Most information on yeasts derives from experiments with the conventional yeasts Saccaromyces cerevisiae and Schizossaccharomyces pombe, the complete nuclear and mitochondrial genome of which has also been sequenced. For all other non-conventional yeasts, investigations are in progress and the rapid development of molecular techniques has allowed an insight also into a variety of non-conventional yeasts.
In this bench manual, over 70 practical protocols using 15 different non-conventional yeast species and in addition several protocols of general use are described in detail. All of these experiments on the genetics, biochemistry and biotechnology of yeasts have been contributed by renowned laboratories and have been reproduced many times. The reliable protocols are thus ideally suited also for undergraduate and graduate practical courses.

Inhalt

Arxula adeninivorans.- I Temperature-Dependent Dimorphism of the Non-Conventional Yeast Arxula adeninivorans.- 2 Expression of Heterologous Genes in Arxula adeninivorans Budding Cells and Mycelia.- Ashbya gossypii.- 3 Effect of Precursor Feeding on Riboflavin Production of Ashbya gossypii and Influence of Threonine Aldolase Overexpression.- Candida maltosa.- 4 Characterization of the 26S-rRNA Gene to Classify an Industrial Strain to be Candida maltosa.- 5 Isolation of a GCN4 Gene Analog and Determination of Its Involvement in Induction of Cycloheximide-Resistant Ribosomal Protein LA1-Q in Candida maltosa.- 6 Dehalogenation During Bio degradation of Chlorophenols by Candida maltosa.- 7 Production of Aromatic Acids During the Oxidation of Phenylalkanes by Candida maltosa.- Debaryomyces hansenii.- 8 Growth of Debaryomyces hansenii in Seawater Culture Medium.- 9 Glycerol and Glucose Dissimilation by Debaryomyces hansenii: Substrate Influence on Growth Rate and Biomass Yield of Yeast.- 10 Induction of GDP from Halotolerant Yeast Debaryomyces hansenii by Salt Stress.- 11 Salt Tolerance in Debaryomyces hansenii. Effect of Sodium on Growth at High Temperature and at Extreme pH.- 12 Preparation of Total DNA from Debaryomyces hansenii.- Hansenula polymorpha.- 13 Life Cycle of Hansenula polymorpha.- 14 Obtention of Auxotrophic Mutants in Hansenula polymorpha.- 15 Transformation by Electroporation of the Yeast Hansenula polymorpha.- 16 Expression of Hansenula polymorpha Nitrate Reductase Under the Control of the MOX1 Promoter.- 17 Promoter Assessment in H. polymorpha Using a lacZ Reporter Gene.- 18 Immuno-Gold Labeling of Invertase in Hansenula polymorpha.- Hortaea werneckii.- 19 Determination of Intra- and Extracellular Concentration of Glycerol in the Halophilic Black Yeast Hortaea werneckii Grown at Different Environmental Salinities.- 20 Regulation of HMGCoA Reductase Activity by Salt Stress in Hortaea werneckii.- 21 Isolation and Identification of Lipids from the Halophilic Black Yeast Hortaea werneckii.- 22 Inhibition of DHN-Melanin Biosynthesis by Tricydazole in Hortaea werneckii.- Kluyveromyces lactis.- 23 Mating, Sporulation, and Tetrad Analysis in Kluyveromyces lactis.- 24 Isolation of Mitochondrial Mutants from Cha pter25GeneDisruptioninthe Yeast Kluyveromyces lactis.- 25 Gene Disruption in the Yeast Kluyveromyces lactis.- 26 Transformation of Kluyveromyces lactis.- 27 Removal of Dominant Markers from the Kluyveromyceslactis Genome Using the Cre/loxP System.- 28 Phenotypic Analysis of the Kluyveromyces lactis Killer Phenomenon.- 29 Genetic Manipulation of the Kluyveromyces lactis Killer Plasmidskl and k2.- 30 Chitin-Binding Capacity of the Zymocin Complex from Kluyveromyces lactis.- 31 Heterologous Protein Production in High Copy Number Vector Systems.- 32 Alcohol Dehydrogenase (ADH) Isozymes in K. lactis: Detection by Activity.- 33 Kluyveromyces lactis Genome Mapping and Sequencing.- Pichia stipitis.- 34 Mating and Segregation in Pichia stipitis.- 35 ADH1 Disruption in P. stipitis by Targeted Restriction Enzyme Mediated Integration.- 36 Examination of Xylose Fermentation in P. stipitis by Means of the Sensomat System.- 37 Regulation of Fermentation and Respiration.- 38 Investigation of Transcriptional Regulation of the Fermentative ADH in Pichia stipitis Using an EGFP Reporter Gene.- 39 Cyanide-Insensitive Respiration.- 40 Freeze Transformation, Plasmid Reisolation and Stability in Pichia stipitis.- Schwanniomyces occidentalis.- 41 Objective Analysis of Medium-Induced Fragility of Schwanniomyces occidentalis.- 42 Subjective Analysis of Medium-Induced Fragility of Schwanniomyces occidentalis.- 43 Analysis of Structural Discontinuities of the Wall of Schwanniomyces occidentalis.- 44 Induction of Amylases in Schwanniomyces occidentalis.- 45 Ethanol Production from Starch by Schwanniomycesoccidentalis.- Trichosporon domesticum.- 46 Characterization of Soluble Cytochrome P450 from Trichosporon domesticum.- Trichosporon montevideense.- 47 Oxidation of the Fungicide Biphenyl by Yeasts of the Genus Trichosporon.- Xanthophyllomyces dendrorhous.- 48 Genetic Complementation Analysis by Protoplast Fusion of Xanthophyllomyces dendrorhous.- 49 Lethal Effect of UV Light and Photoreactivation in Xanthophyllomyces dendrorhous.- 50 Fermentation and Carotenoid Analysis of the Yeast Xanthophyllomyces dendrorhous (Phaffia rhodozyma)..- 51 Metabolic Engineering of the Carotenoid Biosynthetic Pathway in Xanthophyllomyces dendrorhous (Phaffiarhodozyma).- 52 Astaxanthin Biosynthesis in Xanthophyllomyces dendrorhous (Phaffia rhodozyma): Effect of Inhibitors and Low Temperature.- 53 Purification and Characterization of Extrachromosomal Genetic Elements of Double-Stranded RNA (dsRNA) of Xanthophyllomyces dendrorhous.- Yarrowia lipolytica.- 54 Mating, Sporulation and Random Spore Selection by Nystatin in Yarrowia lipolytica.- 55 Induction of the Yeast-Hypha Transition in Yarrowialipolytica.- 56 Tagging of Genes by Insertional Mutagenesis in the Yeast Yarrowia lipolytica.- 57 Detection and Activation of the Yltl Retrotransposon in the Yeast Yarrowia lipolytica.- 58 Flow Cytometry of Yarrowia lipolytica.- 59 A Copper-Regulated Yarrowia lipolytica Vector for the Expression of Heterologous Proteins.- 60 Lipase Assay in Yarrowia lipolytica.- 61 Expression of Mutant Gprlp in Yarrowia lipolytica and Its Effect on Growth on Different Media.- 62 Detection of Biosynthesis and Degradation of Peroxisomes by Different Carbon Sources with a Combined Assay Using a ?-Galactosidase-Gfp-SKL Chimaeric Protein for Labeling of Peroxisomes and the Peroxisomal Isocitrate Lyase as Control.- 63 Induction of Citric Acid/Isocitric Acid and a-Ketoglutaric Acid Production in the Yeast Yarrowia lipolytica.- 64 Regulation of Production of Yarrowia lipolytica Extracellular Ribonuclease and Alkaline Extracellular Protease.- Zygosaccharomyces bailii.- 65 Zygosaccharomyces bailii: A Yeast with a Peculiar Pattern for the Regulation of Acetic Acid Metabolism in the Presence of Glucose.- 66 Transfection of Yeast with Toxin-Encoding dsRNA Viruses of Zygosaccharomyces bailii and Toxin Purification by Receptor-Mediated Affinity Chromatography.- Zygosaccharomyces rouxii.- 67 Heat-Shock-Induced Activation of Trehalase in the Yeast Zygosaccharomyces rouxii.- Miscellaneous Non-Conventional Yeasts.- 68 Linear and Circular Plasmids of Yeasts.- 69 Rapid Method to Extract DNA from Capsulated Yeast Species.- 70 Testing Various Yeasts for Ethanol Production from Xylose.- 71 Use of a Differential Culture Medium for the Enumeration of Zygosaccharomyces bailii, Saccharomyces cerevisiae and Pichia membranifaciens in Wine.- 72 Characterization of Ubiquinone Type (CoQ7-CoQ10) of Non-Conventional Yeasts.- 73 Non-Radioactive Dot Blot DNA Reassociation for Unequivocal Yeast Identification.- 74 Taxonomy Rules and Classification Hints for Non-Taxonomists Working with Unknown or Ill-Identified Yeast Cultures.