Screening of bacterial strains for pectate lyase production and detection of optimal growth conditions for enhanced enzyme activity
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Abstract
In the present study, the pectatelyase production by fifty two bacterial strains isolated from ramie grown soils were studied and the strain RDSM01 showed maximum pectate lyase activity. According to sequence homology of Genbank, the strain RDSM01 was identified as Bacillus subtilis (Genbank Accession No. KX035109). Maximum pectate lyase activity of the strain was observed when 1.5% (v/v) inoculum was added to the growth medium and was incubated for 48 hours at 34-370C and at pH 7.0. The relative activity of the strain was 19% higher when apple pectin was used as carbon source compared to citrus pectin. Maximum enzyme production (149.1 – 153.4 IU/ml) was recorded when ammonium chloride or ammonium sulphate at 0.4% concentration was used as nitrogen source. Thus, B. subtilis strain RDSM01 possessing high pectate lyase activity may be effectively utilized for removal of gum from ramie fibre, which is primarily made of pectin and hemicellulose.
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Keywords
Bacillus subtilis, Enzyme activity, Growth factors, Pectate lyase
References
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Baracat, M. C., Valentin, C., Muchovej, J. J. and Silva, D.O. (1989). Selection of pectinolytic fungi for degumming of natural fibres. Biotechnol Lett., 11: 899-902
Basu, S., Saha, M. N., Chattopadhyay, D. and Chakrabarti, K. (2009). Large-scale degumming of ramie fibre using a newly isolated Bacillus pumilus DKS1 with high pectate lyase activity. J. Ind Microbiol Biotechnol., 236: 239-245
Bhattacharya, S. K. and Paul, N. B. (1976). Susceptibility of ramie with different gum contents to micrfobial damage. Curr. Sci., 45: 417-418
Bhatti, H. N., Ashger, M., Abbas, A., Nawaz, R. and Sheikh, A. (2006). Studies on kinetics and thermostability of a novel acid invertase from Fusarium solani. J. Agril. Food Chem., 54: 4617-4623
Bruhlman, F. (1995). Purification and characterization of an extracellular pectate lyase from an amycolata sp. Appl. Environ. Microbiol., 61: 3580-3585
Das, Suparna, Majumdar, B. and Saha, A. R. (2015). Biodegradation of plant pectin and hemicelluloses with three novel Bacillus pumilus strains and their combined application for quality jute fibre production. Agril. Res., 4: 354-364
Georis, J., Esteves, F. L., Brasscur, J. L., Bougnet, V., Devreese, J. L. B. and Gianotta, F. (2000). An additional aromatic interaction improves the thermostability and thermophilicity of a mesophilic family xylanase: Structural and molecular study. Prot. Sci., 9: 466-475
Henriksson, G., Akin, D. E., Slomczynski, D. and Eriksson, K.E.L. (1999). Production of highly efficient enzymes for flax retting by Rhizomucor pusillus. J. Biotechnol., 68:115-123
Hodkinson, B.P. and Lutzoni, F. (2009). A microbiotic survey of lichen-associated bacteria reveals a new lineage from the Rhizobiales. Symbiosis, 49: 163-180
Hoondal, G. S., Tiwari, R.P., Tiwari, R., Dahiya, N. and Beg, Q. K. (2000). Microbial alkaline pectinases and their applications: a review. Appl. Microbiol. Biotechnol., 9: 409-418
Jayani, R. S., Shukla, S. K. and Gupta, R. (2010). Screening of bacterial strains for polygalacturonase activity: Its production by Bacillus sphaericus (MTCC 7542) Enzyme Res. http://dx.doi.org/10.4061/2010/306785.
Kapoor, M.,Beg,Q.K.,Bhusan, B., Singh,K.,Dadhich, K.S. and Hoondal,G.S. (2001).Application of an alkaline and thermosable polygalacturonase from Bacillus sp. Mgm-ep-2 in degumming of ramie (Boehmeria nirea) and sunnhemp (Crotalaria juncea) vast fibres. Process Biochem., 36:803-807
Kashyap, D. R., Soni, S. K. and Tewari, R. (2003). Enhanced production of pectinase by Bacillus sp. DT7 using solid state fermentation. Bioresource Technol., 88:251-254
Kashyap, D. R., Vohra, P., Soni, S. K. and Tewari, R. (2001). Degumming of buel (Grewia optira) vast fibres by pectinolytic enzyme from Bacillus sp.DT7. Biotecnol.Lett., 23:1297– 1301
Mitra, S., Satya, P. and Mahapatra, B. S. (2013). Allied fibre crops. In: Textbook of Field Crops Production – Commercial Crops Vol. II. Indian Council of Agricultural Research, New Delhi, India, pp. 377-398.
Nasser, W., Chalet, F. and Robert-Baudouy, J. (1990). Purification and characterization of extracellular pectate lyase from Bacillus subtilis. Biochemie., 72: 689-695.
Palaniyappan, M., Vijayagopal, V., Viswanathan, R. and Viruthagiri, T. (2009). Screening of natural substrates and optimization of operating variables on the production of pectinase by submerged fermentation using Aspergillus niger MTCC 281. African J. Biotechnol., 8:682-686
Rajendran, R., Sundaram, S. K., Radhai, R. and Rajapriya, P. (2011). Bioscouring of cotton fabrics using pectinase enzyme its optimization and comparison with conventional scouring process. Pakistan J. Biol. Sci., 14:519-525
Ramesh, M. V. and Lonsane, B.K. (1990). Production of bacterial thermostable ?-amylase by solid-state fermentation: a potential tool for achieving economy in enzyme production and starch hydrolysis. Advances Appl. Microbiol., 35: 1-56
Ray, D.P., Banerjee, P., Mondal, S. B., Satya, P. and Mitra, S. (2014). Ramie degumming through novel chemical process. In: Jute and Allied Fibres: Issues and Strategies. The Indian Natural Fibre Society, Kolkata, India, pp. 15-20.
Sharma, D.C. and Satyanarayana, T. (2012).Biotechnological potential of Agro residues for economical production of thermoalkali stable peectinase by Bacillus pimilus dcsr1 by solid-state fermentation and its efficacy in the treatment of ramie fibres. Enzyme Res., http://dx.doi.org /10.1155/2012/281384
Teixeira, M. F. S., Filho, J. L. L. and Durán, N. (2000). Carbon sources effect on pectinase production from Aspergillus japonicas 586. Brazillian J. Microbiol., 31:286-290
Turner, S., Pryer, K.M., Miao, V.P.W. and Palmer, J. D. (1999). Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J. Eukaryotic Microbiol., 46: 327–338
Uenojo, M. and Pastore, G.M. (2007). Pectinases: Aplicações industriais e perspectivas. Quimica Nova., 30: 388-394
Baracat, M. C., Valentin, C., Muchovej, J. J. and Silva, D.O. (1989). Selection of pectinolytic fungi for degumming of natural fibres. Biotechnol Lett., 11: 899-902
Basu, S., Saha, M. N., Chattopadhyay, D. and Chakrabarti, K. (2009). Large-scale degumming of ramie fibre using a newly isolated Bacillus pumilus DKS1 with high pectate lyase activity. J. Ind Microbiol Biotechnol., 236: 239-245
Bhattacharya, S. K. and Paul, N. B. (1976). Susceptibility of ramie with different gum contents to micrfobial damage. Curr. Sci., 45: 417-418
Bhatti, H. N., Ashger, M., Abbas, A., Nawaz, R. and Sheikh, A. (2006). Studies on kinetics and thermostability of a novel acid invertase from Fusarium solani. J. Agril. Food Chem., 54: 4617-4623
Bruhlman, F. (1995). Purification and characterization of an extracellular pectate lyase from an amycolata sp. Appl. Environ. Microbiol., 61: 3580-3585
Das, Suparna, Majumdar, B. and Saha, A. R. (2015). Biodegradation of plant pectin and hemicelluloses with three novel Bacillus pumilus strains and their combined application for quality jute fibre production. Agril. Res., 4: 354-364
Georis, J., Esteves, F. L., Brasscur, J. L., Bougnet, V., Devreese, J. L. B. and Gianotta, F. (2000). An additional aromatic interaction improves the thermostability and thermophilicity of a mesophilic family xylanase: Structural and molecular study. Prot. Sci., 9: 466-475
Henriksson, G., Akin, D. E., Slomczynski, D. and Eriksson, K.E.L. (1999). Production of highly efficient enzymes for flax retting by Rhizomucor pusillus. J. Biotechnol., 68:115-123
Hodkinson, B.P. and Lutzoni, F. (2009). A microbiotic survey of lichen-associated bacteria reveals a new lineage from the Rhizobiales. Symbiosis, 49: 163-180
Hoondal, G. S., Tiwari, R.P., Tiwari, R., Dahiya, N. and Beg, Q. K. (2000). Microbial alkaline pectinases and their applications: a review. Appl. Microbiol. Biotechnol., 9: 409-418
Jayani, R. S., Shukla, S. K. and Gupta, R. (2010). Screening of bacterial strains for polygalacturonase activity: Its production by Bacillus sphaericus (MTCC 7542) Enzyme Res. http://dx.doi.org/10.4061/2010/306785.
Kapoor, M.,Beg,Q.K.,Bhusan, B., Singh,K.,Dadhich, K.S. and Hoondal,G.S. (2001).Application of an alkaline and thermosable polygalacturonase from Bacillus sp. Mgm-ep-2 in degumming of ramie (Boehmeria nirea) and sunnhemp (Crotalaria juncea) vast fibres. Process Biochem., 36:803-807
Kashyap, D. R., Soni, S. K. and Tewari, R. (2003). Enhanced production of pectinase by Bacillus sp. DT7 using solid state fermentation. Bioresource Technol., 88:251-254
Kashyap, D. R., Vohra, P., Soni, S. K. and Tewari, R. (2001). Degumming of buel (Grewia optira) vast fibres by pectinolytic enzyme from Bacillus sp.DT7. Biotecnol.Lett., 23:1297– 1301
Mitra, S., Satya, P. and Mahapatra, B. S. (2013). Allied fibre crops. In: Textbook of Field Crops Production – Commercial Crops Vol. II. Indian Council of Agricultural Research, New Delhi, India, pp. 377-398.
Nasser, W., Chalet, F. and Robert-Baudouy, J. (1990). Purification and characterization of extracellular pectate lyase from Bacillus subtilis. Biochemie., 72: 689-695.
Palaniyappan, M., Vijayagopal, V., Viswanathan, R. and Viruthagiri, T. (2009). Screening of natural substrates and optimization of operating variables on the production of pectinase by submerged fermentation using Aspergillus niger MTCC 281. African J. Biotechnol., 8:682-686
Rajendran, R., Sundaram, S. K., Radhai, R. and Rajapriya, P. (2011). Bioscouring of cotton fabrics using pectinase enzyme its optimization and comparison with conventional scouring process. Pakistan J. Biol. Sci., 14:519-525
Ramesh, M. V. and Lonsane, B.K. (1990). Production of bacterial thermostable ?-amylase by solid-state fermentation: a potential tool for achieving economy in enzyme production and starch hydrolysis. Advances Appl. Microbiol., 35: 1-56
Ray, D.P., Banerjee, P., Mondal, S. B., Satya, P. and Mitra, S. (2014). Ramie degumming through novel chemical process. In: Jute and Allied Fibres: Issues and Strategies. The Indian Natural Fibre Society, Kolkata, India, pp. 15-20.
Sharma, D.C. and Satyanarayana, T. (2012).Biotechnological potential of Agro residues for economical production of thermoalkali stable peectinase by Bacillus pimilus dcsr1 by solid-state fermentation and its efficacy in the treatment of ramie fibres. Enzyme Res., http://dx.doi.org /10.1155/2012/281384
Teixeira, M. F. S., Filho, J. L. L. and Durán, N. (2000). Carbon sources effect on pectinase production from Aspergillus japonicas 586. Brazillian J. Microbiol., 31:286-290
Turner, S., Pryer, K.M., Miao, V.P.W. and Palmer, J. D. (1999). Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J. Eukaryotic Microbiol., 46: 327–338
Uenojo, M. and Pastore, G.M. (2007). Pectinases: Aplicações industriais e perspectivas. Quimica Nova., 30: 388-394
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Screening of bacterial strains for pectate lyase production and detection of optimal growth conditions for enhanced enzyme activity. (2017). Journal of Applied and Natural Science, 9(1), 370-374. https://doi.org/10.31018/jans.v9i1.1198
