profiling of hyper-cellulase producing strains developed through inter-specific protoplast fusion between Aspergillus nidulans and Aspergillus tubingensis. Applied Biochemistry and Biotechnology. 2013; 169 (2):393-407. http://dx.doi.org/10.1007/s12010-012-9985-0

16. Kaur, J., Chadha, B.S., Kumar, B.A. and Saini, H.S. Purification and characterization of two endoglucanases from Melanocarpus Sp. MTCC 3922. Bioresource Technology. 2007; 98 (1):74-81. http://dx.doi.org/10.1016/j.biortech.2005.11.019

17. Nazir, A., Soni, R., Saini, H.S., Manhas, R.K. and Chadha, B.S. Purification and caharacterization of an endoglucanase from Aspergillus terreus highly active against barley β-glucan and xyloglucan. World Journal of Microbiology and Biotechnology. 2009; 25(7):1189-1197. http://dx.doi.org/10.1007/s11274-009-0001-y

18. Pol, D., Laxman, R.S. and Rao, M. Purification and biochemical characterization of endoglucanase form Penicillium pinophilum MS 20. Indian Journal of Biochemistry and Biophysics. 2012; 49(3):189-194.

19. Begum, F., Absar, N. and Alam, S.M. Purification and characterization of extracellular cellulase from A.oryzae ITCC-4857.01. Journal of Applied Sciences and Research. 2009; 5(10): 1645-1651.

20. Tavers, E.Q de Pinho., Rubini, M.R., Mello-de-Sousa, T.M., Duarte, G.C., Paula de Faria, F., Filho, E.X.F., Kyaw, C.M., Pereira, I.S. and Pocas-Fonseca, M.J. An acidic thermostable recombinant Aspergillus nidulans endoglucanase is active towards distinct agriculture residues. Enzyme Research. 2013; http://dx.doi.org.1155/2013/287343.

21. Vlasenko, E., Schulein, M., Cherry, J. and Xu, F. Substrate specificity of family 5,6,7,9,12 and 45 endoglucanases. Bioresource Technology. 2010; 101(7):2405-2411. http://dx.doi.org/10.1016/j.biortech.2009.11.057

22. Bukhtojarov, F.E, Ustinov, B.B., Salanovich, T.N., Antonov, A.I., Gusakova, V. and Sinitsyn, A.P. Cellulase complex of the fungus Chrysosporium lucknowense: isolation and characterization of endoglucanase and cellobiohydrolases. Biochem. Mosc. 2004; 69(5):542-551. http://dx.doi.org/10.1023/B:BIRY.0000029853.34093.13

23. Luo, H., Yang, J., Yang, P., Li, Huang, H., Shi, P., Bai, Y., Wang, Y., Fan, Y. and Yao, B. Gene cloning and expression of a new acidic family 7 endo-β-1,3-1,4-glucanase from the acidophilic fungus Bispora Sp. MEY-1. Applied Microbiology and Biotechnology. 2010; 85(4):1015-1023. http://dx.doi.org/10.1007/s00253-009-2119-0

24. Van Dyk, J.S. and Pletschke, B.I. A review of lignocelluloses bioconversion using enzymatic hydrolysis and synergistic cooperation between enzymes-factors affecting enzymes, conversion and synergy. Biotechnology Advances. 2012; 30(6):1458-1480. http://dx.doi.org/10.1016/j.biotechadv.2012.03.002

25. Rouvinen, J., Berfors, T., Teeri T., Knowles, J.K. and Jones, T.A. Three dimensional structure of cellobiohydrolase II from Trichoderma reesei. Science. 1990; 249(4967):380-386. http://dx.doi.org/10.1126/science.2377893

26. Adsul, M.G., Bastawde, K.B. and Gokhale, D.V. Biochemical characterization of two xylanases from yeast Pseudozyma hubeiensis producing only xylooligosaccharides. Bioresource Technology. 2009; 100(24):6488-6495. http://dx.doi.org/10.1016/j.biortech.2009.07.064

27. Andersson, A.A.M., Armo, E., Grangeon, E., Fredriksson, H., Andersso, R. and Aman, P. Molecular weight and structure units of (1-3, 1-4)-β-glucans in dough and bread made from hull-less barley milling fractions. Journal of cereal Science. 2004; 40(3):195-204. http://dx.doi.org/10.1016/j.jcs.2004.07.001

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