Impact of Yeast Glycosylation Pathway on Cell Integrity and Morphology

Janik, A.; Juchimiuk, Mateusz; Kruszewska, Joanna S.; Orłowski, Jacek; Pasikowska, Monika; Palamarczyk, Grażyna

doi:10.5772/48102

Cited by 6 publications

(8 citation statements)

References 60 publications

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“…In yeast, the mevalonate pathway is important not only for the biosynthesis of ergosterol, the main sterol presents in cell membranes, but also for generating dolichol phosphate (DolP), a lipid acceptor of sugar residues, particularly mannose to form DolP mannose (DolP Man). This sugar-lipid intermediate is involved in the N- and O-glycosylation at the golgi, including the synthesis of the sugar part of the glycosyl phosphatidyl inositol (PGI) anchor as well as of proteins with internal repeats (Pir-CWP) [ 5 ].…”

Section: Resultsmentioning

confidence: 99%

“…The cell wall of S. cerevisiae represents 15 to 30 % of the dry weight, 25 to 50 % of the cell volume and is largely composed of polysaccharides and proteins [ 4 ]. Four classes of interacting components, including chitin, β 1,3 glucan, β 1,6 glucan, and mannoproteins have been reported [ 5 ]. The cell wall represents a dynamic structure that can adapt to physiological and morphological changes [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

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Hypoxia-elicited impairment of cell wall integrity, glycosylation precursor synthesis, and growth in scaled-up high-cell density fed-batch cultures of Saccharomyces cerevisiae

et al. 2016

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BackgroundIn this study we examine the integrity of the cell wall during scale up of a yeast fermentation process from laboratory scale (10 L) to industrial scale (10,000 L). In a previous study we observed a clear difference in the volume fraction occupied by yeast cells as revealed by wet cell weight (WCW) measurements between these scales. That study also included metabolite analysis which suggested hypoxia during scale up. Here we hypothesize that hypoxia weakens the yeast cell wall during the scale up, leading to changes in cell permeability, and/or cell mechanical resistance, which in turn may lead to the observed difference in WCW. We tested the cell wall integrity by probing the cell wall sensitivity to Zymolyase. Also exometabolomics data showed changes in supply of precursors for the glycosylation pathway.ResultsThe results show a more sensitive cell wall later in the production process at industrial scale, while the sensitivity at early time points was similar at both scales. We also report exometabolomics data, in particular a link with the protein glycosylation pathway. Significantly lower levels of Man6P and progressively higher GDP-mannose indicated partially impaired incorporation of this sugar nucleotide during co- or post-translational protein glycosylation pathways at the 10,000 L compared to the 10 L scale. This impairment in glycosylation would be expected to affect cell wall integrity. Although cell viability from samples obtained at both scales were similar, cells harvested from 10 L bioreactors were able to re-initiate growth faster in fresh shake flask media than those harvested from the industrial scale.ConclusionsThe results obtained help explain the WCW differences observed at both scales by hypoxia-triggered weakening of the yeast cell wall during the scale up.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0542-3) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Hypoxia-elicited impairment of cell wall integrity, glycosylation precursor synthesis, and growth in scaled-up high-cell density fed-batch cultures of Saccharomyces cerevisiae

et al. 2016

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“…At a given concentration of native Dol-P the yeast DPMS exhibited from 8- to 55-fold higher activity. The dolichyl phosphate mannose formed by DPMS is mostly used for protein glycosylation, GPI-anchor formation and the synthesis of the cell wall [35,36,37]. Thus, one could expect a positive correlation between the requirement for dolichyl phosphate mannose for these processes and the speed for its synthesis.…”

Section: Discussionmentioning

confidence: 99%

Poly-Saturated Dolichols from Filamentous Fungi Modulate Activity of Dolichol-Dependent Glycosyltransferase and Physical Properties of Membranes

Gryz

Perlińska-Lenart

Gawarecka

et al. 2019

IJMS

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Mono-saturated polyprenols (dolichols) have been found in almost all Eukaryotic cells, however, dolichols containing additional saturated bonds at the ω-end, have been identified in A. fumigatus and A. niger. Here we confirm using an LC-ESI-QTOF-MS analysis, that poly-saturated dolichols are abundant in other filamentous fungi, Trichoderma reesei, A. nidulans and Neurospora crassa, while the yeast Saccharomyces cerevisiae only contains the typical mono-saturated dolichols. We also show, using differential scanning calorimetry (DSC) and fluorescence anisotropy of 1,6-diphenyl-l,3,5-hexatriene (DPH) that the structure of dolichols modulates the properties of membranes and affects the functioning of dolichyl diphosphate mannose synthase (DPMS). The activity of this enzyme from T. reesei and S. cerevisiae was strongly affected by the structure of dolichols. Additionally, the structure of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) model membranes was more strongly disturbed by the poly-saturated dolichols from Trichoderma than by the mono-saturated dolichols from yeast. By comparing the lipidome of filamentous fungi with that from S. cerevisiae, we revealed significant differences in the PC/PE ratio and fatty acids composition. Filamentous fungi differ from S. cerevisiae in the lipid composition of their membranes and the structure of dolichols. The structure of dolichols profoundly affects the functioning of dolichol-dependent enzyme, DPMS.

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“…The C. albicans cell wall composition is similar to that of S. cerevisiae and contains four classes of interacting components: chitin, β1,3 and β1,6 glucan, and highly glycosylated mannoproteins [50,51].…”

Section: Defective Activity Of Alg13p Influences Cell Wall Structurementioning

confidence: 99%

The role of Alg13 N-acetylglucosaminyl transferase in the expression of pathogenic features of Candida albicans

Niewiadomska

Janik

Perlińska-Lenart

et al. 2017

Biochimica et Biophysica Acta (BBA) - General Subjects

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Impact of Yeast Glycosylation Pathway on Cell Integrity and Morphology

Cited by 6 publications

References 60 publications

Hypoxia-elicited impairment of cell wall integrity, glycosylation precursor synthesis, and growth in scaled-up high-cell density fed-batch cultures of Saccharomyces cerevisiae

Hypoxia-elicited impairment of cell wall integrity, glycosylation precursor synthesis, and growth in scaled-up high-cell density fed-batch cultures of Saccharomyces cerevisiae

Poly-Saturated Dolichols from Filamentous Fungi Modulate Activity of Dolichol-Dependent Glycosyltransferase and Physical Properties of Membranes

The role of Alg13 N-acetylglucosaminyl transferase in the expression of pathogenic features of Candida albicans

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