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Environmental Microbiology / Microbial Diversity  |  Environmental Microbiology

Microbiol. Biotechnol. Lett. 2020; 48(3): 358-365

https://doi.org/10.4014/mbl.1912.12015

Received: December 23, 2019; Accepted: June 8, 2020

Biochemical Characterization of a Novel Alkaline and Detergent Stable Protease from Aeromonas veronii OB3

Laila manni *, Asmae Misbah , Nouhaila Zouine and Samir Ananou

Laboratory of Microbial Biotechnology and Bioactive Molecules,Faculty of Sciences and Techniques, Fez, Morocco

An organic solvent- and bleach-stable protease-producing strain was isolated from a polluted river water sample and identified as Aeromonas veronii OB3 on the basis of biochemical properties (API 20E) and 16S rRNA sequence analysis. The strain was found to hyper-produce alkaline protease when cultivated on fish waste powder-based medium (HVSP, 4080 U/ml). The biochemical properties and compatibility of OB3 with several detergents and additives were studied. Maximum activity was observed at pH 9.0 and 60℃. The crude protease displayed outstanding stability to the investigated surfactants and oxidants, such as Tween 80, Triton X-100, and H2O2, and almost 36% residual activity when incubated with 1% SDS. Remarkably, the enzyme demonstrated considerable compatibility with commercial detergents, retaining more than 100% of its activity with Ariel and Tide (1 h, 40℃). Moreover, washing performance of Tide significantly improved by the supplementation of small amounts of OB3 crude protease. These properties suggest the potential use of this alkaline protease as a bio-additive in the detergent industry and other biotechnological processes such as peptide synthesis

Keywords: Protease, alkaline, Aeromonas veronii, detergent, solvent-stable, fish waste medium

  1. Singh S, Bajaj BK. 2017. Potential application spectrum of microbial proteases for clean and green industrial production. Energy Ecol. Env. 2: 370-386.
    CrossRef
  2. Razzaq A, Shamsi S, Ali A, Ali Q, Sajjad M, Malik A, et al. 2019. Microbial proteases applications. Front. Bioeng. Biotechnol. 7: 110.
    Pubmed KoreaMed CrossRef
  3. Sarrouh B, Santos TM, Miyoshi A, Dias R, Azevedo V. 2012. Up-ToDate insight on industrial enzymes applications and global market. J. Bioproces. Biotech. 4: 002. doi:10.4172/2155-9821.S4-002
    CrossRef
  4. Niyonzima FN, More S. 2014. Detergent-compatible proteases:Microbial production, properties, and stain removal analysis. Prep. Biochem. Biotechnol. 45: 233-258.
    Pubmed CrossRef
  5. Rao MB, Tanksale AM, Ghatge MS, Deshpande VV. 1998. Molecular and biotechnological aspects of microbial proteases. Microbiol. Mol. Biol. Rev. 62: 597-635.
    Pubmed KoreaMed CrossRef
  6. Gupta R, Beg QK, Lorenz P. 2002. Bacterial alkaline proteases:molecular approaches and industrial applications. Appl. Microbiol. Biotechnol. 59: 15-32.
    Pubmed CrossRef
  7. Sharma KM, Kumar R, Panwar S, Kumar A. 2017. Microbial alkaline proteases: Optimization of production parameters and their properties. J. Gen. Eng. Biotechnol. 15: 115-126.
    Pubmed KoreaMed CrossRef
  8. Verma J, Pandey S. 2019. Characterization of partially purified alkaline protease secreted by halophilic bacterium Citricoccus sp. isolated from agricultural soil of northern India. Biocat. Agric. Biotechnol. 17: 605-612.
    CrossRef
  9. Salwan R, Sharma V. 2019. Trends in extracellular serine proteases of bacteria as detergent bioadditive: alternate and environmental friendly tool for detergent industry. Arch. Microbiol. 201:863-877.
    Pubmed CrossRef
  10. Lakshmi BK, Muni Kumar D, Hemalatha KP. 2018. Purification and characterization of alkaline protease with novel properties from Bacillus cereus strain S8. J. Gen. Eng. Biotechnol. 16: 295-304.
    Pubmed KoreaMed CrossRef
  11. Zacaria J, Delamare APL, Costa SOP, Echeverrigaray S. 2010. Diversity of extracellular proteases among Aeromonas determined by zymogram analysis. J. Appl. Microbiol. 109: 212-219.
    Pubmed CrossRef
  12. Laishram S, Pennathur G. 2015. Purification and characterization of a membrane-unbound highly thermostable metalloprotease from Aeromonas Caviae. Arab. J. Sci. Eng. 41: 2107-2116.
    CrossRef
  13. Cho SJ, Park JH, Park SJ, Lim JS, Kim EH, Cho YJ, et al. 2003. Purification and characterization of extracellular temperature-stable serine protease from Aeromonas hydrophila. J. Microbiol. 41: 207-211.
  14. Divakar K, Deepa Arul Priya J, Gautam P. 2010. Purification and characterization of thermostable organic solvent-stable protease from Aeromonas veronii PG01. J. Mol. Catal. B: Enz. 66: 311-318.
    CrossRef
  15. Datta S, Menon G, Varughese B. 2016. Production, characterization, and immobilization of partially purified surfactant-detergent and alkali-thermostable protease from newly isolated Aeromonas caviae. Prep. Biochem. Biotechnol. 47: 349-356.
    Pubmed CrossRef
  16. Thompson JD, Higgins DG, Gibson TJ. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucl. Acids Res. 22: 4673-4680.
    Pubmed KoreaMed CrossRef
  17. Sellami-Kamoun A, Ghorbel-Frikha B, Haddar A, Nasri M. 2011. Enhanced Bacillus cereus BG1 protease production by the use of sardinelle (Sardinella aurita) powder. Ann. Microbiol. 61: 273-280.
    CrossRef
  18. Miller JH. 1972. Experiments in moleculer genetics. pp. 431-435. Cold Spring Harbor, NY: Cold483 Spring Harbor Laboratory Press.
  19. Kembhavi AA, Kulkarni A, Pant A. 1993. Salt-tolerant and thermostable alkaline protease from Bacillus subtilis NCIM No. 64. Appl. Biochem. Biotechnol. 38: 83-92.
    Pubmed CrossRef
  20. Manni L, Jellouli K, Ghorbel-Bellaaj O, Agrebi R, Haddar A, SellamiKamoun A, et al. 2009. An oxidant- and solvent-stable protease produced by Bacillus cereus SV1: Application in the deproteinization of shrimp wastes and as a laundry detergent additive. Appl. Biochem. Biotechnol. 160: 2308-2321.
    Pubmed CrossRef
  21. Maurer KH. 2004. Detergent proteases. Curr. Opi. Biotechnol. 15:330-334.
    Pubmed CrossRef
  22. Haddar A, Bougatef A, Agrebi R, Sellami-Kamoun A, Nasri M. 2009. A novel surfactant-stable alkaline serine-protease from a newly isolated Bacillus mojavensis A21. Purification and characterization. Proc. Biochem. 44: 29-35.
    CrossRef
  23. Uttatree S, Charoenpanich J. 2016. Isolation and characterization of a broad pH- and temperature-active, solvent and surfactant stable protease from a new strain of Bacillus subtilis. Biocat. Agricul. Biotechnol. 8: 32- 38.
    CrossRef
  24. Thakur N, Kumar A, Sharma A, Bhalla TC, Kumar D. 2018. Purification and characterization of alkaline, thermostable and organic solvent stable protease from a mutant of Bacillus sp. Biocat. Agricult. Biotechnol. 16: 217-224.
    CrossRef
  25. Aryaei A, Farhadi A, Moradian F, Mianji GR. 2019. Cloning, expression and characterization of a novel alkaline serine protease gene from native Iranian Bacillus sp.; a producer of protease for use in livestock. Gene 693: 10-15.
    Pubmed CrossRef
  26. Nakasone N, Toma C, Song T, Iwanaga M. 2004. Purification and characterization of a novel metalloprotease isolated from Aeromonas caviae. FEMS Microbiol. Lett. 237: 127-132.
    Pubmed CrossRef
  27. Karunakaran T, Devi BG. 1995. Proteolytic activity of Aeromonas caviae. J. Basic Microbiol. 35: 241-247.
    Pubmed CrossRef
  28. Horikoshi K. 1990. Enzymes of alkalophiles. In: Louis, M.O., St. pp. 275-294. (Ed.), Microbial Enzymes and Biotechnology, second ed. Elsevier Applied Science, Amsterdam.
    CrossRef
  29. Sareen R, Mishra P. 2008. Purification and characterization of organic solvent stable protease from Bacillus licheniformis RSP09-37. Appl.Microbiol. Biotechnol. 79: 399-405.
    Pubmed CrossRef
  30. Toma C, Ichinose Y, Iwanaga M. 1999. Purification and characterization of an Aeromonas caviae metalloprotease that is related to the Vibrio cholera hemagglutinin/protease. FEMS Microbiol. Lett. 170: 237-242.
    Pubmed CrossRef
  31. Hadjidj R, Badis A, Mechri S, Eddouaouda K, Khelouia L, Annane R, et al. 2018. Purification, biochemical, and molecular characterization of novel protease from Bacillus licheniformis strain K7A. Int. J. Biol. Macromol. 114: 1033-1048.
    Pubmed CrossRef
  32. Patel AR, Mokashe NU, Chaudhari DS, Jadhav AG, Patil UK. 2019. Production optimisation and characterisation of extracellular protease secreted by newly isolated Bacillus subtilis AU-2 strain obtained from Tribolium castaneum gut. Biocatal. Agric. Biotechnol. 19: 101-122.
    CrossRef
  33. Iqbal A, Hakim A, Hossain MS, Rahman MR, Islam K, Azim MF, et al. 2018. Partial purification and characterization of serine protease produced through fermentation of organic municipal solid wastes by Serratia marcescens A3 and Pseudomonas putida A2. J. Gen. Eng. Biotechnol. 16: 29-37.
    Pubmed KoreaMed CrossRef
  34. Zhang J, Wang J, Zhao Y, Li J, Liu Y. 2019. Study on the interaction between calcium ions and alkaline protease of Bacillus. Int. J. Biol. Macromol. 124: 121-130.
    Pubmed CrossRef
  35. Lee S, Jang DJ. 2001. Progressive rearrangement of subtilisin Carlsberg into orderly and inflexible conformation with Ca2+ binding. Biophys. J. 81: 2972-2988.
    CrossRef
  36. Rivero O, Anguita J, Paniagua C, Naharro G. 1990. Molecular cloning and characterization of an extracellular protease gene from Aeromonas hydrophila. J. Bacteriol. 172: 3905-3908.
    Pubmed KoreaMed CrossRef
  37. Beg QK, Gupta R. 2003. Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis. Enz. Microb. Technol. 32: 294-304.
    CrossRef
  38. Vulfson EN, Halling PJ, Holland HL. 2001. Methods in Biotechnology:Enzymes in Nonaqueous Solvents. Part II, 532, pp. 241-422.
    CrossRef
  39. Manni L, Ghorbel-Bellaaj O, Jellouli K, Younes I, Nasri M. 2009. Extraction and characterization of chitin, chitosan, and protein hydrolysates prepared from shrimp waste by treatment with crude protease from Bacillus cereus SV1. Appl. Biochem. Biotechnol. 162: 345-357.
    Pubmed CrossRef
  40. Sundus H, Mukhtar H, Nawaz A. 2016. Industrial applications and production sources of serine alkaline proteases: a review. J. Bacteriol. Myc. 3: 191-194.
    CrossRef
  41. Kumar CG, Takagi H. 1999. Microbial alkaline proteases from a bioindustrial viewpoint. Biotechnol. Adv. 17: 561-594.
    CrossRef
  42. Gupta R, Gupta K, Saxena RK, Khan S. 1999. Bleach-stable, alkaline protease from Bacillus sp. Biotechnol. Lett. 21: 135-138.
    CrossRef
  43. Rekik H, Jaouadi NZ, Gargouri F, Bejar W, Frikha F, Jmal, N, et al. 2019. Production, purification and biochemical characterization of a novel detergent-stable serine alkaline protease from Bacillus safensis strain RH12. Int. J. Biol. Macromol. 121: 1227-1239.
    Pubmed CrossRef
  44. Hammami A, Hamdi M, Abdelhedi O, Jridi M, Nasri M, Bayoudh A. 2017. Surfactant- and oxidant-stable alkaline proteases from Bacillus invictae?: Characterization and potential applications in chitin extraction and as a detergent additive. Int. J. Biol. Macromol. 96: 272-281.
    Pubmed CrossRef
  45. Biver S, Portetelle D, Vandenbol M. 2013. Characterization of a new oxidant-stable serine protease isolated by functional metagenomics. Springerplus 2: 410.
    Pubmed KoreaMed CrossRef
  46. Haddar A, Agrebi R, Bougatef A, Hmidet N, Sellami-Kamoun A, Nasri M. 2009. Two detergent stable alkaline serine-proteases from Bacillus mojavensis A21: purification, characterization and potential application as a laundry detergent additive. Bioresour. Technol. 100: 3366-3373.
    Pubmed CrossRef
  47. Sellami-Kamoun A, Haddar A, Ali NEH, Ghorbel-Frikha B, Kanoun S, Nasri M. 2008. Stability of thermostable alkaline protease from Bacillus licheniformis RP1 in commercial solid laundry detergent formulations. Microbiol. Res. 163: 299-306.
    Pubmed CrossRef
  48. Yu P, Huang X, Ren Q, Wang X. 2019. Purification and characterization of a H2O2-tolerant alkaline protease from Bacillus sp. ZJ1502, a newly isolated strain from fermented bean curd. Food Chem. 274: 510-517.
    Pubmed CrossRef
  49. David A, Singh Chauhan P, Kumar A, Angural S, Kumar D, Puri N, Gupta N. 2018. Coproduction of protease and mannanase from Bacillus nealsonii PN-11 in solid state fermentation and their combined application as detergent additives. Int. J. Biol. Macromol. 108: 1176-1184.
    Pubmed CrossRef
  50. Singh J, Batra N, Sobti R. 2001. Serine alkaline protease from a newly isolated Bacillus sp. SSR1. Proc. Biochem. 36: 781-785.
    CrossRef
  51. Banik RM, Prakash M. 2004. Laundry detergent compatibility of the alkaline protease from Bacillus cereus. Microbiol. Res. 159: 135-140.
    Pubmed CrossRef
  52. Jain D, Pancha I, Mishra SK, Shrivastav A, Mishra S. 2012. Purification and characterization of haloalkaline thermoactive, solvent stable and SDS induced protease from Bacillus sp.: a potential additive for laundry detergents. Bioresour. Technol. 115: 228-236.
    Pubmed CrossRef

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