Co-synthesis of kenyacin 404 and heterologous thurincin H enhances the antibacterial activity of Bacillus thuringiensis

Oros-Flores, Zuleyka S; Casados‐Vázquez, Luz E.; Bideshi, Dennis K.; Salcedo‐Hernández, Rubén; Barboza‐Corona, José E.

doi:10.1007/s10529-018-2601-9

Cited by 9 publications

(6 citation statements)

References 28 publications

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“…Nevertheless, similar to chitinases, thurincin H is produced in the vegetative stage and secreted. Another drawback of thurincin H is that it requires 10 genes for its production, including genes for regulation, processing, and immunity (Lee, Churey, et al, 2009; Lee, Gray, et al, 2009; Oros‐Flores et al, 2018). In HD1 WT and HD1 EV strains, thurincin H is regulated by its native promoter, both strains reached the maximum activity ~20,000 U mg −1 at 24–36 h. When the pSTAB promoter was added upstream of the thnA , thnB , thnT and thnI genes, HD1/pSTAB‐ThurH and HD1/pSTAB‐ThurH/pSTAB‐ChiA74 strains, the antibacterial activity was maintained up to ~4000 U mg −1 at 72 h. This activity is lower in comparison with previous hours, but higher in comparison with strains harbouring the native promoter (HD1 WT and HD1 EV).…”

Section: Discussionmentioning

confidence: 99%

“…One of the most studied is thurincin H, which is an antimicrobial peptide of 31 amino acids with the potential to be used as a growth factor (Lee, Churey, et al, 2009; Lee, Gray, et al, 2009). The synthesis, posttranslational modification, and translocation of thurincin H are carried out by proteins encoded by a cluster of ten genes whose expression is controlled by a bidirectional promoter (Casados‐Vázquez et al, 2018; Oros‐Flores et al, 2018). Thurincin H can be used as a growth factor, and its application to leaves or roots has stimulated growth in soybean and corn.…”

Section: Introductionmentioning

confidence: 99%

“…In this work, the native promoter that controls the expression of the structural thurincin H gene (i.e. thnA ) and the genes thnB , thnT and thnI (Casados‐Vázquez et al, 2018; Lee, Churey, et al, 2009; Lee, Gray, et al, 2009; Oros‐Flores et al, 2018) was replaced by a strong‐sporulation promoter (i.e. pcytA‐STAB).…”

Section: Introductionmentioning

confidence: 99%

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Expression of thurincin H, ChiA74 and Cry proteins at the sporulation phase in Bacillus thuringiensis HD1

Rivas

Barboza‐Corona

Massange-Sánchez

et al. 2022

Journal of Applied Microbiology

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Aims:The objective of this study was to produce thurincin H, ChiA74 and Cry proteins together using Bacillus thuringiensis subsp. kurstaki HD1 as a heterologous host. Methods and Results: pSTAB-ThurH and pSTAB-ChiA74 constructs were designed to produce thurincin H and chitinase, respectively, at the sporulation phase.They were transformed into Bt HD1 generating the recombinant strains HD1/ pSTAB-ThurH and HD1/pSTAB-ThurH/pSTAB-ChiA74. Antimicrobial and chitinolytic activity tests were performed with recombinant strains. Both strains were able to produce thurincin H up to 72 h with antibacterial activity of ~4000 U mg −1 .The HD1/pSTAB-ThurH/pSTAB-ChiA74 strain also showed chitinolytic activity of ~23 mU mg −1 at 72 h. All B. thuringiensis strains exhibited crystal formation at 72, and 96 h. In addition, the application of thurincin H in corn seeds increased the germination percentage and root length by 7% and 10%, respectively. Conclusions:We showed that is possible to produce three proteins of biotechnological interest at the sporulation stage in B. thuringiensis, which two of them (thurincin H, and ChiA74) are naturally expressed in the vegetative stage.Significance and Impact of the Study: These results form the basis for developing of a biocontrol and biostimulator product that can be used as an alternative for chemical application.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression of thurincin H, ChiA74 and Cry proteins at the sporulation phase in Bacillus thuringiensis HD1

Rivas

Barboza‐Corona

Massange-Sánchez

et al. 2022

Journal of Applied Microbiology

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“…The following bacteria were used for screening antimicrobial activity against 16 indicator bacteria from a bacterial collection of the Biotechnology and Molecular Microbiology Laboratory at the University of Guanajuato, Mexico (Oros-Flores et al, 2018) (Table 1): E. coli A, Clostridium beijerinckii B, Enterococcus faecium F, E. faecium G, Lactobacillus plantarum LB1, Lactobacillus dextrinicus LB5, Lactobacillus pentosus LB7 and Pediococcus acidilactici LB4. These bacteria were previously isolated from a H 2 P bioreactor fed with untreated wheat straw (Pérez-Rangel et al, 2021).…”

Section: Bacterial Strains and Culture Conditionsmentioning

confidence: 99%

Evaluation of inhibitory compounds produced by bacteria isolated from a hydrogen-producing bioreactor during the self-fermentation of wheat straw

Pérez-Rangel

Valdez‐Vazquez

Martínez-Zavala

et al. 2022

Journal of Applied Microbiology

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Aims:The objective of this study was to evaluate the inhibitory activity of compounds secreted by bacteria isolated from a hydrogen-producing bioreactor to understand how these microorganisms interact in this community. Methods and Results:In vitro inhibitory assays were performed using samples secreted by bacteria subject to different treatments to determine if their inhibitory effect was due to organic acids, non-proteinaceous compounds or bacteriocin-like inhibitory substances (BLIS). Bacterial isolated were suppressed 43%, 30% and 27% by neutralized, precipitated and non-neutralized cell-free supernatants, respectively.Non-hydrogen producers (non-H 2 P) lactic acid bacteria (LAB) (Lactobacillus plantarum LB1, Lactobacillus pentosus LB7, Pediococcus acidilactici LB4) and hydrogen producers (H 2 P) LAB (Enterococcus faecium F) were inhibited by the production of organic acids, non-proteinaceous compounds and BLIS. Meanwhile, the obligate anaerobe H 2 P (Clostridium beijerinckii B) inhibited by the production of non-proteinaceous compounds and BLIS. The presence of BLIS was confirmed when proteolytic enzymes affected the inhibitory activity of secreted proteins in values ranging from 20% to 42%. The BLIS produced by L. plantarum LB1, P. acidilactici LB4, L. pentosus LB7 and E. faecium F showed molecular masses of ~11, 25, 20 and 11 kDa, respectively. Conclusions: It was demonstrated antagonistic interactions between Lactobacillus-Enterococcus and Pediococcus-Enterococcus species, generated by the secretion of organic acids, non-proteinaceous compounds and BLIS. Significance and Impact of the Study:We report the interactions between LAB isolated from hydrogen-producing bioreactors. These interactions might impact the dynamics of the microbial population during hydrogen generation. Our work lays a foundation for strategies that allow controlling bacteria that can affect hydrogen production.

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“…El empleo de estas cepas nos permite generar proteínas recombinantes con interés biotecnológico. Thurincina H es una bacteriocina producida por Bacillus thuringiensis, tiene muchas características favorables para ser considerada una proteína de interés; entre ellas destacan que tiene un amplio espectro de actividad contra diferentes bacterias tanto Gram positivas como Gram negativas, algunos de ellos contaminadores de alimentos; además, esta bacteriocina es activa en pH, que va de muy ácido hasta alcalino (pH 3.0-9.0), una ventaja con respecto a nisina, la cual actúa a pH ácido mayormente (Casados-Vázquez et al, 2017;Lee et al, 2009;Oros-Flores et al, 2018). También es resistente a altas temperaturas (hasta 80 °C) y a la liofilización (datos no publicados).…”

Section: Introductionunclassified

El operón thnRDE de B. thuringiensis le brinda inmunidad a Lactococcus lactis contra thurincina H

et al. 2022

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Lactococcus lactis es una bacteria ácido-láctica de gran importancia para la industria de los alimentos. Es una cepa segura, y por tanto se han diseñado sistemas para la expresión de proteínas recombinantes de interés biotecnológico. Dentro de las proteínas con importancia biotecnológica se encuentra la bacteriocina thurincina H, esta tiene muchas características favorables para ser empleada como bioconservador en alimentos. Se ha reportado que necesita al menos nueve genes para su biosíntesis, dentro de estos están los genes thnRDE que brindan inmunidad a la cepa productora nativa. El objetivo de este trabajo fue integrar estos tres genes a la cepa L. lactis para hacerla resistente a thurincina H y poder producirla en un futuro. Para lograrlo, se clonaron los marcos abiertos de lectura (ORF, por sus siglas en inglés) de los genes thnRDE en un vector para L. lactis, y se transformaron en la cepa sensible. Se logró obtener una cepa de L. lactis resistente a thurincina H que podrá ser empleada para su producción heteróloga segura.

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Co-synthesis of kenyacin 404 and heterologous thurincin H enhances the antibacterial activity of Bacillus thuringiensis

Cited by 9 publications

References 28 publications

Expression of thurincin H, ChiA74 and Cry proteins at the sporulation phase in Bacillus thuringiensis HD1

Expression of thurincin H, ChiA74 and Cry proteins at the sporulation phase in Bacillus thuringiensis HD1

Evaluation of inhibitory compounds produced by bacteria isolated from a hydrogen-producing bioreactor during the self-fermentation of wheat straw

El operón thnRDE de B. thuringiensis le brinda inmunidad a Lactococcus lactis contra thurincina H

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