Yeast Genetics and Biotechnological Applications

Yeasts in Biotechnology, 2019

Microbial Cell Factories, 2014

Yeasts are regarded as the first microorganisms used by humans to process food and alcoholic beverages. The technology developed out of these ancient processes has been the basis for modern industrial biotechnology. Yeast biotechnology has gained great interest again in the last decades. Joining the potentials of genomics, metabolic engineering, systems and synthetic biology enables the production of numerous valuable products of primary and secondary metabolism, technical enzymes and biopharmaceutical proteins. An overview of emerging and established substrates and products of yeast biotechnology is provided and discussed in the light of the recent literature.

Journal of Biotechnology and Biomedical Science, 2019

Yeast as unicellular organism, has shown multiple application due to exhibition of noble ability in its cells. And engineered yeast has found more suitability in bioprocesses application as well as adverse conditions adaptation. Different types of yeast strains showed their best capability to adapt the salt and sugar rich environment with their optimal growth capability. These strains, used as suitable and novel cell factories for production of value added bio-products (via utilization of fermentation processes) and also for different types of bioprocesses. Application of yeast species in biotechnology field, enhanced in current periods, due to conversion of its wild to engineer strain, suitable for bioprocesses utilization and also for different types of biochemical synthesis. Different yeast species identified due to known their genetic, regulatory mechanism and also competitive metabolic pathways. In this regards, different type of engineering approaches (for genetic or pathways ...

Methods in Molecular Biology, 2011

Saccharomyces cerevisiae, commonly known as baker's yeast, is a vital model organism in genetic research and biotechnology, playing a crucial role in both fundamental biological studies and various industrial applications. This report provides a comprehensive overview of the diverse procedures involved in the genetic modification of yeast. It examines common methods such as transformation, including cell wall permeabilization techniques like lithium acetate treatment and enzymatic digestion to form spheroplasts, as well as the types of vectors used to introduce foreign DNA, such as plasmids and integrating vectors. The report further details the selection and screening of genetically modified yeast cells using selectable and reporter genes, and explores advanced genome editing techniques, with a focus on the CRISPR-Cas9 system for precise genetic manipulation. The procedures for verifying successful genetic modification, including PCR and Southern blotting, are also discussed. Furthermore, the report addresses the critical aspects of safety guidelines and ethical considerations associated with yeast genetic modification in both research and industrial settings, particularly concerning food and beverage production. This overview highlights the breadth of techniques available for manipulating the yeast genome, underscoring its importance in advancing scientific discovery and the development of biotechnological products

Biotechnology Techniques, 1996

Yeast chromosomal DNA was prepared under different conditions. Treatment of intact cells with proteinase I( (1 mg/ml) resultes in appropriate electrophoretic karyotypes; when protoplasts were formed III s&r, the presence of both sodium lauroylsarcosine and EDTA was essential. Further, the duration of cell wall lysis (12 h) and the concentrations of lytic enzymes (0.5% snail enzyme and 0.25% Novozym)had to be kept at a minimum.

Gene, 1986

An expression cassette of mouse dihydrofolate reductase (Mdhfr) cDNA under control of the yeast cytochrome c promoter was inserted in a yeast plasmid containing the ARSl sequence. The ARS replicating function was destroyed by BgfiI treatment prior to yeast transformation. Using this linearized plasmid, genomic transformants could be obtained from either laboratory or industrial strains of bakers' yeast based on direct methotrexate (MTX>resistance selection. The entire sequence of the linearized plasmid was integrated by homologous recombination at the ARS region of the host chromosome. The results indicate that repetitive and homologous recombination occurs readily in such transformations. The stability of the constructed integrants was more than 99.95 y0 per generation in non-selective medium, and tandem repeats of up to six copies (i.e., about 44 kb) were not changed even after 30 generations in rich medium. Expression in rich medium of cointegrated, human interleukin 2 cDNA under control of the triose phosphate isomerase promoter was shown by Western blot experiments in both laboratory and industrial yeast strains. Furthermore, a comparison of the transcription efficiency of the Mdhfr gene in the chromosome with that in the plasmid revealed that the efficiency was almost proportional to the number of gene copies, irrespective of the location of the transcription unit. These results show that by using the MTX/Mdhfr dominant selection-amplification system one can construct stable recombinant yeast strains suitable for heterologous gene expression in laboratory as well as in industrial fermentation conditions.

Yeast, 1985

Gene, 1995

An expression system for Saccharomyces cerevisiae (Sc) has been developed which, depending on the chosen vector, allows the constitutive expression of proteins at different levels over a range of three orders of magnitude and in different genetic backgrounds. The expression system is comprised of cassettes composed of a weak CYC1 promoter, the ADH promoter or the stronger TEF and GPD promoters, flanked by a cloning array and the CYC1 terminator. The multiple cloning array based on pBIISK (Stratagene) provides six to nine unique restriction sites, which facilitates the cloning of genes and allows for the directed cloning of cDNAs by the widely used ZAP system (Stratagene). Expression cassettes were placed into both the centromeric and 2~t plasmids of the pRS series [Sikorski and Hieter, Genetics 122 (1989) 19-27; Christianson et al., Gene 110 (1992) 119-122] containing HIS3, TRPI, LEU2 or URA3 markers. The 32 expression vectors created by this strategy provide a powerful tool for the convenient cloning and the controlled expression of genes or cDNAs in nearly every genetic background of the currently used Sc strains.

Current Protocols Essential Laboratory Techniques, 2008