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EH/APR 2011/CBE451

UNIVERSITI TEKNOLOGI MARA FINAL EXAMINATION

COURSE COURSE CODE EXAMINATION TIME

BIOCHEMISTRY AND METABOLIC REGULATION CBE451 APRIL 2011 3 HOURS

INSTRUCTIONS TO CANDIDATES 1. 2. 3. 4. This question paper consists of five (5) questions. Answer ALL questions in the Answer Booklet. Start each answer on a new page. Do not bring any material into the examination room unless permission is given by the invigilator. Please check to make sure that this examination pack consists of: i) ii) the Question Paper an Answer Booklet - provided by the Faculty

DO NOT TURN THIS PAGE UNTIL YOU ARE TOLD TO DO SO

This examination paper consists of 7 printed pages
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QUESTION 1 (P01,C01,C1,C2, C3) a) Use the reaction below to answer the following questions.

R'

R'

H-N-C-C-OH + I I II H H O
Compound A

H-N-C-C-OH I I II H H O
Compound B

^^HaO

R'

R'

I I H-N-C-C-N-C-C-OH I I II I I II H H O H H O
Compound C

i) Identify the functional groups at both ends on compound A and B. ii) Identify the class of biomolecule represented by compound A. iii) Identify the reaction type occurring above. Justify your answer.

(2 marks) (1 mark) (3 marks)

iv) Identify the specific type of polar covalent bond being formed in compound C. (1 mark) b) The membrane transport in cells can be classified into two major groups which are passive transport and active transport. i) Identify the difference between passive transport and active transport. (3 marks) ii) There are two (2) kinds of passive transport. Briefly describe them. (4 marks)

iii) Explain the meaning of cotransport and identify the types of membrane transport which fall in this group. (2 marks)

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c) A production of lactic acid was done by a Bioprocess Engineer at 37C. In the first step of glucose catabolism by glycolysis in Lactobacillus acidophilus, the phosphoryl group transfer reaction is as follow: glucose +ATP +-* glucose-6-phosphate +ADP

It was found that the standard free energy change (AG0) of the reaction is -16.7 kJ/mol. i) Determine the concentration of glucose 6-phosphate in the reaction if the glucose concentration is 1.33 M and the ratio of [ADP] to [ATP] is 10. (3 marks) ii) Calculate the ratio of [ADP] to [ATP] if the ratio of [glucose-6-phosphate] to [glucose] is 8. (2 marks) iii) If the ratio of [ADP] to [ATP] is equal to the ratio of [glucose-6-phosphate] to [glucose] which is 1, determine the Keq and AG0 of the reaction. Estimate the energy released during this condition. Comment on your answer. (5 marks)

QUESTION 2 (P06, C02, C3, C5) a) Figure 1 was obtained by a Bioprocess Engineer for the reaction of amylase enzyme in bioconversion of starch into glucose, where the total enzyme concentration was 0.005 mM. _ . * = Jl | 3 E 'S 1300 -, 1200 1100 TOOO 900 800 700- ( 600 -i \ 500 - / 400 -/ SCO200 100 -

"I

c3 1 2 3 4 5 6 7 8 9
Starch Concentration (mM) Figure 1
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10 11 12 13

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i) Identify the maximum velocity (Vmax) and Michaelis constant, (KM) for this enzyme. (3 marks) ii) Estimate the turnover number (kcat) and the catalytic efficiency for the enzyme. (5 marks) iii) Using the value of Vmax and KM obtained from the graph, calculate the substrate concentration, [S], if the engineer wants the reaction to yield a reaction rate of 2/3
Vmax-

(5 marks) iv) Prove that the initial velocity is 7* Vmax when KM equals the substrate concentration. (4 marks) b) A Bipprocess Engineer, Yusri, planned to produce mannitol from cassava starch. In the bioconversion stage, he used two types of enzymes, glucoamylase and glucose isomerase to produce glucose and fructose. He found that the maximum glucose syrups production using glucoamylase occurred after 24 hours and the production of fructose syrups was reached after 96 hours of reaction with glucose isomerase. Figure 2 and 3 show these enzymes activity against pH and temperature.

100 90 ~80 70 .|60 tJ 50 " 40

Glucoamylase Glucose Isomerase

I 30
* 20

10
~i 1 1 1 r

1 2

9 10 11 12 13

py
Figure 2

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100 -i 90 ^ 80 -

A. y / A V /

s*-\

-*-Glucoamylase \-Glucose \ Isomerase

J70"
1 50 J
5 40 1 30 6 20 H 10 0 0 > 60 /

y
i i i

.
i

/
/
i i i i

^-^
i i i i i

10 20 30 40 50 60 70 80 90 100 110120130 Temperature (C) Figure 3

i)

With the aid of Figure 2 and 3 provided, suggest the environmental conditions that Yusri needs to control for the enzymes to produce higher yield of glucose and fructose. (4 marks) Evaluate the consequences if Yusri increased the temperature from 60C to 80C in the bioconversion process. (4 marks)

ii)

QUESTION 3 (P01,C01,C2, C3, C4) a) Metabolism is the chemical reactions that happen in living organisms to maintain life which allow organisms to grow and reproduce, maintain their structures, and respond to their environments. Metabolism consists of catabolism and anabolism. i) Explain anabolic and catabolic processes in terms of reactions and energy implications. (6 marks) ii) Identify the source of energy being supplied in anabolism. (2 marks) iii) In animal metabolism, glucose is converted to C0 2 and H 2 0. Identify whether the overall process breakdown is anabolism or catabolism. Explain. (3 marks)

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b) The Embden-Meyerhof Pathway is shown in Figure 4.

Glucose

Glucose-6-Phoqshate Phosphog lua>- isomerase

F iuctDS&-Phosphate J6-P

I I
I I

Bankings*

Phosphofructokirtase

FnJctos^1,6-bisphosphate

B-tai

Aldolase

Glvcerafd^T(yde-3-phosphatfi dehydrogenase 1,3-Btsphosphoglycerate Phosphoglyoeratek insse 3-Phosphogtycerate

2-Phosphoglycerate I Ertolase Phosphoatoljyruvate I Pyruvate kinase Pymvate

PhcEphoglycero-mutase

Figure 4 i) Identify the compound produced at X and Y. Name the enzyme being used in this reaction. (3 marks) ii) By stating the enzymes and chemical formulas of the products and reactants, outline the reaction to complete the degradation of glucose from pyruvate to bioethanol. (5 marks) iii) In glucose metabolism, two reactions were coupled to produce a substrate cycle. Two substrate cycles have been identified and one of it were the phosphoryl transfer to fructose-6-phosphate and the hydrolysis of a phosphoryl group from fructose-1,6bisphosphate. Outline another couple of reactions that form a substrate cycle. State the enzymes involved in the reaction. (4 marks)

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QUESTION 4 (P01,C01,C4, C2) a) Describe the fate of NADH and pyruvate in catabolic pathways under aerobic and anaerobic. (8 marks) b) Describe the major similarity and difference between glycolysis and the pentose phosphate pathway in terms of the reaction and product produced. (3 marks) c) The tricarboxylic acid cycle is a series of biochemical reactions that eventually completely oxidize substrates, such as glucose and fatty acids to form C0 2 , H 2 0, and the reduced coenzymes NADH and FADH2. Describe two (2) important roles of the tricarboxylic acid cycle in metabolism. (4 marks)

QUESTION 5 (P01,C01,C1,C2, C6) a) Fatty acids are an important source of energy and adenosine triphosphate (ATP) for many cellular organisms. Excess fatty acids, glucose, and other nutrients can be stored efficiently as fat. Give the reasons why fatty acids are suitable as the energy storage. (4 marks) b) Protein synthesis is sometimes used to refer only to protein translation but more often it refers to a multi-step process, beginning with amino acid synthesis and transcription of nuclear DNA into messenger RNA, which is then used as input for translation. Briefly describe the phases involved in protein synthesis. (5 marks) c) Biochemist always uses heat to increase the rate of certain reactions, for example in the fermentation by microorganism for the production of valuable products. Describe to what extent living organisms could use this method to speed up the metabolic reactions. (2 marks)

END OF QUESTION PAPER

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