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Registro Completo |
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Biblioteca(s): |
Embrapa Agroenergia. |
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Data corrente: |
27/09/2023 |
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Data da última atualização: |
25/10/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
CARDOSO, S. L.; SOUZA, P. M.; RODRIGUES, K.; MOTA, I. de S.; FERREIRA FILHO, E.; FAVARO, L. C. de L.; ARAUJO, F. S.; HOMEM-DE-MELLO, M.; PESSOA, A.; SILVEIRA, D.; BAZZO, Y. M. F.; MAGALHÃES, P. O. |
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Afiliação: |
SAMUEL LEITE CARDOSO, UNIVERSITY OF BRASILIA; PAULA MONTEIRO SOUZA, UNIVERSITY OF BRASILIA; KELLY RODRIGUES, EMBRAPA AGROENERGIA; ISABELLA DE SOUZA MOTA, UNIVERSITY OF BRASILIA; EDIVALDO FERREIRA FILHO, UNIVERSITY OF BRASÍLIA; LEIA CECILIA DE LIMA FAVARO, CNPAE; FELIPE SALDANHA ARAUJO, UNIVERSITY OF BRASILIA; MAURICIO HOMEM-DE-MELLO, UNIVERSITY OF BRASILIA; ADALBERTO PESSOA, UNIVERSITY OF SÃO PAULO; DÂMARIS SILVEIRA, UNIVERSITY OF BRASILIA; YRIS MARIA FONSECA-BAZZO, UNIVERSITY OF BRASILIA; PÉROLA OLIVEIRA MAGALHÃES, UNIVERSITY OF BRASILIA. |
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Título: |
L-Asparaginase type II from Fusarium proliferatum: heterologous expression and in silico analysis. |
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Ano de publicação: |
2023 |
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Fonte/Imprenta: |
Pharmaceutics, v. 15, 2352, 2023. |
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DOI: |
https://doi.org/10.3390/pharmaceutics15092352 |
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Idioma: |
Inglês |
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Conteúdo: |
The search for new drug-producing microorganisms is one of the most promising situations in current world scientific scenarios. The use of molecular biology as well as the cloning of protein and compound genes is already well established as the gold standard method of increasing productivity. Aiming at this increase in productivity, this work aims at the cloning, purification and in silico analysis of L-asparaginase from Fusarium proliferatum in Komagataella phaffii (Pichia pastoris) protein expression systems. The L-asparaginase gene (NCBI OQ439985) has been cloned into Pichia pastoris strains. Enzyme production was analyzed via the quantification of aspartic B-hydroxamate, followed by purification on a DEAE FF ion exchange column. The in silico analysis was proposed based on the combined use of various technological tools. The enzymatic activity found intracellularly was 2.84 IU/g. A purification factor of 1.18 was observed. The in silico analysis revealed the position of five important amino acid residues for enzymatic activity, and likewise, it was possible to predict a monomeric structure with a C-score of 1.59. The production of the enzyme L-asparaginase from F. proliferatum in P. pastoris was demonstrated in this work, being of great importance for the analysis of new methodologies in search of the production of important drugs in therapy. |
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Palavras-Chave: |
In silico analysis. |
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Thesaurus Nal: |
Asparaginase; Fusarium proliferatum; Heterologous gene expression. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/doc/1156949/1/L-Asparaginase-Type-II.pdf
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Marc: |
LEADER 02309naa a2200313 a 4500
001 2156949
005 2023-10-25
008 2023 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.3390/pharmaceutics15092352$2DOI
100 1 $aCARDOSO, S. L.
245 $aL-Asparaginase type II from Fusarium proliferatum$bheterologous expression and in silico analysis.$h[electronic resource]
260 $c2023
520 $aThe search for new drug-producing microorganisms is one of the most promising situations in current world scientific scenarios. The use of molecular biology as well as the cloning of protein and compound genes is already well established as the gold standard method of increasing productivity. Aiming at this increase in productivity, this work aims at the cloning, purification and in silico analysis of L-asparaginase from Fusarium proliferatum in Komagataella phaffii (Pichia pastoris) protein expression systems. The L-asparaginase gene (NCBI OQ439985) has been cloned into Pichia pastoris strains. Enzyme production was analyzed via the quantification of aspartic B-hydroxamate, followed by purification on a DEAE FF ion exchange column. The in silico analysis was proposed based on the combined use of various technological tools. The enzymatic activity found intracellularly was 2.84 IU/g. A purification factor of 1.18 was observed. The in silico analysis revealed the position of five important amino acid residues for enzymatic activity, and likewise, it was possible to predict a monomeric structure with a C-score of 1.59. The production of the enzyme L-asparaginase from F. proliferatum in P. pastoris was demonstrated in this work, being of great importance for the analysis of new methodologies in search of the production of important drugs in therapy.
650 $aAsparaginase
650 $aFusarium proliferatum
650 $aHeterologous gene expression
653 $aIn silico analysis
700 1 $aSOUZA, P. M.
700 1 $aRODRIGUES, K.
700 1 $aMOTA, I. de S.
700 1 $aFERREIRA FILHO, E.
700 1 $aFAVARO, L. C. de L.
700 1 $aARAUJO, F. S.
700 1 $aHOMEM-DE-MELLO, M.
700 1 $aPESSOA, A.
700 1 $aSILVEIRA, D.
700 1 $aBAZZO, Y. M. F.
700 1 $aMAGALHÃES, P. O.
773 $tPharmaceutics$gv. 15, 2352, 2023.
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Registro original: |
Embrapa Agroenergia (CNPAE) |
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 | Acesso ao texto completo restrito à biblioteca da Embrapa Agroenergia. Para informações adicionais entre em contato com cnpae.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Agroenergia. |
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Data corrente: |
12/03/2021 |
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Data da última atualização: |
12/03/2021 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
B - 1 |
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Autoria: |
JASER, SUHAILA K. K.; PERAZZA, C.; FAVARO, L. C. de L.; GOTO, M. A.; OLIVEIRA, A. M. DE; HALLERMAN, E.; HILSDORF, A. W. S. |
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Afiliação: |
SUHAILA K. K. JASER, Universidade de São Paulo; CAIO A. PERAZZA, Universidade de Mogi das Cruzes; LEIA CECILIA DE LIMA FAVARO, CNPAE; MARIANA A. GOTO, Universidade de São Paulo; ALZIRA M. DE OLIVEIRA, Instituto Nacional de Pesquisas na Amazônia; ERIC HALLERMAN, Virginia Polytechnic Institute and State University; ALEXANDRE W. S. HILSDORF, Universidade de Mogi das Cruzes. |
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Título: |
Identification and analysis of a novel microsatellite marker within the growth hormone gene promoter of Colossoma macropomum (Characiformes: Characidae) detected by TAIL-PCR. |
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Ano de publicação: |
2021 |
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Fonte/Imprenta: |
Journal of Applied Ichthyology, 2021. |
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DOI: |
https://doi.org/10.1111/jai.14170 |
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Idioma: |
Inglês |
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Notas: |
First online. |
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Conteúdo: |
Summary: While in all vertebrates, growth hormone (GH) promotes post‐natal growth, in fishes it also affects such metabolic functions as foraging rate, digestion, osmoregulation, and reproduction. The promoter region of the GH gene is an important target for studies of mechanisms regulating its expression, and polymorphisms within the promoter have been associated with performance traits in fishes. We used Thermal Asymmetric Interlaced PCR (TAIL-PCR) to amplify and sequence the 5'-flanking regions of the Colossoma macropomum GH (cmGH) gene. Based on a sequence of 1,038 bp, we designed three specific nested primers (R-290, R-186 and R-26) which were used with shorter arbitrary degenerate primers to amplify the 5'proximal region of the cmGH gene. We identified a tetranucleotide (ATCC)4 microsatellite motif in this region, exhibiting four alleles (118, 122, 126 and 130 bp) within the population study. Genotypes at this locus deviated significantly from Hardy-Weinberg expectations and showed a low level of polymorphism (polymorphic information content = 0.163). High homozygosity (FIS = 0.147) was observed in the overall population. The polymorphism at the microsatellite makes it an important candidate for association studies between the respective genotypes, growth rate and other traits in farmed populations. Such studies may contribute to future breeding programs using marker-assisted selection upon this aquaculturally important species. |
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Palavras-Chave: |
Cachama negra. |
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Thesagro: |
Tambaqui. |
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Thesaurus NAL: |
Breeding; Polymorphism; Tandem repeat sequences. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02355naa a2200277 a 4500
001 2130659
005 2021-03-12
008 2021 bl uuuu u00u1 u #d
024 7 $ahttps://doi.org/10.1111/jai.14170$2DOI
100 1 $aJASER, SUHAILA K. K.
245 $aIdentification and analysis of a novel microsatellite marker within the growth hormone gene promoter of Colossoma macropomum (Characiformes$bCharacidae) detected by TAIL-PCR.$h[electronic resource]
260 $c2021
500 $aFirst online.
520 $aSummary: While in all vertebrates, growth hormone (GH) promotes post‐natal growth, in fishes it also affects such metabolic functions as foraging rate, digestion, osmoregulation, and reproduction. The promoter region of the GH gene is an important target for studies of mechanisms regulating its expression, and polymorphisms within the promoter have been associated with performance traits in fishes. We used Thermal Asymmetric Interlaced PCR (TAIL-PCR) to amplify and sequence the 5'-flanking regions of the Colossoma macropomum GH (cmGH) gene. Based on a sequence of 1,038 bp, we designed three specific nested primers (R-290, R-186 and R-26) which were used with shorter arbitrary degenerate primers to amplify the 5'proximal region of the cmGH gene. We identified a tetranucleotide (ATCC)4 microsatellite motif in this region, exhibiting four alleles (118, 122, 126 and 130 bp) within the population study. Genotypes at this locus deviated significantly from Hardy-Weinberg expectations and showed a low level of polymorphism (polymorphic information content = 0.163). High homozygosity (FIS = 0.147) was observed in the overall population. The polymorphism at the microsatellite makes it an important candidate for association studies between the respective genotypes, growth rate and other traits in farmed populations. Such studies may contribute to future breeding programs using marker-assisted selection upon this aquaculturally important species.
650 $aBreeding
650 $aPolymorphism
650 $aTandem repeat sequences
650 $aTambaqui
653 $aCachama negra
700 1 $aPERAZZA, C.
700 1 $aFAVARO, L. C. de L.
700 1 $aGOTO, M. A.
700 1 $aOLIVEIRA, A. M. DE
700 1 $aHALLERMAN, E.
700 1 $aHILSDORF, A. W. S.
773 $tJournal of Applied Ichthyology, 2021.
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