Research Article | Volume: 4, Issue: 1, Jan-Feb, 2016

Medium Formulation and its optimization for increased protease production by Penicillium sp. LCJ228 and its potential in blood stain removal

V. Benluvankar S. Evelyne Priya J. Joel Gnanadoss   

Open Access   

Published:  Feb 19, 2016

DOI: 10.7324/JABB.2016.40104
Abstract

The production of protease by Penicillium sp. LCJ228 was optimized under submerged fermentation. Nutritional and physical factors that influence protease production were optimized by one factor at a time (OFAT) method in order to achieve high yield of protease. Maximum protease production was obtained on the 4th day in a liquid medium containing glucose (15 g/L), yeast extract (15 g/L), black gram husk (10 g/L) with an initial pH of 10 and an inoculum size of 2 g/L of fungal mycelium. About 2.4 fold increase in protease production was observed in the optimized medium. The maximally yielded crude protease was then precipitated and characterized by SDS-PAGE and native-PAGE. The molecular weight of the ammonium sulphate precipitated protease was ~66 to 70 kDa and native-PAGE showed three isoenzymes. The crude and precipitated protease had the ability to completely remove blood stains on cotton fabric indicating its potential to be used as a stain remover in detergents.


Keyword:     Protease Penicillium sp. submerged fermentation conventional optimization blood stain removal.


Citation:

Benluvankar V, Priya SE and Gnanadoss JJ. Medium Formulation and its optimization for increased protease production by Penicillium sp. LCJ228 and its potential in blood stain removal. J App Biol Biotech. 2016; 4 (01): 020-026. DOI: 10.7324/JABB.2016.40104

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. Shivanand P, Jayaraman G. Production of extracellular protease from halotolerent bacterium Bacillus aguimaris strain VITP4 isolated from kumta coast. Proc Biochem. 2009; 44: 1088-1094.

2. Deng A, Wua J, Zhang Y, Zhang C, Wena T. Purification and characterization of a surfactant -stable high-alkaline protease Bacillus sp. B001. Bioresour Technol. 2010; 101: 7100-7106.

3. Marchand S, Vandriesche G, Cooreviets A, Coudijzer K, Jonghe VD, Dewettinck K. Heterogenisity of heat resistance proteases form milk Pseudomonas sp. Int J Food Microbiol. 2009; 133: 68-77.

4. Ribitscha D, Karla W. Birner-Cruenbergera R, Crubera K. Eiteljoerg I, Remlera P. 2010. C - terminal truncation of a metagenome-derived detergent protease for effective expression in E.coli. J Biotech. 2009; 150: 408-416.

5. Mala B, Rao AM, Deshpande VV. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev. 1998; 62: 597-635.

6. Joo HS, Kumar CG, Park GC, Park SR, Chang CS. Bleach resistant alkaline protease produced by a Bacillus sp isolated from the Korean Polychaete Perriserrula leucophryna. Proc Biochem. 2004; 39:1441-1447.

7. Mei C, Jiang X. A novel surfactant and oxidation stable alkaline protease from Vibrio metschnikovii Dl-33-51. Proc Biochem. 2005; 40: 2167-2172.

8. Barett AJ. Proteolytic enzymes: serine and cysteine peptidases. Methods Enzymol. 1994; 244:1-15.

9. Madan M, Dhillion S, Singh R. Production of alkaline protease by a UV mutant of Bacillus polymyxa. Indian J Microbiol. 2002; 42:155-159.

10. Beg QK, Saxena RK, Gupta R. De-repression and subsequent induction of protease synthesis by Bacillus mojavensis under fed-batch operations. Proc Biochem. 2002; 37: 1103-1109.

11. Fan-Ching Y, Lin IH. Production of acid protease using thin stillage from rice spirit distillery by Aspergillus niger. Enzyme Microb Technol. 1998; 23: 397-402.

12. Oyeleke SB, Egwim EC, Auta SH. Screening of Aspergillus flavus and Aspergillus fumigatus strains for extracellular protease enzyme production. J Microbiol Antimicrob. 2010; 2: 83-87.

13. Gnanadoss JJ, Rebecca R, Jebapriya, GR. Production of protease from Aspergillus niger and Mucor mucedo under submerged and solid state fermentation. Int J Curr Res. 2011; 3(10): 075-078.

14. Farley PC, Ikasari L. Regulation of secretion of Rhizopus oligosporus extra cellular carboxyl proteinases. J Gen Microbiol. 1992; 138: 2539-2544.

15. Chrzanowska J, Kolaczkowska M, Polanowski A. Production of extracellular proteolytic enzymes by various species of Penicillium. Enzyme Microb Technol. 1993; 15:140-143.

16. Ikram-ul-haq, Mukhtar H, Umber, H. Production of protease by Penicillium chrysogenum through optimization of environmental conditions. J Agri Soc Sci. 2006; 2(1): 23-25.

17. Gnanadoss JJ, Devi SK. Optimization of nutritional and culture conditions for improved protease production by Aspergillus nidulans and Aspergillus flavus. J Microbiol Biotech Food Sci. 2015; 4(6): 518-523.

18. Kumar A, Sachdev A, Balasubramanyan SD, Saxena A, Lata K. Optimization of conditions purification and for production of neutral and alkaline protease characterization of the heat stable serine proteinase from species of Bacillus and Pseudomanas. Ind J Microbiol. 2002;42: 233-236.

19. Kumar CG, Takagi H. Microbial alkaline proteases: From a bio-industrial viewpoint. Biotechnol Adv. 1999; 17: 561-594.

20. Rao MB, Tanksale AM, Ghatge MS, Deshpande VV. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev. 1998; 62: 597-635.

21. Sarker PK, Talukdar SA, Deb P, Sayem SM, Mohsina K. Optimization and partial characterization of culture conditions for the production of alkaline protease from Bacillus licheniformis P003. SpringerPlus. 2013; 2: 506.

22. Pangrikar PP, Wadikarl MS, Borde VU, Chavan AM. Production of protease enzyme from rhizospheric fungi of bt and non bt cotton varieties. Biosci Discovery. 2011; 2 (2): 249 -250.

23. Sayem SMA, Alam MJ, Hoq MM. Effect of temperature, pH and metal ions on the activity and stability of alkaline protease from novel Bacillus licheniformis MKZ03. Proc Pakistan Acad Sci. 2006; 43(4): 257-262.

24. Karthick R, Namasivayam S, Sivasubramanian S, Ganesh K. Influence of media on protease production by Beauveria bassiana ( Bals.) Vuil. and their stability towards commercially available detergents, surfactants and enzyme inhibitors. Int J Biol Technol. 2010; 1(1): 78-83.

25. Keay L, Wildi BS. Proteases of the genus Bacillus, I. Neutral proteases. J Biotechnol Bioeng. 1970; 12: 179-212.

26. Lowry OH, Roserbrough NJ, Farr AL, Randall R. Protein measurement with Folin Phenol Reagent. J Biol Chem. 1951; 193: 265-275.

27. Beg QK, Gupta R. Purification and characterization of an oxidation stable, Thiol dependent serine protease from Bacillus mojavensis. Enzyme Microbial Technol. 2003; 32: 294-304.

28. Ellaiah P, Srinivasulu B, Adinarayana K. A review on microbial alkaline proteases. J Sci Ind Res. 2002; 61: 690-704.

29. Shridhar P, Deepti A, Panka P, Tushar B. Production of alkaline protease by Penicillium sp. under SSF conditions and its application to soy protein hydrolysis. Proc Biochem. 2003; 39: 977-981.

30. Abirami VS, Menakshi A, Kanthym K, Bhar R, Mahlingm R. Partial purfication and characterization of an extracellular protease from Penicillium janthinelum and Neurospa crassa. Eur J Exp Biol. 2011; 1(3): 114-123.

31. Oskouie SF, Tabandeh F, Yakhchali B, Eftekhar F. Response surface optimization of medium composition for alkaline protease production by Bacillus clausii. J. Biochem. Engineer. 2008; 39: 37-42.

32. Nochure SV, Roberts MF, Demain AI. True cellulase production by Clostridium thermocellum grown on different carbon sources. Biotechnol Lett. 1993; 15: 641-646.

33. Ire FS, Okolo NBN, Moneke AN,Odibo FJC. Influence of cultivation conditions on the production of a protease from Aspergillus carbonarius using submerged fermentation. Afr J Food Sci. 2011; 5: 353-365.

34. Varela H, Ferrari MD, Belobradjic L. Effect of medium composition on the production by a new Bacillus subtilis isolate of protease with promising unhairing activity. World J Microbiol Biotechnol. 1996; 12: 643-645.

35. Kumara CG, Tiwarib MP, Janya KD. Novel alkaline serine proteases from alkalophilic Bacillus spp.: purification and some properties. Proc Biochem. 1999; 34(5): 441-449.

36. Chellapandi P. Production and preliminary characterization of alkaline protease from Aspergillus flavus and Aspergillus terreus. E-J Chem. 2010; 7(2): 479-482.

37. Srinubabu G, Lokeswari N, Jayaraju K. Screening of nutritional parameters for the production of protease from Aspergillus oryzae. E-J Chem. 2007; 4(2): 208-215.

38. Tremacoldi CR, Carmona EC. Production of extracellular alkaline proteases by Aspergillus clavatus. World J Microbiol Biotechnol. 2005; 21: 169-172.

39. Wang Y, Lee M. Influence of culture and nutritional condition on the production of protease from Thermophilic strain Aspergillus species NTIJ-FC-671. J Chin Agr Chem Soc. 1996; 34: 732-742.

40. Nehra KS, Dhillon S, Chaudhary K, Singh R. Production of alkaline pr1otease by Aspergillus species under submerged and solid state fermentation. Indian J Microbiol. 2002; 42: 43-47.

41. Chauhan B, Gupta R. Application of statistical experimental design for optimization of alkaline protease production from Bacillus sp. RGR-14. Proc Biochem. 2004; 39: 2115-2122.

42. Ashour SA, EL Shore HM, Metwally M, Habib SA. Fungal fermentation of Whey incorporated with certain supplements for the production of protease. Microbios. 1996; 86:59-69.

43. Kirankumar S, Jayashree V, Hanchinalmath Y, Sai S, Debajit B, Prasad T. Optimization and production of alkaline proteases from agro byproducts using a novel Trichoderma Viridiae strain VPG 12, isolated from agro soil. Int Lett Nat Sci. 2014; 9: 77-84.

44. Jenitta XJ, Gnanadoss JJ. Effect of carbon, nitrogen sources and inducers on protease production by Penicillium citrinum LCJ222. Int J Curr Res. 2014. 6(11): 9544-9549.

45. Sandhya C, Sumantha A, Szakacs G, Pandey A. Comparative evaluation of neutral protease production by Aspergillus oryzae in submerged and solid state fermentation. Proc Biochem. 2005; 40: 2689-2694.

46. Paranthaman R, Alagusundaram K, Indhumathi. Production of protease from rice mill wastes by Aspergillus niger in solid state fermentation. World J Agric Sci. 2009; 5(3): 308-312.

47. Palanivel P, Ashokkumar L, Balagunathan R. Production, purification and fibrinolytic characterization of alkaline protease from extremophilic soil fungi. Int J Pharm Bio Sci. 2013; 4(2): (B) 101-110.

48. Coral G, Arikan B, Unaldi MN, Guvenmes H. Purification and characterization of a protease-resistant cellulase from Aspergillus niger. J Fermentation Bioeng. 2002; 39: 122-127.

49. Muthulakshmi CD, Gomathi DG, Kumar G, Ravikumar G, Kalaiselvi M, Uma C. Production, purification and characterization of protease by Aspergillus flavus under solid state fermentation. JJBS 2011; 4: 137-148.

50. Calik P, Calik G, Ozdamar TH. Oxygen transfer effects in serine alkaline protease fermentation by Bacillus licheniformis: Use of citric acid as the carbon source. Enzyme Microb Technol. 1998; 23: 451-461.

51. Ducros E, Ferrari M, Pellegrino M, Raspanti C, Bogni C. Effect of aeration and agitation on the protease production by Staphylococcus aureus mutant RC128 in a stirred tank bioreactor. Bioprocess Biosyst Eng. 2009; 32 (1):143-148.

52. Hwang ST, Wachter C, Schatz G. Protein import into the yeast mitochondrial matrix- A new translocation intermediate between the two mitochondrial membranes. J Bio Chem. 1991; 266: 21083-21089.

53. Malathi S, Chakraborty R. Production of alkaline protease by a new Aspergillus flavus isolate under solid substrate fermentation conditions for use as a depilation agent. Appl Environ Microbiol. 1991; 57: 712-16.

54. Dubey R, Adhikary S, Kumar J, Sinha N. Isolation, production, purification, assay and characterization of alkaline protease enzyme from Aspergillus niger and its compatibility with commercial detergents. Dev Microbiol Mol Bio. 2010; 1: 75-94.

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