Lysis of Yeast Cells Showing Low Susceptibility to Zymolyase

Kitamura, Kumpei; Yamämoto, Yasushi

doi:10.1080/00021369.1981.10864786

Cited by 2 publications

(2 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Apparently, interference with the attack of the enzyme P1 has occurred. Lipids 37) are located in the cell wall that disrupt the enzyme attack 55) . Hence reaction should be possible after extraction of the lipids.…”

Section: Separation Of the Cell Wall Components With Specific Enzymesmentioning

confidence: 99%

Mild enzymatic isolation of mannan and glucan from yeast Saccharomyces cerevisiae

Kath

Kulicke²

1999

Angew. Makromol. Chem.

View full text Add to dashboard Cite

SUMMARY: An enzymatic procedure to isolate polysaccharides from the yeast saccharomyces cerevisiae was developed which would both be environmentally friendly and preserve the native structure of the polymers. Two proteases and three enzyme cocktails were employed.To isolate the cell wall polysaccharides, the cell walls were first separated from the internal components by mechanical cell disruption. The quality of cell disruption was investigated for four types of glass beads with diameters from 0.25 -1.5 mm and monitored via the inner cellular proteins released. With large glass beads (Ø = 0.75 -1.5 mm) both brewer's and baker's yeast cells exhibited more than 90% cell disruption after as little as 12 min, whereas disruption was still incomplete with smaller beads. The isolated cell wall material was purified on microporous membranes, freeze-dried and then incubated with enzymes. Proteases caused protein lysis in the outer cell wall and released soluble mannan. The insoluble glucan was separated by centrifugation and the mannan was purified by dialysis or chromatography.In the digestion with proteases, complete conversion was achieved at concentrations higher than 240 mg g -1 of cell wall material within 26 h. Cell walls from baker's yeast had to undergo a preliminary extraction with petroleum ether. Complete conversion was only possible with one of the enzyme cocktails investigated, and this was achieved after as little as 6 h for cell walls from both baker's and brewer's yeast. The activity of the two other enzyme cocktails investigated was weaker, with brewer's yeast cell walls generally reacting more readily than those of baker's yeast. ZUSAMMENFASSUNG: Im Hinblick auf die Entwicklung umweltfreundlicher Verfahren und den Schutz der nativen Polymerstruktur wurde ein enzymatisches Verfahren zur Isolierung von Polysacchariden aus derHefe Saccharomyces cerevisiae entwickelt. Eingesetzt wurden zwei Proteasen sowie drei Enzymcocktails.Zur enzymatischen Gewinnung der Zellwand-Polysaccharide wurden zunächst die Zellwände durch einen mechanischen Zellaufschluß vom Zellinneren getrennt. Die Aufschlußgüte wurde für vier Glasperlensorten mit Durchmessern von 0,25 -1,5 mm untersucht und anhand der freigesetzten innerzellulären Proteine kontrolliert. Sowohl für Brauereihefezellen als auch Bäckerhefezellen zeigte sich mit großen Glasperlen (Ø = 0,75 -1,5 mm) bereits nach 12 min ein mehr als 90proz. Zellaufschluß, während bei kleineren Glasperlen der Aufschluß noch unvollständig war. Das gewonnene Zellwandmaterial wurde an mikroporösen Membranen gewaschen, gefriergetrocknet und anschließend mit Enzymen umgesetzt. Proteasen zerstörten in der äußeren Zellwand die Proteinmatrix und setzten lösliches Mannan frei. Das unlösliche Glucan wurde durch Zentrifugieren abgetrennt und das Mannan durch Dialyse oder Chromatographie gereinigt.Bei Umsetzung mit Proteasen war vollständiger Umsatz ab einer Konzentration von 240 mg g -1 Zellwandmaterial in 26 h möglich. Zellwände aus Bäckerhefe mußten zuvor mit Petrolether extrahiert werden. Vo...

show abstract

Section: Separation Of the Cell Wall Components With Specific Enzymesmentioning

confidence: 99%

Mild enzymatic isolation of mannan and glucan from yeast Saccharomyces cerevisiae

Kath

Kulicke²

1999

Angew. Makromol. Chem.

View full text Add to dashboard Cite

show abstract

“…The extent of lysis was gradually decreased as incubation of the organisms proceeded in the absence of polyamines (Goldemberg et a[., 1990). Agents which favour the penetration of the glucanase through the mannoprotein, such as mercaptoethanol or digitonin (Kaneko et al, 1973;Kitamura and Yamamoto, 1981;Zlotnik et al, 1984), had some stimulatory action on zymolyase activity against intact cells (Goldemberg et al, 1990). In isolated walls of polyamine-deprived S. cerevisiae 179-5, glucan should be freely accessible to zymolyase; however, the susceptibility to attack was not improved in this case (data not shown).…”

Section: Cell Wall Compositionmentioning

confidence: 99%

Polyamines and cell wall organization in Saccharomyces cerevisiae

Miret

Solari

Barderi

et al. 1992

Yeast

View full text Add to dashboard Cite

Cells of Saccharomyces cerevisiae 179-5, an ornithine decarboxylase mutant (spe-1), showed several ultrastructural abnormalities when cultivated in the absence of polyamines. Besides the appearance of microvacuole-like spaces in the cytoplasm and of deformed nuclei, the most important alterations seemed to be located in the cell wall, which was thicker and of heterogeneous texture, and in the cell membrane, of irregular contour. These modifications could not be evoked by general stress conditions elicited by lack of nutrients. The relative levels of cell wall polysaccharides were altered in polyamine-deprived organisms, giving an envelope with increased mannan and decreased glucan content; this cell wall was incompletely attacked by the lytic enzyme zymolyase. Polyamine depletion led also to some abnormalities in the budding pattern. The above observations suggest the involvement of polyamines in the correct structure and organization of the yeast cell.

show abstract

Lysis of Yeast Cells Showing Low Susceptibility to Zymolyase

Cited by 2 publications

References 3 publications

Mild enzymatic isolation of mannan and glucan from yeast Saccharomyces cerevisiae

Mild enzymatic isolation of mannan and glucan from yeast Saccharomyces cerevisiae

Polyamines and cell wall organization in Saccharomyces cerevisiae

Contact Info

Product

Resources

About