Escolar Documentos
Profissional Documentos
Cultura Documentos
2 Semi-conservative DNA replication 1)Two single strands of the DNA molecule unzip along the line of hydrogen bonds by DNA helicase. 2) Each revealed parent strand acts as a template for a new strand to synthesise. 3)The exposed bases of the parent strand attract free mononucleotides to join each template via complementary base pairing. The enzymes DNA polymerase and DNA ligase join the nucleotides together with hydrogen bonds through condensation reactions to form new DNA strands. 4)The result is two new strands of DNA identical with the original piece. The new molecules automatically coil up into the double helix.
Evidence for DNA semi-conservative replication Meselson and Stahl Two isotopes of nitrogen (as DNA contains nitrogen) heavy nitrogen (15N) and light nitrogen (14N) 1) Samples of bacteria grown one in light nitrogen / other with heavy nitrogen as the bacteria reproduced they took up nitrogen to make nucleotides for new DNA so the nitrogen became part of their DNA 2) Sample of DNA was taken from each batch spun in centrifuge heavy DNA sinks lower 3) Heavy nitrogen bacteria placed in light nitrogen broth left for replication sample taken 4) Semi-conservative new bacterial DNA molecules contain strand of each. The DNA settles between where the light/heavy settled originally. 5) The DNA settle in the middle showing that it has mixture of heavy/light the bacterial DNA replicated semi-conservatively in light nitrogen.
Protein synthesis
RNA is closely related to DNA but it doesnt form complex molecules. The sequence of bases along a strand of RNA is related to the sequence of bases on a small part of DNA in the nucleus different types of RNA take different roles in the process of protein synthesis. Messenger RNA (mRNA) carries information from the DNA in the nucleus out into the cytoplasm. It is formed when a smell length of the DNA double helix unzips. The coding or antisense strand of the DNA acts as a template for the formation of the mRNA. The mRNA then moves out of the nucleus transporting the instruction from the genes to the surface of the ribosomes which are the site of protein synthesis. Transfer RNA (tRNA) is found in the cytoplasm it picks up particular amino acids from the vast numbers always free there. The tRNA molecules each carrying an amino acid line up alongside the mRNA on the surface of the ribosome building up a long chain of amino acids. Peptide links are formed between the amino acids, joining them together to form a polypeptide chain which in turn can be used to form a larger protein. 1) DNA is transcribed to give a length of mRNA 2) tRNA in the cell attaches to specific amino acids 2a) mRNA moves out of the nucleus and becomes engulfed by a ribosome 3) tRNA molecule carrying amino acid lines up against matching mRNA on the ribosome 4) Peptide links are formed between the amino acids brought by the tRNA 5) When the polypeptide is released into the cytoplasm, the tRNA units also unbind and return to the cytoplasm to pick up more amino acids. The ribosome may read the mRNA again. In protein synthesis the information held in the sequence of bases in a gene is translated into a sequence of amino acids in a polypeptide chain.
Activation energy
- Activation energy amount of energy required for chemicals before reaction can start (heat) - Enzymes lower activation energy reactions happen at lower temperatures Enzyme-substrate complex lowers activation energy - If two substrate molecules need to be joined, being attached to the enzyme holds them close together reducing any repulsion between the molecules so they can join more easily. - If the enzyme is catalysing a breakdown reaction, fitting into the active site puts strain on bonds in the substrate so the substrate molecule breaks up more easily.
Lock and key substrate fits into the enzymes active site specific shapes for each Induced fit enzyme and substrate have to fit together in the first place enzyme-substrate
complex changes shape slightly to complete the fit locking the substrate more tightly to the enzyme. The substrate also has to make the active site change shape in a particular way.
Immobilized enzymes
- packed into columns and used over long periods - higher temperatures can be used thermal stability - high operating temperatures increase rate of reaction - product doesnt have to be separated from enzyme saves money - reusability
Enzyme experimentsMeasuring the initial rate of an enzyme-controlled reaction / repeat it / exponential / correlation Measure how fast the product of the reaction appears Catalase > hydrogen peroxide - state what variables are controlled - measure how much oxygen is given off in a gas cylinder - different concentrations of enzyme (state them 10%, 20% etc.) - measure the amount of oxygen given off after certain time period e.g. 1 min - divide the volume of oxygen produced by the time initial rate of reaction - more given off in first minute faster initial rate of reaction.
Rate of reaction
Affected by enzyme concentration - More enzyme molecules more likely a substrate molecule will collide with one and form an enzyme-substrate complex. - If the amount of substrate is limited there comes a point when there are more than enough enzyme molecules to deal with all the available substrate, so adding more enzymes has no further effect. Affected by substrate concentration The enzyme can become saturated all of the active sites are occupied by substrate molecules and a further increase in substrate concentration will not increase the reaction further. This point is called v-max which is the maximum rate of reaction.
Temperature
Higher temperature more kinetic energy > more heat energy more collisions rate of reaction increases. Optimum temperature enzymes catalytic activity is greatest (37.5) in humans. Above this level the enzyme structure begins to break down (denature) (third/fourth structures unravel) High levels - molecular bonds break down in protein permanently altering active site rendering it functionless. Exceptions thermophillic / temperature coefficient rate of reaction at (x+10) / rate of reaction at x
pH level
A lower ph more acidic higher number of hydrogen to form bonds with enzyme protein structure High ph alkaline less bonds form (both affect structure that holds 3D protein structure. - alteration of intermolecular bonds in extremes can denature the active site making it functionless - enzymes have an optimum pH range works most effectively
Inhibition
active site-directed: occupy active site to prevent substrate molecule from binding. non-active site-directed: inhibitors that attach to other parts distorting its shape