Biochemistry Enzyme Project Worksheet

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I am working on an enzyme project, I did the first part of the project and it is in the file below, you can use the references or add new references

enzyme name:

1-to find more information to expand further on those aspects, by reading the published references you had collected before (not just what the three protein databases had provided in paragraphs)

2-For new things listed below, search and find the information. This new information may be either in the references that you had collected before or new ones you need to find from Brenda's database menu and the references from this database

3-You can also use other databases of your choice, beyond the three you had used before.

the steps:

1)Introduction: You already have all of the following information from the previous submission (in the file below). Just reorganize to include ONLY the following aspects in the Introduction section.

Name of the enzyme or recommended name

Systematic name

Alternate names (= synonyms)

E.C. Number

Which Main Class, Subclass, or SubSubClass your enzyme belong to.

In addition to humans, which other organisms it is present in?

Within the eukaryotes, which organs/tissues/cells?

What is the subcellular localization?

The following expectations apply not only to Introduction section, but also for all other sections in your report:

Make sure proper references are cited (author, year) in the paragraph statements. These should be the actual publications. Do not simply cite the Database this time. If one student did not find reference in her specific database last time, your team mates will have it.

Make sure you use correct grammar and spelling. The information you found are paraphrased in your own words, no direct copy and paste for statements. No plagiarism.

When writing the statements, do NOT use first person (I, we, us, our, etc); Likewise, do not use second person either (they, then, their, etc). And do NOT use direct statements; they should all be indirect statements.

For e.g. this is wrong: “We found the enzyme name as “aldolase”. Instead write it as “the enzyme is known as aldolase.

Likewise, it is wrong to state: These researchers found the enzyme in kidney of humans. Instead, state is as “ the enzyme has been found to be present in kidneys in humans” and give the authors name and year.

2)Reactions Catalyzed by the enzyme: (you/your team already have this from previous step-2 of the project. Include it under this title, with actual publications references.)

Which major reaction or substrate it catalyzes (can be humans and other organisms if the enzyme is present in different organisms.

Other reactions or substrates it catalyzes (if any): Give 3 examples if more than 3 are found

Write the reaction equation for the main substrate catalysis

Include the structural diagrams (remember! A good way to insert it in your report is by inserting it in a text box).

Make sure the figure gets a figure number and title. At the end of the title, give the author name and year, to show from which reference this diagram was taken, not just the database).

3)Mechanism and Kinetics of the Main Reaction Catalyzed (for the main reaction you chose in previous section #3):

You must find allof the following new information listed below. If you cannot find it for human enzyme it is OK to include it for other organisms.

And you may have to go back to Brenda site, bring up your enzyme through the E.C. Number, and click on the “Cofactor” and “enzyme-ligand interactions” menu on the left side. You will find the information. But make sure you get those references listed and read them to find all the details needed. Database might not directly give the full information.

oCofactors and metal ions or other compounds the enzyme needs for its function

oActive site and allosteric sites information with regard to the binding of the substrate and cofactors.

oIntermediates formed (if any)

oThe mechanisms of the reaction (for example, if it is oxido-reductase Main class, how did the electron transfer happen, at which parts of the substrate and in the cofactor, and so on……

oAre there any conformational changes to the structure of the enzyme when the substrate binds?

oInclude any diagrams you find from the actual reference articles from these databases, and give figure numbers, description and author, year

oKinetics Details for at least one substrate and at least for one organism (human or another organism, if details are not available for human enzyme)

§In the Brenda site, look under “functional parameters” main menu on the left side of the enzyme page. All the kinetic details links are provided. But in this part of the report we need only:

o Km, Vmax, Kcat Turn over number, specific activity, pH and temperature information

Unformatted Attachment Preview

Basic Biochemistry CHEM 351 Fall 2023 Enzyme Project Term Paper Step-3 Report: Introductory Information Report for 9-Carbonic anhydrase Enzyme. Carbonic anhydrase (EC number 4.2.1.1) is an enzyme that falls under the main class, Lyases in the subclass Carbon-Oxygen Lyases, and in the SubSubClass, Hydro Lyases. EC Tree 4 Lyases 4.2 Carbon-oxygen lyases 4.2.1 Hydro-lyases 4.2.1.1 carbonic anhydrase (BRENDA:EC4.2.1.1) The accepted name is carbonic anhydrase while its systemic name is carbonic acid hydrolyase. It is also known by numerous other names such as; anhydrase, carbonate anhydrase, carbonic acid anhydrase, carbonate hydro-lyase, carbonate hydro-lyase carboxyanhydrase, carbonic anhydrase A, carbonate dehydratase, alkalistable alpha-carbonic anhydrase, alphatype CA and alpha-type carbonic anhydrase, Fisher (2006). Carbonic anhydrase is well-studied and well-known enzyme because it was conserved through many organisms. According to De Oliveira Maciel et al. (2022) various creatures, such as bacteria, plants, and animals, there are three discrete families of cellular automata, namely alpha, beta, and gamma, which are not evolutionarily linked to one another. All Carbonic Anhydrase that has been identified across the animal kingdom are categorized as alpha Carbonic Anhydrase. The principal function of Carbonic Anhydrase is to expedite the fast and reversible transformation between CO2 and bicarbonate ions. The enzymatic reaction is of utmost importance in a wide range of biological processes. CO2 reacts with water in a variety of organisms, including humans, with the assistance of the enzyme Carbonic Anhydrase. The chemical process results in the generation of carbonic acid, which subsequently experiences dissociation into a hydrogen ion and bicarbonate. Carbonic acid → Carbon IV dioxide + Water H2CO3 → CO2(g) + H2O The reason for a reduction in respiratory rate resulting in a decline in pH can be attributed to the following factors. Doyle and Cooper (2018) found that enzyme Carbonic Anhydrase is of significant importance in a range of physiological processes, such as respiration, since it aids in the movement of carbon dioxide from tissues to the lungs for expiration. Apart from catalyzing the reversible hydration of carbon IV oxide to carbonic acid. Carbonic anhydrase catalyzes hydrolysis reaction of many substrates including nitrophenyl acetate Elleby (1999): 1- 2-nitrophenyl acetate + H2O →nitrophenol + acetate + H2O → + 2-3-nitrophenyl acetate + H2O→3-nitrophenol + acetate +H2O→ + 3-4-nitrophenyl propionate + H2O=4-nitrophenol + propionate +H2O→ + Carbonic Anhydrase also aid in the process of producing gastric acid in the stomach. And physiology of all photosynthetic organisms. Cyanobacteria possess carbonic anhydrases that are located inside the carboxysome architecture. The mentioned mechanism enables the conversion of CO2 into HCO3-, which plays a vital role as an essential constituent within the Calvin cycle. The process is a series of metabolic reactions utilized by plants to synthesize organic molecules, such as sugars, by assimilating atmospheric carbon dioxide (Santos Correa et al., 2022). There are 8 distinct classes that evolved independently. Though zinc is the most prevalent cofactor, others have also been identified including; Cd2+, Co2+, Fe2+, and Mn2+. Carbonic Anhydrase exhibits ubiquitous presence inside a wide range of organs and tissues across several organisms. The enzyme is distributed throughout several tissues in the human body, displaying unique isoforms in each tissue. One illustration of this idea is evident in the significant abundance of Carbonic Anhydrase II within erythrocytes, where it fulfills a crucial role in enabling transportation of CO2 and preserving equilibrium of acid-base levels. According to Wang et al. (2020), Carbonic Anhydrase IV is found in the stomach mucosa and has a significant role in the process of gastric acid secretion. Renal structure, ocular structures, and many biological tissues have additional isoforms of the enzyme. These isoforms have specific roles in maintaining pH balance and enabling the transport In mammals homosepines in particular carbonic anhydrase is abundant in most of body the cells such as astrocytoma cell , eyes cell, also blood and brain cella. Haapasalo(2008), Arslan(2002), Hilvo(2007), Juozapaitiene(2016).Inside these cells carbonic anhydrase is present in isoforms that aids in cell functions and regulation in the cytoplasm ,mitochondria and neurocells axons . Hilvo(2007), Carbonic anhydrase is also present in herbuvourse mammals such as Alces alces , Carlsson (1973) In prokaryotes a study conducted by Guilloton et al.'s (1992) aimed to analyze and describe a Carbonic Anhydrase enzyme encoded by the Carbonic Anhydrase I in the Escherichia coli can operon. In a metabolic pathway that includes the Carbonic Anhydrase I gene, which codes for the cyanate enzyme, Carbonic Anhydrase are an essential component. A zinc ion was found in each subunit when the Carbonic Anhydrase enzyme was removed and purified from E. coli strains that overexpressed the Carbonic Anhydrase I gene. In addition, it was found that the enzyme is an oligomer in solution. The enzyme's kinetic properties, such as its sensitivity to sulfonamide and cyanate inhibition, resembled Carbonic Anhydrase in different animals. The E. coli Carbonic Anhydrase’s amino acid sequence shared many similarities with carbonic anhydrase found in plants but little in common with those found in mammals and algae. This induced carbonic anhydrase's primary function appears to be to promote carbon dioxide hydration, hence preventing the depletion of cellular bicarbonate. Carbonic anhydrase is also present in other bacterial phylum’s and strain such as, Bacillus subtilis and mycobacterium tuberculosis. Ramanan (2009) Homology refers to the presence of similarities in both the amino acid sequences and structural features of proteins across a wide range of animal species, suggesting a common evolutionary origin. Carbonic anhydrases demonstrate similarity among their different groups. Nevertheless, there are distinct features observed in the chloroplast carbonic anhydrases of bacteria and plants, as well as those of eukaryotes (Jensen et al., 2020). The Prosite database focuses on a distinct group of Carbonic Anhydrase, namely those present in prokaryotes and plant chloroplasts, exhibiting notable differences from the Carbonic Anhydrase observed in eukaryotes. Carbonic Anhydrase has a ubiquitous function in aiding the hydration of carbon dioxide. However, it is noteworthy that they have undergone divergent evolutionary mechanisms in prokaryotes and plants compared to eukaryotes. • 1. References De Oliveira Maciel, A., Christakopoulos, P., Rova, U., & Antonopoulou, I. (2022). Carbonic anhydrase to boost CO2 sequestration: Improving carbon capture utilization and storage (CCUS). Chemosphere, 299, 134419. 2. Doyle, J., & Cooper, J. S. (2018, October 27). Physiology, Carbon Dioxide Transport. Nih.gov; StatPearls Publishing. 3. Guilloton, M. B., Korte, J. J., Lamblin, A. F., Fuchs, J. A., & Anderson, P. M. (1992). Carbonic anhydrase in Escherichia coli. A product of the cyn operon. The Journal of Biological Chemistry, 267(6), 3731–3734. 4. Jensen, E. M., Maberly, S. C., & Gontero, B. (2020). Insights on the Functions and Ecophysiological Relevance of the Diverse Carbonic Anhydrases in Microalgae. International Journal of Molecular Sciences, 21(8), 2922–2922. 5. Santos Correa, S., Schultz, J., Lauersen, K. J., & Soares Rosado, A. (2022). Natural carbon fixation and advances in synthetic engineering for redesigning and creating new fixation pathways. Journal of Advanced Research. 6. Wang, B., Jiang, H., Wan, X., Wang, Y., Zheng, X., Li, P., Guo, J., Ding, X., & Song, H. (2020). Carbonic anhydrase IV inhibits cell proliferation in gastric cancer by regulating the cell cycle. Oncology Letters. Keilin, D. and Mann, T. Carbonic anhydrase. Nature 144 (1939) 442-443 7. Kannan, K.K., Ramanadham, M. and Jones, T.A. Structure, refinement, and function of carbonic anhydrase isozymes: refinement of human carbonic anhydrase I. Ann. N.Y. Acad. Sci. 429 (1984) 4960. [PMID: 6430186] 8. Murakami, H. and Sly, W.S. Purification and characterization of human salivary carbonic anhydrase. J. Biol. Chem. 262 (1987) 1382-1388. [PMID: 2433278] 9. Iverson, T.M., Alber, B.E., Kisker, C., Ferry, J.G. and Rees, D.C. A closer look at the active site of γclass carbonic anhydrases: high-resolution crystallographic studies of the carbonic anhydrase from Methanosarcina thermophila. Biochemistry 39 (2000) 9222-9231. [PMID: 10924115] 10. Smith, K.S. and Ferry, J.G. Prokaryotic carbonic anhydrases. FEMS Microbiol. Rev. 24 (2000) 335366. [PMID: 10978542] 11. Cronk, J.D., Endrizzi, J.A., Cronk, M.R., O'neill, J.W. and Zhang, K.Y. Crystal structure of E. coli βcarbonic anhydrase, an enzyme with an unusual pH-dependent activity. Protein Sci. 10 (2001) 911922. [PMID: 11316870] 12. Merlin, C., Masters, M., McAteer, S. and Coulson, A. Why is carbonic anhydrase essential to Escherichia coli. J. Bacteriol. 185 (2003) 6415-6424. [PMID: 14563877] 13. Hirakawa Y, Senda M, Fukuda K, Yu HY, Ishida M, Taira M, Kinbara K, Senda T (2021). "Characterization of a novel type of carbonic anhydrase that acts without metal cofactors." BMC Biol 19(1);105. PMID: 34006275 14. Fisher, S.Z.; Tariku, I.; Case, N.M.; Tu, C.; Seron, T.; Silverman, D.N.; Linser, P.J.; McKenna, R.Expression, purification, kinetic, and structural characterization of an alpha-class carbonic anhydrase from Aedes aegypti (AaCA1) (2006), Biochim. Biophys. Acta, 764, 1413-1419.Faridi, S.; Satyanarayana, T. 15. Novel alkalistable alpha-carbonic anhydrase from the polyextremophilic bacterium Bacillus halodurans characteristics and applicability in flue gas CO2 sequestration (2016), Environ. Sci. Pollut. Res. Int., 23, 15236-15249 . 16. Fasseas, M.K.; Tsikou, D.; Flemetakis, E.; Katinakis, P.Molecular and biochemical analysis of the alpha class carbonic anhydrases in Caenorhabditis elegans (2011), Mol. Biol. Rep., 38, 1777-1785. 17. Elder, I.; Han, S.; Tu, C.; Steele, H.; Laipis, P.J.; Viola, R.E.; Silverman, D.N. Activation of carbonic anhydrase II by active-site incorporation of histidine analogs (2004), Arch. Biochem. Biophys., 412, 283-289 18. Wingo, T.; Tu, C.; Laipis, P.J.; Silverman, D.N.The catalytic properties of human carbonic anhydrase IX (2001), Biochem. Biophys. Res. Commun., 288, 666-669. 19. Elleby, B.; Sjoeblom, B.; Lindskog, S.Changing the efficiency and specificity of the esterase activity of human carbonic anhydrase II by site-specific mutagenesis (1999), Eur. J. Biochem., 262, 516-521. 20. Haapasalo, J.; Hilvo, M.; Nordfors, K.; Haapasalo, H.; Parkkila, S.; Hyrskyluoto, A.; Rantala, I.; Waheed, A.; Sly, W.S.; Pastorekova, S.; Pastorek, J.; Parkkila, A.K.Identification of an alternatively spliced isoform of carbonic anhydrase XII in diffusely infiltrating astrocytic gliomas (2008), Neurooncology, 10, 131-138 21. -Arslan, O.; Cakir, Ue.; Ugras, H.I.Synthesis of new sulfonamide inhibitors of carbonic anhydrase (2002), Biochemistry (Moscow), 67, 1273-1276. 22. -Buelbuel, M.; Saracoglu, N.; Kuevrevioglu, Oe.I.; Ciftci, M.Bile acid derivatives of 5-amino-1,3,4thiadiazole-2-sulfonamide as new carbonic anhydrase inhibitors: Synthesis and investigation of inhibition effects (2002), Bioorg. Med. Chem., 10, 2561 23. Hilvo, M.; Supuran, C.T.; Parkkila, S.Characterization and inhibition of the recently discovered carbonic anhydrase isoforms CA XIII, XIV and XV (2007), Curr. Top. Med. Chem., 7, 893-899. 24. Purification, enzymatic activity and inhibitor discovery for recombinant human carbonic anhydrase XIV (2016), J. Biotechnol., 240, 31-42 . 25. Wistrand, P.J.; Lindahl, S.; W�hlstrand, T.Human renal carbonic anhydrase. Purification and properties (1975), Eur. J. Biochem., 57, 189-195. 26. Idrees, D.; Kumar, S.; Rehman, S.A.A.; Gourinath, S.; Islam, A.; Ahmad, F.; Imtaiyaz Hassan, M. Cloning, expression, purification and characterization of human mitochondrial carbonic anhydrase VA (2016), 3 Biotech, 6, 16 . 27. Scozzafava, A.; Supuran, C.T.Hydroxyurea is a carbonic anhydrase inhibitor (2003), Bioorg. Med. Chem., 11, 2241-2246. 28. Carlsson, U.; Hannestad, U.; Lindskog, S.Purification and some properties or erythrocyte carbonic anhydrase from the European moose (1973), Biochim. Biophys. Acta, 327, 515-527. 29. Ramanan, R.; Kannan, K.; Vinayagamoorthy, N.; Ramkumar, K.; Sivanesan, S.; Chakrabarti, T. Purification and characterization of a novel plant-type carbonic anhydrase from Bacillus subtilis (2009), Biotechnol. Bioprocess Eng., 14, 32-37. 30. Kolayli, S.; Karahalil, F.; Sahin, H.; Dincer, B.; Supuran, C.T. Characterization and inhibition studies of an alpha-carbonic anhydrase from the endangered sturgeon species Acipenser gueldenstaedti (2011), J. Enzyme Inhib. Med. Chem., 26, 895-900. 31. Cau, Y.; Mori, M.; Supuran, C.T.; Botta, M. Mycobacterial carbonic anhydrase inhibition with phenolic acids and esters kinetic and computational investigations (2016), Org. Biomol. Chem., 14, 8322-8330 . ‫ـ‬Enzyme Project for Term Paper Basic Biochemistry Create the following Subtitles: • Abstract • Introduction • Reaction(s) Catalyzed • Mechanism and Kinetics of the catalysis See next page for the details expected under each Subtitle 1) Cover page: Create a Cover Page having the information as follows and it should be the first page of your report. • Your university name • College name • Department (Chemistry and Earth Sciences should be the department name) • Course Name and the section you are registered for • Semester • (leave a few lines space and continue as follows) • Name of enzyme selected • Complete names of students and QU ID • Instructor’s name • (Leave a few lines space and continue as follows) • Which databases were used 2) Abstract: Just put this heading and Leave it empty this time (you will work in it for final step submission, not now. 3) Introduction: Your team already has all of the following information from previous submission. Just reorganize to include ONLY the following aspects in the Introduction section. • Name of the enzyme or recommended name • Systematic name • Alternate names (= synonyms) • E.C. Number • Which Main Class, Subclass, SubSubClass your enzyme belongs to. • In addition to humans, which other organisms it is present in? • Within the eukaryotes, which organs/tissues/cells? • What is the subcellular localization? The following expectations apply not only to Introduction section, but also for all other sections in your report: Page 1 of 3 Make sure proper references are cited (author, year) in the paragraph statements. These should be the actual publications. Do not simply cite the Database this time. If one student did not find reference in her specific database last time, your team mates will have it. Make sure you use correct grammar and spelling. The information you found are paraphrased in your own words, no direct copy and paste for statements. No plagiarism. When writing the statements, do NOT use first person (I, we, us, our, etc); Likewise, do not use second person either (they, then, their, etc). And do NOT use direct statements; they should all be indirect statements. For e.g. this is wrong: “We found the enzyme name as “aldolase”. Instead write it as “the enzyme is known as aldolase. Likewise, it is wrong to state: These researchers found the enzyme in kidney of humans. Instead, state is as “ the enzyme has been found to be present in kidneys in humans” and give the authors name and year. 4) Reactions Catalyzed by the enzyme: (you/your team already have this from previous step-2 of the project. Include it under this title, with actual publications references.) • Which major reaction or substrate it catalyzes (can be humans and other organisms if the enzyme is present in different organisms. • Other reactions or substrates it catalyzes (if any): Give 3 examples if more than 3 are found • Write the reaction equation for the main substrate catalysis • Include the structural diagrams (remember! A good way to insert it in your report is by inserting it in a text box). • Make sure the figure gets a figure number and title. At the end of the title, give the author name and year, to show from which reference this diagram was taken, not just the database). 5) Mechanism and Kinetics of the Main Reaction Catalyzed (for the main reaction you chose in previous section #3): You must find allof the following new information listed below. enzyme it is OK to include it for other organisms. If you cannot find it for human And you may have to go back to Brenda site, bring up your enzyme through the E.C. Number, and click on the “Cofactor” and “enzyme-ligand interactions” menu on the left side. You will find the information. But make sure you get those references listed and read them to find all the details needed. Database might not directly give the full information. o Cofactors and metal ions or other compounds the enzyme needs for its function o Active site and allosteric sites information with regard to the binding of the substrate and cofactors. o Intermediates formed (if any) Page 2 of 3 o The mechanisms of the reaction (for example, if it is oxido-reductase Main class, how did the electron transfer happen, at which parts of the substrate and in the cofactor, and so on…… o Are there any conformational changes to the structure of the enzyme when the substrate binds? o Include any diagrams you find from the actual reference articles from these databases, and give figure numbers, description and author, year o Kinetics Details for at least one substrate and at least for one organism (human or another organism, if details are not available for human enzyme) ▪ In the Brenda site, look under “functional parameters” main menu on the left side of the enzyme page. All the kinetic details links are provided. But in this part of the report we need only: o Km, Vmax, Kcat Turn over number, specific activity, pH and temperature information . Page 3 of 3
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Basic Biochemistry CHEM 351
Fall 2023
Enzyme Project Term Paper
Step-3 Report: Introductory Information Report for
9-Carbonic anhydrase Enzyme

Carbonic Anhydrase (EC number 4.2.1.1) is an enzyme that falls under the main class, Lyases
in the subclass Carbon-Oxygen Lyases, and in the SubSubClass, Hydro Lyases.
EC Tree
4 Lyases
4.2 Carbon-oxygen lyases
4.2.1 Hydro-lyases
4.2.1.1 carbonic anhydrase
(BRENDA:EC4.2.1.1)
The accepted name is carbonic anhydrase while its systemic name is carbonic acid hydrolyase. It is also known by numerous other names such as; anhydrase, carbonate anhydrase,
carbonic acid anhydrase, carbonate hydro-lyase, carbonate hydro-lyase carboxyanhydrase,
carbonic anhydrase A, carbonate dehydratase, alkalistable alpha-carbonic anhydrase, alphatype CA and alpha-type carbonic anhydrase, Fisher (2006).

Carbonic Anhydrase is a well-studied and well-known enzyme because it was
conserved through many organisms. According to De Oliveira Maciel et al. (2022) various
creatures, such as bacteria, plants, and animals, there are three discrete families of cellular
automata, namely alpha, beta, and gamma, which are not evolutionarily linked to one
another. All Carbonic Anhydrase that has been identified across the animal kingdom are
categorized as alpha Carbonic Anhydrase.
The principal function of Carbonic Anhydrase is to expedite the fast and reversible
transformation between CO2 and bicarbonate ions. The enzymatic reaction is of utmost
importance in a wide range of biological processes. CO2 reacts with water in a variety of
organisms, including humans, with the assistance of the enzyme Carbonic Anhydrase. The
chemical process results in the generation of carbonic acid, which subsequently experiences
dissociation into a hydrogen ion and bicarbonate.
Carbonic acid → Carbon IV dioxide + Water
H2CO3 → CO2(g) + H2O

The reason for a reduction in respiratory rate resulting in a decline in pH can be
attributed to the following factors. Doyle and Cooper (2018) found that enzyme Carbonic
Anhydrase is of significant importance in a range of physiological processes, such as
respiration, since it aids in the movement of carbon dioxide from tissues to the lungs for
expiration. Apart from catalyzing the reversible hydration of carbon IV oxide to carbonic
acid, Carbonic Anhydrase catalyzes hydrolysis reactions of many substrates, including
nitrophenyl acetate (Elleby, 1999):
1- 2-nitrophenyl acetate + H2O →nitrophenol + acetate

+ H2O →

+

2-3-nitrophenyl acetate + H2O→3-nitrophenol + acetate

+H2O→

+

3-4-nitrophenyl propionate + H2O=4-nitrophenol + propionate

+H2O→

+

Carbonic Anhydrase also aids in the process of producing gastric acid in the stomach
and in the physiology of all photosynthetic organisms. Cyanobacteria possess carbonic
anhydrases that are located inside the carboxysome architecture. The mentioned
mechanism enables the conversion of CO2 into HCO3-, which plays a vital role as an
essential constituent within the Calvin cycle. The process is a series of metabolic reactions
utilized by plants to synthesize organic molecules, such as sugars, by assimilating
atmospheric carbon dioxide (Santos Correa et al., 2022).
There are 8 distinct classes that evolved independently. Though zinc is the most
prevalent cofactor, others have also been identified, including; Cd2+, Co2+, Fe2+, and Mn2+.
Carbonic Anhydrase exhibits ubiquitous presence inside a wide range of organs and
tissues across several organisms. The enzyme is distributed throughout several tissues in the
human body, displaying unique isoforms in each tissue. One illustration of this idea is
evident in the significant abundance of Carbonic Anhydrase II within erythrocytes, where it
fulfills a crucial role in enabling transportation of CO2 and preserving equilibrium of acidbase levels. According to Wang et al. (2020), Carbonic Anhydrase IV is found in the stomach
mucosa and has a significant role in the process of gastric acid secretion. Renal structure,
ocular structures, and many biological tissues have additional isoforms of the enzyme.
These isoforms have specific roles in maintaining pH balance and enabling the transport.
In mammals, homosepines, in particular, carbonic anhydrase is abundant in most of
the body cells such as astrocytoma cell, eye cells, also blood and brain cells (Haapasalo,
2008; Arslan, 2002; Hilvo, 2007; Juozapaitiene, 2016). Inside these cells, carbonic anhydrase
is present in isoforms that aid in cell functions and regulation in the cytoplasm,
mitochondria, and neurocell axons (Hilvo, 2007).
Carbonic anhydrase is also present in herbivorous mammals such as Alces alces
(Carlsson, 1973).
In prokaryotes, a study conducted by Guilloton et al. (1992) aimed to analyze and describe a
Carbonic Anhydrase enzyme encoded by the Carbonic Anhydrase I in the Escherichia coli can

operon. In a metabolic pathway that includes the Carbonic Anhydrase I gene, which codes
for the cyanate enzyme, Carbonic Anhydrases are an essential component. A zinc ion was
found in each subunit when the Carbonic Anhydrase enzyme was removed and purified
from E. coli strains that overexpressed the Carbonic Anhydrase I gene. In addition, it was
found that the enzyme is an oligomer in solution. The enzyme's kinetic properties, such as
its sensitivity to sulfonamide and cyanate inhibition, resembled Carbonic Anhydrase in
different animals.
The E. coli Carbonic Anhydrase’s amino acid sequence shared many similarities with
carbonic anhydrase found in plants but little in common with those found in mammals and
algae. This induced carbonic anhydrase's primary function appears to be to promote carbon
dioxide hydration, hence preventing the depletion of cellular bicarbonate. Carbonic
anhydrase is also present in other bacterial phyla and strains such as Bacillus subtilis and
Mycobacterium tuberculosis (Ramanan, 2009).
Homology refers to the presence of similarities in both the amino acid sequences and
structural features of proteins across a wide range of animal species, suggesting a common
evolutionary origin. Carbonic anhydrases demonstrate similarity among their different
groups. Nevertheless, there are distinct features observed in the chloroplast carbonic
anhydrases of bacteria and plants, as well as those of eukaryotes (Jensen et al., 2020). The
Prosite database focuses on a distinct group of Carbonic Anhydrase, namely those present
in prokaryotes and plant chloroplasts, exhibiting notable differences from the Carbonic
Anhydrase observed in eukaryotes. Carbonic Anhydrase has a ubiquitous function in aiding
the hydration of carbon dioxide. However, it is noteworthy that they have undergone
divergent evolutionary mechanisms in prokaryotes and plants compared to eukaryotes.
Mechanism of Carbonic Anhydrase

Carbonic anhydrase, with synonyms including CA IX, CA II, HCA II, HCA I, CA IX, CA II, HCA IX,
and CA IV, is a pivotal enzyme facilitating the reversible hydration of carbon dioxide to
bicarbonate ions. It relies on essential cofactors like calcium (Ca2+), iron (Fe3+), and zinc
ions (Zn), among others (Fisher, 2006). The enzyme's active site and allosteric sites display
pH-dependent changes, ensuring optimal functionality within a specific pH range, while
conformational adjustments in response to pH fluctuations underscore its dynamic nature
(Doyle and Cooper, 2018; De Oliveira Maciel et al., 2022). The main catalyzed reaction
involves the hydration of carbon dioxide, where cofactors like zinc ions play a crucial role in
facilitating the generation of a hydroxide ion (OH-) (Le Chatelier's principle) (Fisher, 2006).
Carbonic anhydrase exhibits regenerative ability, ensuring efficiency in processing carbon
dioxide rapidly. Allosteric regulation and binding sites significantly influence the enzyme's
overall functionality, providing a precise and controlled catalytic response to varying pH
conditions (Doyle and Cooper, 2018).
The kinetic details of carbonic anhydrase, varying across substrates and organisms, reveal a
pH optimum around 7-7.4 and a pH range for activity spanning from 6.5 to 8.5 in Homo
sapiens. The temperature optimum for the enzyme in humans is typically around 37°C.
Various substrates, including 4-nitrophenyl acetate, 4-nitrophenyl phosphate, CO2, and
HCO3-, exhibit distinct kinetic parameters, such as Km, Vmax, turnover number, and specific
activity, shedding light on the enzyme's efficiency under diverse conditions (BRENDA)..
References
1.

De Oliveira Maciel, A., Christakopoulos, P., Rova, U., & Antonopoulou, I. (2022). Carbonic anhydrase
to boost CO2 sequestration: Improving carbon capture utilization and storage
(CCUS). Chemosphere, 299, 134419.

2.

Doyle, J., & Cooper, J. S. (2018, October 27). Physiology, Carbon Dioxide Transport. Nih.gov;
StatPearls Publishing.

3.

Guilloton, M. B., Korte, J. J., Lamblin, A. F., Fuchs, J. A., & Anderson, P. M. (1992). Carbonic
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