Electrofusion of brewers' yeast protoplasts and enrichment of the fusants using a flow cytometer

Urano, Naoto; Sahara, Hirohisa; Koshino, Shohei

doi:10.1016/0141-0229(93)90172-x

Cited by 10 publications

(3 citation statements)

References 22 publications

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“…The success rate of the hybrid formation mainly depends on the taxonomic proximity of the strains and applied fusion protocol (Peberdy, 1980;Pina et al, 1986;Kavanagh & Whittaker, 1996;Attfield & Bell, 2003). Intraspecific fusion frequencies usually vary from 10 À3 to 10 À4 , while for intergeneric fusions, it can be as low as 10 À6 to 10 À7 (Pina et al, 1986;Urano et al, 1993). Consequently, development of an optimal hybrid selection procedure is a crucial step in order to maximize the chance of achieving the desired genetic combination (further discussed in 'Selection of outcrossed hybrids').…”

Section: Protoplast Fusionmentioning

confidence: 99%

Improving industrial yeast strains: exploiting natural and artificial diversity

Steensels¹,

Snoek²,

Meersman³

et al. 2014

FEMS Microbiol Rev

404

288

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Yeasts have been used for thousands of years to make fermented foods and beverages, such as beer, wine, sake, and bread. However, the choice for a particular yeast strain or species for a specific industrial application is often based on historical, rather than scientific grounds. Moreover, new biotechnological yeast applications, such as the production of second-generation biofuels, confront yeast with environments and challenges that differ from those encountered in traditional food fermentations. Together, this implies that there are interesting opportunities to isolate or generate yeast variants that perform better than the currently used strains. Here, we discuss the different strategies of strain selection and improvement available for both conventional and nonconventional yeasts. Exploiting the existing natural diversity and using techniques such as mutagenesis, protoplast fusion, breeding, genome shuffling and directed evolution to generate artificial diversity, or the use of genetic modification strategies to alter traits in a more targeted way, have led to the selection of superior industrial yeasts. Furthermore, recent technological advances allowed the development of high-throughput techniques, such as ‘global transcription machinery engineering’ (gTME), to induce genetic variation, providing a new source of yeast genetic diversity.

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Section: Protoplast Fusionmentioning

confidence: 99%

Improving industrial yeast strains: exploiting natural and artificial diversity

Steensels¹,

Snoek²,

Meersman³

et al. 2014

FEMS Microbiol Rev

404

288

View full text Add to dashboard Cite

show abstract

“…High electric fields can induce cell membrane breakdown and cell death, and such an electroporation approach can also be used to kill unwanted cells as they pass through a flow cytometer (56). Similarly, flow cytometric and sorting procedures can be of great value in the detection of the heterogeneous, electroporation-mediated uptake of molecules into cells (93,97,126,353,779,780,798) and the assay of electrofusion products (57,454,956,957).…”

Section: Cell Sorting and The Isolation Of High-yielding Strains For mentioning

confidence: 99%

“…Unfortunately, not all strains of interest possess relevant enzyme activities which may be assessed flow cytometrically. Sorting on the basis of the expression of particular surface antigens is straightforward (270,635,930,957), however, and Francisco et al (320) achieved a 10 5 -fold enrichment of strains of E. coli producing a digoxin-binding single-chain antibody fragment by using only two steps. For cell surface antigens, however, magnetic labelling and magnetic cell sorting (634,787) may have a greater throughput than conventional fluorescence-activated cell sorting (930).…”

Section: Cell Sorting and The Isolation Of High-yielding Strains For mentioning

confidence: 99%