Research Article
BibTex RIS Cite
Year 2018, Volume: 7 Issue: 2, 130 - 142, 17.08.2018
https://doi.org/10.18036/aubtdc.322711

Abstract

References

  • [1] Chu WH. Optimization of extracellular alkaline protease production from species of Bacillus. J Ind Microbiol Biot 2007; 34: 241-245. [2] Christanti AD. Isolasi dan Karakterisasi Bakteri Halotoleran pada terasi. MSc, Bogor Agricultural University, Bogor, Indenosia, 2006. [3] DasSarma S, Arora P. Halophiles. In: Encyclopedia of Life Sciences. Nature Publishing Group, 2001. [4] Dewi WK. Pemurnian dan Pencirian Protease dari Isolat Bakteri W-1 yang Dihasilkan oleh Tauco Hitam. Msc, Bogor Agricultural University, Bogor, Indenosia, 2006. [5] Dias DR, Vilela DM, Silvestre MPC, Schwan RF. Alkaline protease from Bacillus sp.isolated from coffee bean grown on cheese whey. World J Microb Biot 2008; 24: 2027-2034. [6] Elbanna K, Ibrahim IM, Revol-Junelles AM. Purification and characterization of haloalkali thermophilic protease from Halobacterium sp. strain HP25 isolated from raw salt, Lake Qarun, Fayoum, Egypt. Extremophiles 2015;19: 763-774. [7] Fitriani S. Partial Purification and Characterization of Protease Enzyme from B19 KUB BPPT CC isolate. MSc, Bogor Agricultural University, Bogor, Indenosia, 2013. [8] Ghafoori H, Askari M, Sarikhan S. Purification and characterization of an extracellular haloalkaline serine protease from the moderately halophilic bacterium, Bacillus iranensis (X5B). Extremophiles 2016; 20:115-123. [9] Gupta S, Sharma P, Dev K, Sourirajan A. Halophilic Bacteria of Lunsu Produce an Array of Industrially Important Enzymes with Salt Tolerant Activity. Biochemistry Research International 2016; http://dx.doi.org/10.1155/2016/9237418 [10] Guven K, Mutlu MB, Gulbandilar A, Cakir P. Occurrence and Characterization of Staphylococcus aureus Isolated From Meat And Dairy Products Consumed In Turkey. J Food Safety 2008; 30: 196-212. [11] Heyndrickx M, Lebbe L, Kersters K, De Vos P, Forsyth G, Logan' NA. Virgibacillus: a new genus to accommodate Bacillus pantothentics Emended description of Virgibacillus pantothenticus. Int J Syst Bacteriol 1998; 48: 99-106. [12] Ibrahim ASS, Al-Salamah AA, El-Badawi YB, El-Tayeb MA, Antranikian G. Detergent, solvent and salt compatible thermoactive alkaline serine protease from halotolerant alkaliphilic Bacillus sp. NPST‑AK15: purification and characterization. Extremophiles 2015; 19: 961-971. [13] Joo HS, Chang CS. Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties. Process Biochem 2005; 40:1263-1270. [14] Karbalaei-Heidari HR, Ziaee AA, Amoozegar MA. Purification and biochemical characterization of a protease secreted by the Salinivibrio sp. strain AF-2004 and its behavior in organic solvents. Extremophiles 2007; 11: 237-243. [15] Karbalaei-Heidari HR, Ziaee AA, Hajighasemi M, Amoozegar MA, Ventosa A. Production, optimization and purification of a novel extracellular protease from the moderately halophilic bacterium Halobacillus karajensis. J Ind Microbiol Biot 2009; 36: 21-27. [16] Laemmli UK. Cleavage of structural proteins during teh assembly of the head of Bateriophage T4. Nature 1970; 7: 680-685. [17] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent.1951; http://www.jbc.org. (Accessed date : 13.04.16). [18] Madern D, Ebel C, Zacca G. Halophilic adaptation of enzymes. Extremophiles 2000; 4: 91-98. [19] Maruthiah T, Immanuel G, Palavesam A. Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci 2015. [20] Mutlu MB. Tuz Gölü Bakterilerinin Karakterizasyonu ve Mevsimsel Dağılımı.PhD, Anadolu Üniversitesi, Eskisehir, Türkiye, 2006. [21] Namwong S, Hiraga K, Takada K, Tsunemi M, Tanasupawat S, Oda K. A halophilic serine proteinase from Halobacillus sp. SR5-3 isolated from fish sauce: purification and characterization. Biosci Biotechnol Biochem 2006; 70:1395-1401. [22] Nawab A, Nimat U, Muhammad Q, Hazir R, Shahid K, Abdul S, Muhammad A. Molecular characterization and growth optimization of halotolerant protease producing Bacillus subtilis Strain BLK‑1.5 isolated from salt mines of Karak, Pakistan. Extremophiles 2016; 20(4):395-402. [23] Olajuyigbe FM, Falade AM. Purification and partial characterization of serine alkaline metalloprotease from Bacillus brevis MWB-01. Bioresources Bioproces 2014; 1:8. [24] Oren A. Industrial and environmental applications of halophilic microorganisms. Environ Technol 2010; 8(31): 825-834. [25] Rao MB, Tanksale AM, Ghatge MS, Desphande VV. Molecular and biotechnological aspects of microbial proteases. Mol Biol Rev 1998; 62(3): 597-635. [26] Shahbazi M, Karbalaei-Heidari HR. A novel low molecular weight extracellular protease from a moderately halophilic bacterium Salinivibrio sp. strain MS-7: production and biochemical properties. Mol Biol Res Comm 2012; 1(2): 45-56. [27] Spring S, Ludwug W, Marquez MC, Ventosa A, Schleifer KH. Halobacillus gen. nov., with Descriptions of Halobacillus litoralis sp. nov. and Halobacillus trueperi sp. nov., and Transfer of Sporosarcina halophila to Halobacillus halophilus comb. nov. Int J Syst Bacteriol (1996). 46(2), 492-496. [28] Steinkraus KH. Handbook of indigenous Fermented Foods. 2nd ed. New York, Marcel Dekker, Inc,1995. [29] Surono IS, Hosono A. Microflora and Their Enzyme Profile in "Terasi" Starter. Biosci. Biotech. Biochem. 1994; 58 (6):1167-1169. [30] Taprig T, Akaracharanya A, Sitdhipol J, Visessanguan W, Tanasupawat S. Screening and characterization of protease-producing Virgibacillus, Halobacillus and Oceanobacillus strains from Thai fermented fish. J App Pharm Sci 2013; 3(02):25-30. [31] Wang ZF, Huang MQ, Zuo XM, Zhou HM. Unfolding, conformational change of active sites and inactivation of creatine kinase in SDS solutions. Biochim Biophys Acta 1995; 1251(2): 109-114. [32] Wu JW and Chen XL. Extracellular metalloproteases from bacteria. Appl Microbiol Biotechnol 2011; 92: 253-262. [33] Xu J, Jiang M, Sun H, He B. An organic solvent-stable protease from organic solvent-tolerant Bacillus cereus WQ9-2: Purification, biochemical properties, and potential application in peptide synthesis. Bioresource Technol 2010;101: 7991-79
  • Reference2
  • Reference3

ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD

Year 2018, Volume: 7 Issue: 2, 130 - 142, 17.08.2018
https://doi.org/10.18036/aubtdc.322711

Abstract














The protease producing bacteria were screened
from Indonesian traditional fermented food, tauco and terasi.
During the study, 4 halophilic protease producers were isolated from tauco and
terasi. Among these isolates, halophilic bacterial isolate TANN 4 was
recorded as the best protease producer. Extracellular protease from isolate
TANN 4 was partially purified using ammonium sulfate precipitation. The
protease was partially purified with final yield of 72.87 % and 25.41 fold
purity. This moderate thermoactive and alkaliphilic protease showed a pH
optimum of 8.0 and temperature optimum was 50 °C. The enzyme was also active at
salt concentrations ranging from 1 to 15 % (w/v), with optimum activity at 1 %
NaCl (w/v). Ethylenediaminetetraacetic acid (EDTA) completely inhibited the
enzyme activity suggesting that it was a metalloprotease. Among metal ions, the
Ca2+, K+ and Mg2+ ions enhanced the activity
of enzyme. The KM and Vmax values exhibited by partially purified
protease were 0.0649 mM and 216.45 U mg−1 using casein as substrate.
The molecular weight was estimated to be 19.8 kDa on SDS PAGE. The enzyme also
fairly stable in Triton X-100, SDS, 1 % commercial detergents (OMO and Ariel)
and 25 % methanol. This enzyme was capable of hydrolyzing casein, hemoglobin
and bovine serum albumin (BSA). Automated ribotyping analysis revealed that 3
isolate (TANN 4, TR 2 and TR 4) resembled Halobacillus trueperi that
exhibited 71, 68 and 69 % similarity respectively, and isolate (TR 1) resembled
Virgibacillus pantothenticus with 64 % similarity. These characteristics
make this halophilic bacterial extracellular metalloprotease seems to be
potentially useful for biotechnological and industrial applications.




References

  • [1] Chu WH. Optimization of extracellular alkaline protease production from species of Bacillus. J Ind Microbiol Biot 2007; 34: 241-245. [2] Christanti AD. Isolasi dan Karakterisasi Bakteri Halotoleran pada terasi. MSc, Bogor Agricultural University, Bogor, Indenosia, 2006. [3] DasSarma S, Arora P. Halophiles. In: Encyclopedia of Life Sciences. Nature Publishing Group, 2001. [4] Dewi WK. Pemurnian dan Pencirian Protease dari Isolat Bakteri W-1 yang Dihasilkan oleh Tauco Hitam. Msc, Bogor Agricultural University, Bogor, Indenosia, 2006. [5] Dias DR, Vilela DM, Silvestre MPC, Schwan RF. Alkaline protease from Bacillus sp.isolated from coffee bean grown on cheese whey. World J Microb Biot 2008; 24: 2027-2034. [6] Elbanna K, Ibrahim IM, Revol-Junelles AM. Purification and characterization of haloalkali thermophilic protease from Halobacterium sp. strain HP25 isolated from raw salt, Lake Qarun, Fayoum, Egypt. Extremophiles 2015;19: 763-774. [7] Fitriani S. Partial Purification and Characterization of Protease Enzyme from B19 KUB BPPT CC isolate. MSc, Bogor Agricultural University, Bogor, Indenosia, 2013. [8] Ghafoori H, Askari M, Sarikhan S. Purification and characterization of an extracellular haloalkaline serine protease from the moderately halophilic bacterium, Bacillus iranensis (X5B). Extremophiles 2016; 20:115-123. [9] Gupta S, Sharma P, Dev K, Sourirajan A. Halophilic Bacteria of Lunsu Produce an Array of Industrially Important Enzymes with Salt Tolerant Activity. Biochemistry Research International 2016; http://dx.doi.org/10.1155/2016/9237418 [10] Guven K, Mutlu MB, Gulbandilar A, Cakir P. Occurrence and Characterization of Staphylococcus aureus Isolated From Meat And Dairy Products Consumed In Turkey. J Food Safety 2008; 30: 196-212. [11] Heyndrickx M, Lebbe L, Kersters K, De Vos P, Forsyth G, Logan' NA. Virgibacillus: a new genus to accommodate Bacillus pantothentics Emended description of Virgibacillus pantothenticus. Int J Syst Bacteriol 1998; 48: 99-106. [12] Ibrahim ASS, Al-Salamah AA, El-Badawi YB, El-Tayeb MA, Antranikian G. Detergent, solvent and salt compatible thermoactive alkaline serine protease from halotolerant alkaliphilic Bacillus sp. NPST‑AK15: purification and characterization. Extremophiles 2015; 19: 961-971. [13] Joo HS, Chang CS. Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties. Process Biochem 2005; 40:1263-1270. [14] Karbalaei-Heidari HR, Ziaee AA, Amoozegar MA. Purification and biochemical characterization of a protease secreted by the Salinivibrio sp. strain AF-2004 and its behavior in organic solvents. Extremophiles 2007; 11: 237-243. [15] Karbalaei-Heidari HR, Ziaee AA, Hajighasemi M, Amoozegar MA, Ventosa A. Production, optimization and purification of a novel extracellular protease from the moderately halophilic bacterium Halobacillus karajensis. J Ind Microbiol Biot 2009; 36: 21-27. [16] Laemmli UK. Cleavage of structural proteins during teh assembly of the head of Bateriophage T4. Nature 1970; 7: 680-685. [17] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent.1951; http://www.jbc.org. (Accessed date : 13.04.16). [18] Madern D, Ebel C, Zacca G. Halophilic adaptation of enzymes. Extremophiles 2000; 4: 91-98. [19] Maruthiah T, Immanuel G, Palavesam A. Purification and Characterization of Halophilic Organic Solvent Tolerant Protease from Marine Bacillus sp. APCMST-RS7 and Its Antioxidant Potentials. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci 2015. [20] Mutlu MB. Tuz Gölü Bakterilerinin Karakterizasyonu ve Mevsimsel Dağılımı.PhD, Anadolu Üniversitesi, Eskisehir, Türkiye, 2006. [21] Namwong S, Hiraga K, Takada K, Tsunemi M, Tanasupawat S, Oda K. A halophilic serine proteinase from Halobacillus sp. SR5-3 isolated from fish sauce: purification and characterization. Biosci Biotechnol Biochem 2006; 70:1395-1401. [22] Nawab A, Nimat U, Muhammad Q, Hazir R, Shahid K, Abdul S, Muhammad A. Molecular characterization and growth optimization of halotolerant protease producing Bacillus subtilis Strain BLK‑1.5 isolated from salt mines of Karak, Pakistan. Extremophiles 2016; 20(4):395-402. [23] Olajuyigbe FM, Falade AM. Purification and partial characterization of serine alkaline metalloprotease from Bacillus brevis MWB-01. Bioresources Bioproces 2014; 1:8. [24] Oren A. Industrial and environmental applications of halophilic microorganisms. Environ Technol 2010; 8(31): 825-834. [25] Rao MB, Tanksale AM, Ghatge MS, Desphande VV. Molecular and biotechnological aspects of microbial proteases. Mol Biol Rev 1998; 62(3): 597-635. [26] Shahbazi M, Karbalaei-Heidari HR. A novel low molecular weight extracellular protease from a moderately halophilic bacterium Salinivibrio sp. strain MS-7: production and biochemical properties. Mol Biol Res Comm 2012; 1(2): 45-56. [27] Spring S, Ludwug W, Marquez MC, Ventosa A, Schleifer KH. Halobacillus gen. nov., with Descriptions of Halobacillus litoralis sp. nov. and Halobacillus trueperi sp. nov., and Transfer of Sporosarcina halophila to Halobacillus halophilus comb. nov. Int J Syst Bacteriol (1996). 46(2), 492-496. [28] Steinkraus KH. Handbook of indigenous Fermented Foods. 2nd ed. New York, Marcel Dekker, Inc,1995. [29] Surono IS, Hosono A. Microflora and Their Enzyme Profile in "Terasi" Starter. Biosci. Biotech. Biochem. 1994; 58 (6):1167-1169. [30] Taprig T, Akaracharanya A, Sitdhipol J, Visessanguan W, Tanasupawat S. Screening and characterization of protease-producing Virgibacillus, Halobacillus and Oceanobacillus strains from Thai fermented fish. J App Pharm Sci 2013; 3(02):25-30. [31] Wang ZF, Huang MQ, Zuo XM, Zhou HM. Unfolding, conformational change of active sites and inactivation of creatine kinase in SDS solutions. Biochim Biophys Acta 1995; 1251(2): 109-114. [32] Wu JW and Chen XL. Extracellular metalloproteases from bacteria. Appl Microbiol Biotechnol 2011; 92: 253-262. [33] Xu J, Jiang M, Sun H, He B. An organic solvent-stable protease from organic solvent-tolerant Bacillus cereus WQ9-2: Purification, biochemical properties, and potential application in peptide synthesis. Bioresource Technol 2010;101: 7991-79
  • Reference2
  • Reference3
There are 3 citations in total.

Details

Primary Language English
Journal Section Articles
Authors

Sarah Fıtrıanı This is me

Kıymet Güven

Publication Date August 17, 2018
Published in Issue Year 2018 Volume: 7 Issue: 2

Cite

APA Fıtrıanı, S., & Güven, K. (2018). ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology, 7(2), 130-142. https://doi.org/10.18036/aubtdc.322711
AMA Fıtrıanı S, Güven K. ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology. August 2018;7(2):130-142. doi:10.18036/aubtdc.322711
Chicago Fıtrıanı, Sarah, and Kıymet Güven. “ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD”. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology 7, no. 2 (August 2018): 130-42. https://doi.org/10.18036/aubtdc.322711.
EndNote Fıtrıanı S, Güven K (August 1, 2018) ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology 7 2 130–142.
IEEE S. Fıtrıanı and K. Güven, “ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD”, Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology, vol. 7, no. 2, pp. 130–142, 2018, doi: 10.18036/aubtdc.322711.
ISNAD Fıtrıanı, Sarah - Güven, Kıymet. “ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD”. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology 7/2 (August 2018), 130-142. https://doi.org/10.18036/aubtdc.322711.
JAMA Fıtrıanı S, Güven K. ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology. 2018;7:130–142.
MLA Fıtrıanı, Sarah and Kıymet Güven. “ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD”. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology, vol. 7, no. 2, 2018, pp. 130-42, doi:10.18036/aubtdc.322711.
Vancouver Fıtrıanı S, Güven K. ISOLATION, SCREENING, PARTIAL PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM HALOPHILIC BACTERIA ISOLATED FROM INDONESIAN FERMENTED FOOD. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology. 2018;7(2):130-42.