DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from <i>Annona Squamosa</i> (L.) and <i>Annona Muricata</i> (L.) Fruits

Lenny Mwagandi Chimbevo; Gibson Kamau Gicharu; Fredrick x Mwamburi Mjomba; Suliman Essuman

doi:doi:10.11648/j.bmb.20210603.14

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DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits

Lenny Mwagandi Chimbevo, Gibson Kamau Gicharu, Fredrick x Mwamburi Mjomba, Suliman Essuman

Published in Biochemistry and Molecular Biology (Volume 6, Issue 3)

Received: 9 July 2021 Accepted: 30 July 2021 Published: 13 August 2021

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Abstract

Some plants metabolites serve as antiprotozoal and antitumour by binding to nuclear enzyme; DNA Topo I affecting DNA function and cell survival. This study was aimed at screening DNA binding interactions and DNA Topo I inhibitory activity of crude extracts from fruits of Annona muricata (L) and Annona squamosa (L) which can form the basis of developing efficacious, safe and low cost antiprotozoal and antitumor agents. Aqueous, Methanolic, Ethyl acetate and Hexane extracts from fruits of two hypothesized antiprotozoal and antitumour plants; Annona muricata (L) and Annona squamosa (L) were screened for DNA-binding interaction and DNA Topo I inhibition. For DNA-methyl green test, 50 µL crude extracts were incubated with 200 μL DNA-methyl green in darkness at 25°C for 24 hours. Absorbance decrease at 650 nm using UV-vis spectrophotometer was calculated as a percentage of untreated DNA-methyl green value whereas with IC₅₀ calculated by regression analysis. For DNA Topo I inhibitory activity, crude extracts were incubated in 10 µg/mL with 0.5 µg of supercoiled pBR322 DNA and 1U of DNA Topo I at 37°C for 2 hours, reaction terminated using stop buffer containing 3% SDS, 60 mM EDTA, 50% glycerol, 0.25% bromophenol blue. Products were determined by electrophoresis on 1% agarose gel in Tris-acetate-EDTA (TAE) running buffer at 65 V/cm for 2 hours. 24 extracts were studied, percentage decrease in absorbance were between 18.14±2.67 - 38.06±1.47 (Aqueous), 17.14±2.67 - 41.01±1.09% (Methanolic), 9.05±1.67 - 20.50±2.01% (Ethyl acetate) and 4.04±1.12 - 10.09±1.39% (Hexane)., IC₅₀ values were between 50 μg/mL – 100 μg/mL (6), 100 μg/mL – 150 μg/mL (8), 150 μg/mL – 200 μg/mL (7) and <200 μg/mL (3). The activity against DNA Topo I mediated relaxation of supercoiled pBR322 DNA at 5 µM, 25 µM and 100 µM observed in 8 hits with percentage decrease in absorbance between 17.14±2.67 - 40.01±1.09% with IC₅₀ between 62.97±3.37 μg/mL - 131.37±10.77 μg/mL. The extracts of A. muricata and A. squamosa showed DNA Topo I inhibitory activities by inhibiting the relaxation of supercoiled DNA pBR322. However, further studies need to be conducted on the purified fractions of aqueous and methanolic extracts.

Published in	Biochemistry and Molecular Biology (Volume 6, Issue 3)
DOI	10.11648/j.bmb.20210603.14
Page(s)	58-66
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2021. Published by Science Publishing Group

Keywords

Topoisomerase I Inhibitors, DNA-Binding, Annona squamosa, Annona muricata

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Lenny Mwagandi Chimbevo, Gibson Kamau Gicharu, Fredrick x Mwamburi Mjomba, Suliman Essuman. (2021). DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits. Biochemistry and Molecular Biology, 6(3), 58-66. https://doi.org/10.11648/j.bmb.20210603.14

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Lenny Mwagandi Chimbevo; Gibson Kamau Gicharu; Fredrick x Mwamburi Mjomba; Suliman Essuman. DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits. Biochem. Mol. Biol. 2021, 6(3), 58-66. doi: 10.11648/j.bmb.20210603.14

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Lenny Mwagandi Chimbevo, Gibson Kamau Gicharu, Fredrick x Mwamburi Mjomba, Suliman Essuman. DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits. Biochem Mol Biol. 2021;6(3):58-66. doi: 10.11648/j.bmb.20210603.14

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@article{10.11648/j.bmb.20210603.14,
  author = {Lenny Mwagandi Chimbevo and Gibson Kamau Gicharu and Fredrick x Mwamburi Mjomba and Suliman Essuman},
  title = {DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits},
  journal = {Biochemistry and Molecular Biology},
  volume = {6},
  number = {3},
  pages = {58-66},
  doi = {10.11648/j.bmb.20210603.14},
  url = {https://doi.org/10.11648/j.bmb.20210603.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bmb.20210603.14},
  abstract = {Some plants metabolites serve as antiprotozoal and antitumour by binding to nuclear enzyme; DNA Topo I affecting DNA function and cell survival. This study was aimed at screening DNA binding interactions and DNA Topo I inhibitory activity of crude extracts from fruits of Annona muricata (L) and Annona squamosa (L) which can form the basis of developing efficacious, safe and low cost antiprotozoal and antitumor agents. Aqueous, Methanolic, Ethyl acetate and Hexane extracts from fruits of two hypothesized antiprotozoal and antitumour plants; Annona muricata (L) and Annona squamosa (L) were screened for DNA-binding interaction and DNA Topo I inhibition. For DNA-methyl green test, 50 µL crude extracts were incubated with 200 μL DNA-methyl green in darkness at 25°C for 24 hours. Absorbance decrease at 650 nm using UV-vis spectrophotometer was calculated as a percentage of untreated DNA-methyl green value whereas with IC50 calculated by regression analysis. For DNA Topo I inhibitory activity, crude extracts were incubated in 10 µg/mL with 0.5 µg of supercoiled pBR322 DNA and 1U of DNA Topo I at 37°C for 2 hours, reaction terminated using stop buffer containing 3% SDS, 60 mM EDTA, 50% glycerol, 0.25% bromophenol blue. Products were determined by electrophoresis on 1% agarose gel in Tris-acetate-EDTA (TAE) running buffer at 65 V/cm for 2 hours. 24 extracts were studied, percentage decrease in absorbance were between 18.14±2.67 - 38.06±1.47 (Aqueous), 17.14±2.67 - 41.01±1.09% (Methanolic), 9.05±1.67 - 20.50±2.01% (Ethyl acetate) and 4.04±1.12 - 10.09±1.39% (Hexane)., IC50 values were between 50 μg/mL – 100 μg/mL (6), 100 μg/mL – 150 μg/mL (8), 150 μg/mL – 200 μg/mL (7) and 50 between 62.97±3.37 μg/mL - 131.37±10.77 μg/mL. The extracts of A. muricata and A. squamosa showed DNA Topo I inhibitory activities by inhibiting the relaxation of supercoiled DNA pBR322. However, further studies need to be conducted on the purified fractions of aqueous and methanolic extracts.},
 year = {2021}
}

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TY  - JOUR
T1  - DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits
AU  - Lenny Mwagandi Chimbevo
AU  - Gibson Kamau Gicharu
AU  - Fredrick x Mwamburi Mjomba
AU  - Suliman Essuman
Y1  - 2021/08/13
PY  - 2021
N1  - https://doi.org/10.11648/j.bmb.20210603.14
DO  - 10.11648/j.bmb.20210603.14
T2  - Biochemistry and Molecular Biology
JF  - Biochemistry and Molecular Biology
JO  - Biochemistry and Molecular Biology
SP  - 58
EP  - 66
PB  - Science Publishing Group
SN  - 2575-5048
UR  - https://doi.org/10.11648/j.bmb.20210603.14
AB  - Some plants metabolites serve as antiprotozoal and antitumour by binding to nuclear enzyme; DNA Topo I affecting DNA function and cell survival. This study was aimed at screening DNA binding interactions and DNA Topo I inhibitory activity of crude extracts from fruits of Annona muricata (L) and Annona squamosa (L) which can form the basis of developing efficacious, safe and low cost antiprotozoal and antitumor agents. Aqueous, Methanolic, Ethyl acetate and Hexane extracts from fruits of two hypothesized antiprotozoal and antitumour plants; Annona muricata (L) and Annona squamosa (L) were screened for DNA-binding interaction and DNA Topo I inhibition. For DNA-methyl green test, 50 µL crude extracts were incubated with 200 μL DNA-methyl green in darkness at 25°C for 24 hours. Absorbance decrease at 650 nm using UV-vis spectrophotometer was calculated as a percentage of untreated DNA-methyl green value whereas with IC50 calculated by regression analysis. For DNA Topo I inhibitory activity, crude extracts were incubated in 10 µg/mL with 0.5 µg of supercoiled pBR322 DNA and 1U of DNA Topo I at 37°C for 2 hours, reaction terminated using stop buffer containing 3% SDS, 60 mM EDTA, 50% glycerol, 0.25% bromophenol blue. Products were determined by electrophoresis on 1% agarose gel in Tris-acetate-EDTA (TAE) running buffer at 65 V/cm for 2 hours. 24 extracts were studied, percentage decrease in absorbance were between 18.14±2.67 - 38.06±1.47 (Aqueous), 17.14±2.67 - 41.01±1.09% (Methanolic), 9.05±1.67 - 20.50±2.01% (Ethyl acetate) and 4.04±1.12 - 10.09±1.39% (Hexane)., IC50 values were between 50 μg/mL – 100 μg/mL (6), 100 μg/mL – 150 μg/mL (8), 150 μg/mL – 200 μg/mL (7) and 50 between 62.97±3.37 μg/mL - 131.37±10.77 μg/mL. The extracts of A. muricata and A. squamosa showed DNA Topo I inhibitory activities by inhibiting the relaxation of supercoiled DNA pBR322. However, further studies need to be conducted on the purified fractions of aqueous and methanolic extracts.
VL  - 6
IS  - 3
ER  -

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Lenny Mwagandi Chimbevo

Department of Pure and Applied Science, School of Applied and Health Sciences, Technical University of Mombasa, Mombasa, Kenya
Gibson Kamau Gicharu

Department of Pure and Applied Science, School of Applied and Health Sciences, Technical University of Mombasa, Mombasa, Kenya
Fredrick x Mwamburi Mjomba

Department of Pure and Applied Science, School of Applied and Health Sciences, Technical University of Mombasa, Mombasa, Kenya
Suliman Essuman

Department of Medical Microbiology, Medical School, Mount Kenya University, Thika, Kenya

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Lenny Mwagandi Chimbevo, Gibson Kamau Gicharu, Fredrick x Mwamburi Mjomba, Suliman Essuman. (2021). DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits. Biochemistry and Molecular Biology, 6(3), 58-66. https://doi.org/10.11648/j.bmb.20210603.14

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ACS Style

Lenny Mwagandi Chimbevo; Gibson Kamau Gicharu; Fredrick x Mwamburi Mjomba; Suliman Essuman. DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits. Biochem. Mol. Biol. 2021, 6(3), 58-66. doi: 10.11648/j.bmb.20210603.14

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AMA Style

Lenny Mwagandi Chimbevo, Gibson Kamau Gicharu, Fredrick x Mwamburi Mjomba, Suliman Essuman. DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits. Biochem Mol Biol. 2021;6(3):58-66. doi: 10.11648/j.bmb.20210603.14

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@article{10.11648/j.bmb.20210603.14,
  author = {Lenny Mwagandi Chimbevo and Gibson Kamau Gicharu and Fredrick x Mwamburi Mjomba and Suliman Essuman},
  title = {DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits},
  journal = {Biochemistry and Molecular Biology},
  volume = {6},
  number = {3},
  pages = {58-66},
  doi = {10.11648/j.bmb.20210603.14},
  url = {https://doi.org/10.11648/j.bmb.20210603.14},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.bmb.20210603.14},
  abstract = {Some plants metabolites serve as antiprotozoal and antitumour by binding to nuclear enzyme; DNA Topo I affecting DNA function and cell survival. This study was aimed at screening DNA binding interactions and DNA Topo I inhibitory activity of crude extracts from fruits of Annona muricata (L) and Annona squamosa (L) which can form the basis of developing efficacious, safe and low cost antiprotozoal and antitumor agents. Aqueous, Methanolic, Ethyl acetate and Hexane extracts from fruits of two hypothesized antiprotozoal and antitumour plants; Annona muricata (L) and Annona squamosa (L) were screened for DNA-binding interaction and DNA Topo I inhibition. For DNA-methyl green test, 50 µL crude extracts were incubated with 200 μL DNA-methyl green in darkness at 25°C for 24 hours. Absorbance decrease at 650 nm using UV-vis spectrophotometer was calculated as a percentage of untreated DNA-methyl green value whereas with IC50 calculated by regression analysis. For DNA Topo I inhibitory activity, crude extracts were incubated in 10 µg/mL with 0.5 µg of supercoiled pBR322 DNA and 1U of DNA Topo I at 37°C for 2 hours, reaction terminated using stop buffer containing 3% SDS, 60 mM EDTA, 50% glycerol, 0.25% bromophenol blue. Products were determined by electrophoresis on 1% agarose gel in Tris-acetate-EDTA (TAE) running buffer at 65 V/cm for 2 hours. 24 extracts were studied, percentage decrease in absorbance were between 18.14±2.67 - 38.06±1.47 (Aqueous), 17.14±2.67 - 41.01±1.09% (Methanolic), 9.05±1.67 - 20.50±2.01% (Ethyl acetate) and 4.04±1.12 - 10.09±1.39% (Hexane)., IC50 values were between 50 μg/mL – 100 μg/mL (6), 100 μg/mL – 150 μg/mL (8), 150 μg/mL – 200 μg/mL (7) and 50 between 62.97±3.37 μg/mL - 131.37±10.77 μg/mL. The extracts of A. muricata and A. squamosa showed DNA Topo I inhibitory activities by inhibiting the relaxation of supercoiled DNA pBR322. However, further studies need to be conducted on the purified fractions of aqueous and methanolic extracts.},
 year = {2021}
}

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TY  - JOUR
T1  - DNA Binding Interactions and DNA Topoisomerase I Inhibition Activities of Crude Extracts from Annona Squamosa (L.) and Annona Muricata (L.) Fruits
AU  - Lenny Mwagandi Chimbevo
AU  - Gibson Kamau Gicharu
AU  - Fredrick x Mwamburi Mjomba
AU  - Suliman Essuman
Y1  - 2021/08/13
PY  - 2021
N1  - https://doi.org/10.11648/j.bmb.20210603.14
DO  - 10.11648/j.bmb.20210603.14
T2  - Biochemistry and Molecular Biology
JF  - Biochemistry and Molecular Biology
JO  - Biochemistry and Molecular Biology
SP  - 58
EP  - 66
PB  - Science Publishing Group
SN  - 2575-5048
UR  - https://doi.org/10.11648/j.bmb.20210603.14
AB  - Some plants metabolites serve as antiprotozoal and antitumour by binding to nuclear enzyme; DNA Topo I affecting DNA function and cell survival. This study was aimed at screening DNA binding interactions and DNA Topo I inhibitory activity of crude extracts from fruits of Annona muricata (L) and Annona squamosa (L) which can form the basis of developing efficacious, safe and low cost antiprotozoal and antitumor agents. Aqueous, Methanolic, Ethyl acetate and Hexane extracts from fruits of two hypothesized antiprotozoal and antitumour plants; Annona muricata (L) and Annona squamosa (L) were screened for DNA-binding interaction and DNA Topo I inhibition. For DNA-methyl green test, 50 µL crude extracts were incubated with 200 μL DNA-methyl green in darkness at 25°C for 24 hours. Absorbance decrease at 650 nm using UV-vis spectrophotometer was calculated as a percentage of untreated DNA-methyl green value whereas with IC50 calculated by regression analysis. For DNA Topo I inhibitory activity, crude extracts were incubated in 10 µg/mL with 0.5 µg of supercoiled pBR322 DNA and 1U of DNA Topo I at 37°C for 2 hours, reaction terminated using stop buffer containing 3% SDS, 60 mM EDTA, 50% glycerol, 0.25% bromophenol blue. Products were determined by electrophoresis on 1% agarose gel in Tris-acetate-EDTA (TAE) running buffer at 65 V/cm for 2 hours. 24 extracts were studied, percentage decrease in absorbance were between 18.14±2.67 - 38.06±1.47 (Aqueous), 17.14±2.67 - 41.01±1.09% (Methanolic), 9.05±1.67 - 20.50±2.01% (Ethyl acetate) and 4.04±1.12 - 10.09±1.39% (Hexane)., IC50 values were between 50 μg/mL – 100 μg/mL (6), 100 μg/mL – 150 μg/mL (8), 150 μg/mL – 200 μg/mL (7) and 50 between 62.97±3.37 μg/mL - 131.37±10.77 μg/mL. The extracts of A. muricata and A. squamosa showed DNA Topo I inhibitory activities by inhibiting the relaxation of supercoiled DNA pBR322. However, further studies need to be conducted on the purified fractions of aqueous and methanolic extracts.
VL  - 6
IS  - 3
ER  -

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