Experiment Analysis: Glycoside Beta Drummin and Glycolysis Pathway

This assignment analyzes an experiment investigating the effects of Glycoside Beta Drummin on glycolysis and cellular metabolism. The experiment explores the conversion of glucose through the pentose phosphate cycle and glycolysis, examining the role of 14C in these pathways and the significance of lactate accumulation. The analysis discusses the potential for enzyme depletion or substrate limitation, and proposes that Glycoside Beta Drummin acts as an inhibitor of enzymatic activity in glycolytic metabolism. The findings indicate that Glycoside Beta Drummin can cause inhibition reactions, likely targeting the production of 3-phosphoglycerate and acting as a competitive inhibitor. Furthermore, the study considers the importance of fructose-1,6-biphosphate in ATP production and its implications for spermatozoa motility, as well as the potential for increased competition for inhibition in crude extracts. The assignment concludes that Glycoside Beta Drummin's inhibition mechanism is competitive, affecting ATP production and sperm motility.

Student No./10001
Experiment: Glycoside Beta Drummin
Assignment (type)
Date
1000 words
Student’s Name
Institutional Affiliation

Student No./10002
Experiment: Glycoside Beta Drummin
1) According to the Embden-Meyerhof pathway, the pentose cycle involves the conversion
of glucose molecules (and fructose molecules) to CO2. Glycolysis involves the conversion of
glucose to produce ATP. 3 pentose phosphates elements are converted to form fructose 6-
phosphate and triose phosphate 1. The 14C are used directly while the fructose 6-P is not
broken down to Glucose. As such, the 14C is used up to complete for the pentose phosphate
cycle due to randomized of 14C metabolism use in the pentose cycle or glycolysis pathway. In
some cases, when the fructose-6-phosphate is equal or in equilibrium with glucose-6-
phosphate, fructose-6-phosphate can be reversed. However, even when the process is
reversed in glycolysis the use of 14C from the substrates. Hence, randomized use of 14C in
glucose-6-phosphate is predicted for metabolism in order to complete the pentose cycle.
2) The significance in lack of accumulation of lactate in the experiment is that no
abnormalities would be experienced during the pentose cycle. Accumulation in lactate would
lead to metabolic dysregulation which would result in reduced or abnormal metabolism of
pentose-phosphate 2. Metabolism would affect the pH balance in pentose-phosphate
metabolism if lactate would be produced in high or excess amounts. The other explanation
would be the incorporation of the lactate-acid cycle. Since the process is anaerobic the use of
CO2 to convert the lactate reduced the amount of carbon (IV) oxide. Nonetheless, as
glycolysis is happening probably the lactate acid cycle is taking place. Therefore, based on
the results of the experiment, the equilibrium in CO2 and O2 at 50% reduced the production
of lactate.
1 Katz, J., Wood, H.G. “The use of glucose-C14 for the evaluation of the pathways of
glucose metabolism.” J. Biol. Chem. 235, 2165–2177, 2010
2 Hoff, J., Støren, Ø., Finstad, A., Wang, E., Helgerud, J. “Increased blood lactate level
deteriorates running economy in world class endurance athletes.” J. Strength Cond.
Res. 30, 1373–1378, 2016.

Student No./10003
3) The enzymes necessary for metabolic activity may have been depleted or were less
compared to the number of available enzymes were lower than substrates 3. Therefore, the
rate of oxygen input in addition carbon (IV) oxide output may have reached an equilibrium.
The isomerization of 14C in substrates as a result of reduction of substrates and enzymes
necessary for glycolysis. Another reason is that substrates may have reduced in amount which
may have resulted in reduced metabolic activity leading to an equilibrium in oxygen input
and carbon (IV) oxide output.
4) The opinion is that glycoside beta drummin may be an inhibitor to the enzymatic activity
in glycolytic metabolism4. Glycolysis heavily relies on enzymatic action which is crucial to
the catalysis of the cycle resulting into the creation of carbon (IV) oxide and intake of
oxygen. As such, inability of the enzymes to induce metabolism, there was a reduced oxygen
intake and carbon (IV) oxide output. The possible explanation is that the binding of the
inhibitor, in this case, glycoside beta drummin, to the substrates, at concentration between 10-
100mM created an imbalance in the enzyme-substrate complex as a result reducing
metabolism.
5) The addition of inhibition to RSW did not alter any results and thus indicates that
glycoside beta drummin, can cause inhibition reaction in the glycolysis process. The plausible
explanation is that glycoside beta drummin contains inhibitor molecules that either bind to
the substrate-enzyme complex or on the enzymes limiting their biochemical functionality 5.
3 Canto, C., Menzies, K.J., Auwerx, J. “NAD+ metabolism and the control of energy
homeostasis: a balancing act between mitochondria and the nucleus.” Cell Metab. 22,
31–53, 2015.
4 Vander Heiden, M.G., DeBerardinis, R.J. “Understanding the intersections between
metabolism and cancer biology.” Cell 168, 657–669, 2017.
5 Vander Heiden, M.G., DeBerardinis, R.J. “Understanding the intersections between
metabolism and cancer biology.”Cell 168, 657–669, 2017.

Student No./10004
6) According to the table (Table 1), the probable site of inhibition is during the production
of 3-phosphoglycerate. In the glycolysis cycle, enzyme triose phosphate dehydrogenase
allocates the hydrogen atom from Phosphate to the reacting agent NAD+ 6. Based on the
table, there is no evidential percentage control compared to products such as glyceraldehyde-
3-phosphate which is marked at >300 while 3-phosphoglyceraldehyde is marked as “not-
detected.” Therefore, the probable explanation is that the enzyme-substrate complex, triose
phosphate dehydrogenase is inhibited to produce glycolysis product 2-glyceraldehyde 3-
phosphate which also limits the production of subsequent product, 2-phosphoenolpyruvate.
7) Fructose-1,6-biphosphate is considered to be an intermediary for the high glycolysis
sustenance and increases ATP (Adenosine Triphosphate) production. Therefore, cells are
programmed to exude high amounts of fructose-1,6-biphosphate as possible in order to
ensure that high amounts of ATP (Adenosine Triphosphate) are produced and the pentose
cycle is maintained. ATP (Adenosine Triphosphate) is an essential by-product that is
necessary in enzyme-mediated processes within the cells including intermediary glycolytic
pathways and other pathways. Additionally, another probable reason is because, with the
limited intake of oxygen, production of fructose-1,6-biphosphate may be essential in
production of ATP as a reservation when the metabolism rates are reducing due to reduced
oxygen intake and increased carbon (IV) oxide output 7.
8) The postulation is that with crude extracts, there is increased competition for inhibition
of the metabolic processes compared to when the enzymes are in their pure forms. The
proposition is that crude extracts may include additional inhibitory factors in addition to the
6 Chung, J.-J., Shim, S.-H., Everley, R.A., Gygi, S.P., Zhuang, X., Clapham, D.E.
“Structurally distinct Ca 2+ signaling domains of sperm flagella orchestrate tyrosine
phosphorylation and motility.” Cell 157, 808–822, 2014.
7 Zhang, C.-S., Hawley, S.A., Zong, Y., Li, M., Wang, Z., Gray, A., Ma, T., Cui, J.,
Feng, J.-W., Zhu, M. “Fructose-1, 6-bisphosphate and aldolase mediate glucose
sensing by AMPK.” Nature 548, 112, 2017.

Student No./10005
glycoside beta drummin. The linear effect is that glycoside beta drummin will compete with
the other inhibitors to seize the enzyme-substrate complex reducing metabolic rates 8. In this
case, beta drummin may compete for the substrate-binding site of the enzymes with the
substrate since the substrate and the inhibitor may bind to the same overlapping sites.
Therefore, competitive inhibition is exhibited.
9) The inhibition mechanism exhibited by glycoside beta drummin is competitive
inhibition. Competitive inhibitors are determined to be substrate-binding site-specific when it
comes to the enzymes. Therefore, in this case, the beta drummin overlaps with the binding
sites on the enzymes creating an enzyme-inhibitor complex. The proposition is that the
substrate is de-linked or displaced from the enzyme. Hence, for each enzyme there is an
inhibitor complex that is formed. The complex may not be as effective since, in the
experiments there is evidence of oxygen uptake and carbon (IV) oxide production- meaning
that glycoside beta drummin is an a highly competitive inhibitor9.
10) The physiological explanation is that glucose or pentose phosphate is linked to the
motility and ATP concentration in spermatozoa 10. The inhibition in metabolism of the
glycolytic pathway by the glycoside beta drummin through competitive inhibition limits the
production of ATP- which is essential in motility and concentration of ATP in spermatozoa.
The consequence is that with reduced amounts of glucose there is reduced motility of
spermatozoa.
8 9.4. Enzyme inhibition mechanisms [WWW Document], n.d. URL
http://elte.prompt.hu/sites/default/files/tananyagok/IntroductionToPracticalBiochemist
ry/ch09s04.html (accessed 3.23.18).
9 9.4. Enzyme inhibition mechanisms [WWW Document], n.d. URL
http://elte.prompt.hu/sites/default/files/tananyagok/IntroductionToPracticalBiochemist
ry/ch09s04.html (accessed 3.23.18).
10 du Plessis, S.S., Agarwal, A., Mohanty, G., Van der Linde, M. Oxidative
phosphorylation versus glycolysis: what fuel do spermatozoa use? Asian J. Androl.
17, 230, 2015.

Student No./10006

Student No./10007
References
9.4. Enzyme inhibition mechanisms [WWW Document], n.d. URL
http://elte.prompt.hu/sites/default/files/tananyagok/IntroductionToPracticalBiochemist
ry/ch09s04.html (accessed 3.23.18).
Canto, C., Menzies, K.J., Auwerx, J. NAD+ metabolism and the control of energy
homeostasis: a balancing act between mitochondria and the nucleus. Cell Metab. 22,
31–53. 2015.

Student No./10008
Chung, J.-J., Shim, S.-H., Everley, R.A., Gygi, S.P., Zhuang, X., Clapham, D.E. Structurally
distinct Ca 2+ signaling domains of sperm flagella orchestrate tyrosine
phosphorylation and motility. Cell 157, 808–822, 2014.
du Plessis, S.S., Agarwal, A., Mohanty, G., Van der Linde, M. Oxidative phosphorylation
versus glycolysis: what fuel do spermatozoa use? Asian J. Androl. 17, 230, 2015.
Hoff, J., Støren, Ø., Finstad, A., Wang, E., Helgerud, J. Increased blood lactate level
deteriorates running economy in world class endurance athletes. J. Strength Cond.
Res. 30, 1373–1378, 2016.
Katz, J., Wood, H.G. The use of glucose-C14 for the evaluation of the pathways of glucose
metabolism. J. Biol. Chem. 235, 2165–2177, 2010.
Vander Heiden, M.G., DeBerardinis, R.J. Understanding the intersections between
metabolism and cancer biology. Cell 168, 657–669, 2017.
Zhang, C.-S., Hawley, S.A., Zong, Y., Li, M., Wang, Z., Gray, A., Ma, T., Cui, J., Feng, J.-
W., Zhu, M. Fructose-1, 6-bisphosphate and aldolase mediate glucose sensing by
AMPK. Nature 548, 112, 2017.