Improvement of protease production by Chryseobacterium indologenes BYK27 and its application in de-colorization of blood on clothes

Subject Areas : Microbial Biotechnology

Yasamin Binabadi ¹ , Arastoo Badoei-dalfard ² , Abdolhamid Namaki-Shoushtari ³

1 - M.Sc., Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.
2 - Associate Professor, Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.
3 - Associate Professor, Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran.

Received: 2017-04-15 Accepted : 2017-05-29 Published : 2018-05-22

Keywords: Optimization, Protease, Organic solvents, Chryseobacterium indologenes,

Abstract :

Protease is one of the most important industrial enzymes occupying nearly 60% of global enzyme sales. Extracellular protease finds numerous applications in industrial processes like in leather tanning, detergents, dairy, brewery as well as meat tenderization industries. In spite of that, the low level of enzyme production is the main challenge of industrial production of enzyme. Therefore, optimization of industrial protease production and its application in blood de-staining were the aims of this study. The sewage samples were cultivated on the skim milk agar. BYK27 isolates with the highest clear halo around the colonies were selected for further studies. Optimization of parameters affecting protease production by Chryseobacterium indologenes BYK27 was studied by Taguchi approach. De-staining ability of protease was also investigated by de-colorization of bloody cotton cloth. The optimal factors for protease production by Ch. indologenes BYK27 were found to be the temperature of 40 ˚C, pH of 9.0, 0.06% yeast extract and 1% glucose supplements. Protease production under optimal condition was found to be 590 (U/ml) which was improved by 63%, as compared to the basal medium. The protease activity and stability were increased 50% by beta-mercaptoethanol but inhibited about 88% by DMF. In addition, BYK27 protease was able to completely de-stain blood  after 20 min of incubation. The results of this study indicate that BYK27 protease has biotechnological potential, specifically in the detergent industry and provision of valuable compounds. 

References:

1. Zambare V, Nilegaonkar S, Kanekar P. A novel extracellular protease from Pseudomonas
   aeruginosa MCM B-327: enzyme production and its partial characterization. New Biotechnol.
   2011; 28 (2), 173-181.
   
   2. Haddar A, Sellami-Kamoun A, Fakhfakh-Zouari N, Hmidet N, Nasri M. Characterization of
   detergent stable and feather degrading serine protease from Bacillus mojavensis A21. Biochem
   Eng J. 2010; 51(1-2): 53-63.
   3. Rao M, Tanksale A, Ghatge M, Deshpande, V. Molecular and biotechnological aspects of
   microbial protease. Microbiol Mol Biol Rev. 1988; 62(3): 597-635.
   4. Anbu P. Characterization of solvent stable extracellular protease from Bacillus koreensis
   (BK-P21A). Int J Biological Macromolecules. 2013; 56: 162-168.
   5. Ghafoor A, Hasnain Sh. Purification and characterization of an extracellular protease from
   Bacillus subtilis EAG-2 strain isolated from ornamental plant nursery. Pol J Microbiol. 2010;
   59(2): 107-112.
   6. Gupta A, Khare SK. A protease stable in organic solvents from solvent tolerant strain of
   Pseudomonas aeruginosa. Bioresour Technol. 2006; 97(15): 1788-1793.
   7. Hajji M, Kanoun S, Nasri M, Gharsallah N. Purification and characterization of an alkaline
   serine-protease produced by a new isolated Aspergillus clavatus ES1. Process Biochem. 2007;
   42(5): 791-797.
   8. Chaudhari P, Chaudhari B, Chincholkar S. Iron containing keratinolytic metallo-protease
   produced by Chryseobacterium gleum. Process Biochem. 2013; 48(1): 144-151.
   9. Lv LX, Sim MH, Li YD, Min J, Feng WH, Guan WJ, Li YQ. Production, characterization and
   application of a keratinase from Chryseobacterium l99 sp. nov. Process Biochem. 2010; 45 (8):
   1236-1244.
   10. Wang S, Yang Ch, Liang T, Yen Y. Optimization of conditions for protease production by
   Chryseobacterium taeanense TKU001. Bioresour Technol. 2008, 99(99): 3700-3707.
   11. Riffel A, Brandelli A, Bellato MC, Souza, HMFG, Eberlin NM, Tavares CAF. Purification and
   characterization of a keratinolytic metalloprotease from Chryseabacterium sp.Kr6. J Biotechnol.
   2007; 128(3): 693-703.
   12. Wang S, Hsu W, Liang T, Yen Y, Wang Ch. Purification and characterization of three novel
   keratinolytic metalloprpteases produced by Chryseobacterium indologenes TKU014 in a shrimp
   shell powder medium. Bioresour Technol. 2008; 99(13): 5679-5686.
   13. Prakasham RS, Subba-Rao Ch, Sreenivas R, Rajesham S, Sarma PN. Optimization of alkaline
   protease production by Bacillus sp. using Taguchi methodology. Appl Biochem Biotechnol.
   2004; 120(2): 133-144.
   14. Beinabadi Y, Namaki-Soushtari AH, Badoei-Dalfard A. Isolation and characterization of
   protease producing Chryseobacterium indologenes strain BYK27 from Kerman, s dairy industry
   sewage. J Cell Mol Res. 2016; 29: 33-47.
   15. Badoei-Dalfard A, Karami Z. Screening and isolation of an organic solvent tolerant-protease
   from Bacillus sp. JER02: Activity optimization by response surface methodology. J Mol
   Catalysis B: Enzymatic. 2013; 89: 15-23.
   16. Badoei-Dalfard A, Karami Z, Ravan H. Purification and characterization of a thermo- and
   
   organic solvent-tolerant alkaline protease from Bacillus sp. JER02. Preparative Biochem
   Biotechnol. 2015; 45: 128-143.
   17. Meena P, Dutt Tripathi A, Srivastava SK, Jha, A. Utilization of agro-industrial waste (wheat
   bran) for alkaline protease production by Pseudomonas aeruginosa in SSF using Taguchi
   (DOE) methodology. Biocatal Agric Biotechnol. 2013; 2(3): 210-216.
   18. Badoei-Dalfard A, Amiri-Bahrami M, Riahi-Madvar A, Karami Z, Ebrahimi M A. Isolation,
   identification and characterization of organic solvent tolerant protease from Bacillus sp.
   DAF-01. Biological J Microorganism. 2012; 1(2): 37-48.
   19. Ward OP. Proteolytic enzymes. Comprehensive Biotechnol. 1985; 8: 789-818.
   20. Sarkar PK, Cook PE, Owens JD. Bacillus fermentation of soybeans. World J
   Microbiol Biotechnol. 1993; 9(3): 295-299.
   21. Sinagh S, Sinagh S, Tripathi V, Grag S. Purification, characterization and secondary structure
   elucidation of a detergent stable, halotolerant, thermoalkaline protease from Bacillus cereus
   SIU1. Process Biochem. 2012; 47(10): 1479-1487.
   22. Venil CK, Lakshmanaperumalsamy P. Taguchi experimental design for medium optimization
   for enhanced protease production by Bacillus subtilis HB04. J Sci Technol. 2009; 2: 54-59.
   23. Ghaemi Oskouie SF, Tabandeh F, Yakhchali B, Eftekhar F. Enhancement of alkaline protease
   production by Bacillus clausii using Taguchi experimental design. Afr J Biotechnol. 2007; 6
   (22): 2559-2564.
   24. Raut GR, Chakraborty S, Chopade BA, Kokare CR. Isolation and characterization of organic
   solvent stable protease from alkaliphilic marine Saccharopolyspora species. Ind J Geo-Mar Sci.
   2013; 42 (1): 131-138.
   25. Annamalai N, Veeramuthu Rajeswari M, Kumar Sahu S, Balasubramanian Th. Purification
   and characterization of solvent stable, alkaline protease from Bacillus firmus CAS 7 by
   microbial conversion of marine wastes and molecular mechanism underlying solvent stability.
   Process Biochem. 2014; 49(6): 1012-1019.
   26. Paul T, Das A, Mandal AK, Halder S, Jana A, Maity Ch, Kumar DasMohapatra, P, Pati RB,
   Mondal CK. An efficient cloth cleaning properties of a crude keratinase combined with
   detergent: towards industrial viewpoint. J Clean Prod. 2014; 66(1): 672-684.
   27. Ubba-Rao Ch, Sathish T, Ravichandra P, Prakasham RS. Characterization of thermos- and
   detergent stable serine protease from isolated Bacillus circulans and evaluation of eco-friendly
   applications. Process Biochem. 2009; 44(3): 262-268.