Characterization of thermophilic bacteria from Ie Seum Hot Springs, Aceh Besar, Indonesia as producers of protease enzyme
##plugins.themes.bootstrap3.article.sidebar##
Issue
Section
Articles
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
##plugins.themes.bootstrap3.article.main##
Abstract
Abstract. Sabaria E, Yasmin Y, Ismail YS, Bessania MA, Putri I, Fitri L. 2024. Characterization of thermophilic bacteria from Ie Seum Hot Springs, Aceh Besar, Indonesia as producers of protease enzyme. Biodiversitas 25: 1867-1874. Thermophilic bacteria are microorganisms that can survive in high-temperature environments exceeding 75°C. The Ie Seum Hot Springs in Aceh Besar, Indonesia are known to be a place where these thermophilic bacteria inhabit. In such extreme conditions, proteins and enzymes are often denatured, making these bacteria of interest due to their ability to adapt and produce enzymes such as proteases. Proteases are called proteolytic enzymes that can be applied both in the economic and medical fields. This study aimed to isolate and identify thermophilic bacteria species that have the highest potential for protease production in Ie Seum Hot Spring, Aceh Besar by analyzing the 16S rRNA gene and characterizing them using biochemical tests. Based on the result, seven thermophilic bacterial isolates, namely BT1, BT2, BT3, BT4, BT5, BT6, and BT7 were obtained, each showing different colony characteristics. The biochemical tests also revealed differences metabolic activity of each isolate. Of these isolates, BT4 exhibited the highest proteolytic index (4.65). Additionally, the 16S rRNA gene sequence analysis showed that the BT4 strain belongs to the genus Bacillus and has high similarities to Bacillus sp., B. licheniformis and B. sonorensis. These findings suggest that BT4 has significant potential as a source of thermophilic protease enzymes.
##plugins.themes.bootstrap3.article.details##
References
Alameri F, Tarique M, Osaili T, Obaid R, Abdalla A, Masad R, Al-Sbiei A, Cabezudo MF, Liu SQ, Al-Ramadi A, Ayyash M. 2022. Lactic acid bacteria isolated from fresh vegetable products: potential probiotic and postbiotic characteristics including immunomodulatory effects. Microorganisms. 10: 389. DOI: 10.3390/microorganisms10020389
Alghamdi S. 2022. Isolation and identification of the oral bacteria and their characterization for bacteriocin production in the oral cavity. Saudi J. Biol. Sci. 29: 318–323. DOI: 10.1016/j.sjbs.2021.08.096
Alrumman S, Mostafa YSM, Al-Qahtani S, Taha THT. 2018. Hydrolytic enzyme production by thermophilic bacteria isolated from Saudi hot springs. Open Life Sci. 13: 470–480. DOI: 10.1515/biol-2018-0056
Ardhi A, Sidauruk AN, Suraya N. 2020. Molecular identification of amylase-producing thermophilic bacteria isolated from Bukit Gadang Hot Spring, West Sumatra, Indonesia. Biodiversitas. 21: 994–1000. DOI: 10.13057/biodiv/d210319
Artha OA, Sudarno, Pramono H, Sari LA. 2019. Identification of extracellular enzyme-producing bacteria (proteolytic, cellulolytic, and amylolytic) in the sediment of extensive ponds in Tanggulrejo Gresik. IOP Conf. Ser.: Earth Environ. Sci. DOI: 10.1088/1755-1315/236/1/012003
Calogero R, Rizzo C, Arcadi E. Stipa MG, Consoli P, Romeo T, Battaglia P. 2022. Isolation and identification of luminescent bacteria in deep sea marine organisms from Sicilian waters (Mediterranean Sea). J. Mar. Sci. Eng. 10: 1113. 10.3390/jmse10081113
Das S, Nurunnabi TR, Parveen R, Mou AN, Emdadul MD, Islam MD, Rahman SMM. 2019. Isolation and characterization of indole acetic acid producing bacteria from rhizosphere soil and their effect on seed germination. Int J Curr Microbiol Appl Sci. 8: 1237–1245. DOI: 10.20546/ijcmas.2019.803.146
Dhayalan A, Velramar B, Govindasamy B, Ramalingam KR, Dilipkumar A, Pachiappan P. 2022. Isolation of a bacterial strain from the gut of the fish, Systomus sarana, identification of the isolated strain, optimized production of its protease, the enzyme purification, and partial structural characterization. J. Genet. Eng. & Biotechnol. 20:24. DOI: 10.1186/s43141-022-00299-3
Do T, Page JE, Walker S .2020. Uncovering the activities, biological roles, and regulation of bacterial cell wall hydrolases and tailoring enzymes. J. Biol. Chem. 295: 3347–3361. DOI: 10.1074/jbc.rev119.010155
Ejaz U, Muhammad S, Hashmi IA, Ali FI, Sohail M .2020. Utilization of methyltrioctylammonium chloride as new ionic liquid in pretreatment of sugarcane bagasse for production of cellulase by novel thermophilic bacteria. J. Biotech. 317: 34–38. DOI: 10.1016/j.jbiotec.2020.04.013
Fachrial E, Krisdianilo V, Harmileni, Lister INE, Nugroho TT, Saryono .2021. Isolation, characterization, activity test and molecular identification of thermophilic bacteria producing proteases from Dolok Tinggi Raja Natural Hot Springs, North Sumatra, Indonesia. Biodiversitas. 22: 1725–1732. 10.13057/biodiv/d220416
Feizabadi F, Sharifan A, Tajabadi N. 2021. Isolation and identification of lactic acid bacteria from stored Apis mellifera honey. J. Apic. Res. 60: 421–426. DOI: 10.1080/00218839.2020.1765490
Ginting EL, Kemer K, Wullur S, Uria AR. 2019. Identification of proteolytic thermophiles from Moinit Coastal Hot-Spring, North Sulawesi, Indonesia. Geomicrobiol. J. 37: 50–58. DOI: 10.1080/01490451.2019.1662524
Ginting EL. 2020. Screening and characterization of proteolytic thermophiles bacteria from Moinit Coastal Hot-Spring, North Sulawesi. J. Ilm. Platax. 8: 118-125. DOI: 10.35800/jip.8.1.2020.28628
Hadi SN, Widiyawati I, Dewi PS, Kartini. 2019. Isolation and characterization of buprofezin tolerant bacteria from rhizosfer of paddy at Marginal Land of Banyumas Regency. IOP Conf. Ser.: Earth Environ Sci. 255: 0–9. DOI: 10.1088/1755-1315/255/1/012009
Hamdani S, Asstiyani N, Astriany D, Singgih M, Ibrahim S . 2019. Isolation and identification of proteolytic bacteria from pig sludge and protease activity determination. IOP Conf. Ser.: Earth Environ. Sci. 1-7. DOI: 10.1088/1755-1315/230/1/012095
Harlé O, Falentin H, Niay J, Valence F, Courselaud C, Chuat V, Maillard MB, Guédon E, Deutsch SM, Thieery A. 2020. Diversity of the metabolic profiles of a broad range of lactic acid bacteria in soy juice fermentation. Food Microbiology. 89: 103410. DOI: 10.1016/j.fm.2019.103410
Ifandi S, Alwi M .2018. Isolation of thermophilic bacteria from Bora Hot Springs in Central Sulawesi. Biosaintifika. 10: 291–297. DOI: 10.15294/biosaintifika.v10i2.14905
Jitpakdee J, Kantachote D, Kanzaki H, Nitoda T. 2021. Selected probiotic lactic acid bacteria isolated from fermented foods for functional milk production: Lower cholesterol with more beneficial compounds. LWT-Food Sci Technol. 135: 110061. DOI: 10.1016/j.lwt.2020.110061
Kadaikunnan S, Rejiniemon SS, Khaled JM, Alharbi NS, Mothana R .2015. In-vitro antibacterial, antifungal, antioxidant and functional properties of Bacillus amyloliquefaciens. Ann. clin. microbiol. 14: 1–11. DOI: 10.1186/s12941-015-0069-1
Kasi PD, Suhaeni, Sasa. 2021. Karakterisasi morfologis isolat bakteri termofilik dari sumber Air Panas Pincara. Indigenous Biologi?: Jurnal Pendidikan dan Sains Biologi. 3: 51–56. DOI: https://doi.org/10.33323/indigenous.v3i2.40
Khalila R, Fitri L, Suhartono S. 2020. Isolation and characterization of thermophilic bacteria as cellulolytic enzyme producer from the Hot Spring of Ie Seuum Aceh Besar, Indonesia. Microbiol Indones. 14: 4. https://doi.org/10.5454/mi.14.1.4
Khosravi F, Khaleghi M, Naghavi H. 2021. Screening and identification of cellulose-degrading bacteria from soil and leaves at Kerman province, Iran. Arch. Microbiol. 204: 88. DOI: 10.1007/s00203-021-02713-9
Lischer K, Putra ABRD, Guslianto BW, Avilla F, Sitorus SG, Nugraha Y, Sarmoko. Short communication: The emergence and rise of indigenous thermophilic bacteria exploration from hot springs in Indonesia. Biodiversitas. 21: 5474–5481. DOI: 10.13057/biodiv/d211156
Logan NA, Vos P De. 2015. Bergey's Manual of Systematics of Archaea and Bacteria. Springer, New York.
Majidah R, Lubis SS, Harahap D. 2023. Potensi enzim selulase dari bakteri termofilik di Kawasan Wisata Ie Uum Kabupaten Aceh Besar. Kenanga. 3: 25-34. DOI: 10.22373/kenanga.v..i
Marasco R, Gazzillo M, Campolattano N, Sacco M, Muscariello L. 2022. Isolation and identification of lactic acid bacteria from natural whey cultures of buffalo and cow milk. Foods. 11: 233. DOI: 10.3390/foods11020233
Marathe SK, Vashistht MA, Prashanth A, Parveen N, Chakborty S, Nair SS. 2018. Isolation, partial purification, biochemical characterization and detergent compatibility of alkaline protease produced by Bacillus subtilis, Alcaligenes faecalis and Pseudomonas aeruginosa obtained from sea water samples. J. Genet. Eng. & Biotechnol. 16: 39–46. Doi: 10.1016/j.jgeb.2017.10.001
Masi C, Gemechu G, Tafesse M. 2021. Isolation, screening, characterization, and identification of alkaline protease-producing bacteria from leather industry effluent. Ann. Microbiol. 71:24. DOI: 10.21203/rs.3.rs-158491/v1
Nnolim NE, Nwodo UU. 2020. Bacillus sp. CSK2 produced thermostable alkaline keratinase using agro-wastes: keratinolytic enzyme characterization. BMC Biotechnol. 20: 1–14. DOI: 10.1186/s12896-020-00659-2
Octarya Z, Nugroho TT, Nurulita Y, Saryono. 2022. Molecular identification, GC-MS analysis of bbioactive compounds and antimicrobial activity of thermophilic bacteria derived from West Sumatra Hot-Spring Indonesia. Hayati J Biosci. 29: 549–561. DOI: 10.4308/hjb.29.4.549-561
Pandey A, Dhakar K, Sharma A (2015) Thermophilic bacteria that tolerate a wide temperature and pH range colonize the Soldhar (95°C) and Ringigad (80°C) hot springs of Uttarakhand, India. Ann. Microbiol. 65: 809–816. DOI: 10.1007/s13213-014-0921-0
Raveendran S, Parameswaran B, Ummalyma SB, Abraham A., Mathew AK., Madhavan A., Rebello S & Pandey A. (2018). Applications of microbial enzymes in food industry. Food Technol Biotech. 56, 16. DOI: 10.17113/ftb.56.01.18.5491
Runtuboi DYP, Gunaedi T, Simonapendi M, Pakpahan NNL (2018) Isolasi dan identifikasi bakteri termofilik dari sumber air panas di moso distrik muara tami kota jayapura provinsi papua. J. biol. Papua. 10: 68–73. DOI: 10.31957/jbp.474
Sanam MUE, Detha AIR, Rohi NK. 2022. Detection of antibacterial activity of lactic acid bacteria, isolated from Sumba mare's milk, against Bacillus cereus, Staphylococcus aureus, and Escherichia coli. J. adv. vet. anim. res.9: 53–58. DOI: 10.5455/javar.2022.i568
Sandoval BA, Hyster TK. 2020. Emerging strategies for expanding the toolbox of enzymes in biocatalysis. Curr Opin Chem Biol. 55: 45–51. DOI: 10.1016/j.cbpa.2019.12.006.
Sang P, Liu SQ, Yang LQ. 2020. New insight into mechanisms of protein adaptation to high temperatures: A comparative molecular dynamics simulation research of thermophilic and mesophilic subtilisin-like serine proteases. Int. J. Mol. Sci. 21: 3128. DOI: 10.3390/ijms21093128
Saryono, Novianty R, Suraya N, Piska F, Devi S, Pratiwi NW, Ardhi A. 2022. Molecular identification of cellulase-producing thermophilic fungi isolated from Sungai Pinang Hot Spring, Riau Province, Indonesia. Biodiversitas 23: 1457–1465. DOI:10.13057/biodiv/d230333
Sauka DH, Peralta C, Pérez MP, Onco MI, Fiodor A, Caballero J, Caballero P, Berry G, Valle EED, Palma L. 2022. Bacillus toyonensis biovar Thuringiensis: A novel entomopathogen with insecticidal activity against lepidopteran and coleopteran pests. Biocontrol. DOI: 10.1016/j.biocontrol.2022.104838
Sharma M, Gat Y, Arya S, Kumar V, Panghal A, Kumar. 2019. A Review on microbial alkaline protease: an essential tool for various industrial approaches. Ind. Biotechnol. 15: 69–78. DOI: 10.1089/ind.2018.0032
Suleiman AD, Rahman NA, Yusof HM, Shariff FM, Yasid NA. 2020. Effect of cultural conditions on protease production by a thermophilic GeoBacillus thermoglucosidasius SKF4 isolated from Sungai Klah Hot Spring Park, Malaysia. Molecules 25: 1–14. https://doi.org/10.3390/molecules25112609
Song P, Zhang X, Wang S, Xu W, Wang F, Fu R, Wei F. 2023. Microbial proteases and their applications. Front Microbiol. 14: 1236368. DOI: 10.3389/fmicb.2023.1236368.
Tuysuz E, Gonul-Baltaci N, Omeroglu MA, Adiguzel A, Taskin M, Ozkan H. 2020. Co-production of amylase and protease by locally isolated thermophilic bacterium AnoxyBacillus rupiensis T2 in sterile and non-sterile media using waste potato peels as substrate. Waste and Biomass Valorization. 11: 6793–6802. DOI: 10.1007/s12649-020-00936-3
Vanniyasingam J, Kapilan R, Vasantharuba S. 2019. Isolation and characterization of potential probiotic lactic acid bacteria isolated from cow milk and milk products. Agrieast 13: 32-43. DOI: /10.4038/agrieast.v13i1.62
Wang WY, Kong WL, Liao YCZ, Zhu LH. 2022. Identification of Bacillus velezensis SBB and its antifungal effects against Verticillium dahliae. J. Fungi. 8: 1-18. DOI: 10.3390/jof8101021
Azizah, A., Purwatiningsih, P., Wiyono, H. T., & Muzakhar, K. 2020. Morphological and biochemical characteristic of endosymbiont cellulolytic bacteria from gut of Hypothenemus hampei Ferr. and its enzyme activity. Proceeding of AIP Conference Proceedings. Universitas Sebelas Maret, Surakarta, 7 July 2020. [Indonesian]. DOI: 10.1063/5.0030576
Zhang Q, Xue C, Owens G, Chen Z. 2022. Isolation and identification of 17?-estradiol degrading bacteria and its degradation pathway. J. Hazard. Mater. DOI: 10.1016/j.jhazmat.2021.127185
Zuraidah Z, Wahyuni D, Astuty E. 2020. Karakteristik morfologi dan uji aktivitas bakteri termofilik dari kawasan wisata Ie Seuum (air panas). Jurnal Ilmu Alam dan Lingkungan. 11: 40-47. DOI: 10.20956/jal.v11i2.10200.
Most read articles by the same author(s)
- LENNI FITRI, MOHAMMAD ADZANNIE BESSANIA, NADIA SEPTI, SUHARTONO SUHARTONO , Isolation and characterization of soil actinobacteria as cellulolytic enzyme producer from Aceh Besar, Indonesia , Biodiversitas Journal of Biological Diversity: Vol. 22 No. 11 (2021)
- FAUZIAH FAUZIAH, MAULINASARI, ESSY HARNELLY, YULIA SARI ISMAIL, LENNI FITRI, Toxicity test of rose periwinkle (Catharanthus roseus) leaves endophytic bacteria using Brine Shrimp Lethality Test (BSLT) method , Biodiversitas Journal of Biological Diversity: Vol. 23 No. 1 (2022)
- SRI RAHAYU, LENNI FITRI, YULIA SARI ISMAIL, Short communication: Endophytic actinobacteria isolated from ginger (Zingiber officinale) and its potential as a pancreatic lipase inhibitor and its toxicity , Biodiversitas Journal of Biological Diversity: Vol. 20 No. 5 (2019)
- LENNI FITRI, Diversity of endophytic actinobacteria isolated from medicinal plants and their potency as pancreatic lipase inhibitor , Biodiversitas Journal of Biological Diversity: Vol. 18 No. 3 (2017)
- FADHLIAH AMFAR, LENNI FITRI, SUHARTONO, Molecular identification of a new isolate of actinobacteria ATIS61 and characterization of the protease activities , Biodiversitas Journal of Biological Diversity: Vol. 22 No. 3 (2021)
- LENNI FITRI, KARTINI AMELIA PUTRI, SUHARTONO, YULIA SARI ISMAIL, Short Communication: Isolation and characterization of thermophilic actinobacteria as proteolytic enzyme producer from Ie Seuum Hot Spring, Aceh Besar, Indonesia , Biodiversitas Journal of Biological Diversity: Vol. 20 No. 10 (2019)
- LENNI FITRI, BETTY MAULIYA BUSTAM, Screening of antimicrobial producing strains isolated from the soil of grassland rhizosphere in Pocut Meurah Intan Forest Park, Seulawah, Aceh Besar , Biodiversitas Journal of Biological Diversity: Vol. 11 No. 3 (2010)
- LENNI FITRI, YEKKI YASMIN, FAUZIAH, DWI ANDRI SEPTIANI, SUHARTONO, Characterization of BSL6 isolates isolated from honeybee hive and to determine its antibacterial activity , Biodiversitas Journal of Biological Diversity: Vol. 21 No. 10 (2020)
- YEKKI YASMIN, LENNI FITRI, FAUZIAH, FITRI WASLIYAH, Isolation of bacteria from Apis cerana hive, their antibacterial potency and cytotoxicity , Biodiversitas Journal of Biological Diversity: Vol. 20 No. 9 (2019)