James P. McDonald Week 3
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Biological Terms
- Permease: "General term for a membrane protein that increases the permeability of the plasma membrane to a particular molecule, by a process not requiring metabolic energy." [[1]]
- Isomerase: "An enzyme that converts molecules into their positional isomers." [[2]]
- Oligonucleotides: "Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesised to match a region where a mutation is known to occur, and then used as a probe (oligonucleotide probes)." [[3]]
- Dehydrogenase: "Enzyme that oxidizes a substrate by transferring hydrogen to an acceptor that is either NAD/NADP or a flavin enzyme. An enzyme that is used to remove hydrogen from its substrate, which is used in the cytochrome (hydrogen carrier) system in respiration to produce a net gain of ATP." [[4]]
- Synthetase: "Enzymes of class 6 in the e classification, catalyse synthesis of molecules, their activity being coupled to the breakdown of a nucleotide triphosphate." [[5]]
- Biosynthetic: "Relating to or produced by biosynthesis." [[6]]
- Glutamate: "Major fast excitatory neurotransmitter in the mammalian central nervous system." [[7]]
- Glutamine: "A crystalline amino acid occurring in proteins; important in protein metabolism. One of the 20 amino acids that are commonly found in proteins." [[8]]
- GAP1: "General amino acid permease, a gene found in Saccharomyces cerevisiae." [[9]]
- PUT4: "Proline permease, a gene found in Saccharomyces cerevisiae." [[10]]
Outline
Introduction
- Saccharomyces cerevisiae was grown in various ammonia concentrations and the effects on the growth was observed.
- A single dilution rate was using with a range of different ammonia concentrations.
- The ammonia concentrations were varied to observe its effects on gene expression and enzyme activities.
- The main result of the study was that nitrogen metabolism is dependent on ammonia concentration, not its flux.
Significance
- Ammonia is the prefferred growth source of Saccharomyces cerevisiae as it results in faster growth.
- Nitrogen metabolism is regulated by gene expression and enzyme activity.
- Previous research seems to show that ammonia concentration itself is the most important factor in nitrogen metabolism.
- But, in these previous studies the cultures have differed in ammonium flux, leaving flux as the possible key factor.
- This experiment uses cultures with the same level flux, only the ammonium concentrations fed in are different.
Methods
Physiological Parameters
- Saccharomyces cerevisiae SU32 was grown in continuous cultures
- They were fed with different ammonia concentrations: 29, 44, 61, 66, 78, 90, 96, 114, 118 mM.
- Contained a fixed glucose concentration at 100 mM.
- Had a dilution rate of 0.15h-1.
- Biomass and residual ammonia concentration were measured at the different ammonia concentrations.
- The ammonium flux was calculated using the biomass, ammonia concentration, and the residual ammonia concentration.
- The respiratory quotient was calculated using the measured values of CO2 production and O2 consumption at the different ammonia concentrations.
- Alpha-ketoglutarate, glutamate, and glutamine concentrations were measured and the different ammonia concentrations.
Northern Analyses
Enzyme Activities
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