Screening Flocculant Yeast Mutants Higher Ethanol Tolerance
•Download as PPTX, PDF•
1 like•223 views
1. The study screened the flocculant yeast Saccharomyces cerevisiae NCYC 1195 for tolerance to high ethanol concentrations using gamma ray radiation mutagenesis. 2. Mutant yeast strains Y7 and Y8 showed higher tolerance to ethanol (5% concentration) compared to the wild type (2.5% concentration). 3. Mutant Y8 demonstrated 15.9% higher cell growth and sugar consumption compared to the wild type yeast under growth profile testing.
Recommended
Recommended
More Related Content
Similar to Screening Flocculant Yeast Mutants Higher Ethanol Tolerance
Similar to Screening Flocculant Yeast Mutants Higher Ethanol Tolerance (20)
More from Global R & D Services
More from Global R & D Services (20)
Recently uploaded
Recently uploaded (20)
Screening Flocculant Yeast Mutants Higher Ethanol Tolerance
- 1. SCREENING OF FLOCCULANT SACCHAROMYCES CEREVISIAE (NCYC-1195) FOR HIGH TOLERANCE OF ETHANOL CONCENTRATION Harsojo National Nuclear Technology Agency (BATAN) Jakarta - Indonesia Edwin Yulian E. Brawijaya University Malang - Indonesia Agustin K. Wardani Brawijaya University Malang - Indonesia ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore
- 2. Background ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore
- 3. Background Concentrations more than 8% (v/v) : -increasing membrane fluidity -interfere on cell homeostasis and several metabolic pathways ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Increasing yeast tolerance ability in the presence of ethanol is a necessary. Ethanol-tolerant yeast would have enhanced ethanol productivity and yields.
- 4. ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Background Mutagenesis research by Mehdikhani, P. et al. (2011) showed that gamma ray 60Co can increase yeast tolerance on high ethanol concentration and high temperature during fermentation Increasing the ethanol tolerance of yeast can be done by mutation technique. One of the mutation technique is random mutagenesis by gamma ray 60Co. Its have ability to do self-floculating → easy separation → reduce cost production. Flocculant S. cerevisiae (NCYC 1195)
- 5. ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Aim of The Study Increase the tolerance ability of flocculant Saccharomyces cerevisiae (NCYC 1195) to high concentration of ethanol Determine gamma ray 60Co dose that give highest cell lethality to flocculant Saccharomyces cerevisiae (NCYC 1195)
- 6. Methodology ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore S. cerevisiae NCYC 1195 Ethanol-tolerant Mutant
- 7. Reseacrh Step ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore • Gamma ray treatment • Isolation survival coloniesRadiation • Ethanol-tolerant screening • Isolation ethanol-tolerant mutantScreening • Determine the viable cell count • Determine the sugar consumption Growth Profile test
- 8. ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Radiation 3 x 108 cell/ml 0 KGy 0,6 KGy 1,4 KGy 0,8 KGy 1 KGy 1,2 KGy Incubation 24 hours at 30OC
- 9. Ethanol-tolerant Screening ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Day 1 Day 2 Day 3 Day 4 0.1 ml 108 cell/ml 5 ml YPG broth 2.5 % ethanol (v/v) Day 5
- 10. ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Growth Profile Test 100 ml YPG Broth medium : 10 % glucose 2 % peptone 1% yeast extract 30 OC and 120 rpm At shaker waterbath for 72 hours 5 % inoculum
- 11. ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Results Dose (KGy) Survival Colonies (CFU/ml) 0 44 x 106 0.6 26 0.8 2 1 0 1.2 0 1.4 0 Dose (KGy) Number of Screened Colonies Mutant 0.6 26 4 0.8 2 2 Table 1. Number of survival colonies by different doses of gamma ray 26 survival colonies obtained after gamma ray treatments at 0.6 KGy dose 2 survival colonies obtained after gamma r treatments at 0.8 KGy dose No survival colonies obtained excess 1 KGy dose 1. 2. Table 2. Number of screened colonies and ethanol-tolerance 4 colonies (Y6a, Y6b, Y6c and Y6d) from 0.6 KGy dose that could survive in the presence of 2.5 % (v/v) ethanol concentration 2 colonies (Y7 and Y8) from 0.8 KGy dose that could survive in the presence of 5 % (v/v) ethanol concentration Mutant Y7 and Y8 show higher tolerance of ethanol concentration than the WT. Mutant Y7 and Y8 = 5% (v/v) ethanol concentration WT = 2.5% (v/v) ethanol concentration
- 12. ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Growth Profile -20 0 20 40 60 80 100 120 140 0 12 24 36 48 60 72 0 2 4 6 8 10 12 Cells(106CFU/ml) Time (h) TotalResidualSugars(g/100ml) ▬▲▬ Viable cell growth of Mutant Y8 ▬■▬ Viable cell growth of WT ▲ Sugar consumption of Mutant Y8 ■ Sugar consumption of Mutant Y8
- 13. The present study shows that : 1. Flocculant Saccharomyces cerevisiae (NCYC 1195) have highest cell lethality by gamma irradiation treatments at 0.6-0.8 KGy. 2. Two mutants, Y7 and Y8, have higher ethanol tolerance (5 % ethanol concentration (v/v)) than the wild type (2.5 % ethanol concentration (v/v)). 3. Mutant Y8 display 15.9% higher numbers of viable cell than the wild type. Conclusion ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore
- 14. In the next study, we will determine the optimum fermentation condition for ethanol production and investigate the stability of mutants ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore Future Research
- 15. Thank you for your kind attention ‘International Conference on Advance in Healthcare and Life Sciences’ 8-9 November 2014, Singapore
Editor's Notes
- Good afternoon, thank for the time. My name edwin….., I come from brawijaya university, indonesia. Today I wil present my research about screening…..
- Ethanol has been regarded as alternative energy source, because its environmental energy and renewable energy. Ethanol produce by fermentation, ethanol fermentation is done by yeast
- During fermentation, yeast are exposed to various stress, that is ethanol, sugar, temperature and by-product. Among the stress, ethanol is considered to be major stress that responsible for decreasing ethanol production
- The aim of this study is determine…… and increase……..
- We use S.cerevisiae NCYC 1195 for the strains. To increase the ethanol tolerance ability of this strain, we use gamma ray treatment. Gamma ray have high energy to change the phsycal, chemical, and biologic properties of the irradiated material.
- Table 1 show the number of …………. There was 26 survival……………, there was 2 survival and no survival…………. Table 2 show the number of screened…….., we screened 28 colonies from gamma ray treatments. The 26 colonies was caried out from 0.6 KGy dose and 2 colonies was caried out from 0.8 Kgy. We obtained 4 mutants (Y6a……) that could survive in the presence of 2,5 % ethanol and 2 mutant (Y7 and Y8) that could survive in the presence of 5% ethanol, while the wild type have tolerance ability to 2,5 % ethanol concentration
- This figure display the growth profile of mutant Y8 and WT. the dashed line with filled triangle is the viable cell count of mutant Y8, while the dashed line with filled square is the viable cell count of WT. the mutant Y8 produce 15.9% higher viable cell than the WT. the highest viable cell count obtained at 48 hours. The solid line with filled triangle is sugar consumption by mutant Y8 and the solid line with filled square is sugar consumption by WT. The graph show that sugar consumption of mutant Y8 is lower than WT.