Codon Optimization is Required to Express Fluorogenic Reporter Proteins in Lactococcus lactis.

Lactococcus lactis is a Gram-positive bacterium used to produce fermented foods and heterologous proteins. Its Nisin-controlled gene expression system stands out for its versatility and safety. However, the lower GC content in its genome may lead to some limitations in protein production.

The Perpetual Vector Mosquito Threat and Its Eco-Friendly Nemeses.

Mosquitoes are the most notorious arthropod vectors of viral and parasitic diseases for which approximately half the world's population, ~4,000,000,000, is at risk. Integrated pest management programs (IPMPs) have achieved some success in mitigating the regional transmission and persistence of these diseases. However, as many vector-borne diseases remain pervasive, it is obvious that IPMP successes have not been absolute in eradicating the threat imposed by mosquitoes.

The chitin-binding domain of Bacillus thuringiensis ChiA74 inhibits gram-negative bacterial and fungal pathogens of humans and plants.

The chitinase ChiA74 is synthesized by Bacillus thuringiensis and possesses a modular organization composed of four domains. In the C-terminal of the enzyme is located the chitin-binding domain (CBD), which has not been isolated as a single unit or characterized. Here, we aimed to isolate the ChiA74's CBD as a single unit, determine the binding properties, and evaluate its antimicrobial and hemolytic activities.

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

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.

Thurincin H Is a Nonhemolytic Bacteriocin of Bacillus thuringiensis with Potential for Applied Use.

Thurincin H, a bacteriocin produced by Bacillus thuringiensis, exhibits antibacterial activity against Gram-positive and Gram-negative bacteria. While much is known about its expression and antimicrobial spectrum, its hemolytic property has yet to be established. In this study, thurincin H was produced in a plasmid-free acrystalliferous strain of B.

The thnI gene is not required for thurincin H biosynthesis or immunity.

Thurincin H is a bacteriocin produced by Bacillus thuringiensis, it is encoded in a group of ten genes, most of which have been characterized experimentally or by homology. However, the activity of the thnI gene encoding a 95 amino acid ORF remains unknown. In this work, the thnI gene was cloned under the regulation of two promoters and transformed into a sensitive strain to determine if it acts as an immunity protein.

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

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.

The duo Clostridium and Lactobacillus linked to hydrogen production from a lignocellulosic substrate.

The study aimed to identify interspecies interactions within a native microbial community present in a hydrogen-producing bioreactor fed with two wheat straw cultivars. The relationships between the microbial community members were studied building a canonical correspondence analysis and corroborated through in vitro assays. The results showed that the bioreactor reached a stable hydrogen production of ca.

Analysis of virulence traits of isolated from bovine mastitis in semi-intensive and family dairy farms.

Background: is one of the main microorganisms that causes bovine mastitis, and its well-known virulence characteristics and interactions with the environment are used to aid the design of more efficient therapies.

Objectives: To determine whether the virulence traits, such as antibiotic resistance and biofilm-forming and internalization abilities, of isolated from bovine mastitis are related to dairy production system types.

Methods: The study was performed in the Mexican states of Guanajuato and Michoacan.

Agave: a natural renewable resource with multiple applications.

Agaves are a group of succulent plants that thrive in arid or semiarid environments. Indeed, genes associated with their resilience are a potential resource for genetic engineering of other agronomically important crops grown in adverse climates. Agave is mainly used for the production of distilled (spirits) and non-distilled alcoholic beverages, including tequila, mezcal, bacanora, raicilla, and pulque, all of which have special connections to Mexican history and culture, and contribute to the Mexican economy.

Improving the yields of thurincin H in a native producer strain.

Thurincin H is a bacteriocin produced by Bacillus thuringiensis with activity against a wide range of bacteria, including Gram positive and Gram negative. Disadvantages of producing thurincin H in B. thuringiensis is the low production level in the native strain probably due to the highly regulated mechanism of biosynthesis.

Chitinolytic activity of phylogenetically diverse Bacillus cereus sensu lato from natural environments.

Bacillus cereus s.l., Gram-positive endospore-forming bacilli, persist ubiquitously in different natural habitats and play various ecological roles.

Class I defensins (BraDef) from broccoli (Brassica oleracea var. italica) seeds and their antimicrobial activity.

The objective of this study was to determine whether seeds of Brassica oleracea var. italica (i.e.

Chitinases of : Phylogeny, Modular Structure, and Applied Potentials.

The most important bioinsecticide used worldwide is and its hallmark is a rich variety of insecticidal Cry protein, many of which have been genetically engineered for expression in transgenic crops. Over the past 20 years, the discovery of other insecticidal proteins and metabolites synthesized by , including chitinases, antimicrobial peptides, vegetative insecticidal proteins (VIP), and siderophores, has expanded the applied value of this bacterium for use as an antibacterial, fungicidal, and nematicidal resource. These properties allow us to view not only as an entity for the production of a particular metabolite, but also as a multifaceted microbial factory.

New bacteriocin-like substances produced by Streptomyces species with activity against pathogens.

Streptomyces spp. are Gram-positive bacteria well-known for their ability to produce antibiotics and other metabolites, but few studies on bacteriocins produced by these bacteria have been reported. We tested eight Streptomyces strains against different pathogenic bacteria, and selected S.

The crystal structure of the chitinase ChiA74 of Bacillus thuringiensis has a multidomain assembly.

There is no structural information about any chitinase synthesized by Bacillus thuringiensis, the most successful microbial insect larvicide used worldwide. In this study, we solved the 3D structure of the chitinase ChiA74 at 2.26 Å.

Expression of ChiA74∆sp and its truncated versions in Bacillus thuringiensis HD1 using a vegetative promoter maintains the integrity and toxicity of native Cry1A toxins.

Our objective was to determine whether a recombinant chitinase ChiA74∆sp of Bacillus thuringiensis and its truncated versions (ChiA74∆sp-60, ChiA74∆sp-50) could be produced in B. thuringiensis HD1 with no detrimental effect on the size and insecticidal activity of the native bipyramidal Cry crystal. chiA-p, the promoter used to drive chitinase gene expression, was active during vegetative growth of CryB.

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

Objectives: To develop a recombinant strain of Bacillus thuringiensis that synthesizes two bacteriocins that enhance the antibacterial potency of the strain and that could have applied clinical and industrial value.

Results: We cloned the thurincin H cluster into the pHT3101 vector by assembling two genetic cassettes harboring genes for the synthesis, modification, immunity and transport of thurincin H. This construct was used to transform a thurincin H-sensitive strain of B.

Antibiotic susceptibility of Staphylococcus aureus isolated from subclinical bovine mastitis cases and in vitro efficacy of bacteriophage.

Staphylococcus aureus is an opportunistic pathogen that may cause severe infections in livestock, and represents the major cause of mastitis in dairy cows. Currently, instead of using antibiotics, new strategies are sought to reduce this clinical health problem. The aim of this study was to determine the efficacy of phage therapy to kill S.

Regulator ThnR and the ThnDE ABC transporter proteins confer autoimmunity to thurincin H in Bacillus thuringiensis.

The structural gene that encodes thurincin H, a bacteriocin produced by Bacillus thuringiensis, is harboured in a genetic cluster (thnP, E, D, R, A1, A2, A3, B, T, I) that controls its synthesis, modification, secretion and autoimmunity. The specific genes in the cassette that confer immunity in B. thuringiensis to thurincin H are unknown.

Characterization and Identification of Cryptic Biopeptides in (Wangenh K. Koch) Storage Proteins.

The objective of this research was to identify and characterize the encoded peptides present in nut storage proteins of . It was found, through in silico prediction, proteomic analysis, and MS spectrometry, that bioactive peptides were mainly found in albumin and glutelin fractions. Glutelin was the major fraction with ~53% of the nut storage proteins containing at least 21 peptides with different putative biological activities, including antihypertensives, antioxidants, immunomodulators, protease inhibitors, and inhibitors of cell cycle progression in cancer cells.

Antimicrobial activity of broccoli (Brassica oleracea var. italica) cultivar Avenger against pathogenic bacteria, phytopathogenic filamentous fungi and yeast.

Aims: The objective of this study was to show whether the edible part of broccoli has antibacterial and antifungal activity against micro-organism of importance in human health and vegetable spoilage, and to test if this effect was partially due to antimicrobial peptides (AMPs).

Methods And Results: Crude extracts were obtained from florets and stems of broccoli cultivar Avenger and the inhibitory effect was demonstrated against pathogenic bacteria (Bacillus cereus, Staphylococcus xylosus, Staphylococcus aureus, Shigella flexneri, Shigella sonnei, Proteus vulgaris), phytopathogenic fungi (Colletotrichum gloeosporioides, Asperigillus niger) and yeasts (Candida albicans and Rhodotorula sp.).

Role of the C-terminal and chitin insertion domains on enzymatic activity of endochitinase ChiA74 of Bacillus thuringiensis.

ChiA74 has modular structure that includes a secretion signal peptide (sp) sequence, and catalytic (CD), chitin insertion (CID), fibronectin type-III (FnIII) and chitin binding (CBD) domains. We described for the first time the existence of a putative CID in ChiA74. Mature ChiA74 lacking its sp sequence (rChiA74Δsp, ∼70kDa) and two truncated versions, rChiA74Δsp-60, rChiA74Δsp-50 lacking, respectively, CBD and CDB-FnIII were produced.

Recombinant Bacillus thuringiensis subsp. kurstaki HD73 strain that synthesizes Cry1Ac and chimeric ChiA74∆sp chitinase inclusions.

In this study, the endochitinase chiA74 gene lacking its secretion signal peptide sequence (chiA74∆sp) was fused in frame with the sequence coding for the C-terminal crystallization domain and transcription terminator of cry1Ac. The chimeric gene was expressed under the strong pcytA-p/STAB-SD promoter system in an acrystalliferous CryB strain of Bacillus thuringiensis and B. thuringiensis subsp.