Antarctic yeasts as a source of L-asparaginase: Characterization of a glutaminase-activity free L-asparaginase from psychrotolerant yeast Leucosporidium scottii L115
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , , , |
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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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 |
|
_version_ |
1808128800938000384 |