常见4种微生态制剂菌种产淀粉酶的比较

目的 探索常见4种微生态制剂菌种地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌和纳豆芽胞杆菌产生淀粉酶的能力,以筛选和研制动物微生态制剂和饲料添加剂的使用菌种。方法 将各菌种接于淀粉酶试验培养基,培养后滴加稀碘溶液,观察透明圈,判定产酶能力。结果 地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌和纳豆芽胞杆菌都能产生淀粉酶,以蜡样芽胞杆菌产生的淀粉酶较多。结论 4种常见芽胞杆菌产淀粉酶能力依次为：蜡样芽胞杆菌＞纳豆芽胞杆菌＞枯草芽胞杆菌＞地衣芽胞杆菌。     

福矛高温大曲中芽孢杆菌16S rDNA-RFLP及系统发育分析

目的:从福建建瓯黄华山酿酒有限公司高温大曲中分离出89株芽孢杆菌,通过初步筛选鉴定并进行微生物多样性研究.方法:对其16S rDNA进行PCR - RFLP分析和系统发育研究.结果:初步筛选得到的18株芽孢杆菌被HhaⅠ和MspⅠ酶切聚类分为四大组.通过系统发育分析样品中有6株Bacillus subtilis,4株Bacillus cereus,2株Bacillus sonorensis,2株Bacillus licheniformis,以及Bacillus pumilus、Bacillus oleronius、Bacillus coagulans和Bacillus thuringiensis各1株.结论:研究显示该高温大曲中可培养芽孢杆菌具有微生物多样性.     

Cloning of the genes controlling the biosynthesis of bacitracin in Bacillus licheniformis

The DNA fragment from bacitracin-producing Bacillus licheniformis strain is cloned on pMX39 vector plasmid in Bacillus subtilis cells. Bacillus subtilis cells carrying the cloned fragment inhibit the growth of bacitracin-sensitive tester strain. The observed inhibition of growth is due to the production by Bacillus subtilis of bacteriocin substance that is identified as bacitracin by TLC-chromatography. In contrast to the data published earlier it is shown that Bacillus subtilis can in fact produce the small amounts of bacitracin. Introduction of the cloned Bacillus licheniformis DNA into Bacillus subtilis cells stimulates this bacitracin production. The restriction site map of the Bacillus licheniformis chromosomal region bearing the cloned fragment is constructed.     

Production of diarrheal enterotoxins and other potential virulence factors by veterinary isolates of bacillus species associated with nongastrointestinal infections

With the exceptions of Bacillus cereus and Bacillus anthracis, Bacillus species are generally perceived to be inconsequential. However, the relevance of other Bacillus species as food poisoning organisms and etiological agents in nongastrointestinal infections is being increasingly recognized. Eleven Bacillus species isolated from veterinary samples associated with severe nongastrointestinal infections were assessed for the presence and expression of diarrheagenic enterotoxins and other potential virulence factors. PCR studies revealed the presence of DNA sequences encoding hemolysin BL (HBL) enterotoxin complex and B. cereus enterotoxin T (BceT) in five B. cereus strains and in Bacillus coagulans NB11. Enterotoxin HBL was also harbored by Bacillus polymyxa NB6. After 18 h of growth in brain heart infusion broth, all seven Bacillus isolates carrying genes encoding enterotoxin HBL produced this toxin. Cell-free supernatant fluids from all 11 Bacillus isolates demonstrated cytotoxicity toward human HEp-2 cells; only one Bacillus licheniformis strain adhered to this test cell line, and none of the Bacillus isolates were invasive. This study constitutes the first demonstration that Bacillus spp. associated with serious nongastrointestinal infections in animals may harbor and express diarrheagenic enterotoxins traditionally linked to toxigenic B. cereus.     

学生食堂餐厨垃圾中淀粉的生物利用菌株筛选

目的探索地衣芽胞杆菌、枯草芽胞杆菌和蜡样芽胞杆菌分解大学生食堂厨余中淀粉的能力,以筛选和研制餐厨垃圾生物降解的使用菌种。方法将各菌种接于淀粉酶试验培养基,培养后滴加碘溶液,观察透明圈,判定产淀粉酶能力;收集大学生食堂的厨余,观察三种细菌在不同接种量（5%、10%、15%、20%、25%）、不同接种时间（24 h、48 h、72 h）及不同菌株配伍方式下发酵淀粉的能力。结果地衣芽胞杆菌、枯草芽胞杆菌、蜡样芽胞杆菌都能产生淀粉酶,以地衣芽胞杆菌产生的淀粉酶较多,其次为枯草芽胞杆菌,蜡样芽胞杆菌较少,三种细菌分解厨余中淀粉的最佳接种量都为15%-20%,最佳发酵时间为48 h,枯草芽胞杆菌和地衣芽胞杆菌各7.5%的接种量混合配伍发酵效果最佳。结论可采用枯草芽胞杆菌和地衣芽胞杆菌各7.5%的接种量混合配伍发酵学生食堂的厨余中的淀粉。     

曾候乙墓穴木椁微生物的分离与鉴定

从曾侯乙墓中椁室、东椁室、西椁室和北椁室分别取样,经平板划线培养,分离,纯化共获得典型的菌株16株。将16株分别涂片镜检,生理生化分析鉴定,结果证明芽孢杆菌属11株,其中苏云金变种1株,地衣芽孢杆菌1株,巨大芽孢杆菌1株,球形芽孢杆菌5株,蜡状芽孢杆菌2株,多粘芽孢杆菌1株。其余菌株微杆菌属4株,黄色杆菌属黄杆菌1株。     

Genetic map of the Bacillus stearothermophilus NUB36 chromosome.

A circular genetic map of Bacillus stearothermophilus NUB36 was constructed by transduction with bacteriophage TP-42C and protoplast fusion. Sixty-four genes were tentatively assigned a cognate Bacillus subtilis gene based on growth response to intermediates or end products of metabolism, cross-feeding, accumulation of intermediates, or their relative order in a linkage group. Although the relative position of many genes on the Bacillus stearothermophilus and Bacillus subtilis genetic map appears to be similar, some differences were detected. The tentative order of the genes in the Bacillus stearothermophilus aro region is aspB-aroBAFEC-tyrA-hisH-(trp), whereas it is aspB-aroE-tyrA-hisH-(trp)-aroHBF in Bacillus subtilis. The aroA, aroC, and aroG genes in Bacillus subtilis are located in another region. The tentative order of genes in the trp operon of Bacillus stearothermophilus is trpFCDABE, whereas it is trpABFCDE in Bacillus subtilis.     

芽胞杆菌在防治水稻稻瘟病中的应用及生防机制

芽胞杆菌具有人畜安全、不污染环境、病原菌不易产生抗药性、抗逆性强和促进植物生长等优点,是稻瘟病防治上的重要生防菌。芽胞杆菌的生防机制主要包括竞争作用、拮抗作用和诱导抗病性。芽胞杆菌定殖在水稻植株上,产生抗菌活性物质抑制稻瘟病菌的生长,诱导水稻产生抗病性,对水稻植株具有促生作用,可以挽回水稻产量损失。芽胞杆菌可以制备生防制剂用来防治我国南方稻区和北方稻区的稻瘟病危害,在水稻产业的可持续发展中对稻瘟病的生物防治具有指导意义。本文主要综述芽胞杆菌在防治水稻稻瘟病中的应用研究、芽胞杆菌在防治水稻稻瘟病中的生防机制、影响稻瘟病生防芽胞杆菌防效的因素。     

Effect of Carbon Source on the Antimicrobial Activity of the Air Flora

Summary In an attempt to screen for air flora producing new potent antimicrobial substances, Bacillus megaterium NB-3, Bacillus cereus NB-4, Bacillus cereus NB-5, Bacillus subtilis NB-6 and Bacillus circulans NB-7, were isolated and were found to be antagonistic to bacteria and/or fungi. Production of antimicrobial substances by the bacterial strains was greatly influenced by variation of carbon sources. Glycerol strongly enhanced the antimicrobial activity of strains NB-3 and NB-6, whereas glucose increased the antimicrobial activity of strains NB-4 and NB-5. The maximum antibiotic yield of NB-7 was achieved with fructose as a carbon source. Starch (Bacillus megaterium NB-3), maltose (Bacillus cereus NB-5), glycerol (Bacillus circulans NB-7), arabinose, ribose (Bacillus cereus NB-4) and arabinose, fructose, glucose, ribose and sucrose (Bacillus subtilis NB-6) repressed the production of antimicrobial substances by the respective bacterial strains.     

Effects of plant growth promoting rhizobacteria (PGPR) on rooting and root growth of kiwifruit (Actinidia deliciosa) stem cuttings

The effects of plant growth promoting rhizobacteria (PGPR) on the rooting and root growth of semi-hardwood and hardwood kiwifruit stem cuttings were investigated. The PGPR used were Bacillus RC23, Paenibacillus polymyxa RC05, Bacillus subtilis OSU142, Bacillus RC03, Comamonas acidovorans RC41, Bacillus megaterium RC01 and Bacillus simplex RC19. All the bacteria showed indole-3-acetic acid (IAA) producing capacity. Among the PGPR used, the highest rooting ratios were obtained at 47.50% for semi-hardwood stem cuttings from Bacillus RC03 and Bacillus simplex RC19 treatments and 42.50% for hardwood stem cuttings from Bacillus RC03. As well, Comamonas acidovorans RC41 inoculations indicated higher value than control treatments. The results suggest that these PGPR can be used in organic nursery material production and point to the feasibility of synthetic auxin (IBA) replacement by organic management based on PGPR.     

枯草芽胞杆菌基因组删减对异源酶表达影响的研究进展

枯草芽胞杆菌作为一种遗传背景清晰、基因编辑成熟的革兰氏阳性菌,是多种重要工业酶的生产宿主。随着转录组、蛋白质组、代谢组等多组学测序和分析技术的发展,通过合理设计简化枯草芽胞杆菌基因组,减少细胞内冗余的调控和代谢网络,使得细胞更精简且便于控制,展现出了枯草芽胞杆菌作为异源酶表达宿主细胞的应用潜力。本文简要综述了枯草芽胞杆菌基因组删减的研究进展,归纳了必需基因的确定方法,重点介绍了枯草芽胞杆菌通过删减基因组提升异源酶表达的研究进展及删减策略,充分展示了枯草芽胞杆菌基因组删减在构建异源酶表达底盘细胞中的重要作用。     

Sigma 29-like protein is a common sporulation-specific element in bacteria of the genus Bacillus.

A monoclonal antibody specific for an antigenic determinant on the Bacillus subtilis sporulation-induced sigma factor sigma 29 reacted with proteins similar in size to sigma 29 in extracts of sporulating Bacillus licheniformis, Bacillus amyloliquifaciens, Bacillus cereus, Bacillus natto, and Bacillus pumilus but not in extracts prepared from vegetatively growing cultures of these bacteria. These results indicate that RNA polymerase modifications, initially described for B. subtilis, are likely to be common among sporulating Bacillus spp. and that at least some of the specific modifications that are observed in sporulating B. subtilis are conserved among members of this genus.     

Presence of N6-methyladenine in GATC sequences of Bacillus popilliae and Bacillus lentimorbus KLN2.

Nine strains of Bacillus popilliae and Bacillus lentimorbus KLN2 contain N6-methyladenine in GATC sequences, as determined by using the restriction enzymes MboI and DpnI. Among eight other Bacillus species examined, all, except one strain of Bacillus brevis (ATCC 9999), lacked adenine methylation in GATC. A methylase with Escherichia coli dcm site specificity was not present in any of the Bacillus species studied.     

Whole-genome sequences of Bacillus subtilis and close relatives

We sequenced four strains of Bacillus subtilis and the type strains for two closely related species, Bacillus vallismortis and Bacillus mojavensis. We report the high-quality Sanger genome sequences of B. subtilis subspecies subtilis RO-NN-1 and AUSI98, B. subtilis subspecies spizizenii TU-B-10(T) and DV1-B-1, Bacillus mojavensis RO-H-1(T), and Bacillus vallismortis DV1-F-3(T).     

Mapping and cloning of the gene coding for beta-glucanase from various bacilli

Beta-glucanase gene from Bacillus subtilis 168 has been mapped by bacteriophage pBS1 transduction technique between sacA and purA genes. The stimulating effect of pleiotropic mutations pap, amyB and sacUh on beta-glucanase production in Bacillus subtilis and Bacillus amyloliquefaciens has been described. Beta-glucanase gene from Bacillus amyloliquefaciens has been cloned ona Charon 4A vector. Expression of the gene in E. coli cells depended on the orientation of the cloned DNA on a pBR322 vector plasmid. Maximal enzymatic activity was registered in periplasm. Beta-glucanase gene was recloned in Bacillus subtilis cells. Bacillus subtilis strain, harbouring pBG1, produces 500 times more beta-glucanase as compared with the wild type strain of Bacillus subtilis.     

坚强芽孢杆菌β-淀粉酶基因的核苷酸序列分析

淀粉水解酶广泛用于淀粉加工业中,何秉旺等在选育产耐热β-淀粉酶菌株中得到一株坚强芽孢杆菌（Bacillusfirmus）725,该菌株产生的淀粉酶有较好的热稳定性,水解淀粉的主要产物为麦芽糖。自然菌株产生的淀粉酶往往是多种淀粉酶的混合,为进一步研究该菌株产生的淀粉酶的性质和在工业上应用的可能性,分离了三个淀粉酶基因,在大肠杆菌中克隆和表达[1]。其中重组质粒pBA150产生的淀粉酶的淀粉水解产物主要是麦芽糖［1］。β-淀粉酶（EC.3．2.1．2）水解淀粉的主要产物是麦芽糖,工业上可用于生产高麦芽糖浆,近年来又有β-淀粉酶用于啤酒工业的报道[2]。本文报道重组质粒pBA150的β-淀粉酶基因的序列分析及推导出的氨基酸序列同己知β-淀粉酶的氨基酸序列比较。     

芽胞杆菌B13功能的初步研究

用选择性培养基从土壤中分离到芽胞杆菌B13。结果表明：芽胞杆菌B13能够同时溶解无机磷和分解有机磷,并且芽胞杆菌B13活菌及其发酵液能够抑制烟草青枯病菌和黄曲霉的生长;共培试验证明芽胞杆菌B13能够抑制烟草疫霉的生长;盆栽试验证明芽胞杆菌B13对烟草青枯病具有一定的防治效果。可以进一步将其开发出集解养分与抗病于一体的微生物活体制剂。     

Differential cell surface properties of vegetative Bacillus

Aims:  The genus Bacillus encompasses a wide range of species which display varying pathogenic abilities. The hydrophobicity of a range of Bacillus species was determined to evaluate the correlation between bacterial hydrophobicity and pathogenicity.
Methods and Results:  Bacterial adhesion to hydrocarbon assays were used to determine the hydrophobicity of various Bacillus species. Significant differences in the hydrophobicity of vegetative Bacilli were found. Specifically, vegetative Bacillus anthracis or Bacillus thuringiensis cells were highly hydrophobic whereas Bacillus cereus or Bacillus subtilis were only slightly hydrophobic using this test. Cell adhesion assays using A549 or J774 cells were used to demonstrate a correlation between the bacterial hydrophobicity profiles with the ability to adhere to the mammalian cell lines.
Conclusions:  The ability of Bacillus species to adhere to mammalian cell lines correlates with the hydrophobicity of the bacteria and also correlates with the relative pathogenicity of some of the Bacillus species tested.
Significance and Impact of the Study:  This work suggests that study of the physical-chemical properties of vegetative cells could inform future approaches for the rapid identification and discrimination of potentially pathogenic Bacilli .     

DNA-methylases from different strains of Bacillus subtilis and Bacillus natto

DNA-methyltransferase activity has been detected in some of Bacillus subtilis and Bacillus natto strains. Two strains of Bacillus subtilis exhibited DNA-cytosine methyltransferase activity, and the strains of Bacillus natto exhibited DNA-adenine methyltransferase activity. A possible effect of DNA-methyltransferase specificity on transformation efficiency is discussed.     

Comparative sequence analyses on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov.

Comparative 16S rRNA (rDNA) sequence analyses performed on the thermophilic Bacillus species Bacillus acidocaldarius, Bacillus acidoterrestris, and Bacillus cycloheptanicus revealed that these organisms are sufficiently different from the traditional Bacillus species to warrant reclassification in a new genus, Alicyclobacillus gen. nov. An analysis of 16S rRNA sequences established that these three thermoacidophiles cluster in a group that differs markedly from both the obligately thermophilic organisms Bacillus stearothermophilus and the facultatively thermophilic organism Bacillus coagulans, as well as many other common mesophilic and thermophilic Bacillus species. The thermoacidophilic Bacillus species B. acidocaldarius, B. acidoterrestris, and B. cycloheptanicus also are unique in that they possess omega-alicylic fatty acid as the major natural membranous lipid component, which is a rare phenotype that has not been found in any other Bacillus species characterized to date. This phenotype, along with the 16S rRNA sequence data, suggests that these thermoacidophiles are biochemically and genetically unique and supports the proposal that they should be reclassified in the new genus Alicyclobacillus.