Investigation of the Best Saccharomyces cerevisiae Growth Condition

Salari, Roshanak; Salari, Rosita

doi:10.19082/3592

Cited by 70 publications

(47 citation statements)

References 14 publications

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“…Decreasing growth of S. cerevisiae occurred after 48 hours of fermentation, where at 72 hours fermentation the absorbance value decreased to 0.176. This absorbance value represents the number of S. cerevisiae cells, the higher absorbance value the higher number of S. cerevisiae cells obtained (Salari and Salari, 2017). Fig.…”

Section: Resultsmentioning

confidence: 99%

The Utilization of Vegetable and Fruit Wastes for Saccharomyces cerevisieae Cell Wall Based β-Glucan Production with Antioxidant Activity

Utama¹,

Irena²,

Lembong³

et al. 2020

Acta Univ. Agric. Silvic. Mendelianae Brun.

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The study aims to determine the number of mass and antioxidant activity of β-glucan extracted from S.cerevisieae which grown on vegetable and fruit wastes. The method used is experimental with descriptive analysis which consisted of 3 treatments namely banana waste, papaya waste, and napa cabbage waste as fermentation medium, repeated thrice. Fermentation medium was made by mixing waste with water with the ratio of 1:2. Ten percent (w/v) of S.cerevisieae was inoculated and incubated for 48 hours, at 27 ℃. Extraction of β-glucan was carried out using acid-alkaline methods and antioxidant activity was tested by DPPH (2,2-Diphenyl-1-picrylhydrayl) method and the microstructure of β-glucan is determined by Scanning Electrone Microscope. The result showed that the best medium in producing β-glucan was papaya waste which resulting 19.094 g β-glucan mass, with radical scavenging activity of 20.71 % and globular diameter of 533 μm.

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Section: Resultsmentioning

confidence: 99%

The Utilization of Vegetable and Fruit Wastes for Saccharomyces cerevisieae Cell Wall Based β-Glucan Production with Antioxidant Activity

Utama¹,

Irena²,

Lembong³

et al. 2020

Acta Univ. Agric. Silvic. Mendelianae Brun.

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“…Comparative growth assessment of WT, ydr131cΔ, atg1Δ and ydr131cΔatg1Δ was carried out both on solid and liquid media as mentioned in the (S harma et al 2019) and (S alari and S alari 2017). For comparative growth assessment on solid media each strains was streaked on YPD + agar plates and incubated for 36-48 hrs at 30° C. For comparative growth kinetics of strains in the liquid media, optical density measurement at 600nm for 14 hrs was carried out.…”

Section: Methodsmentioning

confidence: 99%

Deletion of Autophagy geneATG1and F-box motif encoding geneYDR131Ctogether leads to Synthetic Growth Defects and Flocculation behaviour inSaccharomyces cerevisiae

Shoket

Parvez

Sharma

et al. 2020

Preprint

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Running title: GeneticInteractions between YDR131C and ATG1 shows ubiquitination and 12 autophagy pathway cross talk governing growth fitness 13 Abstract: 20 F-box motif encoding YDR131C is functionally uncharacterized gene which forms the complex 21 with the SCF-E3 ligase. The F-box motif containing proteins are involved in substrate 22 recruitment for the ubiquitination and subsequent degradation through 26S proteasome. 23 Autophagy gene, ATG1 (ULK1in human) is a well conserved serine-threonine kinase, required 24 for vesicle formation and cytoplasm to vacuole targeting pathway. Atg1p forms the complex 25 with Atg13p and Atg17p during autophagy. The understanding of crosstalk between ubiquitin 26 and autophagy pathways is crucial for synthetic lethality screen and drug targeting. Here we have 27 conducted the study for genetic interaction between uncharacterized YDR131C and ATG1 gene 28 representing both specific and non-specific protein degradation pathways. The single and double 29 gene knockout strains of YDR131Cand ATG1 genes were constructed in the BY4741 genetic 30 background and analysed for growth fitness. The strains were also evaluated for cellular growth 31 response in presence of hydroxyurea (HU), methyl methane sulfonate (MMS), and hydrogen 32 peroxide (H 2 O 2 ) stress causing agents by spot assay. The ydr131cΔatg1Δ showed the synthetic 33 growth defect phenotype with floc formation in rich medium which showed floc disruption in 34 presence of EDTA. The ydr131cΔatg1Δ cells showed the sensitivity to stress agents HU, MMS, 35 and H 2 O 2 when compared with ydr131cΔ, atg1Δ, and WT cells.. Based on the observations, we 36 report that YDR131C and ATG1 functions in parallel pathways for growth fitness and cellular 37 growth response to stress agents. Interestingly this study also revealed the crosstalk between 38 ubiquitination and autophagy pathways. The defects in both the pathways could lead to synthetic 39 growth defects which may have implication for the precision medicine initiatives. 40 41 107 2019). Briefly Wild type (BY4741) and deletion derivatives (WT, ydr131cΔ, atg1Δ and 108 ydr131cΔatg1Δ) were grown to log phase (OD 600 0.8-1.0) and equal number of cell were serially 109 6 diluted. From each dilution a 3µl aliquot was spotted onto agar plates containing YPD, YPD + 110 stress causing agents such as a hydroxyurea (200mM), MMS (0.035%) and hydrogen peroxide 111 (4mM). The plates were incubated at 30°C for 2-3 days and imaged. 112 113 Fluorescence Microscopy for Cell Wall and Nuclear status 114To compare the cell wall status of WT, ydr131cΔ, atg1Δ, and ydr131cΔatg1Δ cells, Calcofluor 115 white staining assay described in (PRINGLE 1991; PREECHASUTH et al. 2015; SHARMA et al. 116 2019) was adopted. Briefly, WT and mutant strains were grown overnight at 30ºC and next day 117 transferred to fresh culture in 1:10 ratio. Cells were grown to log phase and collected by 118 centrifugation. Further, cells were suspended in 100µl of solution containing Calcofluor white 119 (50 μg/ml...

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“…Growth assessment of the WT and mutants was carried out as mentioned (17). Growth kinetics of strains was analyzed for 14 hrs by measuring optical density at 600nm using TOSHVIN UV-800 SHIMADZU spectrophotometer.…”

Section: Methodsmentioning

confidence: 99%

Loss of F-box Motif Encoding Gene SAF1 and RRM3 Together Leads to Synthetic Growth Defect and Sensitivity to HU, MMS inS.cerevisiae

Sharma

Verma

Bairwa

2019

Preprint

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The analysis of the nucleus using DAPI stain indicated increased percentage of cells showing 126 multi-nuclei phenotype in saf1∆rrm3∆ cellsin comparison to saf1∆, rrm3∆, and WT ( Figure 127 2A). The multi-nuclei phenotype indicated the defect in the nuclear migration.. 128The saf1∆rrm3∆cells showed increased Ty1 retro-transposition 129 Ty1 elements are class of retro-transposons in the S. cerevisiae genome which remain in the 130 dormant state due the host encoded genetic factors. In the absence of RRM3, the frequency 131 of Ty1retro-transposition goes up as reported previously (15, 16). In this study the single 132 gene mutant saf1∆showed nearly 8 fold whereas the rrm3∆ showed the 85fold change in 133 His+ prototroph formation in comparison to WT. However the absence of both the genes 134 (saf1∆rrm3∆) leads to increased frequency (~ 180-fold) of Ty1-retro transposition in the 135 comparison to WT cells. (Figure 3 A, B). 136 Loss of SAF1 and RRM3 leads to HU and MMS sensitivity 137 The hydroxyurea (HU) and methyl methanesulfonate (MMS) act as replication checkpoint 138 inhibitor and DNA damaging agent respectively. The Rrm3 mutant reported to have slightly 139 sensitive phenotype in the 200mM hydroxyurea (22). With regard to SAF1 mutant the data is 140 not available. The semi-quantitative growth assay by spot analysis on YPD medium and 180 Manjithya, NCBS, India for strains and plasmids.

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Investigation of the Best Saccharomyces cerevisiae Growth Condition

Cited by 70 publications

References 14 publications

The Utilization of Vegetable and Fruit Wastes for Saccharomyces cerevisieae Cell Wall Based β-Glucan Production with Antioxidant Activity

The Utilization of Vegetable and Fruit Wastes for Saccharomyces cerevisieae Cell Wall Based β-Glucan Production with Antioxidant Activity

Deletion of Autophagy geneATG1and F-box motif encoding geneYDR131Ctogether leads to Synthetic Growth Defects and Flocculation behaviour inSaccharomyces cerevisiae

Loss of F-box Motif Encoding Gene SAF1 and RRM3 Together Leads to Synthetic Growth Defect and Sensitivity to HU, MMS inS.cerevisiae

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