Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis
Autor(a) principal: | |
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Data de Publicação: | 2005 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Medical and Biological Research |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005001100004 |
Resumo: | Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1) in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB) formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS) that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa) is the smallest non-bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis. |
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Brazilian Journal of Medical and Biological Research |
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Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactisLactococcus lactisAntifungal proteinSecretionStreptomyces tendae antifungal protein 1Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1) in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB) formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS) that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa) is the smallest non-bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis.Associação Brasileira de Divulgação Científica2005-11-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005001100004Brazilian Journal of Medical and Biological Research v.38 n.11 2005reponame:Brazilian Journal of Medical and Biological Researchinstname:Associação Brasileira de Divulgação Científica (ABDC)instacron:ABDC10.1590/S0100-879X2005001100004info:eu-repo/semantics/openAccessFreitas,D.A.Leclerc,S.Miyoshi,A.Oliveira,S.C.Sommer,P.S.M.Rodrigues,L.Correa Junior,A.Gautier,M.Langella,P.Azevedo,V.A.Le Loir,Y.eng2005-10-26T00:00:00Zoai:scielo:S0100-879X2005001100004Revistahttps://www.bjournal.org/https://old.scielo.br/oai/scielo-oai.phpbjournal@terra.com.br||bjournal@terra.com.br1414-431X0100-879Xopendoar:2005-10-26T00:00Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)false |
dc.title.none.fl_str_mv |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
title |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
spellingShingle |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis Freitas,D.A. Lactococcus lactis Antifungal protein Secretion Streptomyces tendae antifungal protein 1 |
title_short |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
title_full |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
title_fullStr |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
title_full_unstemmed |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
title_sort |
Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis |
author |
Freitas,D.A. |
author_facet |
Freitas,D.A. Leclerc,S. Miyoshi,A. Oliveira,S.C. Sommer,P.S.M. Rodrigues,L. Correa Junior,A. Gautier,M. Langella,P. Azevedo,V.A. Le Loir,Y. |
author_role |
author |
author2 |
Leclerc,S. Miyoshi,A. Oliveira,S.C. Sommer,P.S.M. Rodrigues,L. Correa Junior,A. Gautier,M. Langella,P. Azevedo,V.A. Le Loir,Y. |
author2_role |
author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Freitas,D.A. Leclerc,S. Miyoshi,A. Oliveira,S.C. Sommer,P.S.M. Rodrigues,L. Correa Junior,A. Gautier,M. Langella,P. Azevedo,V.A. Le Loir,Y. |
dc.subject.por.fl_str_mv |
Lactococcus lactis Antifungal protein Secretion Streptomyces tendae antifungal protein 1 |
topic |
Lactococcus lactis Antifungal protein Secretion Streptomyces tendae antifungal protein 1 |
description |
Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1) in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB) formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS) that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa) is the smallest non-bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis. |
publishDate |
2005 |
dc.date.none.fl_str_mv |
2005-11-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005001100004 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2005001100004 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0100-879X2005001100004 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica |
publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica |
dc.source.none.fl_str_mv |
Brazilian Journal of Medical and Biological Research v.38 n.11 2005 reponame:Brazilian Journal of Medical and Biological Research instname:Associação Brasileira de Divulgação Científica (ABDC) instacron:ABDC |
instname_str |
Associação Brasileira de Divulgação Científica (ABDC) |
instacron_str |
ABDC |
institution |
ABDC |
reponame_str |
Brazilian Journal of Medical and Biological Research |
collection |
Brazilian Journal of Medical and Biological Research |
repository.name.fl_str_mv |
Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC) |
repository.mail.fl_str_mv |
bjournal@terra.com.br||bjournal@terra.com.br |
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1754302934195109888 |