AhmadWeek3: Difference between revisions
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==Part 1: Experimenting with different levels of ammonia== | |||
*<i> Saccharomyces cerevisae</i>, a type of yeast, uses ammonia as its primary nitrogen source | *<i> Saccharomyces cerevisae</i>, a type of yeast, uses ammonia as its primary nitrogen source | ||
**Past research has shown that ammonia concentration affects growth | **Past research has shown that ammonia concentration affects growth | ||
Line 7: | Line 8: | ||
**After 61 mM, however, there was no significant increase in biomass | **After 61 mM, however, there was no significant increase in biomass | ||
**Formula to calculate ammonia flux: [dilution x (input ammonia concentration - residual ammonia concentration)/biomass] | **Formula to calculate ammonia flux: [dilution x (input ammonia concentration - residual ammonia concentration)/biomass] | ||
** | **At a level above 44 mM, oxygen consumption was equal to carbon dioxide production | ||
*Ketoglutarate, Glutamate, Glutamine concentrations as ammonia concentrations were increased | |||
**Ketoglutarate: Decreased | |||
**Glutamate: Increased | |||
**Glutamine: Increased | |||
*Figure 1 | |||
**X axis: NH4 concentration | |||
**Y axis: | |||
***A: Residual NH4 concentration | |||
***B: 02 consumption/CO2 production | |||
***C: Ketoglutarate, Glutamate, Glutamine concentrations | |||
==Part 2: Gene Expression== | |||
*Northern Analyses are RNA analyses that were done to see how the changes in nitrogen affect nitrogen-regulated genes | |||
*Figure 2 shows gene expression in response to different levels of NH4 concentrations | |||
**X axis: NH4 concentration | |||
**Y axis: Gene expression | |||
*Permease-encoding genes: | |||
**GAP1 | |||
**PUT4 | |||
*Biosynthetic genes | |||
**ILV5 | |||
**HIS4 | |||
**GDH1 | |||
**GLN1 | |||
==Part 3: Enzymes== | |||
*Figure 3 shows enzyme activity at different NH4 concentrations | |||
**X axis: NH4 concentration | |||
**Y axis: Enzyme activity | |||
*Enzymes Studied: | |||
**NADPH | |||
**NAD-DGH | |||
**GS Transferase | |||
*NADPH activity consistently decreased with increased NH4 | |||
*NAD-GDH activity consistently increased with increased NH4 | |||
*GS Transferase and Synthetase decreased slightly until 60 mM, but then leveled off with increased NH4 | |||
*The conclusion of the paper is that that the nitrogen metabolism of <i> S. cerevisiae </i> is regulated by ammonia concentration | |||
**The species may have an ammonia sensor | |||
==Word Definitions== | |||
#[http://www.biology-online.org/dictionary/Flux Flux]: "The total amount of a quantity passing through a given surface per unit time." | |||
#[http://www.biology-online.org/dictionary/Permease 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." | |||
#[http://www.biology-online.org/dictionary/Oligonucleotide Oligonucleotide]: "Linear sequence of up to 20 nucleotides joined by phosphodiester bonds." | |||
#[http://www.biology-online.org/dictionary/Gram-negative Gram-negative]:"This is a primary characteristic of bacteria that have a cell wall composed of a thin layer of peptidoglycan covered by an outer membrane of lipoprotein and lipopolysaccharide containing endotoxin." | |||
#[http://www.biology-online.org/dictionary/Gram-positive Gram-positive]:"This is a primary characteristic of bacteria whose cell wall is composed of a thick layer of peptidologlycan containing teichoic and lipoteichoic acid complexed to the peptidoglycan." | |||
#[http://www.biology-online.org/dictionary/Glutamate Glutamate]:"Major fast excitatory neurotransmitter in the mammalian central nervous system." | |||
#[http://www.biology-online.org/dictionary/Glutamine Glutamine]:"A crystalline amino acid occurring in proteins; important in protein metabolism.One of the 20 amino acids that are commonly found in proteins." | |||
#[http://www.biology-online.org/dictionary/Isomerase Isomerase]:"An enzyme that converts molecules into their positional isomers." | |||
#[http://www.biology-online.org/dictionary/Synthetase Synthetase]:"Enzymes of class 6 in the e classification, catalyse synthesis of molecules, their activity being coupled to the breakdown of a nucleotide triphosphate." | |||
#[http://www.biology-online.org/dictionary/Metabolite Metabolite]:"Any substance produced by metabolism or by a metabolic process." | |||
{{Salman Ahmad}} |
Latest revision as of 22:22, 7 February 2013
Part 1: Experimenting with different levels of ammonia
- Saccharomyces cerevisae, a type of yeast, uses ammonia as its primary nitrogen source
- Past research has shown that ammonia concentration affects growth
- In this paper, the researches wanted to know if ammonia flux, and not concentration, is the limiting factor
- Physiological Parameters
- S. cerevisae was first grown in cultures with different levels of ammonia, but fixed glucose concentrations
- Increases in the ammonia concentration from 29 to 61 mM showed a change in biomass from 4.9 to 8.2 g/L
- After 61 mM, however, there was no significant increase in biomass
- Formula to calculate ammonia flux: [dilution x (input ammonia concentration - residual ammonia concentration)/biomass]
- At a level above 44 mM, oxygen consumption was equal to carbon dioxide production
- Ketoglutarate, Glutamate, Glutamine concentrations as ammonia concentrations were increased
- Ketoglutarate: Decreased
- Glutamate: Increased
- Glutamine: Increased
- Figure 1
- X axis: NH4 concentration
- Y axis:
- A: Residual NH4 concentration
- B: 02 consumption/CO2 production
- C: Ketoglutarate, Glutamate, Glutamine concentrations
Part 2: Gene Expression
- Northern Analyses are RNA analyses that were done to see how the changes in nitrogen affect nitrogen-regulated genes
- Figure 2 shows gene expression in response to different levels of NH4 concentrations
- X axis: NH4 concentration
- Y axis: Gene expression
- Permease-encoding genes:
- GAP1
- PUT4
- Biosynthetic genes
- ILV5
- HIS4
- GDH1
- GLN1
Part 3: Enzymes
- Figure 3 shows enzyme activity at different NH4 concentrations
- X axis: NH4 concentration
- Y axis: Enzyme activity
- Enzymes Studied:
- NADPH
- NAD-DGH
- GS Transferase
- NADPH activity consistently decreased with increased NH4
- NAD-GDH activity consistently increased with increased NH4
- GS Transferase and Synthetase decreased slightly until 60 mM, but then leveled off with increased NH4
- The conclusion of the paper is that that the nitrogen metabolism of S. cerevisiae is regulated by ammonia concentration
- The species may have an ammonia sensor
Word Definitions
- Flux: "The total amount of a quantity passing through a given surface per unit time."
- 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."
- Oligonucleotide: "Linear sequence of up to 20 nucleotides joined by phosphodiester bonds."
- Gram-negative:"This is a primary characteristic of bacteria that have a cell wall composed of a thin layer of peptidoglycan covered by an outer membrane of lipoprotein and lipopolysaccharide containing endotoxin."
- Gram-positive:"This is a primary characteristic of bacteria whose cell wall is composed of a thick layer of peptidologlycan containing teichoic and lipoteichoic acid complexed to the peptidoglycan."
- Glutamate:"Major fast excitatory neurotransmitter in the mammalian central nervous system."
- Glutamine:"A crystalline amino acid occurring in proteins; important in protein metabolism.One of the 20 amino acids that are commonly found in proteins."
- Isomerase:"An enzyme that converts molecules into their positional isomers."
- Synthetase:"Enzymes of class 6 in the e classification, catalyse synthesis of molecules, their activity being coupled to the breakdown of a nucleotide triphosphate."
- Metabolite:"Any substance produced by metabolism or by a metabolic process."
==Assignments==
- Class Journal Week 1
- Journal Assignment Week 2
- Class Journal Week 2
- Journal Assignment Week 3
- Class Journal Week 3
- Journal Assignment Week 4
- Class Journal Week 4
- Journal Assignment Week 5
- Class Journal Week 5
- Journal Assignment Week 6
- Class Journal Week 6
- Journal Assignment Week 8
- Class Journal Week 8
- Journal Assignment Week 9
- Class Journal Week 9
- Journal Assignment Week 11
- Class Journal Week 11
- Journal Assignment Week 12
- Class Journal Week 12
- Journal Assignment Week 13
- Class Journal Week 13
- Assignment Week 13
- Journal Assignment Week 14
- Class Journal Week 14
- Assignment Week 14