首页期刊导航|The Journal of General and Applied Microbiology
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The Journal of General and Applied Microbiology
Institute of Applied Microbiology, University of Tokyo
The Journal of General and Applied Microbiology

Institute of Applied Microbiology, University of Tokyo

0022-1260

The Journal of General and Applied Microbiology/Journal The Journal of General and Applied Microbiology
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    From"Bacillus subtilis conference"to"Conference on functional genomics of Gram-positive bacteria"

    Ken-ichi Yoshida
    2页
    查看更多>>摘要：Bacillus subtilis is a microorganism that was described first in 1835(Ehrenberg,1835).It was once named Vibrio subtilis but later in 1872 renamed to its current scientific name(Cohn,1872).Its Japanese name Ko-so-kin derives from the fact that the bacterium is often isolated from hay.One of its subspecies,B.subtilis var.natto,is used to produce traditional fermented soybean food Natto,and it is a microorganism with that Japanese people are familiar particularly.In addition,degradative enzymes secreted by B.subtilis are useful in industrial processes and detergents for household use.These facts make B.subtilis one of the most representatively beneficial microorganisms.The whole genomic sequence of B.subtilis was determined in 1997 by an international collaboration of European countries and Japan(Kunst et al.,1997).Since then,B.subtilis has been one of the model microorganisms studied in basic genetic research and applied to construction of beneficial strains that display desired traits.Furthermore,as one of the organisms with the richest accumulation of knowledge on genetic and biochemical functions,it has been indispensable as a template to understand the other microorganisms that used to be poorly understood.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    Bacillus subtilis 168 as a unique platform enabling synthesis and dissemination of genomes

    Mitsuhiro Itaya
    9页
    查看更多>>摘要：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.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    Full Paper Galactolipids from Arabidopsis thaliana can replace the function of glucolipids in Bacillus subtilis

    Manami KawakamiSatoshi Matsuoka
    8页
    查看更多>>摘要：Arabidopsis thaliana monogalactosyldiacylglycerol synthase 1(AtMGDl)and digalactosyldiacylglycerol synthase 2(AtDGD2)genes introduced into a Bacillus subtilis chromosome with disrupted galE,which encodes UDP-glucose 4-epimerase,enabled the mutant to produce monogalactosyldiacylglycerol.When galE mutant cells are cultivated in galactose containing medium they show abnormal morphology.This phenotype is correlated with a decrease in the amount of glucolipids.Nucleoids of the ugtP and galE mutants were stained by propidium iodide,which does not permeate intact cell membranes,whereas nucleoids of wild type and of a pssA mutant we examined were not stained.Expression of the AtMGDl gene in a ugtP galE double mutant restored cell membrane integrity.Expression of galactolipid synthase genes from a multi-copy plasmid,pDGHisN4,allowed higher production of galactolipids.Activation of the extracytoplasmic function sigma factors SigM,SigV,and SigX,in the ugtP mutant was decreased by expression of AtMGDl,and SigX activity was strongly repressed when both AtMGDl and AtDGD2 genes were expressed in the mutant.We conclude that the number of sugars that bind to diacylglycerol-rather than the exact sugar species-is important for glycolipid function in B.subtilis.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    Analysis of cell death in Bacillus subtilis caused by sesquiterpenes from Chrysopogon zizanioides(L.)Roberty

    Yu ShinjyoNaoya MidorikawaTakashi Matsumoto
    9页
    查看更多>>摘要：Recently,the antibacterial effects of essential oils have been investigated in addition to their therapeutic purposes.Owing to their hydrophobic nature,they are thought to perturb the integrity of the bacterial cell membrane,leading to cell death.Against such antibiotic challenges,bacteria develop mechanisms for cell envelope stress responses(CESR).In Bacillus subtilis,a gram-positive sporulating soil bacterium,the extracytoplasmic function(ECF)sigma factor-mediated response system plays a pivotal role in CESR.Among them,σ~M is strongly involved in response to cell envelope stress,including a shortage of available bactoprenol.Vetiver essential oil,a product of Chrysopogon zizanioides(L.)Roberty root,is also known to possess bactericidal activity.oM was exclusively and strongly induced when the cells were exposed to Vetiver extract,and depletion of multi-ECF sigma factors(AsigM,AsigfV,AsigX,and AsigV)enhanced sensitivity to it.From this quadruple mutant strain,the suppressor strains,which restored resistance to the bactericidal activity of Vetiver extract,emerged,although attempts to obtain resistant strains from the wild type did not succeed.Whole-genome resequencing of the suppressor strains and genetic analysis revealed inactivation of xseB orpnpA,which code for exode-oxyribonuclease or polynucleotide phosphorylase,respectively.This allowed the quadruple mutant strain to escape from cell death caused by Vetiver extract.Composition analysis suggested that the sesquiterpene,khusimol,might contribute to the bactericidal activity of the Vetiver extract.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    New Bacillus subtilis vector,pSSp,as genetic tool for site-specific integration and excision of cloned DNA,and prophage elimination

    Shota SuzukiSachie OsadaDaisuke Imamura
    8页
    查看更多>>摘要：Site-specific recombination(SSR)systems are employed in many genetic mobile elements,including temperate phages,for their integration and excision.Recently,they have also been used as tools for applications in fields ranging from basic to synthetic biology.SPp is a temperate phage of the Siphoviridae family found in the laboratory standard Bacillus subtilis strain 168.SPβ encodes a ser-ine-type recombinase,SprA,and recombination directionality factor(RDF),SprB.SprA catalyzes recombination between the attachment site of the phage,attP,and that of the host,attB,to integrate phage genome into the attB site of the host genome and generate attL and attR at both ends of the prophage genome.SprB works in conjunction with SprA and switches from attBlattP to attL/R recombination,which leads to excision of the prophage.In the present study,we took advantage of this highly efficient recombination system to develop a site-specific integration and excision plasmid vector,named pSSp.It was constructed using pUC plasmid and the SSR system components,attP,sprA and sprB of SPp.pSSp was integrated into the attB site with a significantly high efficiency,and the resulting pSSp-integrated strain also easily eliminated pSSp itself from the host genome by the induction of SprB expression with xylose.This report presents two applications using pSSp that are particularly suitable for gene complementation experiments and for a curing system of SPp prophage,that may serve as a model system for the removal of prophages in other bacteria.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    Visualization and characterization of spore morphogenesis in Paenihacillus polymyxa ATCC39564

    Kimihiro AbeHiroko KatoYuta Hasegawa
    8页
    查看更多>>摘要：Paenibacillus polymyxa is a spore-forming Gram-positive bacterial species.Both its sporulation process and the spore properties are poorly understood.Here,we investigated sporulation in P.polymyxa ATCC39564.When cultured at 37℃ for 24 h in sporulation medium,more than 80% of the total cells in the culture were spores.Time-lapse imaging revealed that cellular morphological changes during sporulation of P.polymyxa were highly similar to those of B.subtilis.We demonstrated that genetic deletion of spoOA,sigE,sigF,sigG,or sigK,which are highly conserved transcriptional regulators in spore-forming bacteria,abolished spore formation.In P.polymyxa,spoOA was required for cell growth in sporulation medium,as well as for the initiation of sporulation.The sigE and sigF mutants formed abnormal multiple asymmetric septa during the early stage of sporulation.The sigG and sigK mutants formed forespores in the sporangium,but they did not become mature.Moreover,fluorescence reporter analysis confirmed compartment-specific gene expression of spoIID and spoVFA in the mother cell and spoIIQ and sspF in the forespore.Transmission electron microscopy imaging revealed that P.polymyxa produces multi-layered endospores but lacking a balloon-shaped exosporium.Our results indicate that spore morphogenesis is conserved between P.polymyxa and B.subtilis.However,P.polymyxa genomes lack many homologues encoding spore-coat proteins that are found in B.subtilis,suggesting that there are differences in the spore coat composition and surface structure between P.polymyxa and B.subtilis.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    Functional analysis of a gene cluster for putative bacteriocin deduced from the genome sequence of Aeribacillus pallidus PI8

    Kyosuke KitaSanako YoshidaShu Ishikawa
    8页
    查看更多>>摘要：Bacteriocins are a large family of peptides synthesized ribosomally by a variety of bacterial species.The genome of one of the thermophilic Gram-positive bacteria,Aeribacillus pallidus PI8,was found to possess an operon comprising five genes possibly involved in the production of a putative bacteriocin that was named pcnABCDE for the production of"pallidocyclicin."This study investigated the function of the pen operon experimentally.The heterologous expression of the entire pen operon from the plasmid was toxic to Escherichia coli but not to Bacillus subtilis.However,when the entire pen operon was expressed constitutively,even the growth of B.subtilis was impaired,and at least pcnA was implied to serve as the precursor of pallidocyclicin.In addition,a strain of B.subtilis expressing the entire pen operon from the plasmid showed toxicity to another thermophilic species,Geobacillus kaustophilus,at elevated temperatures,whereas another strain lacking pcnE alone from the pen operon lost the toxicity,suggesting that pcnE might be involved in the biosynthesis of pallidocyclicin when it is produced in B.subtilis.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    Novel heat shock response mechanism mediated by the initiation nucleotide of transcription

    Aya SatoMisaki TakamatsuSatona Kobayashi
    14页
    查看更多>>摘要：Among SigA-dependent promoters in Bacillus subtilis,we compared the nucleotide sequences of heat shock responding and non-responding promoters.Chimeric promoter experiments revealed that the heat shock response could be ascribed to the initiation nucleotide(iNTP)of the transcription.Our in vivo reporter assay results indicated that a full response was achieved using GTP,a reduced response was observed using ATP,and no additional expression was observed using UTP or CTP.We then investigated the in vitro transcription assay in more detail.Enhanced transcription that was dependent upon the iNTP was observed when heat treatment was administered during the pre-initia-tion period.We next analyzed the efficiency of open complex formation using potassium permanganate footprinting,and our results revealed an increase in the ratio of open complex formation at elevated temperatures.Based on this,we suggest that the overall intensification of transcription at high temperatures was derived from the high efficiency of open complex formation together with the high affinity of RNA polymerase(RNAP)for the initiation nucleotide GTP.To determine if this mechanism observed in B.subtilis RNAP is common among bacterial species,we performed similar experiments using Escherichia coli RNAP.Our results indicated that E.coli RNAP also exhibited both temperature-and iNTP-dependent enhancement of transcription.Although the temperature ranges and the ratios of enhancement are somewhat different,the overall heat shock response mechanism mediated by the iNTP of transcription appears to be conserved among bacterial RNAP.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    3-Amino-4-hydroxybenzoic acid production from glucose and/or xylose via recombinant Streptomyces lividans

    Satoko Niimi-NakamuraHideo KawaguchiKouji Uematsu
    8页
    查看更多>>摘要：The aromatic compound 3-amino-4-hydroxybenzoic acid(3,4-AHBA)can be employed as a raw material for high-performance industrial plastics.The aim of this study is to produce 3,4-AHBA via a recombinant Streptomyces lividans strain containing gril and griH genes derived from Streptomyces griseus using culture medium with glucose and/or xylose,which are the main components in lignocel-lulosic biomass.Production of 3,4-AHBA by the recombinant S.lividans strain was successful,and the productivity was aifected by the kind of sugar used as an additional carbon source.Metabolic profiles revealed that L-aspartate-4-semialdehyde(ASA),a precursor of 3,4-AHBA,and coenzyme NADPH were supplied in greater amounts in xylose medium than in glucose medium.Moreover,cultivation in TSB medium with a mixed sugar(glucose/ xylose)was found to be effective for 3,4-AHBA production,and optimal conditions for efficient production were designed by changing the ratio of glucose to xylose.The best productivity of 2.70 g/L was achieved using a sugar mixture of 25 g/L glucose and 25 g/L xylose,which was 1.5 times higher than the result using 50 g/L glucose alone.These results suggest that Streptomyces is a suitable candidate platform for 3,4-AHBA production from lignocel-lulosic biomass-derived sugars under appropriate culture conditions.
      原文链接:
    • NSTL
    • Microbiol Res Foundation

    The Journal of General and Applied Microbiology Instructions for Contributors(revised August 1,2017)

    The Journal of General and Applied Microbiology Group
    3页
    查看更多>>摘要：1.THE JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY(JGAM)is issued by Applied Microbiology,Molecular and Cellular Biosciences Research Foundation.JGAM is a bimonthly,international journal devoted to the publication of original articles in general and applied microbiology.Original papers(Full Papers and Short Communications)and Invited Reviews written in English are considered for publication.Papers already published,in press or under consideration elsewhere are not accepted.
      原文链接:
    • NSTL