* Corresponding Author
Bioprospecting and Molecular Identification of Amylase and Cellulase Producing Thermophilic Bacteria from Sediment of Nglimut Hot Springs, Kendal Regency
https://doi.org/10.22146/jtbb.86756 Anto Budiharjo(1*), Dyah Wulandari(2), Jauhara Shabrina(3), Risa Arum Mawarni(4), Anand Reyna Maulana(5), Nurhayati Nurhayati(6), Wijanarka Wijanarka(7), Laksmi Hartajanie(8), Lindayani Lindayani(9)
(1) Biotechnology Study Program, Faculty of Science and Mathematics, Diponegoro University, Semarang, Indonesia 50219; Molecular and Applied Microbiology Laboratory, Central Laboratory of Research and Services – Diponegoro University Jl. Prof. Soedarto, SH., Tembalang, Semarang, Indonesia 58275
(2) Department of Food Technology, Faculty of Agricultural Technology – Soegijapranata Catholic University (SCU), Semarang, Indonesia 50219
(3) Biotechnology Study Program, Faculty of Science and Mathematics, Diponegoro University, Semarang, Indonesia 50275
(4) Biology Department, Faculty of Science and Mathematics, Diponegoro University, Semarang, Indonesia 50275
(5) Molecular and Applied Microbiology Laboratory, Central Laboratory of Research and Services – Diponegoro University Jl. Prof. Soedarto, SH., Tembalang, Semarang, Indonesia 58275
(6) Biotechnology Study Program, Faculty of Science and Mathematics, Diponegoro University, Semarang, Indonesia 50275
(7) Biology Department, Faculty of Science and Mathematics, Diponegoro University, Semarang, Indonesia 50275
(8) Department of Food Technology, Faculty of Agricultural Technology – Soegijapranata Catholic University (SCU), Semarang, Indonesia 50219
(9) Department of Food Technology, Faculty of Agricultural Technology – Soegijapranata Catholic University (SCU), Semarang, Indonesia 50219
(*) Corresponding Author
Abstract
The utilisation of enzymes in the industry has brought numerous benefits and advantages to production processes. Enzymes serve as biocatalysts, efficiently catalyzing reactions and hydrolysis in biochemical processes. However, there are challenges in applying enzymes in the industry, particularly concerning enzyme stability. The obstacle encountered in the production processes involving industrial enzyme applications is the low stability of enzymes when used at high temperatures. Heat-sensitive enzymes undergo damage or denaturation. Thermophilic microorganisms are chosen because they hold the potential to produce thermophilic enzymes. The thermophilic enzymes exhibit better heat stability compared to other enzymes, making them an effective alternative for future industrial production processes. This study aims to isolate thermotolerant bacteria from Nglimut Hot Spring sediment, screen for cellulase- and amylase-producing isolates, and molecularly identify the best isolate using 16S rRNA barcode. The results show that 22 bacterial isolates were found in the sediment of a hot spring; TS-14 was the best isolate in producing amylase, with the highest average amylolytic index of 2.38, whereas TS-15 had the highest cellulolytic index of 2.11. Based on 16S rRNA identification, TS-14 showed an homological identity of 79% with Bacillus amyloliquefaciens, while TS-15 had a 100% homological identity with Bacillus licheniformis. These results were important as the first step of screening bacterial potential to produce thermophilic enzymes that could be applied in the downstream processing in future industrial and biotechnology companies.
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Akihary, C.V. & Kolondam, B.J., 2020. Pemanfaatan Gen 16S rRNA Sebagai Perangkat Identifikasi Bakteri untuk Penelitian-Penelitian di Indonesia. PHARMACON, 9(1), pp.16-22, doi: 10.35799/pha.9.2020.27405
Akour, R.Y., 2019. Isolation and Screening of Thermophilic α-amylase Producing Bacteria from Hot Springs in Southern Region of Saudi Arabia. Multi-Knowledge Electronic Comprehensive Journal For Education & Science Publications (MECSJ), 16, pp.1-21.
Barus, A.P.S., Ameliana, L. & Nurahmanto, D., 2016. Optimasi Suhu dan Lama Pemanasan dalam Pembentukan Kompleks Inklusi Glibenklamid-β-Siklodekstrin dengan Metode Sealed-Heating (Optimization Temperature and Heating Time Formation of Inclusion Complexes Glibenclamide-β-Cyclodextrin by Sealed-Heating Metho. e-Journal of Health Library, 4(3), pp.471-478.
Butet et al., 2019. Validation of Sea Urchin Species Based on 16s rRNA Molecular Markers from Bantul and Purworejo Waters. Journal of Tropical Fisheries Management, 3(2), pp.28-35, doi: 10.29244/jppt.v3i2.30434
Choi, Y.W., Hodgkiss, I.J. & Hyde, K.D., 2005. Enzyme production by endophytes of Brucea javanica. J Agric Technol, 1, pp.55-66.
de Souza, P.M. & de Oliveira Magalhães, P. 2010. Application of microbial α-amylase in industry - A review. Brazilian Journal of Microbiology, 41(4), pp.850-861, doi: 10.1590/S1517-83822010000400004
Emianto, Y.B. & Aribowo, Y., 2011. Geochemical Study of Geothermal Fluid in Nglimut Geothermal Prospect Area, G. Ungaran, Limbangan District, Kendal Regency, Central Java. Engineering, 32(3), pp.230-233.
Fajriani Bunga, Anto Budiharjo & Sri Pujiyanto. 2018. Isolasi dan Identifikasi Molekuler Bakteri Antagonis Terhadap Vibrio parahaemolyticus Patogen pada Udang Litopenaeus vannamei dari Produk Probiotik dan Sedimen Mangrove di Rembang. Jurnal Biologi, 7(1), pp.52-63.
Fitriani, S., 2013. Purifikasi Parsial dan Karakterisasi Enzim Protease dari Isolat B19 KUB BPPT CC. Institut Pertanian Bogor.
Gislin, D. et al., 2018. Antibacterial Activity of Soil Bacteria Isolated from Kochi, India and their Molecular Identification. Journal of Genetic Engineering and Biotechnology, 16(2), pp.287-294. doi: 10.1016j.jgeb.2018.05.010
Gray, M. J. et al., 2014. Polyphosphate is a primordial chaperone. Molecular cell, 53(5), pp.689-699. doi: 10.1016/j.molcel.2014.01.012
Karim, A. et al., 2015. Hyper production of cellulose degrading endo (1, 4) β-d-glucanase from Bacillus licheniformis KIBGE-IB2. Journal of Radiation Research and Applied Sciences, 8(2), pp.160-165, doi: 10.1016j.jrras.2014.06.004
Khalil, A. 2011. Isolation and Characterization of Three Thermophilic Bacterial Strains (Lipase, Cellulose and Amylase Producers) from Hot Springs in Saudi Arabia. African Journal of Biotechnology, 10(44), pp.8834-4439. doi: 10.5897/AJB10.1907
Khalila, R., Fitri, L. & Suhartono, 2020. Isolation and Characterization of a Thermophilic Bacteria as Cellulolytic Enzyme Producer from Hot Springs of Ie Seuum Aceh Besar, Indonesia. Microbiology Indonesia, 14(1), pp.25-33. doi: 10.5454mi.14.1.4
Konsula, Z. & Liakopoulou-Kyriakides, M., 2004. Hydrolysis of starches by the action of an -amilase from Bacillus subtilis. Process Biochemistry, 39(11), pp.1745–1749. doi: 10.1016/j.procbio.2003.07.003
Kwasna, H., Bateman, G.L. & Ward, E., 2008. Determining Species Diversity of Microfungal Communities in Forest Tree Roots By Pure-Culture Isolation and DNA Sequencing. Applied Soil Ecology, 40, pp.44-56. doi: 10.1016j.apsoil.2008.03.005
Lim, G., Tan, T.K. & Rahim, N.A., 1987. Variations in amilase and protease activities among Rhizopus isolates. MIRCEN Journal, 3, pp.319-322. doi: 10.1007/BF00933585
Mahestri, L., Harpeni, E. & Setyawan, A., 2021. Isolation and Screening of Amylum and Protein Solving Thermophilic Bacteria from Way Panas Kalianda Hot Spring, South Lampung. Jurnal Perikanan dan Kelautan, 26(3), pp.161-168. doi: 10.31258jpk.26.3.161-168
Mawati, S.D., Harpeni, E. & Fidyandini, H.P., 2021. Screening of Potential Amylolytic Thermophilic Bacteria from Way Belerang Kalianda Hot Spring, South Lampung. Journal of Aquatropica Asia, 6(1), pp.1-6. doi: 10.33019aquatropica.v6i1.2458
Mohammad, B.T. et al., 2017. Isolation and Characterization of a Thermophilic Bacteria from Jordanian Hot Springs : Bacillus licheniformis and Thermomonas hydrothermalis Isolates as Potential Producers of Thermostable Enzymes. International Journal of Microbiology, 2017, 6943952. doi: 10.1155/2017/6943952
Naresh, S. et al., 2019. Isolation and Partial Characterisation of Thermophilic Cellulolytic Bacteria from North Malaysian Tropical Mangrove Soil. Tropical Life Sciences Research, 30(1), pp.123–147. doi: 10.21315/tlsr2019.30.1.8
Ningsih, D.R., Rastuti, U. & Kamaludin, R., 2012. Karakterisasi Enzim Amilase dari Bakteri Bacillus amyloliquefaciens. Prosiding Seminar Nasional: Pengembangan Sumber Daya Pedesaan dan Kearifan Lokal Berkelanjutan II, pp.39-45.
Noer, S. 2021. Identifikasi Bakteri secara Molekular Menggunakan 16S rRNA. Edu Biologia, 1(1), pp.1-6.
Octarya, Z., Syukur, S. & Purwati RN, E., 2011. Skrining dan Identifikasi Bakteri Termofilik Penghasil Selulase dan Amilase dari Sumber Air Panas Bukit Kili Solok Sumatera Barat dengan Analisis Gen 16s Rrna. Photon: Jurnal Sains dan Kesehatan, 2(3), pp.37-44. doi: 10.37859/jp.v2i1.125
Panda, M.K. et al., 2013. Isolation and Characterization of a Thermophilic Bacillus sp. with Protease Activity Isolated from Hot Spring of Tarabalo, Odisha, India. Iran J Microbiol., 5(2), pp.159-165.
Poedjiadi, A. 2006. Fundamentals of Biochemistry Revised Edition, Jakarta: UI Press.
Reddy NS, Nimmagadda A and Rao KR. 2003. An overview of themicrobial α - Amylase family. African Journal of Biotechnology, 2, pp.645-648, doi: 10.5897/AJB2003.000-1119
Riyadi, I. 2008. The Potential of Bioprospection Management for Indonesia's Economic Growth. Journal of Agricultural Research and Development, 27(2).
Rori, C.A., Kandou, F.E.F. & Tangapo, A.M., 2020. Extracellular enzyme activity of endophytic bacteria from mangrove Avicennia marina. Journal of Bios Logos, 10(2), pp.48-55.
Satrimafitrah, P. et al., 2020. Thermostable amylase activity produced by thermopHilic bacteria isolated from Pulu Hotspring, Central Sulawesi. J. Phys.: Conf. Ser., 1434, 012034. doi: 10.1088/1742-6596/1434/1/012034
Shaikh, N.M. et al., 2013. Isolation and Screening of Cellulolytic Bacteria Inhabiting Different Environment and Optimization of Cellulase Production. Universal Journal of Environmental Research and Technology, 3(1), pp.39-49.
Shanmugapriya, K. et al., 2012. Isolation, Screening and Partial Purification of Cellulase from Cellulase Producing Bacteria. International Journal of Advanced Biotechnology and Research, 3(1), pp.509-514.
Shofa, A.F., Hariyanti & Wahyudi, P., 2019. Penggunaan DNA Mitokondria Sebagai Penanda Sumber Gelatin Sediaan Gummy Dengan Teknik Polymerase Chain Reaction dan Sekuensing DNA. Jurnal Sains Farmasi & Klinis, 6(1), pp.25-31, doi: 10.25077/jsfk.6.1.25-31.2019
Sianturi, D.C., 2008. Isolasi Bakteri dan Uji Aktivitas Amilase Termofil Kasar dari Sumber Air Panas Penen Sibirubiru Sumatera Utara. University of North Sumatra.
Simpson, M.G., 2006. Plant Systematic, California: Academic Press.
Soeka, Y.S., Rahmansyah M. & Sulistianti, 2015. Optimasi Enzim α-Amilase dari Bacillus amyloliquefaciens O1 yang Diinduksi Substrat Dedak Padi dan Karboksimetilselulosa. Jurnal Biologi Indonesia, 11(2), pp.259-266. doi: 10.14203/jbi.v11i2.2200
Suganthi, C. et al., 2015. Insight on Biochemical Characteristics of Thermotolerant Amilase Isolated from ExtremopHile Bacteria Bacillus vallismortis TD6 (HQ992818). Microbiology, 84, pp.210–218. doi: 10.1134/S0026261715020162
Telle, S. et al., 2011. Molecular phylogenetic analysis of Peronosclerospora (Oomycetes) reveals cryptic species and genetically distinct species parasitic to maize. European Journal of Plant Pathology, 130(4), pp.521-528. doi: 10.1007/s10658-011-9772-8
Turner, P., Mamo, G. & Karlsson, E.N., 2007. Potential and utilization of thermophiles and thermostable enzymes in biorefining. Microbial cell factories, 6, 9. doi: 10.1186/1475-2859-6-9
Ullah, I. et al., 2021. Identification and Characterization of Thermophilic Amylase Producing Bacterial Isolates from The Brick Kiln Soil. Saudi Journal of Biological Sciences, 28(1), pp.970–979. doi: 10.1016/j.sjbs.2020.11.017
Widyadnyana, et al., 2015. Identifikasi Bakteri Asam Laktat Isolat 9A dari Kolon Sapi Bali sebagai Probiotik melalui Analisis Gen 16S rRNA. Jurnal Sains Veteriner, 33(2), pp.228-233. doi: 10.22146/jsv.17923
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(2), pp.40 - 47. doi: 10.5454/mi.14.1.4
DOI: https://doi.org/10.22146/jtbb.86756
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