Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis

Detalhes bibliográficos
Autor(a) principal: Freitas,D.A.
Data de Publicação: 2005
Outros Autores: 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.
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.
id ABDC-1_5f2121883ac5b840b906b6f6b5c33dbd
oai_identifier_str oai:scielo:S0100-879X2005001100004
network_acronym_str ABDC-1
network_name_str Brazilian Journal of Medical and Biological Research
repository_id_str
spelling 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
_version_ 1754302934195109888