This thesis is focused on design and implementation web application it based on the requirements and analysis of the current situation using technologies PHP and MySQL. It is specification system for planning rides, final exams and related functions in driving school

Charles Denby (3437777)

Edward Baidoo (3437768)

George Wang (682337)

Jay Keasling (272439)

Sarah Rodriguez (3437771)

T. Vu (3437774)

Microbial Cell Factories

PubMed

Sarah Rodriguez

Charles M. Denby

T. Van Vu

Edward E. K. Baidoo

George Wang

Jay D. Keasling

Crossref

ATP citrate lyase mediated cytosolic acetyl-CoA biosynthesis increases mevalonate production in Saccharomyces cerevisiae

BackgroundWith increasing concern about the environmental impact of a petroleum based economy, focus has shifted towards greener production strategies including metabolic engineering of microbes for the conversion of plant-based feedstocks to second generation biofuels and industrial chemicals. Saccharomyces cerevisiae is an attractive host for this purpose as it has been extensively engineered for production of various fuels and chemicals. Many of the target molecules are derived from the central metabolite and molecular building block, acetyl-CoA. To date, it has been difficult to engineer S. cerevisiae to continuously convert sugars present in biomass-based feedstocks to acetyl-CoA derived products due to intrinsic physiological constraints-in respiring cells, the precursor pyruvate is directed away from the endogenous cytosolic acetyl-CoA biosynthesis pathway towards the mitochondria, and in fermenting cells pyruvate is directed towards the byproduct ethanol. In this study we incorporated an alternative mode of acetyl-CoA biosynthesis mediated by ATP citrate lyase (ACL) that may obviate such constraints.ResultsWe characterized the activity of several heterologously expressed ACLs in crude cell lysates, and found that ACL from Aspergillus nidulans demonstrated the highest activity. We employed a push/pull strategy to shunt citrate towards ACL by deletion of the mitochondrial NAD(+)-dependent isocitrate dehydrogenase (IDH1) and engineering higher flux through the upper mevalonate pathway. We demonstrated that combining the two modifications increases accumulation of mevalonate pathway intermediates, and that both modifications are required to substantially increase production. Finally, we incorporated a block strategy by replacing the native ERG12 (mevalonate kinase) promoter with the copper-repressible CTR3 promoter to maximize accumulation of the commercially important molecule mevalonate.ConclusionBy combining the push/pull/block strategies, we significantly improved mevalonate production. We anticipate that this strategy can be used to improve the efficiency with which industrial strains of S. cerevisiae convert feedstocks to acetyl-CoA derived fuels and chemicals

Rodriguez, Sarah

Denby, Charles M

Van Vu, T

Baidoo, Edward EK

Wang, George

Keasling, Jay D

eScholarship - University of California

Additional file 3: Figure S2. Growth curves of engineered strains pertaining to Fig.Â 4. Strains were grown in low [(NH4)2SO4]

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MOESM3 of ATP citrate lyase mediated cytosolic acetyl-CoA biosynthesis increases mevalonate production in Saccharomyces cerevisiae

Tovaryš, Ondřej

National Repository of Grey Literature

Development of Supporting Software for Driving School

Additional file 2: Figure S1. Intracellular concentrations of a panel of key related metabolites of depicted strains grown in low nitrogen. *Oxaloacetate was not detected, as it is very labile

MOESM2 of ATP citrate lyase mediated cytosolic acetyl-CoA biosynthesis increases mevalonate production in Saccharomyces cerevisiae

Additional file 5: Figure S4. Growth curves of wild type cultures grown with various mevalonate concentrations added to media at time zero (A) and measurement of mevalonate concentrations in broth after various mevalonate concentrations added to cultures at time zero (B)

MOESM5 of ATP citrate lyase mediated cytosolic acetyl-CoA biosynthesis increases mevalonate production in Saccharomyces cerevisiae

Additional file 6: Figure S5. Comparison of intracellular and extracellular amounts of mevalonate found in the 1Â mL of culture broth. Strain shown is the S. cerevisiae host strain JBEI-10688 harboring the pMvaES plasmid

MOESM6 of ATP citrate lyase mediated cytosolic acetyl-CoA biosynthesis increases mevalonate production in Saccharomyces cerevisiae

Additional file 4: Figure S3. Growth of S. cerevisiae IDH1Δ/2Δ and IDH1Δ/2Δ complemented with IDH1 and IDH2 from Y. lipolytica on dextrose (fermentable) and acetate (non-fermentable)

MOESM4 of ATP citrate lyase mediated cytosolic acetyl-CoA biosynthesis increases mevalonate production in Saccharomyces cerevisiae

https://escholarship.org/uc/item/2hn464rc

Development of Supporting Software for Driving School

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eScholarship - University of California

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