Insights into Asparaginase from Endophytic Fungus Lasiodiplodia theobromae: Purification, Characterization and Antileukemic Activity

Hani A. Moubasher; Bassem A. Balbool; Yosra A. Helmy; Amnah Mohammed Alsuhaibani; Ahmed A. Atta; Donia H. Sheir; Ahmed M. Abdel-Azeem

doi:10.3390/ijerph19020680

Insights into Asparaginase from Endophytic Fungus Lasiodiplodia theobromae: Purification, Characterization and Antileukemic Activity

Hani A. Moubasher
, Bassem A. Balbool
, Yosra A. Helmy
, Amnah Mohammed Alsuhaibani
, Ahmed A. Atta
, Donia H. Sheir
, Ahmed M. Abdel-Azeem

Veterinary Science

Producción científica: Article › revisión exhaustiva

34 Citas (Scopus)

Resumen

Endobiotic fungi are considered as a reservoir of numerous active metabolites. Asparaginase is used as an antileukemic drug specially to treat acute lymphoblastic leukaemia. The presented study aims to optimize the media conditions, purify, characterize, and test the antileukemic activity of the asparaginase induced from Lasiodiplodia theobromae. The culture medium was optimized using an experiment designed by The Taguchi model with an activity ranging from 10 to 175 IU/mL. Asparaginase was induced with an activity of 315 IU/mL. Asparaginase was purified with a specific activity of 468.03 U/mg and total activity of 84.4 IU/mL. The purified asparaginase showed an approximate size of 70 kDa. The purified asparaginase showed an optimum temperature of 37 °C and an optimum pH of 6. SDS reduced the activity of asparaginase to 0.65 U/mL while the used ionic surfactants enhanced the enzyme activity up to 151.92 IU/mL. The purified asparaginase showed a Km of 9.37 µM and Vmax of 127.00 µM/mL/min. The purified asparaginase showed an IC50 of 35.2 ± 0.7 IU/mL with leukemic M-NFS-60 cell lines and CC50 of 79.4 ± 1.9 IU/mL with the normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for metabolites such as asparaginase, provides an opportunity to develop a facile, eco-friendly, costeffective, and rapid synthesis of antileukemic drugs, which have the potential to be used as alternative and reliable sources for potent anticancer agents.

Idioma original	English
Número de artículo	680
Publicación	International Journal of Environmental Research and Public Health
Volumen	19
N.º	2
DOI	https://doi.org/10.3390/ijerph19020680
Estado	Published - ene 1 2022

Nota bibliográfica

Funding Information:
Acknowledgments: We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress.

Funding Information:
We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress.

Publisher Copyright:
© 2022 by the authors. Submitted for possible open access.

Financiación

Acknowledgments: We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress. We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress.

Financiadores	Número del financiador
Botany and Microbiology Department, Faculty of Science
Taif University	TURSP-2020/22

ODS de las Naciones Unidas

Este resultado contribuye a los siguientes Objetivos de Desarrollo Sostenible

Good health and well being

ASJC Scopus subject areas

Pollution
Public Health, Environmental and Occupational Health
Health, Toxicology and Mutagenesis

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10.3390/ijerph19020680

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Moubasher, H. A., Balbool, B. A., Helmy, Y. A., Alsuhaibani, A. M., Atta, A. A., Sheir, D. H., & Abdel-Azeem, A. M. (2022). Insights into Asparaginase from Endophytic Fungus Lasiodiplodia theobromae: Purification, Characterization and Antileukemic Activity. International Journal of Environmental Research and Public Health, 19(2), Artículo 680. https://doi.org/10.3390/ijerph19020680

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abstract = "Endobiotic fungi are considered as a reservoir of numerous active metabolites. Asparaginase is used as an antileukemic drug specially to treat acute lymphoblastic leukaemia. The presented study aims to optimize the media conditions, purify, characterize, and test the antileukemic activity of the asparaginase induced from Lasiodiplodia theobromae. The culture medium was optimized using an experiment designed by The Taguchi model with an activity ranging from 10 to 175 IU/mL. Asparaginase was induced with an activity of 315 IU/mL. Asparaginase was purified with a specific activity of 468.03 U/mg and total activity of 84.4 IU/mL. The purified asparaginase showed an approximate size of 70 kDa. The purified asparaginase showed an optimum temperature of 37 °C and an optimum pH of 6. SDS reduced the activity of asparaginase to 0.65 U/mL while the used ionic surfactants enhanced the enzyme activity up to 151.92 IU/mL. The purified asparaginase showed a Km of 9.37 µM and Vmax of 127.00 µM/mL/min. The purified asparaginase showed an IC50 of 35.2 ± 0.7 IU/mL with leukemic M-NFS-60 cell lines and CC50 of 79.4 ± 1.9 IU/mL with the normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for metabolites such as asparaginase, provides an opportunity to develop a facile, eco-friendly, costeffective, and rapid synthesis of antileukemic drugs, which have the potential to be used as alternative and reliable sources for potent anticancer agents.",

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note = "Funding Information: Acknowledgments: We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress. Funding Information: We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress. Publisher Copyright: {\textcopyright} 2022 by the authors. Submitted for possible open access.",

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AU - Moubasher, Hani A.

AU - Balbool, Bassem A.

AU - Helmy, Yosra A.

AU - Alsuhaibani, Amnah Mohammed

AU - Atta, Ahmed A.

AU - Sheir, Donia H.

AU - Abdel-Azeem, Ahmed M.

N1 - Funding Information: Acknowledgments: We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress. Funding Information: We appreciate and thank Taif University for the financial support for Taif University Re-searchers Supporting Project (TURSP-2020/22), Taif University, Taif, Saudi Arabia. We would like to present their deep appreciation to Prof. Dr. Abdel Ghaffar Abu El Saoud, Botany and Microbiology Department, Faculty of Science, Suez Canal University for his support and help during the data analysis of the results. Also, to the members of Saint Katherine Protectorate for their support during the samples collection in the early stages of the research work progress. Publisher Copyright: © 2022 by the authors. Submitted for possible open access.

PY - 2022/1/1

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N2 - Endobiotic fungi are considered as a reservoir of numerous active metabolites. Asparaginase is used as an antileukemic drug specially to treat acute lymphoblastic leukaemia. The presented study aims to optimize the media conditions, purify, characterize, and test the antileukemic activity of the asparaginase induced from Lasiodiplodia theobromae. The culture medium was optimized using an experiment designed by The Taguchi model with an activity ranging from 10 to 175 IU/mL. Asparaginase was induced with an activity of 315 IU/mL. Asparaginase was purified with a specific activity of 468.03 U/mg and total activity of 84.4 IU/mL. The purified asparaginase showed an approximate size of 70 kDa. The purified asparaginase showed an optimum temperature of 37 °C and an optimum pH of 6. SDS reduced the activity of asparaginase to 0.65 U/mL while the used ionic surfactants enhanced the enzyme activity up to 151.92 IU/mL. The purified asparaginase showed a Km of 9.37 µM and Vmax of 127.00 µM/mL/min. The purified asparaginase showed an IC50 of 35.2 ± 0.7 IU/mL with leukemic M-NFS-60 cell lines and CC50 of 79.4 ± 1.9 IU/mL with the normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for metabolites such as asparaginase, provides an opportunity to develop a facile, eco-friendly, costeffective, and rapid synthesis of antileukemic drugs, which have the potential to be used as alternative and reliable sources for potent anticancer agents.

AB - Endobiotic fungi are considered as a reservoir of numerous active metabolites. Asparaginase is used as an antileukemic drug specially to treat acute lymphoblastic leukaemia. The presented study aims to optimize the media conditions, purify, characterize, and test the antileukemic activity of the asparaginase induced from Lasiodiplodia theobromae. The culture medium was optimized using an experiment designed by The Taguchi model with an activity ranging from 10 to 175 IU/mL. Asparaginase was induced with an activity of 315 IU/mL. Asparaginase was purified with a specific activity of 468.03 U/mg and total activity of 84.4 IU/mL. The purified asparaginase showed an approximate size of 70 kDa. The purified asparaginase showed an optimum temperature of 37 °C and an optimum pH of 6. SDS reduced the activity of asparaginase to 0.65 U/mL while the used ionic surfactants enhanced the enzyme activity up to 151.92 IU/mL. The purified asparaginase showed a Km of 9.37 µM and Vmax of 127.00 µM/mL/min. The purified asparaginase showed an IC50 of 35.2 ± 0.7 IU/mL with leukemic M-NFS-60 cell lines and CC50 of 79.4 ± 1.9 IU/mL with the normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for metabolites such as asparaginase, provides an opportunity to develop a facile, eco-friendly, costeffective, and rapid synthesis of antileukemic drugs, which have the potential to be used as alternative and reliable sources for potent anticancer agents.

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KW - Asparaginase

KW - Endophytic fungi

KW - Induction

KW - Metabolites1

KW - Purification

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JO - International Journal of Environmental Research and Public Health

JF - International Journal of Environmental Research and Public Health

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M1 - 680

ER -

Insights into Asparaginase from Endophytic Fungus Lasiodiplodia theobromae: Purification, Characterization and Antileukemic Activity

Resumen

Nota bibliográfica

Financiación

ODS de las Naciones Unidas

ASJC Scopus subject areas

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