1) What is the mechanism by which enzymes catalyze reactions?

A. They lower the reaction energy by stabilizing the [ES]* transition state.
B. They increase substrate concentration to more effectively bind substrate to
product.
C. They introduce energy from ATP into the reaction.
D. They remove the product so that the reaction is not product-inhibited.
E. They deactivate inhibitors to the reaction.
2)
A.
B.
C.
D.
E.

LDH is short for:

Lactose dehalogenase
Lithium dehydrogenase
Lactate dehydrogenase
Lithium dehalogenase
Lethal dose hydronium

3) During the enzyme kinetics lab, you used a _____________ to directly monitor
the progress of the reaction.
A. balance
B. spectrophotometer
C. stopwatch
D. microscope
E. balance with a stopwatch
4)
A.
B.
C.
D.
E.

During the kinetic studies of LDH, what was [S]?

[lactate]
[NADH]
[LDH]
[pyruvate]
[oxamate]

5) What is a PCR
A. Polyunsaturase Chain reaction
B. Polysaturase Chain reaction
C. Polymerase Chain Reaction
D. Pectinase Chain Reaction
E. None of the Above
6) What step of PCR is where primers go and attach to DNA strand
A. Annealing
B. Extension
C. Denaturation
D. Heating
E. Cooling
7) How many hydrogen bonds are present between A-T base pair

A.
B.
C.
D.
E.

One
Two
Three
Four
None of the above

8) In Gel electrophoresis DNA migrates from

A. Cathode to Anode
B. Anode to Cathode
C. Positive Electrode to Negative Electrode
D. Negative Electrode to Positive Electrode
E. Both A & D
F. Both B & D
9) What is the Charge carried by a DNA strand?
A. Positive
B. Negative
C. No Charge
D. Electrically neutral
E. None of the above
10) In gel electrophoresis DNA strand are separated on the basis of
A. Size
B. Shape
C. Charge
D. A&C
E. A&B
11) 1/10 diluted sample returns a spectrophotometric absorbance reading of 0.8.
Applying this value to a standard curve suggests that this absorbance equates to a
concentration of 5mg/ml. What is the concentration of the original undiluted
sample?
A. 50 mg/ml
B. 0.5mg/ml
C. 0.08
D. 0.8
E. 5mg/ml
12) What was used as a stationary phase in Affinity Chromatography
A. Sephadex
B. Agarose
C. Cibacron Blue
D. Tris buffer
E. Sephadex buffer
13) What is the absorbance max for NADH?

A.
B.
C.
D.
E.

340nm
380nm
590nm
560nm
None of the above

14) LDH is what type of enzyme

A. Oxidase
B. Hydrolase
C. Reductase
D. Oxidoreductase
E. Lyase
Short Answer Questions
1) Using the following data obtained from an LDH kinetic study, plot a MichaelisMenten curve on the graph paper given below, and indicate the approximate K M
and Vmax values for this enzyme. Be sure to label the x-axis and y-axis properly.
(10 points )

Tube
Blank
1
2
3
4
5

[Pyruvate]
(mM)
0.0
0.2
0.4
0.6
0.8
1.0

[NADH]/min
(mM/min)
0.00
0.06
0.13
0.18
0.21
0.22

2) Using the following data, plot a Michaelis-Menten curve in the graph area below.
Label the x- and y-axes correctly, and indicate the approximate Km and Vmax
values on the plot. Absorption co-efficient of NADH at 340nm is 6220 M-1cm-1
(10points)
Tube

Enzyme(L
)

Blank
1
2
3
4
5

50
50
50
50
50
50

3mM
NADH(L
)
100
100
100
100
100
100

14mM
Buffer(L)
pyruvate(L)
0
20
40
80
160
320

850
830
810
770
690
530

Substrate
Concn
(mM)

V0 (A340 V0
(mM
-1
min )
NADH ml-1
min-1)
0
0.06
0.13
0.18
0.21
0.22

3) AFFINITY CHROMOTOGRAPHY
True/False: Dehydrogenases are broadly known as oxidoreductases because they transfer
electrons from one molecule to another.(2points)
Why did we use Cibacron Blue in the column? Explain briefly how this aided in isolating
LDH. (4 points)
What technique did you use to identify the rate at which NAD+ was converted to NADH
and what did that tell you about your fractions?(4 points)
4) Write in short sentences about following terms
a.
b.
c.
d.
e.
f.
g.
h.
i.

Affinity chromatography
Two factors of Enzyme kinetics.
GMO
Gel electrophoresis
PCR
DNA
Promoter of a gene
LDH
Molisch Test