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2012 | 59 | 2 | 225-229
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Locally isolated yeasts from Malaysia: Identification, phylogenetic study and characterization

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Yeasts are a convenient platform for many applications. They have been widely used as the expression hosts. There is a need to have a new yeast expression system to contribute the molecular cloning demands. Eight yeast isolates were screened from various environment sources and identified through ribosomal DNA (rDNA) Internal Transcribed Spacer (ITS). Full sequence of the rDNA ITS region for each isolate was BLASTed and phylogenetic study was constructed by using MEGA4. Among the isolates, isolate WB from 'ragi' (used to ferment carbohydrates) could be identified as a new species in order Saccharomycetales according to rDNA ITS region, morphology and biochemical tests. Isolate SO (from spoiled orange), RT (rotten tomato) and RG (different type of 'ragi') were identified as Pichia sp. Isolates R1 and R2, S4 and S5 (from the surrounding of a guava tree) were identified as Issatchenkia sp. and Hanseniaspora sp., respectively. Geneticin, 50 µg/mL, was determined to be the antibiotic marker for all isolates excepted for isolates RT and SO which used 500 µg/mL and 100 µg/mL Zeocin, respectively. Intra-extracellular proteins were screened for lipolytic activity at 30°C and 70°C. Thermostable lipase activity was detected in isolates RT and R1 with 0.6 U/mg and 0.1 U/mg, respectively. In conclusion, a new yeast-vector system for isolate WB can be developed by using phleomycin or geneticin as the drugs resistance marker. Moreover, strains RT and R1 can be investigated as a novel source of a thermostable lipase.
Physical description
  • Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • Faculty of Science, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
  • Akada F (2002) Genetically modified industrial yeast ready for application. J Biosci Bioeng 94: 536-544.
  • Fadiloglu S, Erkmen O (2002) Effects of carbon and nitrogen sources on lipase production by Candida rugosa. Turkish J Eng Env Sci 26: 249-254.
  • Felsenstein J (1985) Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39: 783-791.
  • García-Martos P, Hernández-Molina JM, Galán F, Ruiz-Henestrosa JR, García-Agudo R, Palomo MJ, Mira J (1999) Isolation of Hanseniaspora uvarum (Kloeckera apiculata) in humans. Mycopathologia 144: 73-75.
  • Gellissen G, Hollenberg CP (1997) Application of yeasts in gene expression studies: a comparison of Saccharomyces cerevisiae, Hansenula polymorpha and Kluyveromyces lactis - a review. Gene 190: 87-97.
  • Gellissen G, Kunze G, Gaillardin C, Cregg JM, Berardi E, Veenhuis M, van der Klei I (2005) New yeast expression platforms based on methylotrophic Hansenula polymorpha and Pichia pastoris and on dimorphic Arxula adeninivorans and Yarrowia lipolytica - A comparison. FEMS Yeast Res 5: 1079-1096.
  • Hadfield C, Raina KK, Shashi-Menon K, Mount RC (1993) The expression and performance of cloned genes in yeasts. Mycol Res 97: 897-944.
  • Hashida-Okado T, Ogawa A, Kato I, Takesako K (1998) Transformation system for prototrophic industrial yeasts using the AUR1 gene as a dominant selection marker. FEBS Letters 425: 117-122.
  • Kaster KR, Burgett SG, Ingolia TD (1984) Hygromycin B resistance as dominant selectable marker in yeast. Curr Genet 8: 353-358.
  • Korabecna M (2007) The Variability in the Fungal Ribosomal DNA (ITS1, ITS2, and 5.8 S rRNA Gene): Its Biological Meaning and Application in Medical Mycology. In Communicating Current Research and Educational Topics and Trends in Applied Microbiology, Méndez-Vilas A ed, pp 783-787. Formatex 2007.
  • Kwon D, Rhee J (1986) A simple and rapids colorimetric method for determination of free fatty acids for lipase assay. J Am Oil Chem Soc 63: 89-92.
  • Madzak C, Gaillardin C, Beckerich JM (2004) Heterologous protein expression and secretion in the non-conventional yeast Yarrowia lipolytica: a review. J Biotechnol 109: 63-81.
  • Medlin L, Elwood HJ, Stickel S, Sogin ML (1988) The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions. Gene 71: 491-499.
  • Millerioux Y, Clastre M, Simkin AJ, Courdavault V, Marais E, Sibirny AA, Noël T, Crèche Jl, Giglioli-Guivarc'h N, Papon N (2011) Drug-resistant cassettes for the efficient transformation of Candida guilliermondii wild-type strains. FEMS Yeast Res 11: 457-463.
  • Pramateftaki PV, Lanaridis P, Typas MA (2000) Molecular identification of wine yeasts at species or strain level: a case study with strains from two vine-growing areas of Greece. J Appl Microbiol 89: 236-248.
  • Romanos MA, Scorer CA, Clare JJ (1992) Foreign gene expression in yeast: a review. Yeast 8: 423-488.
  • Saitou N, Nei M (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406-425.
  • Shi XL, Feng MQ, Shi J, Shi ZH, Zhong J, Zhou P (2007) High-level expression and purification of recombinant human catalase in Pichia pastoris. Protein Expr Purif 54: 24-29.
  • Steinborn G, Wartmann T, Gellissen G, Kunze G (2007) Construction of an Arxula adeninivorans host-vector system based on trp1 complementation. J Biotechnol 127: 392-401.
  • Suh SO, Kurtzman CP, Lachance MA (2006) Phylogenetics of Saccharomycetales, the ascomycete yeasts. Mycologia 98: 1006-1017.
  • Sunga AJ, Tolstorukov I, Cregg JM (2008) Posttransformational vector amplification in the yeast Pichia pastoris. FEMS Yeast Res 8: 870-876.
  • Tamura K, Nei M, Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Nat Acad Sci USA 101: 11030-11035.
  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24: 1596-1599.
  • Thanh NV, Dao AH, Lachance MA (2003) Issatchenkia hanoiensis, a new yeast species isolated from frass of the litchi fruit borer Conopomorpha cramerella Snellen. FEMS Yeast Res 4: 113-117.
  • Vakhlu J, Kour A (2006) Yeast lipases: enzyme purification, biochemical properties and gene cloning. Electron J Biotechnol 9.
  • Voronovsky AY, Rohulya OV, Abbas CA, Sibirny AA (2009) Development of strains of the thermotolerant yeast Hansenula polymorpha capable of alcoholic fermentation of starch and xylan. Metab Eng 11: 234-242.
  • Yamazaki T, Nakagawa Y, Hayakawa M, Iimura Y (2005) Phylogenetic position of the yeast strain Saccharomycodes sinensis IFO10111T. J Gen Appl Microbiol 51: 35-39.
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