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Journal of Bacteriology May 2023Next to Escherichia coli, Bacillus subtilis is the most studied and best understood organism that also serves as a model for many important pathogens. Due to its ability... (Review)
Review
Next to Escherichia coli, Bacillus subtilis is the most studied and best understood organism that also serves as a model for many important pathogens. Due to its ability to form heat-resistant spores that can germinate even after very long periods of time, B. subtilis has attracted much scientific interest. Another feature of B. subtilis is its genetic competence, a developmental state in which B. subtilis actively takes up exogenous DNA. This makes B. subtilis amenable to genetic manipulation and investigation. The bacterium was one of the first with a fully sequenced genome, and it has been subject to a wide variety of genome- and proteome-wide studies that give important insights into many aspects of the biology of B. subtilis. Due to its ability to secrete large amounts of proteins and to produce a wide range of commercially interesting compounds, B. subtilis has become a major workhorse in biotechnology. Here, we review the development of important aspects of the research on B. subtilis with a specific focus on its cell biology and biotechnological and practical applications from vitamin production to concrete healing. The intriguing complexity of the developmental programs of B. subtilis, paired with the availability of sophisticated tools for genetic manipulation, positions it at the leading edge for discovering new biological concepts and deepening our understanding of the organization of bacterial cells.
Topics: Bacillus subtilis; Biotechnology; Spores, Bacterial
PubMed: 37140386
DOI: 10.1128/jb.00102-23 -
Biological Chemistry Nov 2020
Topics: Bacillus subtilis; Escherichia coli; Humans
PubMed: 32918804
DOI: 10.1515/hsz-2020-0229 -
Journal of Bacteriology Nov 2019Reproduction in the bacterial kingdom predominantly occurs through binary fission-a process in which one parental cell is divided into two similarly sized daughter... (Review)
Review
Reproduction in the bacterial kingdom predominantly occurs through binary fission-a process in which one parental cell is divided into two similarly sized daughter cells. How cell division, in conjunction with cell elongation and chromosome segregation, is orchestrated by a multitude of proteins has been an active area of research spanning the past few decades. Together, the monumental endeavors of multiple laboratories have identified several cell division and cell shape regulators as well as their underlying regulatory mechanisms in rod-shaped and , which serve as model organisms for Gram-negative and Gram-positive bacteria, respectively. Yet our understanding of bacterial cell division and morphology regulation is far from complete, especially in noncanonical and non-rod-shaped organisms. In this review, we focus on two proteins that are highly conserved in Gram-positive organisms, DivIVA and its homolog GpsB, and attempt to summarize the recent advances in this area of research and discuss their various roles in cell division, cell growth, and chromosome segregation in addition to their interactome and posttranslational regulation.
Topics: Bacillus subtilis; Bacterial Proteins; Cell Division; Cell Proliferation; Chromosome Segregation; Protein Processing, Post-Translational
PubMed: 31405912
DOI: 10.1128/JB.00245-19 -
Current Issues in Molecular Biology 2021The cell wall of is a rigid structure on the outside of the cell that forms the first barrier between the bacterium and the environment, and at the same time maintains... (Review)
Review
The cell wall of is a rigid structure on the outside of the cell that forms the first barrier between the bacterium and the environment, and at the same time maintains cell shape and withstands the pressure generated by the cell's turgor. In this review, the chemical composition of peptidoglycan, teichoic and teichuronic acids, the polymers that comprise the cell wall, and the biosynthetic pathways involved in their synthesis will be discussed, as well as the architecture of the cell wall. has been the first bacterium for which the role of an actin-like cytoskeleton in cell shape determination and peptidoglycan synthesis was identified and for which the entire set of peptidoglycan synthesizing enzymes has been localised. The role of the cytoskeleton in shape generation and maintenance will be discussed and results from other model organisms will be compared to what is known for . Finally, outstanding questions in the field of cell wall synthesis will be discussed.
Topics: Bacillus subtilis; Biosynthetic Pathways; Cell Wall; Cytoskeleton; Peptidoglycan; Uronic Acids
PubMed: 33048060
DOI: 10.21775/cimb.041.539 -
Journal of Applied Microbiology Jun 2021Increasing demands for bioactive compounds have motivated researchers to employ micro-organisms to produce complex natural products. Currently, Bacillus subtilis has... (Review)
Review
Increasing demands for bioactive compounds have motivated researchers to employ micro-organisms to produce complex natural products. Currently, Bacillus subtilis has been attracting lots of attention to be developed into terpenoids cell factories due to its generally recognized safe status and high isoprene precursor biosynthesis capacity by endogenous methylerythritol phosphate (MEP) pathway. In this review, we describe the up-to-date knowledge of each enzyme in MEP pathway and the subsequent steps of isomerization and condensation of C5 isoprene precursors. In addition, several representative terpene synthases expressed in B. subtilis and the engineering steps to improve corresponding terpenoids production are systematically discussed. Furthermore, the current available genetic tools are mentioned as along with promising strategies to improve terpenoids in B. subtilis, hoping to inspire future directions in metabolic engineering of B. subtilis for further terpenoid cell factory development.
Topics: Alkyl and Aryl Transferases; Bacillus subtilis; Biosynthetic Pathways; Butadienes; Erythritol; Hemiterpenes; Industrial Microbiology; Metabolic Engineering; Sugar Phosphates; Terpenes
PubMed: 33098223
DOI: 10.1111/jam.14904 -
Methods in Molecular Biology (Clifton,... 2022Bacillus subtilis is a widely studied Gram-positive bacterium that serves as an important model for understanding processes critical for several areas of biology...
Bacillus subtilis is a widely studied Gram-positive bacterium that serves as an important model for understanding processes critical for several areas of biology including biotechnology and human health. B. subtilis has several advantages as a model organism: it is easily grown under laboratory conditions, it has a rapid doubling time, it is relatively inexpensive to maintain, and it is nonpathogenic. Over the last 50 years, advancements in genetic engineering have continued to make B. subtilis a genetic workhorse in scientific discovery. In this chapter, we describe methods for traditional gene disruptions, use of gene deletion libraries from the Bacillus Genetic Stock Center, allelic exchange, CRISPRi, and CRISPR/Cas9. Additionally, we provide general materials and equipment needed, strengths and limitations, time considerations, and troubleshooting notes to perform each method. Use of the methods outlined in this chapter will allow researchers to create gene insertions, deletions, substitutions, and RNA interference strains through a variety of methods custom to each application.
Topics: Bacillus subtilis; Bacterial Proteins; CRISPR-Cas Systems; Gene Editing; Genetic Engineering; Humans
PubMed: 35583738
DOI: 10.1007/978-1-0716-2233-9_11 -
Microbial Cell Factories Sep 2020Due to its clear inherited backgrounds as well as simple and diverse genetic manipulation systems, Bacillus subtilis is the key Gram-positive model bacterium for studies... (Review)
Review
Due to its clear inherited backgrounds as well as simple and diverse genetic manipulation systems, Bacillus subtilis is the key Gram-positive model bacterium for studies on physiology and metabolism. Furthermore, due to its highly efficient protein secretion system and adaptable metabolism, it has been widely used as a cell factory for microbial production of chemicals, enzymes, and antimicrobial materials for industry, agriculture, and medicine. In this mini-review, we first summarize the basic genetic manipulation tools and expression systems for this bacterium, including traditional methods and novel engineering systems. Secondly, we briefly introduce its applications in the production of chemicals and enzymes, and summarize its advantages, mainly focusing on some noteworthy products and recent progress in the engineering of B. subtilis. Finally, this review also covers applications such as microbial additives and antimicrobials, as well as biofilm systems and spore formation. We hope to provide an overview for novice researchers in this area, offering them a better understanding of B. subtilis and its applications.
Topics: Bacillus subtilis; Bacterial Proteins; Biocompatible Materials; Biological Products; Enzymes; Gene Expression Regulation, Bacterial; Genetic Engineering; Industrial Microbiology; Recombinant Proteins; Vitamins
PubMed: 32883293
DOI: 10.1186/s12934-020-01436-8 -
Molecular Microbiology Apr 2021Both isomeric forms of alanine play a crucial role in bacterial growth and viability; the L-isomer of this amino acid is one of the building blocks for protein...
Both isomeric forms of alanine play a crucial role in bacterial growth and viability; the L-isomer of this amino acid is one of the building blocks for protein synthesis, and the D-isomer is incorporated into the bacterial cell wall. Despite a long history of genetic manipulation of Bacillus subtilis using auxotrophic markers, the genes involved in alanine metabolism have not been characterized fully. In this work, we genetically characterized the major enzymes involved in B. subtilis alanine biosynthesis and identified an alanine permease, AlaP (YtnA), which we show has a major role in the assimilation of D-alanine from the environment. Our results provide explanations for the puzzling fact that growth of B. subtilis does not result in the significant accumulation of extracellular D-alanine. Interestingly, we find that in B. subtilis, unlike E. coli where multiple enzymes have a biochemical activity that can generate alanine, the primary synthetic enzyme for alanine is encoded by alaT, although a second gene, dat, can support slow growth of an L-alanine auxotroph. However, our results also show that Dat mediates the synthesis of D-alanine and its activity is influenced by the abundance of L-alanine. This work provides valuable insights into alanine metabolism that suggests that the relative abundance of D- and L-alanine might be linked with cytosolic pool of D and L-glutamate, thereby coupling protein and cell envelope synthesis with the metabolic status of the cell. The results also suggest that, although some of the purified enzymes involved in alanine biosynthesis have been shown to catalyze reversible reactions in vitro, most of them function unidirectionally in vivo.
Topics: Alanine; Amino Acid Transport Systems; Bacillus subtilis; Bacterial Proteins; Biosynthetic Pathways; Membrane Transport Proteins; Transaminases
PubMed: 33155333
DOI: 10.1111/mmi.14640 -
Transcription Aug 2021The low G + C Gram-positive bacteria represent some of the most medically and industrially important microorganisms. They are relied on for the production of food and... (Review)
Review
The low G + C Gram-positive bacteria represent some of the most medically and industrially important microorganisms. They are relied on for the production of food and dietary supplements, enzymes and antibiotics, as well as being responsible for the majority of nosocomial infections and serving as a reservoir for antibiotic resistance. Control of gene expression in this group is more highly studied than in any bacteria other than the Gram-negative model Escherichia coli, yet until recently no structural information on RNA polymerase (RNAP) from this group was available. This review will summarize recent reports on the high-resolution structure of RNAP from the model low G + C representative Bacillus subtilis, including the role of auxiliary subunits and , and outline approaches for the development of antimicrobials to target RNAP from this group.
Topics: Bacillus subtilis; Bacterial Proteins; DNA-Directed RNA Polymerases; Gram-Positive Bacteria; Transcription, Genetic
PubMed: 34403307
DOI: 10.1080/21541264.2021.1964328 -
The Journal of General and Applied... Sep 2022Bacillus subtilis Marburg 168 is a unique platform for genome engineering and genome synthesis. Genome scale DNA sequences can be synthesized by repeated integration of...
Bacillus subtilis Marburg 168 is a unique platform for genome engineering and genome synthesis. Genome scale DNA sequences can be synthesized by repeated integration of small DNA segments in the B. subtilis genome. The small DNA segments are collectively called dominos, and should cover the target genome. The B. subtilis strains which have been designed for use in the domino method are collectively called BGM: Bacillus subtilis Genome for Manipulation. The BGM system has been used to produce various genomes in the B. subtilis genome. The synthesized genomes have been demonstrated to be stably maintained as part of the B. subtilis genome. Instability of the synthesized genomes have been observed in genomes with Guanine plus Cytosine contents much higher or lower than that of BGM. The largest synthesized genome produced using this approach to date is that from Synecchosystis PCC6803, a photosynthetic microbe with a genome size of about 3.5 Mbp. The domino method depends on transformation, using the natural competence of B. subtilis. An alternative DNA uptake system, conjugational transfer, has been studied for the past 20 years. A self-transmissible plasmid named pLS20 has been used for the transfer and delivery of large amounts of DNA between B. subtilis. The BGM system is a unique platform for handling very large amounts of DNA from synthesis to dissemination to other cells, and has broad applications in research and practice.
Topics: Bacillus subtilis; DNA; Genome, Bacterial; Plasmids
PubMed: 35491108
DOI: 10.2323/jgam.2021.12.001