PRODUCTION, CHARACTERIZATION AND PURIFICATION OF ALKALINE PROTEASE FROM ALCALIGENES sp., AND ITS APPLICATION IN DETERGENT INDUSTRY

Authors

  • AISHWARYA.M SASTRA University
  • SWATI KUMARI
  • ARVIND SIVASUBRAMANIAN
  • MEENAKSHI SUNDARAM MUTHURAMAN

Abstract

Proteases are enzymes well known for their wide range industrial applications such as food, detergent, pharmaceutical etc. The aim of the present work was to study the production, purification and application of alkaline protease from the strain Alcaligenes sp. (MTCC 9730). Batch and fed batch fermentation were used as the production strategies. The optimum temperature and pH for the production of enzyme was found to be 55°C and pH 9 respectively. Ca2+ increased the activity, Cu2+ and Mg2+ mildly inhibited and by Zn2+ strongly inhibited the activity. The protease enzyme was purified using Ammonium sulphate precipitation and maximum activity of 60 U/ml was found at 70percent saturation. Alkaline protease is known to be an active ingredient of the commercial detergents. The enzyme was used to study its compatibility with other detergents and its ability to remove the protein stain.

 

Keywords: Alkaline protease, Alcaligenes sp, fermentation, purification, detergent.

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References

Akcan N, Uyar F., Production of extracellular alkaline protease from Bacillus subtilis RSKK96 with solid state fermentation. Eurasia J Biosci., 2011; 5: 64-72.

Kumar PPK, Mathivanan V, Karunakaran M, Renganathan S and Sreenivasan RS., Studies on the effects of pH and incubation period on protease production by Bacillus spp. using groundnut cake and wheat bran. Indian Journal of Science Technology., 2008;4:1-4.

Gouda MK, Optimization and purification of alkaline proteases produced by marine Bacillus sp. MIG newly isolated from eastern harbor of Alexandria. Polish Journal of Microbiology., 2006; 55: 119-126.

Kumar CG, Takagi H., Microbial alkaline proteases: From a bioindustrial viewpoint, Biotechnology Advances., 1999; 17:561–594.

Hodgson J. The changing bulk biocatalyst market. Biotechnol 1994 (12):789–90.

International Union of Biochemistry and Molecular Biology, Enzyme Nomenclature. New York: Academic Press, 1992.

Aunstrup K, Outtrup H, Andersen O, Damnmann C. Proteases from alkalophilic Bacillus species. In: Terui G, editor. Fermentation Technology Today. Osaka. Society of Fermentation Technology of Japan., 1972; 299–305.

Kuddus M, Ramteke, PW., Production optimization of an extracellular cold-active alkaline protease from Stenotrophomonas maltophilia MTCC 7528 and its application in detergent industry ; Afr. J. Microbiol. Res., 2011; 7:809-816.

Sameh H. Soror, V. Verma, Ren Rao, Shafaq Rasool, S. Koul, G. N. Qazi, John Cullum., A cold-active esterase of Streptomyces coelicolor A3(2): from genome sequence to enzyme activity. J Ind Microbiol Biotechnol 2007; 34:525–531

Siva Muthuprakash, K. M.; Jayanthi Abraham, P. A comparative analysis of protease producing microbes isolated from tannery effluent. International journal of science and nature, 2011; 2(1):110-113.

Sarath, G.,et al., Proteolytic Enzymes: A Practical Approach, IRL Press, Oxford, England, 1989.

Ganesh Kumar, et al., Purification of extracellular acid protease and analysis of fermentation metabolites by Synergistes sp. utilizing proteinaceous solid waste from tanneries, Bioresource Technology, 2008, 2364–2372.

Shahanara, B, Characterization of an intracellular protease from Pseudomonas aeruginosa, Pakistan Journal of Medical Sciences., 2007; 23: 227-232.

Titilayo Olufunke Femi-Ola and Desmond Olayinka Oladokun: Partial purification and characterization of a thermostable alkaline protease from Lactobacillus brevis: Malaysian Journal of Microbiology, 2012; 8(1): 1-5.

Adinarayana, K.et al., Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS Pharmaceutical Science Technician 4, 2003, 56-63.

Tabandeh,F et al., Fed-batch fermentation of Bacillus sp. L2 for alkaline protease production using different feeding strategies, The 11th Iranian chemical engineering congress(ICHEC11), November 2006.

Baghel VS, et al., Psychrotrophic proteolytic bacteria from cold environment of Gangotri glacier, Western Himalaya, India. Enz. Microb. Technol., 2005, 36: 654-659.

Deutscher MP, Guide to protein purification: Methods in Enzymology. Academic Press, New York, 2005.

Devi,K.M.et al., Purification, characterization of alkaline protease enzyme from native isolate of Aspergillus niger and its compatibility with commercial detergents, Indian Journal of Science and Technology, 2008, 1, 7-13.

Nehra,K.S.etal., Production and characterization of alkaline protease from Aspergillus species and its compatibility with commercial detergents. Asian Journal of Microbiology Biotechnology and Environmental Science, 2004, 6, 67-72.

Published

01-10-2013

How to Cite

AISHWARYA.M, . S. KUMARI, A. SIVASUBRAMANIAN, and M. SUNDARAM MUTHURAMAN. “PRODUCTION, CHARACTERIZATION AND PURIFICATION OF ALKALINE PROTEASE FROM ALCALIGENES sp., AND ITS APPLICATION IN DETERGENT INDUSTRY”. Asian Journal of Pharmaceutical and Clinical Research, vol. 6, no. 4, Oct. 2013, pp. 152-5, https://innovareacademics.in/journals/index.php/ajpcr/article/view/516.

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