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Journal ArticleDOI

Analysis of factors responsible for the regeneration to intact cells from sphaeroplasts of Saccharomyces cerevisiae

01 Jan 1994-Bioprocess Engineering (Springer-Verlag)-Vol. 10, Iss: 1, pp 15-20
TL;DR: The regeneration of Saccharomyces cerevisiae cells from its sphaeroplasts were found to be influenced by a number of factors, including malt-extractglucose-yeast extract-peptone medium and growth medium.
Abstract: The regeneration of Saccharomyces cerevisiae, NCIM 3288, cells from its sphaeroplasts were found to be influenced by a number of factors. The most suitable conditions of regeneration were also dependent on growth medium, that is, using malt-extractglucose-yeast extract-peptone (MGYP) medium: mannitol 0.7 M, pH 6.5, 30 °C and using yeast extract-peptone-glucose (YPG) medium: sucrose 0.7 M, pH 5.0 and 30 °C. The maximum regeneration frequency was observed in YPG medium.
Citations
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Journal ArticleDOI
TL;DR: An attempt to produce ethanol from a cellulosic substrate by a single-stage process using intergeneric hybrids obtained from Trichoderma reesei QM 9414 and Saccharomyces cerevisiae NCIM 3288 fusants is described, showing the highest synthesis of ethanol from filter paper cellulose.

10 citations

Journal ArticleDOI
01 Jan 2012
TL;DR: In this article, the ability of Rhizopus oryzae and rhizopus microsporus strains to produce fumaric acid on glycerol as the sole carbon source in the medium was evaluated.
Abstract: Rhizopus oryzae and Rhizopus microsporus strains were screened for their ability to produce fumaric acid on glycerol as the sole carbon source in the medium. After seven days of stationary culture, fumaric acid was assayed by HPLC analysis, and maximum concentrations of 0.3% (w/v) and 0.33% (w/v) were recorded. Protoplast fusion was used to improve fumaric acid production. A selective medium for the fusant culture was composed on the basis of biochemical differences between parental strains, as examined using the Biolog FF MicroPlate TM Fungi Identification Test. Double fusion rounds led to a 1.46-fold increase in fumaric acid productivity relative to the parental strains. Individual Rhizopus fusants demonstrated a various ability to produce fumaric acid from 2.0% (w/v) of glycerol, with the most effective ones producing from 0.2 to 0.27 g@ g !1 of this acid. To date, no studies have been carried out to improve fumaric acid production by Rhizopus with the use of glycerol as the only carbon source in the medium.

10 citations

Journal Article
TL;DR: Individual Rhizopus fusants demonstrated a various ability to produce fumaric acid from 2.0% (w/v) of glycerol, with the most effective ones producing from 0.2 to 0.27 g@ g !
Abstract: Rhizopus oryzae and Rhizopus microsporus strains were screened for their ability to produce fumaric acid on glycerol as the sole carbon source in the medium. After seven days of stationary culture, fumaric acid was assayed by HPLC analysis, and maximum concentrations of 0.3% (w/v) and 0.33% (w/v) were recorded. Protoplast fusion was used to improve fumaric acid production. A selective medium for the fusant culture was composed on the basis of biochemical differences between parental strains, as examined using the Biolog FF MicroPlate TM Fungi Identification Test. Double fusion rounds led to a 1.46-fold increase in fumaric acid productivity relative to the parental strains. Individual Rhizopus fusants demonstrated a various ability to produce fumaric acid from 2.0% (w/v) of glycerol, with the most effective ones producing from 0.2 to 0.27 g@ g !1 of this acid. To date, no studies have been carried out to improve fumaric acid production by Rhizopus with the use of glycerol as the only carbon source in the medium.

9 citations

References
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Journal ArticleDOI
TL;DR: The results suggest that the life-span of limiting mRNA molecules coding for the proteins of the wall matrix is about 30 min while the turnover of 1,3-β-glucan synthases is relatively low.
Abstract: SUMMARY: While lomofungin at 20 μg ml-1 effectively halted the synthesis of ribonucleic acids in protoplasts of Saccharomyces cerevisiae, the onset of biogenesis of new wall on the protoplast surface was not impaired. The synthesis of mannan-protein wall matrix continued for about 30 min after addition of lomofungin while the formation of 1,3-β-glucan networks was not influenced even after prolonged incubation. These results suggest that the life-span of limiting mRNA molecules coding for the proteins of the wall matrix is about 30 min while the turnover of 1,3-β-glucan synthases is relatively low.

9 citations

Journal ArticleDOI
TL;DR: The method of protoplast immobilization and subsequent isolation from the gel is described in detail, and the morphology of the cell wall regeneration and morphology of reversion to the cell forms correspond to protoplasts development in gelatin or agar gels.
Abstract: Yeast protoplasts may regenerate the cell wall and revert to cells if immobilized in a 2%–5% Ca-alginate gel and cultured in an osmotically stabilized medium. The method of protoplast immobilization and subsequent isolation from the gel is described in detail. The reversion yield is dependent of the actual gel concentration, gel shape (beads vs. sheets) and of a medium molarity, and it may be up to 90%. The morphology of the cell wall regeneration and morphology of reversion to the cell forms correspond to protoplast development in gelatin or agar gels.

9 citations

Journal Article
TL;DR: The preparation and regeneration of protoplasts and spheroplasts are described as a prerequisite to hybrid formation.
Abstract: The yeast Pachysolen tannophilus is capable of the ethanolic fermentation of the aldopentose D-xylose. The possibility of increasing ethanol productivity by constructing intraspecific and interspecific hybrids by the fusion of protoplasts or spheroplasts is under investigation. This paper describes the preparation and regeneration of protoplasts and spheroplasts as a prerequisite to hybrid formation

6 citations

Journal ArticleDOI
TL;DR: Regeneration of protoplasts of Bacillus coagulans was optimized by using low lysozyme concentrations and glycerol as the osmotic support and transfer of plasmids pAB224 and pUB110, using either whole cells or protoplast transformation was not achieved, despite using a variety of conditions.
Abstract: Regeneration of protoplasts of Bacillus coagulans was optimized by using low lysozyme concentrations and glycerol as the osmotic support. Protoplasts formed from cells grown at higher temperatures were thermostable and capable of regeneration at 55°C. Transfer of plasmids pAB224 and pUB110, using either whole cells or protoplast transformation was not achieved, despite using a variety of conditions. However, plasmid transfer was achieved by fusion with B. subtilis protoplasts containing plasmid pAB224.

4 citations

Journal ArticleDOI
TL;DR: Critical analysis of various factors affecting the formation of sphaeroplasts from Saccharomyces cerevisiae were showed, including age, cell age in liquid medium, and other factors.
Abstract: Efficient synthesis of large numbers of viable sphaeroplast from Saccharomyces cerevisiae has been found to be influenced by a number of factors. In this case, Trichoderma harzianum, NCIM 1185, culture filtrate has been used to prepare sphaeroplast from Saccharomyces cerevisiae, NCIM 3288. A method has been devised to isolate large number of viable sphaeroplast from the cell. Detailed analysis of various factors affecting the formation of sphaeroplasts from Saccharomyces cerevisiae has not yet been reported. This study showed critical analysis of various factors which influenced sphaeroplast formation. Most suitable conditions were: Age of the organism in slant — 1 d, cell age in liquid medium — 24 h, time of incubation of cell with 0.3% β-mercaptoethanol — 30 min, level of lytic ezyme concentration — 79.2 ml, concentration of cell (dry wt. equivalent) — 0.1262 g, time of contact with lytic enzyme — 25 min, temperature of sphaeroplast formation — 30 °C, phosphate buffer — 25 mM of pH 6.5 and KCl as osmotic stabilizer — 0.7 M.

3 citations