UNIVERSITEIT VAN DIE VRYSTAAT

UNIVERSITY OF THE FREE STATE

HOOFKAMPUS / MAIN CAMPUS

BOCE2626
DEPARTEMENT MIKROBIESE, BIOCHEMIESE & VOEDSELBIOTEGNOLOGIE
DEPARTMENT OF MICROBIAL, BIOCHEMICAL & FOOD BIOTECHNOLOGY
SEMESTER TOETS / SEMESTER TEST – 12 October 2017
ASSESSOR(E)/ASSESSOR(S): 1. Dr. ED Cason
TYD / TIME: 1 ½ UUR / HOURS PUNTE / MARKS: 50

Question 1 / Vraag 1 [15]

1.1 In the induced-fit model of substrate binding to enzymes / In die geïnduseerde

passing model van substraat binding in ensieme
a. the substrate changes its conformation to fit the active site / die
substraat verander sy konformasie om die aktiewe setel te pas
b. the active site changes its conformation to fit the substrate / die
aktiewe setel verander sy konformasie om die substraat te pas
c. there is a conformational change in the enzyme when the substrate
binds / daar is 'n konformasie verandering in die ensiem wanneer die
substraat bind
d. there is aggregation of several enzyme molecules when the substrate
binds / daar is aggregasie van verskeie ensiem molekules wanneer die
substraat bind
1.2 The KM of hexokinase for glucose = 0.15 mM and for fructose, K M = 1.5 mM.
Which is the preferred substrate? / Die KM van hexokinase vir glukose = 0,15
mM en fruktose, KM = 1,5 mM. Wat is die voorkeur substraat?
a. Glucose. / Glukose.
b. Fructose. / Fruktose.
c. Neither substrate is preferred over the other. / Geen substraat word
verkies bo die ander nie.
d. You cannot tell from the data given. / Jy kan nie sê van die data gegee.
e. None of these answers is correct. / Nie een van hierdie antwoorde is
korrek.

1.3 A velocity curve (V vs. [S]) for a typical allosteric enzyme will be / 'n Snelheid
kurwe (V teen [S]) vir 'n tipiese allosteriese ensiem sal wees
a. a rectangular hyperbola. / 'n reghoekige hiperbool.
b. a sigmoid curve. / 'n sigmoidale kurwe.
c. a straight line. / 'n reguit lyn.
 d. a parabola. / 'n parabool.

1.4 A transition-state analog is likely to bind to an enzyme / 'n Oorgang-staat

analoog is geneig om 'n ensiem te bind
a. more tightly than the substrate. / stywer as die substraat.
b. less tightly than the substrate. / minder styf as die substraat.
c. about as tightly as the substrate. / omtrent so styf as die substraat.
d. at a site other than the catalytic site. / op 'n ander plek as die
katalitiese setel.

1.5 Which of the following statements about coenzymes is true? / Watter van die
volgende stellings oor koënsieme is waar?
a. They are commonly derived from vitamins. / Hulle word algemeen
verkry vanaf vitamiene.
b. They bind to the active site on specific types of enzymes./ Hulle bind
aan die aktiewe setel of spesifieke tipes ensieme.
c. They can be metal ions, such as Zn(II)./ Hulle kan metaalione soos
Zn(II) wees.
d. NAD+ and FAD are both examples of coenzymes. / NAD+ and FAD is
beide voorbeelde van koënsieme
e. All of these statements are true./ Al hierdie stellings is waar.

1.6 The reaction process catalyzed by pyruvate dehydrogenase requires all of the
following, except _______. / Die reaksie proses gekataliseer deur piruvaat
dehidrogenase benodig almal behalwe een van die volgende ________
a. TPP
b. FMN
c. CoASH
d. NAD+
e. Lipoamide / Lipoamied
f. All of the above / Al die bogenoemde

1.7 In what form does the product of glycolysis enter the TCA cycle? / In watter
vorm gaan die produk van glikolise die TCA siklus binne?
a. Acetyl-CoA / Asetiel-KoA
b. Pyruvate / Piruvaat
c. Glucose / Glukose
d. NADH

1.8 Which of the following enzymes does not take part in the TCA cycle? / Watter
van die volgende ensieme is nie deel van die TCA siklus nie?
a. Citrate synthase / Sitraat sintase
b. Iso-citrate dehydrogenase / Iso-sitraat dehidrogenase
c. Pyruvate dehydrogenase / Piruvaat dehidrogenase
d. Malate dehydrogenase / Malaat dehidrogenase

1.9 The coenzymes, NAD+ and FAD+, carry electrons to the _____. / Die
koënsieme, NAD+ en FAD+, dra elektrone na die _____.
a. Nucleus / Selkern
b. Citric Acid cycle / Sitroensuursiklus
 c. Plasma membrane / Plasma membraan
d. Electron transport system / Elektron vervoerstelsel

1.10 Cells only need a small amount of NAD+ because each NAD+ can be used to
transport electrons over and over. / Selle het slegs 'n klein hoeveelheid NAD+
nodig omdat elke NAD+ gebruik kan word om elektrone oor en oor te vervoer.
a. True / Waar
b. False / Vals

1.11 The energy released by the electron transport system produces ATP by ____.
/ Die energie wat vrygestel word deur die electron vervoerstelsel produseer
ATP deur ____.
a. Photophosphorylation / Fotofosforilering
b. Substrate-level phosphorylation / Substraatvlak fosforilering
c. Oxidative phosphorylation / Oksidatiewe fosforilering

1.12 Which one is not one of the main proteins in the electron transport chain? /
Watter een is nie een van die hoof proteïene in die elektronvervoerketting
nie?
a. NADH dehydrogenase / NADH dehidrogenase
b. Cytochrome bc1 complex / Sitochroom bc1 kompleks
c. Cytochrome oxidase / Sitochroomoksidase
d. Citrate synthase / Sitraatsintase
e. Succinate Dehydrogenase / Suksinaat Dehidrogenase

1.13 ATP synthesis and mitochondrial electron transport are coupled by: / ATP
sintese en die mitochondriale elektronvervoer is gekoppel deur:
a. Protonmotive force / Protonmotiewe kragte
b. High energy intermediates / Hoë energie intermediêre
c. Proton gradient / Protongradiënt
d. Both A and C are correct / Beide A en C is korrek
e. All of the above are correct / Al die bogenoemde is korrek

1.14 Place the following reaction intermediates of the citric acid cycle in
chronological order. (i. Malate; ii. Fumarate; iii. Succinyl CoA; iv. Citrate; v. -
Ketoglutarate). / Sorteer die volgende reaksie intermediêre van die
sitroensuur siklus in kronologiese volgorde. (i. Malaat; ii. Fumaraat; iii.
Suksiniel KoA; iv. Sitraat; v. -Ketoglutaraat).
a. i, ii, iv, v, iii
b. iv, v, iii, i, ii
c. iv, i, v, iii, ii
d. v, iii, i, ii, iv
e. iv, v, iii, ii, i
 1.15 There are several steps during the oxidation of NADH in which the EO' is
sufficient for ATP synthesis. These are: / Gedurende die oksidasie van NADH
is daar verskeie stappe waarvan die EO' genoegsaam is vir ATP sintese.
Hulle is:
a. Complexes / Komplekse I, II & III
b. Complexes / Komplekse I, III & V
c. Complexes / Komplekse II, III & IV
d. Complexes / Komplekse III, IV & V
e. None of the above / Geen van die bogenoemde

Question 2 / Vraag 2 [8]

Draw up a balance sheet to summarise the ATP synthesis from ONLY the reactions from the
glycolysis for the oxidation of one molecule of Glucose. No structure formulae are required,
only the various reactions enzyme names, co-enzymes and ATP yield for each reaction. /
Stel ‘n balansstaat saam om ‘n opsomming van die ATP sintese alleenlik vanaf die glikoliese
se reaksies vir die oksidasie van een molekuul glukose te illustreer. Geen struktuurformules
word benodig nie, slegs die verskeie reaksies, ensiemname, koënsieme en ATP uitset van
elke reaksie.

Reaction / Reaksie Enzyme / NADH + H+ ATP formed or used Total amount of ATP
Ensiem ATP gevorm of gebruik Totale hoeveelheid ATP
(A converted to B) Hydrolase Amount / Convert to ATP via
Hoeveelheid oxidative Phoshorylation

Reaction Enzyme NADH ATP

GLYCOLYSIS

Glucose  Glc-6-Phosphate Hexokinase -1

Fructose-6-P  Fru-1,6-BisPhosphate Phosphofructokinase 1 -1

Glyceraldehyde-3-P  Glyald-1,3-BP Gly – 3 P dehydrogenase +2 3 or 5

Glyald-1,3-BP  Glycerate-3-P Phohoglycerate kinase +2

PhosphoEnolPyruvate  Pyruvate Pyruvate kinase +2

Total 5 or 7

Question 3 / Vraag 3 [11]

Fructose-2,6-bisphosphate is an important allosteric regulator of both glycolysis and

gluconeogenesis. A bifunctional enzyme controls both the synthesis and breakdown
of fructose-2,6-bisphosphate using phosphorylation as a control mechanism. /
Fruktose-2,6-bisfosfaat is ‘n belangrike allosteriese reguleerder van beide glukose
en glukoneogenese. ‘n Bifunksionele ensien beheer beide die sintese en afbraak
van fructose-2,6-bisfosfaat deur gebruik te maak van fosforulasie as ‘n beheer
meganisme.

3.1 What enzymatic group of enzymes are responsible for the phosphorylation of
some substrates? / Watter ensimatiese groep van ensieme is verantwoordelik
vir die fosforulasie van sommige substrate? (1)

Kinases

3.2 Which enzymes in glycolysis and gluconeogenesis are allosterically controlled

by fructose-2,6-bisphosphate levels? / Watter ensieme in glikolise en
glukoneogenese word allosteries beheer deur fruktose-2,6-bisfosfaat? (2)

Phosphofructokinase and Fructose-1,6-bisposphatase

3.3 Discuss how phosphorylation is used to control the levels of fructose-2,6-

bisphosphate. Include the names of the enzymes that form part of the
bifunctional enzyme that will be affected. Also include how the levels of
Fructose-2,6-bisphosphate will affect the activity of the enzymes listed in 3.2
above. / Beskryf hoe fosforulasie gebruik word om die vlakke van fruktose-
2,6-bisfosfaatte beheer. Sluit in die name van die ensieme wat deel uitmaak
van die bifunksionele ensiem wat geaffekteer gaan word. Sluit ook in hoe die
vlakke van fruktose-2,6-bisfosfaat die aktiwiteit van die ensieme genoem in
3.2, hier bo, sal affekteer. (8)

The synthesis of Fru-2,6-P2 is performed through a bifunctional enzyme containing

both PFK-2 (1) and FBPase-2 (1), which is dephosphorylated (1), allowing the PFK-2
portion to phosphorylate fructose 6-phosphate using ATP (1). The breakdown of Fru-
2,6-P2 is catalyzed by the phosphorylation of the bifunctional enzyme (1), which
allows FBPase-2 to dephosphorylate fructose 2,6-bisphosphate to produce fructose
6-phosphate and Pi (1).

High levels of F-2,6-BP activates Phosphofructokinase (1) and inhibits Fructose-1,6-

bisphosphatase (1)OR REVERSE.

Question 4 / Vraag 4 [5]

The Pentose Phosphate Pathway is an alternative pathway for glucose metabolism

and differs from glycolysis in several ways. / Die Pentose Fosfaat Baan is ‘n
alternatiewe metaboliese baan vir glikose metabolisme en verskil van glikolise in
verskeie maniere.

4.1 During glycolysis ATP production is important, however, during the Pentose
Phospahte Pathway, this is not the case. What are the most important
products produced in this pathway? / Tydens glikolise is ATP produksie baie
belangrik, maar tydens die Pentose Fosfaat Baan is dit nie die geval nie. Wat
is die mees belangrikste produkte gemaak in die baan? (2)

5 Koolstof Suikers / Ribolose-5-fosfaat

NADPH
4.2 Which reactions will result in the formation of the products listed in 4.1?
Include ONLY substrates, products and enzymes. / Watter reaksies sal lei tot
die vorming van die produkte gelys in 4.1? Sluit SLEGS in die substrate,
produkte en ensieme. (6 x ½ =3)

Glucose-6-phosphate -> 6-Phosphogluconate

Enz: Glucose-6-Phosphate dehydrogenase
6-phosphogluconate -> Ribolose-5-phosphate
Enz: 6-Phosphogluconate dehydrogenase

Question 5 / Vraag 5 [11]

5.1 Apart from Acetyl-CoA what other product is obtained when a fatty acid
containing an odd number of carbon atoms is catabolised? / Benewens
asetiel-CoA watter ander produk word ook verkry wanneer ‘n vetsuur met ‘n
onewe getal koolstofatome gekataboliseer word? (1)

Propionyl-CoA

5.2 The product mentioned in 4.1 is carboxylated to which molecule that then
undergoes rearrangement to which intermediate of the citric acid cycle? / Die
produk genoem in 4.1 word gekarboksileer na watter molekuul wat dan
herrangskikking ondergaan tot watter intermediêr van die sitroensuursiklus?
(2)
Methyl malonyl-CoA & Succinyl-CoA

5.3 An acyl-CoA needs to cross the inner mitochondrial membrane to undergo -

oxidation. Briefly discuss how the acyl-CoA enters the mitochondrion
referencing the the enzyme(s) and transporter(s) that will be involved in this
process. / ‘n Asiel-KoA moet die binneste mitochondriale membraan oorsteek
om -oksidasie te ondergaan. Beskryf kortliks hoe die Acyl-CoA die
mitochondrion binne gaan, verwys na die ensiem(e) en vervoerder(s) wat by
hierdie proses betrokke sal wees. (6 x ½ =3)

 Acyl-CoA crossed the outer membrane, and the Acyl group from Acyl-CoA is
then transferred to carnitine. (1)
[Carnitine acyltransferase/ carnitine palmitoyltransferase (CPT-I)] (1)
 Acyl-Carnitine is then carried across the inner mitochondrial membrane. (1)
[Carnitine translocase] (1)
 The acyl group is transferred back to CoA-SH. (1)
[Carnitine acyltransferase/ carnitine palmitoyltransferase (CPT-II)] (1)

5.4 The first step in cholesterol synthesis is the synthesis of mevalonate. List the
other four major steps involved. / Die eerste stap in cholesterol sintese is die
sintese van mevalonaat. Lys die ander vier hoofstappe betrokke by die
proses. (4 x ½ =2)

• Synthesis of isopentenyl-PPi
• Synthesis of squalene
• Cyclization of squalene to lanosterol
• Lanosterol conversion to cholesterol