ISOLASI KHAMIR DARI BATANG TANAMAN TEBU DAN IDENTIFIKASINYA BERDASARKAN SEKUENS INTERNAL TRANSCRIBED SPACER
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Abstract
Khamir fermentatif yang digunakan dalam industri pangan, kesehatan dan energi perlu dieksplorasi untuk mengetahui potensinya. Tujuan penelitian ini adalah untuk memperoleh isolat khamir fermentatif dari batang tebu dan untuk kemudian diidentifikasi secara morfologi, biokimia dan molekuler. Isolasi khamir epifit dan endofit dilakukan dengan metode cawan sebar dari air rendaman tebu dan jus tebu pada media potato dextrose agar (PDA) dan yeast-glucose-peptone (YGP). Identifikasi morfologi berdasarkan pengamatan makroskopis dan mikroskopis. Identifikasi biokimia menggunakan uji fermentasi karbohidrat dan uji media glukosa 50%. Isolat terpilih diidentifikasi molekuler menggunakan Internal Transcribed Spacer (ITS). Hasil isolasi memperoleh 7 isolat khamir. Satu isolat terpilih (Ed 1B) didapatkan dan memiliki ciri-ciri koloni bulat, putih krem, mengkilap, timbul, bergelombang, bentuk sel ovoid dengan diameter sel 4,74 µm, memiliki budding cell, mampu memfermentasi glukosa dan sukrosa, tidak memfermentasi laktosa, serta tumbuh pada media glukosa 50%. Hasil BLAST menunjukkan bahwa isolat Ed 1B memiliki homologi 99% dengan Kodamaea ohmeri.
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References
Afshan N, Jan M, Hamid M, Nawaz B, Jabeen D, Ashraf A, Alam S (2017) Isolation and characterization of indigenous yeast species from yoghurt and sugarcane juice for production of bio-ethanol. PSM Microbiol 2:9-14
Boyd AR, Gunasekera TS, Attfield PV, Simic K, Vincent SF, Veal DA (2003) A flow-cytometric method for determination of yeast viability and cell number in a brewery. FEMS Yeast Res 3:11-16. doi: 10.1111/j.1567-1364.2003.tb00133.x
Bubnová M, ZemanÄíková J, Sychrová H (2014) Osmotolerant yeast species differ in basic physiological parameters and in tolerance of non-osmotic stresses. Yeast 31:309-321. doi: 10.1002/yea.3024
Dharmayanti NLPI (2011) Filogenetika molekuler: Metode taksonomi organisme berdasarkan sejarah evolusi. Wartazoa 21:1-10
Desjardins P, Conklin D (2010) NanoDrop microvolume quantitation of nucleic acids. J Visualized Exp 45:e2565. doi: 10.3791/2565
Ekasari TWD, Retnoningsih A, Widianti T (2012) Analisis keanekaragaman kultivar pisang menggunakan penanda PCR-RFLP pada Internal Transcribed Spacer (ITS) DNA ribosom. J MIPA 35:21-30
Elias I, Lagergren J (2009) Fast neighbor joining. Theor Comput Sci 410:1993-2000. doi: 10.1016/j.tcs.2008.12.040
Federhen S (2015) Type material in the NCBI Taxonomy Database. Nucleic Acids Res 43: D1086-98 doi: 10.1093/nar/gku1127
Gana NHT, Mendoza BC, Monsalud RG (2014) Isolation, screening and characterization of yeasts with amyloytic, lipolytic, and proteolytic activities from the surface of Philippine bananas (Musa spp.). Philippine J Sci 143:81-87
Giri S, Kindo MJ (2015) Evaluation of five phenotypic tests in the identification of candida species. Nat J Lab Med 4:13-18. doi: njlm/2015/13492:2057
Guo C, Zhao C, He P, Lu D, Shen A, Jiang N (2006) Screening and characterization of yeasts for xylitol production 101:1096-104. J Appl Microbiol. doi: 10.1111/j.1365-2672.2006.02994.x
Hidayat N, Wignyanto, Sumarsih S, Putri, AI (2016) Mikologi Industri. UB Press, Surabaya
Hidayat T, Kusumawaty D, Kusdianti E, Yati DD, Muchtar AA, Mariana D (2008) Analisis filogenetik molekuler pada Phyllanthus niruri L. (Euphorbiaceae) menggunakan urutan basa DNA daerah Internal Transcribed Spacer (ITS). J Matematika dan Sains 13:16-21
Hikmatyar MF, Royani JI, Dasumiati (2015) Isolasi dan amplifikasi DNA keladi tikus (Thyponium Flagelliform) untuk identifikasi keragaman genetik. J Bioteknol Biosains Indones 2:42-48. doi: 10.29122/jbbi.v2i2.507
Intan RMT, Cholil A, Sulistyowati L (2014) Potensi antagonis jamur endofit dan khamir pada tanaman pisang (Musa accumunata) terhadap jamur Mycosphaerella musicola penyebab penyakit bercak kuning sigatoka. J HPT 2:110-118
Isci B, Yildirim HK, Altindisli A (2014) Evaluation of methods for DNA extraction from must and wine. J Inst Brew 120:238–243. doi: 10.1002/jib.129
Jimoh SO, Ado SA, Ameh JB, Whong CM (2012) Osmotolerance and fermentative pattern of brewer’s yeast. World J Life Sci Med Res 2:59
Khattab SMR, Hadi AMA, Dahab NFA, Atta OM (2016) Isolation, characterization, and identification of yeasts associated with foods from Assiut City, Egypt. BMRJ 13:1-10. doi: 10.9734/BMRJ/2016/24170
Koff O, Samagaci L, Goualie B, Niamke S (2017) Diversity of yeasts involved in cocoa fermentation of six major cocoa-producing regions in Ivory Coast. Eur Sci J 13:496-516. doi: 10.19044/esj.2017.v13n30p496
Korabecna M (2003) The variability in the fungal ribosomal DNA (ITS1, ITS2, and 5.8 S rRNA Gene): Its biological meaning and application in medical mycology. Commun Curr Res Educ Top Trends in Appl Microbiol 2:783-787
Kurtzman CP, Fell (2011) The yeast a taxonomy study. Biodiversity and Ecophysiology of Yeast. Springer- verlag, Berlin
Lamey P, Selemi M, Vandamme AM (2009) The Phylogenetic Handbook:A Practical Approach to Phylogenetic Analysus and Hypothesis Testing. Cambrige University Press, UK
Lasmini T (2016) Isolasi dan identifikasi khamir penghasil asam indol asetat dari rhizosfer anggrek tanah Pecteilis susannae (L.) Rafin. J Ipteks Terapan 9:261-268. doi: 10.22216/jit.2015.v9i4.556
Masoud W, Cesar L B, Jespersen L, Jakobsen M (2004) Yeast involved in fermentation of Coffea arabica in East Africa determined by genotyping and by direct denaturating gradient gel electrophoresis. Yeast 21:549–556. doi: 10.1002/yea.1124
Mycobank (2018) Kodamaea ohmeri. http://www.mycobank.org/BioloMICS. Diakses 31 Agustus 2018
Obasi BC, Whong CMZ., Ado SA, Abdullahi IO (2014) Isolation and identification of yeast associated with fermented orange juice. Int J Eng Sci 3:64-69
Obasi BC, Whong CMZ, Ado SA, Abdullahi IO (2017) Leavening ability of some wild yeasts and the mutant species isolated from fermented orange juice in bakery product (bread). FUW Trends In Sci Tech J 2:596-608
Okwulehie, Cyriacus I, Alfred, Kingsley N (2010) Fungi associated with deterioration of sour-sop (Anona muricata. Linn) fruits In Abia State, Nigeria. Afr J Microbiol Res 4:143-146
Purnamasari MI, Prihatna C, Gunawan AW, Suwanto A (2012) Isolasi dan identifikasi secara molekuler Ganoderma spp. yang berasosiasi dengan penyakit busuk pangkal batang di kelapa sawit. J Fitopatologi Indones 8:9-15. doi: 10.14692/jfi.8.1.9
Rada AH, Kasaie Z (2017) A comparative study on different methods for the evaluation of baker’s yeast bioactivity. Int J Food Prop 20:100–106. doi: 10.1080/10942912.2016.1141297
Rahmansyah, Kanti A (1999) Isolat-isolat khamir dari minuman tradisional laru Di NTT. Berita Biologi 4:255-263. doi: 10.14203/beritabiologi.v4i5.1244
Rashid ANMMO, Dash BK, Chowdhury NA (2013) Exploration of potential baker’s yeast from sugarcane juice:Optimation and evaluation. Pak J Biol Sci 16:617-623. doi: 10.3923/pjbs.2013.617.623
Ren A, Ishida A, Akiyama Y (2013) Assessing statistical reliability of phylogenetic trees via a speedy double bootstrap method. Mol Phylogenet Evol 67:429-435. doi: 10.1016/j.ympev.2013.02.011
Santoso, Yahya, Suryaningtyas NH, Rahayu KS (2015) Deteksi mikrofilaria Brugia malayi pada nyamuk Mansonia spp dengan pembedahan dan metode PCR di Kabupaten Tanjung Jabung Timur. Aspirator 7:29-35. doi: 0.22435/aspirator.v7i1.3728.29-35
Sari RMT, Saputro TB, Muhibuddin A (2016) Uji potensi fermentasi etanol yeast tanah yang diisolasi dari metode budidaya SDN di daerah Batu Jawa Timur. J Sains Seni ITS 5:2337-352. doi: 10.12962/j23373520.v5i2.20601
Sharma S, Arora A, Sharma P, Singh S, Nain L, Paul D (2018) Notable mixed substrate fermentation by native Kodamaea ohmeri strains isolated from Lagenaria siceraria flowers and ethanol production on paddy straw hydrolysates. Chem Cent J 12:8. doi: 10.1186/s13065-018-0375-8
Stamps JA, Yang LH, Morales VM, Mills KLB (2012) Drosophila regulate yeast density and increase yeast community similarity in a natural substrate. Plos One 7: e42238.
doi: 10.1371/journal.pone.0042238
Sumerta IN, Kanti A (2017) Keragaman jenis khamir penghasil etanol yang diisolasi dari makanan fermentasi di kepulauan Riau. J Biol Indones 13:61-69. doi: 10.14203/jbi.v13i1.3096
Supriatna (2008) Melestarikan Alam Indonesia. Yayasan Obor Indonesia, Jakarta
Suryaningsih V, Ferniah RS, Kusdiyantini E (2018) Karakteristik morfologi, biokimia, dan molekuler isolat khamir IK-2 hasil isolasi dari jus buah sirsak (Annona muricata L.). J Akademika Biol 7:18-25
Tam HM, Diep CN (2014) Isolation, characterization and identification of endophytic bacteria in sugarcane (Saccharum spp. L.) cultivated on soils of the Dong Nai province, Southeast of Vietnam. Am J Life Sci 2:361-368. doi: 10.11648/j.ajls.20140206.16
Tesfaw A, Assefa F (2014) Current trends in bioethanol production by Saccharomyces cerevisiae: Substrate, inhibitor reduction, growth variables, coculture and immobilization. International Scholarly Res Notices. doi: 10.1155/2014/532852
Tikka C, Osuru HP, Atluri N, Raghavulu PCV, Yellapu NK, Mannur IS, Prasad UV, Aluru S, Varma N, Bhaskar M (2013) Isolation and characterization of ethanol tolerant yeast strains. Bioinformation 9:421-425. doi: 10.6026/97320630009421
Widiastutik N, Alami NH (2014) Isolasi dan Identifikasi Yeast dari Rhizosfer Rhizopora mucronata Wonorejo. J Sains dan Seni ITS 3:2337-3520. doi: 10.12962/j23373520.v3i1.5612
Yamada Y, Suzuki T, Matsuda M, Mikata K (1995) The phylogeny of Yamadazyma ohmeri (Etchells Et Bell) Billon-Grand based on the partial sequences of 18S And 26S ribosomal RNAs: The proposal of Kodamaea Gen. Nov. (Saccharomycetaceae). Biosci Biotechnol Biochem 59:1172-1174. doi: 10.1271/bbb.59.1172