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Flashcards in lecture 18 enzymes Deck (18):
1

role of enzymes in biochemical rxn

start or speed up rxn of conversion of reactants to products

2

relationship btw substrate and enzyme active site

active site of enzyme binds to substrate to undergo chemical run

3

3 ways progress of enzyme catalyzed rxn is monitored

disappearance of substrate, appearance of product, conversion of cofactor

4

what is happening in lag phase

run rate is zero but then substrate start binding to active site, rxn rate incr. once [ES] constant measurements taken

5

what is happening in linear phase

all active sites occupied. [ES] constant. depends only on [enzyme]. independent of [substrate] zero order kinetics

6

what is happening in plateau phase

rxn rate declines as substrate consumed, becomes dependent on [substrate], and becomes non linear. first-order kinetic

7

diff btw cont and fixed time measurement of an enzyme analyte

cont- multiple point measurement
fixed- single point, must know time over which ran is linear

8

why fixed-time measurement enzyme assay only be performed once rate of enzyme reaction is known?

discontinuous, needs to be in the linear phase to be valid

9

why hexokinase method for glucose measurement called coupled enzyme rxn

the first enzyme rxn produces the substrate for the second enzyme rxn

10

compare enzyme or substrate is rate limiting

k

11

compare measuring serum enzyme using reagent substrate vs measuring serum substrate using enzyme reagent

d

12

j

what is EcoRI DNA restriction digest

13

cuts each DNA strand by hydrolyzing phosphodiester linkage betn each G-A, leaving two ‘sticky-ends’ that are complementary

what is EcoRI DNA restriction digest

14

joins dsDNA end, phophosphodiester linkages can reform, recreating the EcoRI restriction site

what is DNA ligase

15

-Taq DNA polymerase
-DNA sample to be tested
-Reaction buffer
-MgCl2
-dNTP (deoxynucleotides, G, A, T, and C)
-Oligonucleotide primers

what are components of PCR amplication

16

h

hexokinase method

17

h

glucose oxidase method

18

s

glucose electrode method