Screening and Characterization of Proteolytic Thermophiles Bacteria from Moinit Coastal Hot-Spring, North Sulawesi

Elvy Like Ginting

doi:10.35800/jip.8.1.2020.28628

Screening and Characterization of Proteolytic Thermophiles Bacteria from Moinit Coastal Hot-Spring, North Sulawesi

Elvy Like Ginting

Abstract

Thermophilic bacteria have been recognized as the ideal producers of thermostable enzymes and therefore they have a long time become an interesting target for research. Thermophilic proteolytic bacteria from Moinit coastal hot-spring North Sulawesi Indonesia have been isolated and some of them, have been identified and reported. In this research, we reported another eleven thermophilic proteolytic bacteria from Moinit coastal hot-spring, North Sulawesi, Indonesia which designated as 1a, 1b, 2a, 3a, 3b, 4a, 11a, 11b, 16a, 16b, and 16c. Those thermophilic proteolytic bacteria had different colony and cell morphology characteristics, and biochemical characteristics. Bacterial isolate 1a, 1b, 2a, 16a and 16c were Gram negative-bacilli, 3a, 3b and 11b were Gram positive-bacilli, 4a, 11a and 16b were Gram positive-coccus. These bacterial isolates could produce proteolytic activities at 55^oC. This showed their ability to produce thermostable protease extracellularly which has the potential to explore biotechnological industries.

Keywords: Moinit, protease, screening, thermophilic

ABSTRAK

Bakteri termofilik dikenal sebagai produsen enzim termostabil dan menjadi target yang menarik untuk diteliti. Bakteri proteolitik termofil dari sumber air panas pantai Moinit Sulawesi Utara telah berhasil diisolasi dan bahkan 5 isolat bakteri telah diidentifikasi dan dilaporkan. Dalam tulisan ini, diuraikan sebelas isolat bakteri proteolitik termofil lainnya dalam kemampuannya menghasilkan protease dan karakteristik bakteri tersebut. Kesebelas bakteri adalah 1a, 1b, 2a, 3a, 3b, 4a, 11a, 11b, 16b, 16b, 16b, dan 16c yang menunjukkan karakteristik morfologi koloni dan sel bakteri, serta karakteristik biokimia yang berbeda. Isolat 1a, 1b, 2a, 16a, 16c adalah Gram negatif-basil, isolat 3a, 3b, 11b adalah Gram positif-basil, dan isolat 4a, 11a,16b adalah Gram positif-kokus. Keseluruhan isolat bakteri ini memiliki kemampuan aktivitas proteolitik pada 55^oC. Hal ini menunjukkan kemampuan bakteri tersebut dalam menghasilkan protease termostabil ekstraseluler yang berpotensi untuk dieksplorasi dalam pemanfaatannya dalam bidang industri bioteknologi.

Kata kunci: Moinit, protease, penapisan, termofilik

Keywords

Moinit, protease, penapisan, termofilik

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References

Baehaki, A., Rinto, Budiman, A. 2011 Isolation and characterization of protease from bacterial Swamp Land Indralaya. J. Teknol and Food Industry 22(1):37–42.

Berquist, P.L., Love, D.R., Croft, J.E., Streiff, M.B., Daniel, R.M., Morgan, W.H. 1987. Genetics and potential biotechnological applications of thermophilic and extremely thermophilic microorganisms. Biotech. Gen. Eng. Rev. 5: 199–237.

Cappuccino, J.G., Sherman, N. 2010. Microbiology: A laboratory manual. Ninth Edition. New York: Addison – Wesley Publishing Company.

Dias, D.R., Vilela, D.M., Silvestre, M.P.C., Schwan, R.F. 2008. Alkaline protease from Bacillus sp. isolated from coffee bean grown on cheese whey. World J. Microbiol. Biotechnol. 24:2027–2034.

Dwidjoseputro, D. 1990. Dasar-dasar mikrobiologi. Penerbit Djambatan Jakarta.

Hadioetomo, R. S. 1993. Mikrobiologi dasar dalam praktek teknik dan prosedur dasar Laboratorium. PT Gramedia. Jakarta.

Friedman, S. M. 1992. Thermophilic microorganisms. Encyclop. Microbial. 4:217–229.

Frock, A.D., Notey, J.S., Kelly, R.M. 2010. The genus Thermotoga: Recent developments. Environ Technol. 31(10): 1169–1181.

Ginting, E.L, Kemer, K., Wullur, S., Uria, A.R. 2019. Identification of proteolytic thermophiles from Moinit coastal hot-spring, North Sulawesi, Indonesia. Geomicrobiology Journal 37(1): 50–58.

Holt, J. G. N. R., Krieg. Ph, A., Sneath. J. T., Staley, Williams. S. T. 1994. Bergey’s manual of determination bacteriology. Williams and Wilkins. Baltomore. 475 hal.

Jawetz, E., Melnick, J.L., Adelberg, E.A. 2001. Mikrobiologi kedokteran. Edisi 2. Penerbit Buku Kedokteran. Jakarta.

Kristjansson, J. K. 1991. Thermophilic bacteria. CRC Press. Florida.

Lakshmi, B.K.M., Ratnasri, P.V., Devi, K.A., Hemalatha, K.P.J. 2014. Screening, optimization of producing and partial characterization of alkaline protease from haloalkaliphilic Bacillus sp. IJRET: International Journal of Research in Engineering and Technology 3(2): 435–443.

Lay, W. B. 1994. Analisis mikroba di laboratorium. PT Raja Grafindo Persada, Jakarta.

Madigan, Marthinko, Stahl, Clark. 2012. Biology of microorganisms. Pearson Education, Inc. San Francisco, CA 94111.

Ninghoujam D. S., Kshetri P. 2010 A thermostable alkaline protease from a moderately halo-alkalithermotolerant Bacillus subtilis strain SH1. Australian J.Basic Appl Sci. 4: 5126–5134.

Panda, M.K., Sahu, M.K., Tayung, K. 2012. Isolation and characterization of a thermophilic Bacillus sp. with protease activity isolated from hot spring of Tarabalo, Odisha, India. Iranian Journal of Microbiol. 5(2): 159–165.

Pathak, A.P., Rathod, M.G. 2014. Cultivable bacterial diversity of terrestrial thermal spring of Unkeshwar, India. J Biochem. 5(4): 814–818.

Pelczar, M. J. Jr, Chan, E. C. S. 1998, Dasar- dasar mikrobiologi 2. Cetakan 1. Penerbit Universitas Indonesia, Jakarta.

Pollitt, E. J. G., Diggle, S. P. 2017. Defining motility in the Staphylococci. Cellular and Molecular Life Sciences, 74(16): 2943–2958.

Purwoko, C. 2007. Fisiologi mikroba. Edisi I. PT Bumi Akasara, Jakarta.

Rai, S.K., Roy, J.K., Mukherjee, A.K. 2010. Characterization of a detergent-stable alkaline protease from a novel thermophilic strain Paenibacillus tezpurensis sp. Nov. AS-S24-II. Appl. Micrbiol. Biotechnol. 85: 1437–1450.

Schelegel, S.G. 1993. General microbiology. Crambridge University Press. Great Britain. 688p.

Synowiecki, J. 2010. Some applications of thermophiles and their enzymes for protein processing. Afr. J. Biotechnol. 9: 7020–7025.

Swithers, K.S., DiPippo, J.L, Bruce,D.C., Detter, C., Tapia, R., Han, S., Saunders, E., Foodwin, L., Han, J., Woyke, T., Pitluck S., Pennacchio, L., Nolan, M., Mikhailova, N., Lykidis, A., Land, M., Brettin, T., Stetter, K.O., Nelson, K.E., Gogarten, J.P., Noll, K.M. 2011. Genome sequence of Thermotoga sp. strain RQ2, a hyperthermophilic bacterium isolated from a geothermal heated region of sea seafloor near Ribeira Quente, the Azores. Journal of Bacteriologi 193 : 5869–5870.

Van den Burg, B. 2003. Extremophiles as source for novel enzymes. Curr. Opin. Microbiol. 6: 213–218.

Ward, O. P. 1983. Proteinase. In microbial enzyme and biotechnology. W. M. Fogarty. Applied Science Publisher. New York.

Wilson, P. Remigio, Z. 2012. Production and characterization of protease enzyme Produced by a novel moderate thermophilic bacterium (EP1001) isolated from an alkaline hotspring, Zimbabwe. African Journal of Microbiology Research 6(27): 5542–5551.

Xu, H., Han, D., Xu, Z. 2015. Expression of heterologous cellulases in Thermotoga sp. strain RQ2. BioMed Research International 2015:11.

Zilda, D.S., Harmayani, E., Widada, J., Asmara, W., Irianto, H.E., Patantis, G., Fawzya, Y.N. 2012. Screening of thermostable protease producing microorganisms isolated from Indonesia Hotspring. Squalen 7(3): 105–114.

DOI: https://doi.org/10.35800/jip.8.1.2020.28628