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Hongwei Liu, Shuli Yin, Likang An, Genwei Zhang, Huicai Cheng, Yanhua Xi, Guanhui Cui, Feiyan Zhang, Liping Zhang
Bacillus subtilis BSD-2, isolated from cotton (Gossypium spp.), had strong antagonistic activity to Verticillium dahlia Kleb and Botrytis cinerea. We sequenced and annotated the BSD-2 complete genome to help us the better use of this strain, which has surfactin, bacilysin, bacillibactin, subtilosin A, Tas A and a potential class IV lanthipeptide biosynthetic pathways.
July 20, 2016: Journal of Biotechnology
Alhosna Benjdia, Alain Guillot, Benjamin Lefranc, Hubert Vaudry, Jérôme Leprince, Olivier Berteau
AlbA is a radical SAM enzyme catalyzing the formation of three unusual thioether bonds in the antibiotic subtilosin A. We demonstrate here that AlbA catalyzes direct Cα H-atom abstraction and likely contains three essential [4Fe-4S] centers. This leads us to propose novel mechanistic perspectives for thioether bond catalysis by radical SAM enzymes.
May 7, 2016: Chemical Communications: Chem Comm
Paul M Himes, Scott E Allen, Sungwon Hwang, Albert A Bowers
Sactipeptides are peptide-derived natural products that are processed by remarkable, radical-mediated cysteine sulfur to α-carbon coupling reactions. The resulting sactionine thioether linkages give rise to the unique defined structures and concomitant biological activities of sactipeptides. An E. coli heterologous expression system, based on the biosynthesis of one such sactipeptide, subtilosin A, is described and this expression system is exploited to probe the promiscuity of the subtilosin A sactionine bond-forming enzyme, AlbA...
June 17, 2016: ACS Chemical Biology
Poochit Nonejuie, Rachelle M Trial, Gerald L Newton, Anne Lamsa, Varahenage Ranmali Perera, Julieta Aguilar, Wei-Ting Liu, Pieter C Dorrestein, Joe Pogliano, Kit Pogliano
Although most clinically used antibiotics are derived from natural products, identifying new antibacterial molecules from natural product extracts is difficult due to the complexity of these extracts and the limited tools to correlate biological activity with specific molecules. Here, we show that bacterial cytological profiling (BCP) provides a rapid method for mechanism of action determination on plates and in complex natural product extracts and for activity-guided purification. We prepared an extract from Bacillus subtilis 3610 that killed the Escherichia coli lptD mutant and used BCP to observe two types of bioactivities in the unfractionated extract: inhibition of translation and permeablization of the cytoplasmic membrane...
May 2016: Journal of Antibiotics
Harsh Mathur, Mary C Rea, Paul D Cotter, Colin Hill, R Paul Ross
The sactibiotics are a recently designated subclass of bacteriocins that contain characteristic cysteine sulphur to α -carbon linkages mediated through post-translational modifications. They are a relatively small subclass of bacteriocins compared to the most thoroughly studied lantibiotics. The sactibiotics that have been extensively studied thus far are thuricin CD, subtilosin A, thurincin H, and propionicin F. Despite their recent discovery, there have already been significant advances made in the study of sactibiotics, most notably the discovery of the narrow spectrum anti-Clostridium difficile sactibiotic, thuricin CD...
2015: Current Protein & Peptide Science
Xiaoming Liu, Jae Yong Lee, Seon-Ju Jeong, Kye Man Cho, Gyoung Min Kim, Jung-Hye Shin, Jong-Sang Kim, Jeong Hwan Kim
A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80°C but reduced by 50% after 15 min at 90°C. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature...
September 2015: Journal of Microbiology and Biotechnology
Ammar Algburi, Anna Volski, Michael L Chikindas
The purpose of this study was to evaluate the ability of clindamycin and metronidazole to synergize with natural antimicrobials against biofilms of bacterial vaginosis (BV)-associated Gardnerella vaginalis. Minimum bactericidal concentrations for biofilm cells (MBCs-B) were determined for each antimicrobial. The MBCs-B of lauramide arginine ethyl ester (LAE), subtilosin, clindamycin and metronidazole were 50, 69.5, 20 and 500 μg mL(-1), respectively. A checkerboard assay and isobologram were used to analyze the type of interactions between these antimicrobials...
July 2015: Pathogens and Disease
John A Latham, Anthony T Iavarone, Ian Barr, Prerak V Juthani, Judith P Klinman
Pyrroloquinoline quinone (PQQ) is a product of a ribosomally synthesized and post-translationally modified pathway consisting of five conserved genes, pqqA-E. PqqE is a radical S-adenosylmethionine (RS) protein with a C-terminal SPASM domain, and is proposed to catalyze the formation of a carbon-carbon bond between the glutamate and tyrosine side chains of the peptide substrate PqqA. PqqD is a 10-kDa protein with an unknown function, but is essential for PQQ production. Recently, in Klebsiella pneumoniae (Kp), PqqD and PqqE were shown to interact; however, the stoichiometry and KD were not obtained...
May 15, 2015: Journal of Biological Chemistry
Veronica L Cavera, Anna Volski, Michael L Chikindas
Bacterial vaginosis (BV) is a common, recurrent vaginal infection linked to increased chances of preterm delivery, incidence of sexually transmitted infections and fertility problems. BV is caused by a shift of the vaginal ecosystem from predominately Lactobacillus to a multispecies Actinomyces biofilm with the most common representatives identified as Gardnerella vaginalis and Prevotella spp. Current treatments have been associated with increased resistance as well as negative effects on healthy microbiota...
June 2015: Probiotics and Antimicrobial Proteins
Alicja Daszczuk, Yonathan Dessalegne, Ismaêl Drenth, Elbrich Hendriks, Emeraldo Jo, Tom van Lente, Arjan Oldebesten, Jonathon Parrish, Wlada Poljakova, Annisa A Purwanto, Renske van Raaphorst, Mirjam Boonstra, Auke van Heel, Martijn Herber, Sjoerd van der Meulen, Jeroen Siebring, Robin A Sorg, Matthias Heinemann, Oscar P Kuipers, Jan-Willem Veening
Here, we developed a cell-based biosensor that can assess meat freshness using the Gram-positive model bacterium Bacillus subtilis as a chassis. Using transcriptome analysis, we identified promoters that are specifically activated by volatiles released from spoiled meat. The most strongly activated promoter was PsboA, which drives expression of the genes required for the bacteriocin subtilosin. Next, we created a novel BioBrick compatible integration plasmid for B. subtilis and cloned PsboA as a BioBrick in front of the gene encoding the chromoprotein amilGFP inside this vector...
December 19, 2014: ACS Synthetic Biology
Nalisa Khochamit, Surasak Siripornadulsil, Peerapol Sukon, Wilailak Siripornadulsil
The antimicrobial activity and probiotic properties of Bacillus subtilis strain KKU213, isolated from local soil, were investigated. The cell-free supernatant (CFS) of a KKU213 culture containing crude bacteriocins exhibited inhibitory effects on Gram-positive bacteria, including Bacillus cereus, Listeria monocytogenes, Micrococcus luteus, and Staphylococcus aureus. The antibacterial activity of the CFS precipitated with 40% ammonium sulfate (AS) remained even after treatment at 60 and 100 °C, at pH 4 and 10 and with proteolytic enzymes, detergents and heavy metals...
January 2015: Microbiological Research
V M Quintana, N I Torres, M B Wachsman, P J Sinko, V Castilla, M Chikindas
AIMS: In this study, we evaluated the antiviral activity of subtilosin, a cyclical peptide isolated from Bacillus amyloliquefaciens, against herpes simplex virus type 2 (HSV-2) in cell cultures and we investigated subtilosin mode of action. METHODS AND RESULTS: We determined, using a virus yield inhibition assay, that noncytotoxic concentrations of subtilosin inhibit HSV-2 replication in Vero cell cultures. Subtilosin strongly inhibited extracellular and total virus production even when it was added at 8 h postinfection indicating that not only virus release but also viral particle formation is impeded by the antiviral peptide...
November 2014: Journal of Applied Microbiology
Andrey V Karlyshev, Vyacheslav G Melnikov, Michael L Chikindas
In this report, we present a draft sequence of Bacillus subtilis KATMIRA1933. Previous studies demonstrated probiotic properties of this strain partially attributed to production of an antibacterial compound, subtilosin. Comparative analysis of this strain's genome with that of a commercial probiotic strain, B. subtilis Natto, is presented.
2014: Genome Announcements
Sujata Sundara Rajan, Veronica L Cavera, Xiaoping Zhang, Yashveer Singh, Michael L Chikindas, Patrick J Sinko
Current treatment options for bacterial vaginosis (BV) have been shown to be inadequate at preventing recurrence and do not provide protection against associated infections, such as that with HIV. This study examines the feasibility of incorporating the antimicrobial peptide subtilosin within covalently cross-linked polyethylene glycol (PEG)-based hydrogels for vaginal administration. The PEG-based hydrogels (4% and 6% [wt/vol]) provided a two-phase release of subtilosin, with an initial rapid release rate of 4...
May 2014: Antimicrobial Agents and Chemotherapy
In-Cheol Yeo, Nam Keun Lee, Byung Wook Yang, Young Tae Hahm
Bacillus subtilis SC-8 produces an antibiotic that has narrow antagonistic activity against bacteria in the Bacillus cereus group. In B. cereus group bacteria, peptide-activating PlcR (PapR) plays a significant role in regulating the transcription of virulence factors. When B. subtilis SC-8 and B. cereus are co-cultured, PapR is assumed to stimulate antibiotic production by B. subtilis SC-8. To better understand the effect of PapR on this interspecies interaction, the global transcriptome profile of B. subtilis SC-8 was analyzed in the presence of PapR...
January 2014: Applied Biochemistry and Biotechnology
Andrew D Frock, Clemente I Montero, Sara E Blumer-Schuette, Robert M Kelly
The genome of the hyperthermophilic bacterium Thermotoga maritima encodes numerous putative peptides/proteins of 100 amino acids or less. While most of these open reading frames (ORFs) are transcribed during growth, their corresponding physiological roles are largely unknown. The onset of stationary phase in T. maritima was accompanied by significant morphological changes and upregulation of several ORFs located in the TM1298-TM1336 genome locus. This region contains putative HicAB toxin-antitoxin pairs, hypothetical proteins, radical S-adenosylmethionine (SAM) enzymes, and ABC transporters...
November 2013: Applied and Environmental Microbiology
Fernanda Leal Leães, Renata Voltolini Velho, Danielle Gregorio Gomes Caldas, Juliana Venturini Pinto, Siu Mui Tsai, Adriano Brandelli
Temperature and pH are key factors influencing the production of antimicrobial peptides. In this work, qRT-PCR methodology was used to demonstrate the effect of these two variables on sboA (subtilosin A) and ituD (iturin A) expression in Bacillus sp. P11, an isolate from aquatic environment of the Amazon. Bacillus sp. P11 was incubated in BHI broth for 36 h at 30, 37 and 42 °C, and the pH values were 6.0, 7.4 and 8.0. The production of subtilosin A and iturin A was confirmed by mass spectrometry. The sboA expression increased 200-fold when the initial pH was 8...
July 2013: Antonie Van Leeuwenhoek
Nicolás I Torres, Katia Sutyak Noll, Shiqi Xu, Ji Li, Qingrong Huang, Patrick J Sinko, Mónica B Wachsman, Michael L Chikindas
In the present study the antiviral properties of the bacteriocin subtilosin against Herpes simplex virus type 1 (HSV-1) and the safety and efficacy of a subtilosin-based nanofiber formulation were determined. High concentrations of subtilosin, the cyclical antimicrobial peptide produced by Bacillus amyloliquefaciens, were virucidal against HSV-1. Interestingly, at non-virucidal concentrations, subtilosin inhibited wild type HSV-1 and aciclovir-resistant mutants in a dose-dependent manner. Although the exact antiviral mechanism is not fully understood, time of addition experiments and western blot analysis suggest that subtilosin does not affect viral multiplication steps prior to protein synthesis...
March 1, 2013: Probiotics and Antimicrobial Proteins
Renata Voltolini Velho, Ana Paula Basso, Jeferson Segalin, Luis Fernando Costa-Medina, Adriano Brandelli
This report demonstrates the usefulness of PCR for the genes spaS and sboA as a means of identifying Bacillus strains with a potential to produce subtilin and subtilosin A. One collection strain and five Bacillus spp. isolated from aquatic environments in the Amazon basin were screened by PCR using primers for sboA and spaS designed specifically for this study. The sequences of the PCR products showed elevated homology with previously described spaS and sboA genes. Antimicrobial peptides were isolated from culture supernatants and analyzed by mass spectrometry...
March 2013: Genetics and Molecular Biology
Clarisse S Compaoré, Dennis S Nielsen, Labia I I Ouoba, Torben S Berner, Kristian F Nielsen, Hagrétou Sawadogo-Lingani, Bréhima Diawara, Georges A Ouédraogo, Mogens Jakobsen, Line Thorsen
Bikalga is a Hibiscus sabdariffa seed fermented condiment widely consumed in Burkina Faso and neighboring countries. The fermentation is dominated by Bacillus subtilis group species. Ten B. subtilis subsp. subtilis (six isolates) and Bacillus licheniformis (four isolates) isolated from traditional Bikalga were examined for their antimicrobial activity against a panel of 36 indicator organisms including Gram-positive and Gram-negative bacteria and yeasts. The Bacillus spp. isolates showed variable inhibitory abilities depending on the method used...
April 1, 2013: International Journal of Food Microbiology
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