胨冻样芽孢杆菌(Bacillus mucilagininosus)的分离、筛选及其发酵培养基研究

本文报道了胨冻样芽孢杆菌 (Bacillusmucilaginosus)的分离、筛选及其对小麦苗期生长的作用 ,并对胨冻样芽孢杆菌的发酵培养基进行了优化 .结果表明 ,胨冻样芽孢杆菌 (B .mucilaginosus)生长的最适培养基 (g/L)为 :蔗糖 1 4、Na2 HPO4 1 .5、FeCl3 0 .0 0 2、CaCO3 1 .5、酵母膏 1 0 ,自来水 1 0 0 0ml、pH7.0～ 7.5;有机氮和无机铵态氮抑制胨冻样芽孢杆菌的生长 ;胨冻样芽孢杆菌液体培养物对小麦苗期生长具有促进作用 .     

响应面法优化芽孢杆菌LJ-7发酵产酯酶条件

为了获得海洋芽孢杆菌LJ-7发酵产酯酶的最佳条件,采用响应面法对其发酵条件进行了优化。首先,通过单因素试验选出对酯酶产量影响较显著的3个因素,即发酵温度、初始pH和发酵时间。在单因素试验的基础上,采用Box-Benhnken中心组合方法进行三因素三水平的试验设计,以酶活为响应值,利用响应面分析法进行进一步优化,确定最佳发酵条件为:发酵时间42.81h,发酵温度29.40℃,pH为6.21,此时预测的酯酶酶活为24.91 U/mL。在此最佳条件下,平行试验测得实际酶活为24.63U/mL,达到理论预测值的95%以上。该模型较好地预测了实际发酵情况,得到的优化条件具有实际应用价值。     

基于矿物晶体结构的铝土矿细菌浸矿机制研究

采用环状芽孢杆菌CGMCC12对5种含不同结构硅酸盐矿物的铝土矿样进行了生物脱硅研究.选用铝土矿中常见的橄榄石、高岭石、钾长石及河南铝土矿作为细菌溶蚀作用对象,采用电镜及XRD衍射技术,研究细菌对它们的分解作用差异.结果表明:浸矿12d,细菌对橄榄石、高岭石及石英型铝土矿脱硅作用显著,铝硅比(A/S)分别从7.12,6.11,2.89提高到16.15,11.61,13.56,而对钾长石型铝土矿及河南铝土矿脱硅效果较差,A/S分别从3.69和4.90仅提高到4.64和7.30;细菌对试验用硅酸盐矿物的溶蚀破坏作用由易至难依次为橄榄石、高岭石、钾长石;在细菌作用下,橄榄石向水镁石与石英转化,钾长石向伊利石转化,高岭石向水铝石与石英转化;多种矿物同时存在的情况下,细菌对不同晶体结构的硅酸盐矿物的分解有一定的选择性,对较易分解的矿物破坏作用较快,细菌会优先选择分解含Fe,Ca等生命元素的赤铁矿与方解石等矿物.     

苏云金杆菌生长的结构动力学模型

考察了苏云金杆菌(Bt)的间歇及流加培养.根据对Bt间歇发酵实验现象的分析和讨论,提出了一个基于Williams分室模型的四组分结构模型,并从实验数据确定了各组分的初值.该模型能较好地拟合Bt间歇发酵实验数据.从间歇发酵获得的模型参数推算出流加培养曲线与实验值也基本吻合.     

压电基因传感器检测芽孢杆菌靶序列研究

在石英晶体微天平(QCM)上用生物素亲和素法和自组装法2种不同的方法固定寡核苷酸探针,构建压电基因传感器,对芽孢杆菌靶序列进行实时检测。结果表明:生物素亲和素法固定探针效果更好,靶序列浓度为0.05~0.5μmol/L时,有很好的线性关系,线性回归方程为Y=93.88X+10.88(R=0.989 0,N=6,P<0.001),非线性误差为±4.7%。该传感器特异性较好,能够识别错配3个碱基的序列。     

Bt新菌株GGD-4的特性研究与杀虫晶体蛋白基因鉴定

从连云港地区的土壤中分离得到一株苏云金芽孢杆菌新菌株GGD-4,对此菌株的个体形态、生理生化特点进行了研究,并通过PCR-RFLP鉴定体系分析了此菌株中cry基因类型,为进一步利用该菌株奠定了基础.     

苏云金芽孢杆菌cry1Aa核心毒素基因转化烟草及其杀虫活性

将苏云金芽孢杆菌cry1Aa的C端编码区截短,获得编码N端609个氨基酸约1.8kb的核心毒素基因,将其构建到植物表达载体转化烟草,研究直接表达的活性ICP核心毒素被昆虫取食后的杀虫效果.T1代种子发芽的抗生素抗性分离和PCR检测证实cry1Aa核心毒素基因整合到了烟草基因组,Western 杂交检测表明转化烟草能够正常表达大小为75.6kD的核心毒素蛋白.生物测定结果表明,转化烟草对初孵棉铃虫平均有高于60%的致死率,对初孵甜菜夜蛾平均达到70%的致死率,最高致死率均可达到90%以上,并对昆虫的生长发育有明显的抑制作用.     

纳豆枯草杆菌的筛选和纳豆激酶发酵条件优化

纳豆激酶是由纳豆枯草杆菌产生的一种具有纤溶活性的丝氨酸蛋白酶.针对目前常用菌株的产酶活力较低、不能完全满足要求的问题,采用酪蛋白平板初筛摇瓶复筛的筛选策略,从纳豆中筛选获得了一株高产纳豆激酶的菌株,该菌株在筛选培养基上进行摇瓶培养,发酵液中纳豆激酶的酶活达到970 IU/mL.用Plackett Burman方法对影响液体发酵的各因素的效应进行了评价,并筛选出了有显著效应的四个主要因素:胰蛋白胨浓度、种龄、发酵温度和Na2HPO4浓度;在此基础上,用最陡爬坡路径逼近最大产酶条件区域;最后通过中心组合实验及响应面分析优化了纳豆枯草杆菌液体发酵的条件.通过在初始发酵条件和优化发酵条件下的对比研究,液体发酵液中纳豆激酶浓度从1 005 IU/mL提高到1 314 IU/mL,表明响应面法优化可成功地用于纳豆枯草杆菌液体发酵的条件优化.     

Development and evaluation of intestinal targeted mucoadhesive microspheres of Bacillus coagulans

Background: Intestinal targeted mucoadhesive microsphere of probiotics may provide numerous associated health benefits.

Aim: To develop mucoadhesive microspheres that will deliver viable probiotic cells into gut protectively against harsh environmental conditions of stomach for extended period.

Materials and methods: Core mucoadhesive microspheres of Bacillus coagulans were prepared using hypromellose, following coacervation and phase separation technique and were then coated with hypromellose phthalate to achieve their site-specific release. Microspheres were evaluated for percent yield, entrapment efficiency, surface morphology, particle size and size distribution, flow property, swelling property, mucoadhesion property by the in vitro wash-off and the ex vivo mucoadhesive strength tests, in vitro release profile and release kinetic, in vivo probiotic activity, and stability. The values for kinetic constant and regression coefficient of model-dependent approaches and the difference factor, the similarity factor, and the Rescigno index of model-independent approaches were determined for accessing and comparing in vitro performance.

Results: Microsphere formulation batches have percent yield value between 56.26% and 69.13% and entrapment efficiency value between 66.95% and 77.89%. Microspheres were coarser with spherical shape having mean particle size from 28.03 to 48.31 μm. In vitro B. coagulans release profile follows zero-order kinetics and depends on the grade of hypromellose and the B. coagulans-to-hypromellose ratio. Experimental microspheres rendered adequate stability to B. coagulans at room temperature.

Conclusion: Microspheres had delivered B. coagulans in simulated intestinal condition following zero-order kinetics, protectively in simulated gastric condition, exhibiting appreciable mucoadhesion in intestinal condition, which could be useful to achieve site-specific delivery for extended period.     

Innovative hydrogen release from sodium borohydride hydrolysis using biocatalyst-like Fe2O3 nanoparticles impregnated on Bacillus simplex bacteria

For the first time in this innovative study, microorganisms such as Bacillus simplex bacteria, mostly used in biological activity studies, are used as a bio-supporter agent of iron to release hydrogen from sodium borohydride hydrolysis at 25.0 ± 0.1 °C. The goal is to investigate thoroughly sodium borohydride hydrolysis catalyzed by Fe₂O₃ nanoparticles impregnated on microorganism such as Bacillus simplex (BS) bacteria (Fe₂O₃@BS NPs) known with strong antibacterial properties, which makes innovative them a candidate for hydrolysis reaction. This study was focused on the preparation, identification, and catalytic use of biocatalyst-like Fe₂O₃@BS NPs for hydrogen release from the sodium borohydride hydrolysis at 25.0 ± 0.1 °C. The characterization results made after and before hydrolysis reaction using by SEM/SEM-EDX, FT-IR, XRD, UV–vis, XPS, DLS, ELS Zeta potential, ESR, and TEM techniques reveal the formation of highly active, stable, durable, and long-lived biocatalysts-like Fe₂O₃@BS NPs.