SELEKSI DAN IDENTIFIKASI BAKTERI ENDOFIT POTENSIAL PENGHASIL ENZIM PROTEASE DARI TAMAN NASIONAL GUNUNG HALIMUN - (The Selection and Identification of Potential Endophyte Bacteria as Protease Enzyme Producer from Halimun Mount National Park)

Ruth Melliawati, Rohmattusolihat Rohmattusolihat, Nuryati Nuryati, Nanik Rahmani, Yopi Yopi

Abstract


Endophytic bacteria have an equal chance to bacteria that live outside the plant tissue as potential bacteria. The selection has done towards 326 bacterial endophyte isolates. This research aimed to find and identify proteolytic potential isolates. The proteolytic selection of endophytic bacteria had done using solid skim milk. The capability of endophytic bacteria to agglomerate milk was tested using liquid skim milk which incubated for 7 days at room temperature. Enzyme production of four selected isolates was made through fermentation in GYS medium. The results showed that 86 isolates have proteolytic potential. Isolate HL.29B.63 had highest protease enzymes activity (65.918 U/mL). Medium optimization was able to increase the enzyme activity into 89.94% (125.04 U/mL). The analysis used 16s rDNA showed that isolate HL.29B.63 was Bacillus amyloliquefacient subs. plantarum strain FZB42.

Keywords: endophytic bacteria, fermentation, identification, protease, selection

 

ABSTRAK

Bakteri endofit mempunyai peluang yang sama dengan bakteri yang hidup diluar jaringan tanaman sebagai bakteri potensial. Seleksi dilakukan terhadap 326 isolat bakteri endofit. Tujuan penelitian ini adalah mencari isolat yang berpotensi proteolitik dan mengidentifikasinya. Seleksi proteolitik terhadap bakteri endofitik menggunakan skim milk padat. Uji kemampuan bakteri endofitik dalam menggumpalkan susu menggunakan medium skim milk cair yang diinkubasi selama 7 hari pada suhu ruang. Produksi enzim terhadap empat isolat terseleksi dilakukan melalui fermentasi dalam medium GYS. Hasilnya menunjukkan bahwa 86 isolat mempunyai potensi proteolitik. Isolat HL.29B.63 mempunyai aktif enzim protease tertinggi (65,918 U/mL). Optimasi medium dapat meningkatkan aktivitas enzim sebesar 89,94% (125,04 U/mL). Analisis menggunakan 16s rDNA menunjukkan bahwa isolat HL.29B.63 adalah Bacillus amyloliquefaciens subs. plantarum strain FZB42.

Kata kunci: bakteri endofit, fermentasi, identifikasi, protease, seleksi


Keywords


endophytic bacteria; fermentation; identification; protease; selection

Full Text:

PDF

References


Adiarti, Retno. (2013). Aktivitas Bakteri Endofit Batang Mangrove Avicennia marina Sebagai Penghasil Antibiotik. Skripsi, Pustaka Ilmiah. Penerbit Unpad.

Arima, K., Yu J., Iwasaki, S. & Tamura, G. (1968). Milk-clotting enzim from microorganism: Purification and crystallization of Mucor rennin from Mucor pusillus var. Lindt. Applied Microbiology, 16(11), 1747-1733.

Fathoni, A., Muhammad, I., Praptiwi, Antonius, H.C. & Andria, A. (2013). Skrining dan isolasi metabolit aktif antibakteri kultur jamur endofit dari tumbuhan Albertisia papuana Becc. Berita Biologi, 12(3), 307-314.

Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39, 783-791.

Gupta, R., Q.K. Beg & P. Lorenz. (2002). Bacterial alkaline proteases: molecular approaches and industrial applications. Appl. Microbiol Biotechnol, 59, 15-32.

Hanson, R.S. & Philips, J.A. (1981). Manual of methods for general bacteriology. American Society for Microbiol, Wanshington DC. p.359-360.

Joseph, B. & M.R. Priya. (2011). Bioactive compounds from endophytes and their potential in pharmaceutical effect: a review. Am J. Biochem. Mol. Biol., 1, 291-309.

Konno, K., Hirayama, C., Nakamura, M., Tateishi, K., Tamura, Y., Hattori, M. & Kohno, K. (2004). Papain protects papaya trees from herbivorous insects: role of cystein proteases in latex. Plant J., 37, 370-378.

Melliawati, R., Dian, N.W. & Apridah, C.D. (2006). Pengkajian bakteri endofit penghasil senyawa bioaktif untuk proteksi tanaman. Journal of Biological Diversity, 7(3), 221-224.

Melliawati, R. & Harni. (2009). Senyawa antibakteri Escherichia coli ATCC 35218 dan Staphylococcus aureus ATCC 25923 dari kapang endofit Taman Nasional Gunung Halimun. Jurnal Natur Indonesia, 12(1), 21-27.

Melliawati, R., Apridah, C.D. & Yopi. (2015). Seleksi bakteri asam laktat sebagai penghasil enzim protease. Pros. Sem. Nas. Masy. Biodiv. Indon., 1(2), 184-188.

Miles, L.A., Lopera, C.A., Gonzales, S., de Garcia, M.C.C., Franco, A.E. & Restrepo, S. (2012). Exploring the biocontrol potential of fungal endophytes from an Andean Colombian Paramo ecosystem. Biocontrol, 57, 697-710.

Najafi, M.F. & Deobagkar, D. (2005). Potential application of protease isolated from Pseudomonas aeruginosa PD100. Electron J. Biotechnol., 8, 203.

Naiola, E. & Nunuk, W. (2007). Semi purifikasi dan karakterisasi enzim protease Bacillus sp. Berk. Penel. Hayati, 13, 51-56.

Nurhasanah, A. (2010). Produksi enzim protease dari bakteri Lactobacilli untuk sediaan bahan baku suplemen kesehatan. http://km.ristek.go.id/index. php/klasifikasi/detail/20381/. Diakses tanggal 23 November 2015.

Rocha, M.V., Romanini, D., Nerli, B.B. & Tubio, G. (2012). Pancreatic serine protease-extraction by affinity partition using a free triazine dye. Int. J. Biol. Macromol, 50, 303-309.

Rani, K., Rana, R. & Datt, S. (2012). Review on lates overview of proteases. Int. J. Curr. Lofe Sci., 2, 12-18.

Saran, S., Isar, J. & Saxena, R.K. (2007). A modified method for the detection of microbial proteases on agar plates using tannic acid. J. Boichem. Bioph. Meth., 70, 697-699.

Sukara, E. (1987). Production of single cell protein from cassava by microfungi. Thesis. Department of Microbiology University of Queensland.

Saitou, N. & Nei, M. (1987). The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4, 406-425.

Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution, 30, 2725-2729. http://blast.ncbi.nlm.nih.gov/Blast.cgi.

Ueda, M., Kubo, T., Miyatake, K. & Nakamura, T. (2007). Purification and characterization of fibrinolytic alkaline protease from Fusarium sp. BLB. App.l Microbiol Biotechnol., 74, 331-338.

Zhang, H.W., Song, Y.C. & Tan, R.X. (2006). Biology and chemistry of endophytes. Nat. Prod. Rep., 23, 753-771.




DOI: http://dx.doi.org/10.36974/jbi.v7i2.707

Refbacks

  • There are currently no refbacks.


BIOPROPAL Industri

Published by :

Institute for Industrial Research and Standardization (Baristand Industri) in Pontianak

Agency for Industrial Research and Development, Ministry of Industry 

Jl. Budi Utomo No. 41 Pontianak, West Kalimantan, Indonesia

Tel / Fax : +62 561 881393, 881533

email      : biopropal.industri@gmail.com

 

BIOPROPAL Industri indexed in: 

Hasil gambar untuk gambar doajHasil gambar untuk gambar google scholar

RJI Main logo