Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115

Detalhes bibliográficos
Autor(a) principal: Sánchez-Moguel, Ignacio
Data de Publicação: 2023
Outros Autores: Costa-Silva, Tales A., Pillaca-Pullo, Omar S., Flores-Santos, Juan Carlos, Freire, Rominne Karla Barros, Carretero, Gustavo, da Luz Bueno, Júlia, Camacho-Córdova, David I., Santos, João H.P.M., Sette, Lara Durães [UNESP], Pessoa-Jr, Adalberto
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1016/j.procbio.2023.03.003
http://hdl.handle.net/11449/249053
Resumo: Microorganisms from extreme environments, such as the Antarctic ecosystems, have a great potential to produce enzymes with novel characteristics. Within this context, L-asparaginase (ASNase) obtained from yeast species has been poorly studied. In this study, yeasts isolated from samples collected at Admiralty Bay (King George Island, Antarctica) were tested to produce ASNase. From an initial screening of 40 strains, belonging to 13 different species, Leucosporidium scottii L115 produced an ASNase activity (LsASNase activity: 6.24 U g-1 of dry cell weight) with the lowest glutaminase activity. The LsASNase was purified 227-fold, with a specific activity of 137.01 U mg-1 at 37 °C, without glutaminase activity. Moreover, the maximum enzyme activity was observed at pH 7.5 and at a temperature of 55 °C. The enzyme is a multimer of 462 kDa, presenting a single band of 53 kDa molecular mass in reduced conditions; after PGNase F treatment, a single band of 45 kDa was observed. The enzymatic kinetic evaluation revealed an allosteric regulation of the enzyme and the kinetic parameters were determined at 37 °C, pH 7.0 as substrate affinity constant, K0.5 = 233 μM, kcat = 54.7 s−1 and Hill coefficient, nH = 1.52, demonstrating positive cooperativity by the enzyme and the substrate. This is the first study to report L. scottii as a source of glutaminase-activity free L-asparaginase, an acute lymphoblastic leukemia drug feature suitable for the treatment of asparagine synthetase negative cancer cells.
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spelling Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115Antarctic ecosystemsCold-adapted yeastL-asparaginaseLeucosporidium scottiiLeukemiaPsychrotolerant yeastMicroorganisms from extreme environments, such as the Antarctic ecosystems, have a great potential to produce enzymes with novel characteristics. Within this context, L-asparaginase (ASNase) obtained from yeast species has been poorly studied. In this study, yeasts isolated from samples collected at Admiralty Bay (King George Island, Antarctica) were tested to produce ASNase. From an initial screening of 40 strains, belonging to 13 different species, Leucosporidium scottii L115 produced an ASNase activity (LsASNase activity: 6.24 U g-1 of dry cell weight) with the lowest glutaminase activity. The LsASNase was purified 227-fold, with a specific activity of 137.01 U mg-1 at 37 °C, without glutaminase activity. Moreover, the maximum enzyme activity was observed at pH 7.5 and at a temperature of 55 °C. The enzyme is a multimer of 462 kDa, presenting a single band of 53 kDa molecular mass in reduced conditions; after PGNase F treatment, a single band of 45 kDa was observed. The enzymatic kinetic evaluation revealed an allosteric regulation of the enzyme and the kinetic parameters were determined at 37 °C, pH 7.0 as substrate affinity constant, K0.5 = 233 μM, kcat = 54.7 s−1 and Hill coefficient, nH = 1.52, demonstrating positive cooperativity by the enzyme and the substrate. This is the first study to report L. scottii as a source of glutaminase-activity free L-asparaginase, an acute lymphoblastic leukemia drug feature suitable for the treatment of asparagine synthetase negative cancer cells.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Consejo Nacional de Ciencia y TecnologíaDepartment of Biochemical and Pharmaceutical Technology School of Pharmaceutical Sciences University of São Paulo, São PauloCentro de Investigación en Biodiversidad para la Salud Universidad Privada Norbert WienerDepartment of Biochemistry Chemistry Institute University of São Paulo, São PauloMarine Biotechnology Division Almirante Paulo Moreira Institute of Studies of the Sea, RJDepartment of General and Applied Biology Biosciences Institute São Paulo State University, São PauloCenter for Natural and Human Sciences Federal University of ABC, São PauloDepartment of General and Applied Biology Biosciences Institute São Paulo State University, São PauloFAPESP: 2013/08617–7FAPESP: 2013/19584–2FAPESP: 2019/23620–0Consejo Nacional de Ciencia y Tecnología: 298596Universidade de São Paulo (USP)Universidad Privada Norbert WienerAlmirante Paulo Moreira Institute of Studies of the SeaUniversidade Estadual Paulista (UNESP)Federal University of ABCSánchez-Moguel, IgnacioCosta-Silva, Tales A.Pillaca-Pullo, Omar S.Flores-Santos, Juan CarlosFreire, Rominne Karla BarrosCarretero, Gustavoda Luz Bueno, JúliaCamacho-Córdova, David I.Santos, João H.P.M.Sette, Lara Durães [UNESP]Pessoa-Jr, Adalberto2023-07-29T14:01:05Z2023-07-29T14:01:05Z2023-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article121-132http://dx.doi.org/10.1016/j.procbio.2023.03.003Process Biochemistry, v. 129, p. 121-132.1359-5113http://hdl.handle.net/11449/24905310.1016/j.procbio.2023.03.0032-s2.0-85150424621Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengProcess Biochemistryinfo:eu-repo/semantics/openAccess2023-07-29T14:01:05Zoai:repositorio.unesp.br:11449/249053Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:22:36.682386Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
title Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
spellingShingle Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
Sánchez-Moguel, Ignacio
Antarctic ecosystems
Cold-adapted yeast
L-asparaginase
Leucosporidium scottii
Leukemia
Psychrotolerant yeast
title_short Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
title_full Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
title_fullStr Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
title_full_unstemmed Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
title_sort Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
author Sánchez-Moguel, Ignacio
author_facet Sánchez-Moguel, Ignacio
Costa-Silva, Tales A.
Pillaca-Pullo, Omar S.
Flores-Santos, Juan Carlos
Freire, Rominne Karla Barros
Carretero, Gustavo
da Luz Bueno, Júlia
Camacho-Córdova, David I.
Santos, João H.P.M.
Sette, Lara Durães [UNESP]
Pessoa-Jr, Adalberto
author_role author
author2 Costa-Silva, Tales A.
Pillaca-Pullo, Omar S.
Flores-Santos, Juan Carlos
Freire, Rominne Karla Barros
Carretero, Gustavo
da Luz Bueno, Júlia
Camacho-Córdova, David I.
Santos, João H.P.M.
Sette, Lara Durães [UNESP]
Pessoa-Jr, Adalberto
author2_role author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Universidad Privada Norbert Wiener
Almirante Paulo Moreira Institute of Studies of the Sea
Universidade Estadual Paulista (UNESP)
Federal University of ABC
dc.contributor.author.fl_str_mv Sánchez-Moguel, Ignacio
Costa-Silva, Tales A.
Pillaca-Pullo, Omar S.
Flores-Santos, Juan Carlos
Freire, Rominne Karla Barros
Carretero, Gustavo
da Luz Bueno, Júlia
Camacho-Córdova, David I.
Santos, João H.P.M.
Sette, Lara Durães [UNESP]
Pessoa-Jr, Adalberto
dc.subject.por.fl_str_mv Antarctic ecosystems
Cold-adapted yeast
L-asparaginase
Leucosporidium scottii
Leukemia
Psychrotolerant yeast
topic Antarctic ecosystems
Cold-adapted yeast
L-asparaginase
Leucosporidium scottii
Leukemia
Psychrotolerant yeast
description Microorganisms from extreme environments, such as the Antarctic ecosystems, have a great potential to produce enzymes with novel characteristics. Within this context, L-asparaginase (ASNase) obtained from yeast species has been poorly studied. In this study, yeasts isolated from samples collected at Admiralty Bay (King George Island, Antarctica) were tested to produce ASNase. From an initial screening of 40 strains, belonging to 13 different species, Leucosporidium scottii L115 produced an ASNase activity (LsASNase activity: 6.24 U g-1 of dry cell weight) with the lowest glutaminase activity. The LsASNase was purified 227-fold, with a specific activity of 137.01 U mg-1 at 37 °C, without glutaminase activity. Moreover, the maximum enzyme activity was observed at pH 7.5 and at a temperature of 55 °C. The enzyme is a multimer of 462 kDa, presenting a single band of 53 kDa molecular mass in reduced conditions; after PGNase F treatment, a single band of 45 kDa was observed. The enzymatic kinetic evaluation revealed an allosteric regulation of the enzyme and the kinetic parameters were determined at 37 °C, pH 7.0 as substrate affinity constant, K0.5 = 233 μM, kcat = 54.7 s−1 and Hill coefficient, nH = 1.52, demonstrating positive cooperativity by the enzyme and the substrate. This is the first study to report L. scottii as a source of glutaminase-activity free L-asparaginase, an acute lymphoblastic leukemia drug feature suitable for the treatment of asparagine synthetase negative cancer cells.
publishDate 2023
dc.date.none.fl_str_mv 2023-07-29T14:01:05Z
2023-07-29T14:01:05Z
2023-06-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1016/j.procbio.2023.03.003
Process Biochemistry, v. 129, p. 121-132.
1359-5113
http://hdl.handle.net/11449/249053
10.1016/j.procbio.2023.03.003
2-s2.0-85150424621
url http://dx.doi.org/10.1016/j.procbio.2023.03.003
http://hdl.handle.net/11449/249053
identifier_str_mv Process Biochemistry, v. 129, p. 121-132.
1359-5113
10.1016/j.procbio.2023.03.003
2-s2.0-85150424621
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Process Biochemistry
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 121-132
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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