Yeast Penghasil Lipase dan Karakterisasi Parsial Morfologinya

Bodhi Dharma; Eggi Mallisa; Linda Oktavianingsih

doi:10.46638/jmi.v6i1.180

Yeast Penghasil Lipase dan Karakterisasi Parsial Morfologinya

Bodhi Dharma^(1*), Eggi Mallisa⁽²⁾, Linda Oktavianingsih⁽³⁾

(1) Jurusan Biologi. Fakultas Matematika dan Ilmu Pengetahuan Alam (FMIPA). Universitas Mulawarman.
(2) Program Studi Biologi, Jurusan Biologi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Mulawarman.
(3) Jurusan Biologi. Fakultas Matematika dan Ilmu Pengetahuan Alam (FMIPA). Universitas Mulawarman.
(*) Corresponding Author

Abstract

Pengisolasian strain-strain yeast penghasil enzim lipase dilakukan dari Limbah Tandan Kosong Kelapa Sawit (TKKS). Yeast diisolasi dengan menggunakan medium Yeast Extract Peptone Dextrose (YEPD) dengan tambahan chloramphenicol. Strain yeast disampling dari tiga lokasi yang berbeda berdasarkan umur TKKS berada ditempat pabrik pengolahan (1, 3, dan 30 hari). Setelah strain yeast berhasil disolasi, kemudian dilakukan skrining menggunakan medium Tributyrin Agar (TBA) pada strain yeast selama 48 jam pada suhu 30^oC. Enzymatic Index (EI) dihitung berdasarkan perbandingan diameter zona bening (halo) yang terbentuk dan diameter koloni. Terhadap lima strain yeast yang memiliki nilai EI tertinggi dilakukan pengamatan morfologi koloni serta pengamatan sel yaitu tipe budding dan ukuran sel. Konstruksi klaster Euclidean (dendrogram) menggunakan algoritma UPGMA (Unweighted Pair Group Method with Arithmetical averages) dilakukan berdasarkan data skoring karakteristik fenotip dengan tujuan untuk mengetahui similaritas antara lima strain yeast terseleksi. Hasil penelitian menunjukan bahwa kelima strain yeast potensial yang diisolasi menghasilkan enzim lipase dengan nilai EI bervariasi yaitu strain C.4.3.1 dengan EI = 4,47; C.3.3.2 dengan EI = 4,14; C.3.1.1 dengan EI = 3,72; D.3.3.1 dengan EI = 3,59; dan C.1.2.2 dengan EI = 3,43. Dendrogram yang dihasilkan menunjukan adanya perbedaan jarak disimilaritas antar strain. Terdapat dua klaster besar yaitu klaster pertama (node 1) yaitu C.3.1.1 dan C.1.2.2, klaster kedua yang terdiri dari dua subklaster yaitu node 3 (strain C.4.3.1), dan node 2 (strain C.3.3.2 dan D.3.3.1).

Keywords

POEFB, lipolitik-yeast, skrining, fenotip, kluster

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References

Anagnostakis S.L., & Hankin L. (1975). Use of Selective Media to Detect Enzyme Production by Microorganisms in Food products. J Milk Food Technol. 38(10):570-572.

Badan Pusat Statistik. (2008). Buletin Statistik Perdagangan Luar Negeri, Import. ISSN: 0216.3668. CV Lintama Printing. Hal: 9.

Bharathi D., Rajalakshmi G., & Komathi S. (2018). Optimization and Production of Lipase Enzyme from Bacterial Strains Isolate from Petrol Spilled Soil. Sci. J. King Saud Univ. Hal: 1-4.

Boekhout, T., & Kurtzman, C. P. (1996) Chap. 1 Principles and Methods Used in Yeast Classification, and an Overview of Currently Accepted Yeast Genera. In: Nonconventional Yeasts in Biotechnology (K. Wolf ed.). Berlin: Springer-Verlag.

Ciafardini G, Zullo, B. A., Iride, A. (2006) Lipase Production by Yeast from Virgin Olive Oil. Food Microbiol. 23(1): 60-67.

Dini, N. (2017) Produksi dan Karakterisasi Lipase dari Lactobacillus plantarum SK (5) Asal Bekasam. (Skripsi). Bogor: Institut Pertanian Bogor (IPB)

Florencio C., Couri S., & Farinas C.S. (2012) Correlation Between Agar Plate Screening and Solid-State Fermentation for the Prediction of Cellulase Production by Trichoderma Strain. Enzyme Research 2012 (Article ID 793708, 7 pages). Doi:10.1155/2012/793708

Goncalves, F. A. G., Colen, G., Takahasi, J. A. (2014) Yarrowia lipolytica and Its Multiple Applications in The Biotechnological Industry. Scientific World Journal. 7.

Gopinath, S.C.B., Hilda A., & Anbu P. (2005) Extracellular Enzymatic Activity Profiles in Fungi Isolated from Oil-Rich Environment. Mycoscience. 46(2): 119-126.

Griebeler, N., Polloni A.E., Remonatto D., Arbter F., Vardanega R., Cechet J.L., Luccio M.D., de Oliveira D., Treichel H., Cansian R.L., Rigo E., Nonow J.L. (2009) Isolation and Screening of Lipase-Producing Fungi with Hydrolytic Activity. Food and Bioprocess Technology. 4(4): 578-586.

Jumiati, Bintari S.H., & Mubarok I. (2012) Isolasi dan Identifikasi Khamir Secara Morfologi di Tanah Kebun Wisata Pendidikan Universitas Negeri Semarang. Biosantifika. 4(1): 27-35.

Kamaladevi B., Prabhavathi P., Sankareswaran M., Anbalagan S., Radhakrishnan N., & Prabhu D. (2014). Screening and Medium Optimization of Lipase Producing Bacteria from Saltpan. Research Journal of Chemistry and Environment Sciences. 2(2): 72-77.

Kurtzman, C.P. & Fell J.W. (2011). The Yeast: A Taxonomic Study. 5th ed. Amsterdam: Elsevier. Pp: 21-44.

Musa H., Han P.C., Kasim F.H., Gopinath S.C.B. & Ahmad M.A. (2017). Turning Oil Palm Empty Fruit Bunch Waste into Substrate for Optimal Lipase Secretion on Solid State Fermentation by Trichoderma Strains. Process Biochemistry. (17): 10.

Ngara T.R. & Zhang H. (2018) Recent Advances in Function-based Metagenomic Screening. Genomics Proteomics & Bioinformatics. Vol. 16: 405-415.

Nurdini, A.L. (2010) Penapisan Bakteri Lipolitik Asal Fruktosfer Kelapa Sawit. (Skripsi). Bogor: Institut Pertanian Bogor.

Obire O., & Hakam I.O. (2015). Evaluation of The Microbiological Quality of Palm Fruits in The Various Stages of Palm Oil Production. Science and Technology. 2(2): 313-323.

Ohnuki S., Okada H., Friedrich A., Kanno Y., Goshima T., Hasuda H., Inahashi M., Okazaki N., Tamura H., Nakamura R., Hirata D., Fukuda H., Shimoi H., Kitamoto K., Watanabe D., Schacherer J., Akao T., & Ohya Y. (2017) Phenotypic Diagnosis of Lineage and Differentiation During Sake Yeast Breeding, G3 Genes Genomes Genetics. 7(8): 2807–2820.

Paludo M.P., de Oliveire K.S.D, Trevisol T.C., de Medeiros Burkert J.F. (2018) Isolation of Lipase Producing Yeast from Industrial Oily Residues in Different Culture Media. IntJ.Cur.MicribioLApp.Sci. 7(1): 2348-2362.

Nigam P.S.nee & Pandey A. (1999) Solid-State Fermentation for The Production of Industrial Enzymes. Current Science. 77(1): 149-162.

Patra, J.K., Das G., Das S.K. & Thatoi H. (2020) A Practical Guide to Environmental Biotechnology. Singapore: Springer. pp: 99-104.

Pera, L. M., C.M. Romero., M.D. Baigori & G.R. Castro (2006) Catalytic Properties of lipase Extracts from Aspergillus niger. Food Technol.Biotechnol. 48: 33-37.

Rahayu Y.P. (2018) Uji Aktivitas Lipase dan Biosurfaktan dari Bakteri Keratinolitik. (Tesis). Medan: Universitas Sumatera Utara.

Rai B., Shrestha A., Sharma S., Joshi J. (2014) Screening, Optimization and Process Scale up for Pilot Scale Production of Lipase by Aspergillus niger. Biomedicine and Biotechnology 2(3) 54-59.

Romero, C.M., Pera L.M., & Loto F. (2012) Purification of an Organic Solvent Tolerant Lipase from Aspergillus niger MYA 135 and Its Application in Ester Synthesis. Biocatal. Agric. Biotechnol. 1(1): 25-31.

Ruusuvuori, P., Lin J., Scott A.C., Tan Z., Sorsa S., Kallio A., Nykter M., Yli-Harja O., Shmulevich I., Dudley A.M., (2014) Quantitative Analisis of Colony Morphology in Yeast. BioTechniques. 5:18-27.

Sharma R. Chsti Y. Banerjee U.C. (2001) Research Review Paper Production Purification Characterization and Application of Lipases. Biotechnol Advanced. 19(8): 627-662.

Swandi, M. K., Periadnadi, Nurmiati (2015) Isolasi Bakteri Pendegradasi Limbah Cair Industri Minyak Sawit. Jurnal Biologi Universitas Andalas. 4(1): 71-76.

Thomas P., Sekhar A.C., Upreti R., Mujawar M.M., & Sadiq S.P. 2015. Optimization of Single Plate Serial Dilution Spotting (SP-SDS) with Sample Anchoring as an Assured Method for Bacterial and Yeast CFU Enumeration and Single Colony Isolation from Diverse Sample. Biotech Reports. Pp: 1-33.

Trismilah, Sumaryono W., Malik A., & Sadikin M. (2014) Isolasi dan Karakterisasi Protease Serupa Tripsin (PTS) dari Lactobacillus plantarum FNCC 0270. Jurnal Ilmu Kefarmasian Indonesia. 12 (1): 57-66

Viana N.C., Portugal, C., & da Cruz S.H. (2017). Morphophysiological and molecular characterization of wild yeast isolates from industrial ethanol process. African Journal of Microbiology Research, 11(37), 1422-1430.

Yarrow, D. (1998) Methods for The Isolation, Maintenance, and Identification of Yeast. In: Kurtzman, C.P. & J.W. Fell. (eds.). 1998. The Yeast: A Taxonomic Study 4th ed. Amsterdam: Elsevier. Pp: 69-100.

DOI: http://doi.org/10.46638/jmi.v6i1.180