枯草芽孢杆菌对Cu2+的吸附及菌体表面基团分析

以枯草芽孢杆菌(Bacillus subtilis)为生物吸附剂,讨论了其对Cu2+的生物吸附规律,并通过酸碱滴定这一表面分析手段,结合相关软件及傅里叶变换红外光谱(FTIR)分析了菌体表面主要的基团种类及数量.结果表明,枯草芽孢杆菌对Cu2+吸附的最佳条件是:pH为6、吸附时间为24 h,菌悬液用量和Cu2+初始浓度...     

Biodegradation of triazine herbicide metribuzin by the strain Bacillus sp. N1

By enrichment culturing of soil contaminated with metribuzin, a highly efficient metribuzin degrading bacterium, Bacillus sp. N1, was isolated. This strain grows using metribuzin at 5.0% (v/v) as the sole nitrogen source in a liquid medium. Optimal metribuzin degradation occurred at a temperature of 30ºC and at pH 7.0. With an initial concentration of 20 mg L^?1, the degradation rate was 73.5% in 120 h. If the initial concentrations were higher than 50 mg L^?1, the biodegradation rates decreased as the metribuzin concentrations increased. When the concentration was 100 mg L^?1, the degradation rate was only 45%. Degradation followed the pesticide degradation kinetic equation at initial concentrations between 5 mg L^?1 and 50 mg L^?1. When the metribuzin contaminated soil was mixed with strain N1 (with the concentration of metribuzin being 20 mg L^?1 and the inoculation rate of 10¹¹ g^?1 dry soil), the degradation rate of the metribuzin was 66.4% in 30 days, while the degradation rate of metribuzin was only 19.4% in the control soil without the strain N1. These results indicate that the strain N1 can significantly increase the degradation rate of metribuzin in contaminated soil.     

Bacillus thuringiensis field applications: Effect of nozzle type,drop size,and application timing on efficacy against gypsy moth

Abstract

In a series of recent studies, we evaluated the influence of delivery systems, drop size and application timing on the efficacy of aerially applied Bacillus thuringiensis against gypsy moth infestations. Use of different nozzle systems including Micronair, Flat Fan or Twin Jet, did not appear to result in significant differences in Bt coverage efficiency, foliage protection or population reduction. Nor was there any significant difference in population reduction when Bt was applied at two different drop sizes with volume median diameters of 110 and 163 μm. The efficacy of different formulations on larval populations were similar when used against the younger 1st and 2nd instar but differed when treatment was delayed until the population matured to the 3rd and 4th instar stages.     

具有降解亚硝酸盐活性的解淀粉芽孢杆菌的分离与安全性分析

从养殖污泥中分离筛选了一株优良的亚硝酸盐降解菌YX01,经ARIS 2X细菌自动鉴定系统以及16SrRNA系统发育分析,该菌株被鉴定为解淀粉芽孢杆菌(Bacillus am yloliquefaciens,GenBank登录号为JX649949),其16S rRNA序列与基因库中 芽孢杆菌属菌株的16S rRNA序列有99％～100％的同源性,而且与解淀粉芽孢杆菌菌株LD5(GenBank登录号:GQ853414)的亲缘关系最近.该亚硝酸盐降解菌YX01在浓度为2.0×107～2.0×109cfu/L时对小球藻(Chlorella sp.)生长有促进作用;在浓度大于2.0×1011cfu/L时对小球藻生长具有显著抑制作用;亚硝酸盐降解菌YX01对小球藻的半数抑制浓度(IC50)大于2.0×1011cfu/L.亚硝酸盐降解菌YX01对大型蚤(Daphnia magna)的48h-半数致死浓度(LC50)大于2.0×1011cfu/L,对草鱼(Ctenopharyngodon idellus)的96h-LC50大于2.0×1011cfu/kg.经分析,亚硝酸盐降解菌YX01为无毒菌株.     

Identification and comprehensive evaluation of a novel biocontrol agent Bacillus atrophaeus JZB120050

Abstract

Bacillus spp. have long been used as biocontrol agents because of their efficient broad-spectrum antimicrobial activity. We identified a novel strain of Bacillus atrophaeus, named JZB120050, from soil. B. atrophaeus JZB120050 had a strong inhibitory effect against Botrytis cinerea and many other phytopathogens. Gas chromatography-mass spectrometry showed that B. atrophaeus JZB120050 produced many secondary metabolites, such as alkanes, alkenes and acids; some of which were related to pathogen inhibition. Enzyme activity analysis showed that B. atrophaeus JZB120050 secreted cell-wall-degrading enzymes, including chitinase, glucanase and protease, which degraded fungal cell walls. Both the novel glucanase gene bglu and chitinase gene chit1 were cloned and heterologously expressed in Escherichia coli and the products showed strong enzyme activity. In addition, B. atrophaeus JZB120050 secreted siderophores and formed a significant biofilm. Future studies should focus on these antimicrobial factors to facilitate widespread application in the field of agricultural biocontrol.     

利用玉米浸泡液制备苏云金杆菌生物杀虫剂

以玉米淀粉生产过程中的浸泡液作为原料培养基,在5L发酵罐中培养苏云金杆菌生物杀虫剂,并以常规黄豆粉培养基为对照,考察了苏云金杆菌在玉米浸泡液中的生长代谢状况(包括菌体形态、菌数增长与芽孢形成)以及48h发酵液的生物毒效.研究表明,无需添加其它成分及前处理,玉米浸泡液所含营养成分即可满足苏云金杆菌生长需求,其培养的总活菌数及芽孢数在27h可达最大值1.51 ×109CFU·mL-1和1.41×109CFU·mL-1,分别比常规培养基高出59%和85%,芽孢形成与晶体释放提前约9～12h;36h扫描电镜(SEM)观察表明,发酵36h浸泡液中大部分芽孢囊已经自溶,游离出卵圆形芽孢与菱形的晶体;而此时常规培养基正处于芽孢形成后期,只有极少数的游离晶体与芽孢;48h发酵终点的生物毒效结果显示,以浸泡液为培养基的苏云金杆菌发酵液毒效(891.51 IU·μL-1)比常规培养基高89%.本试验为浸泡液的再利用提供了一条崭新的途径,同时降低了杀虫剂的生产成本.     

一株芽孢杆菌在低氮源浓度培养基中的生长与氨氮去除特性

对菌株YB3进行了16S rRNA基因序列进化分析,并分别以NH_4Cl、NaNO_2、NaNO_3、尿素和蛋白胨为单一氮源,配制了5种低氮源浓度培养基,研究YB3在与养殖水体相近营养水平条件下的生长与氨氮去除特性.结果显示,菌株YB3属于蜡样芽孢杆菌(Bacullis cereus),在5种培养基中均能够生长,菌悬液(吸光度OD600为1.0)接种量为1.0%(v/v)时,OD600由0.010增长到0.100~0.117.在NH_4Cl培养基中,YB3的氨氮去除速率为1.23 mg·L~(-1)·d~(-1),去除率为93.5%.在尿素、蛋白胨等有机氮源培养基中,YB3将首先导致氨氮的积累,累积倍数分别为51.69和3.38,之后开始去除,去除速率为1.56和0.29 mg·L~(-1)·d~(-1),去除率为93.7%和26.8%.结果也表明,提高YB3接种量至8.0%(v/v),可以使蛋白胨培养基氨氮累积倍数下降至2.02,去除速率提高至1.07 mg·L~(-1)·d~(-1),去除率最终达到98.4%.NaNO_2和NaNO_3培养基中均未检测到氨氮,而NH_4Cl、尿素和蛋白胨培养基中也未检测到NO_2~--N和NO_3~--N,表明YB3的硝化、亚硝化和反硝化作用均不强烈,去除氨氮的同时将不会造成NO_2~--N和NO_3~--N等的大量积累.本文为菌株YB3在养殖水体调控与净化中的应用研究提供了实验基础和理论支持.     

酸性含硒含镉废水低温生物强化处理技术研究

本研究将枯草芽孢杆菌接种于活性污泥中,系统考察了酸性含硒含镉废水的低温生物强化处理效果.研究发现,活性污泥联合枯草芽孢杆菌对硒、镉的去除效率(Se:94.9%,Cd:99.1%)高于单一枯草芽孢杆菌(B菌)、混合枯草芽孢杆菌及活性污泥.影响因素试验结果表明,在20℃、pH 4.0、甲醇为碳源时生物强化硒、镉去除效率较高.利用SBR反应器处理酸性(pH 4.0)含硒(3.7～10.0 mg·L~(-1))含镉(5.0～11.2 mg·L~(-1))废水,在低温(8.0～10.8℃)条件下运行80个周期,硒、镉和COD的平均去除率分别为97.4%、90.7%和95.0%.红外光谱分析表明,酰胺基、芳香族C—H伸缩健在硒、镉去除中起到主要作用.透射电镜与能谱分析证实了生物强化污泥对硒、镉去除之后形成纳米颗粒物.通过高通量测序分析微生物群落结构,发现生物强化污泥中富集了Pseudomonas(假单胞菌属)、Dechloromonas(脱氯单胞菌)等典型硒还原菌.     

一株芘的高效降解菌的选育及其降解性能研究

旨在为多环芳烃污染环境的生物修复提供微生物资源和科学依据,从某焦化厂曝气池的活性污泥中分离纯化出一株降解芘的优势菌株B-1,该菌株可在以芘为惟一碳源的培养基中生长繁殖,能利用芘的最高浓度为130 mg/L左右.经形态学观察、生理生化实验及l6S rDNA序列分析,初步判断菌株B-1为芽胞杆菌属.投菌量、芘初始浓度、pH...     

Enhancement of MK-7 synthesis by engineered strains of Bacillus subtilis natto that overexpress men genes and by increasing available dissolved oxygen北大核心CSCD

In recent years, researchers have discovered novel physiological functions of vitamin K2. In addition to promoting blood clotting, it can prevent osteoporosis and cardiovascular disease and is expected to treat some tumors and Parkinson’s disease. Bacillus subtilis natto is a probiotic that has been approved by the U.S. Food and Drug Administration for use as a bioproducer of vitamin K2. Its product’s main form is MK-7, which has a long half-life in the human body and high bioavailability. Bacillus subtilis natto displays great potential for large-scale biological preparation of vitamin K2. In this study, the Sipizizen method of Bacillus subtilis transformation was optimized to make it suitable for molecular transformation of Bacillus subtilis natto. Vectors for overexpression of all 8 genes involved in the menadione synthetic pathway were constructed, and changes in MK-7 fermentation yield after transformation of Bacillus subtilis natto were investigated. Three enzymes were found to exert significant effects on MK-7 synthesis, namely isopentenyltransferase (MenA), 1,4-dihydroxy-2-naphthoyl-CoA synthase (MenB), and nornaphthoquinone methyltransferase (MenG). A modified strain (BN-pABG) with higher MK-7 productivity was obtained by concerted overexpression of menA, menB, and menG. In a 5 L bioreactor, MK-7 synthesis was further enhanced by optimizing oxygen supply. The final yield of MK-7 from the modified strain was 62.21 mg/L, 1.26 times higher than that of the original strain. These results show that combined overexpression of menA, menB, and menG strongly promotes MK-7 synthesis by Bacillus subtilis natto, and optimizing the oxygen supply conditions also promotes more robust MK-7 synthesis. © 2022 Authors. All rights reserved.