Pilot-scale biopesticide production by Bacillus thuringiensis subsp. kurstaki using starch industry wastewater as raw material

Pilot-scale Bacillus thuringiensis based biopesticide production (2000 L bioreactor) was conducted using starch industry wastewater (SIW) as a raw material using optimized operational parameters obtained in 15 L and 150 L fermenters. In pilot scale fermentation process the oxygen transfer rate is a major limiting factor for high product yield. Thus, the volumetric mass transfer coefficient (K_La) remains a tool to determine the oxygen transfer capacity [oxygen utilization rate (OUR) and oxygen transfer rate (OTR)] to obtain better bacterial growth rate and entomotoxicity in new bioreactor process optimization and scale-up. This study results demonstrated that the oxygen transfer rate in 2000 L bioreactor was better than 15 L and 150 L fermenters. The better oxygen transfer in 2000 L bioreactor augmented the bacterial growth [total cell (TC) and viable spore count (SC)] and delta-endotoxin yield. Prepared a stable biopesticide formulation for field use and its entomotoxicity was also evaluated. This study result corroborates the feasibility of industrial scale operation of biopesticide production using starch industry wastewater as raw material.     

表面活性剂对Bacillus pumilus strain Bap9降解苯并[a]芘的影响

研究了非离子型表面活性剂Triton X-100（TX-100）和Tween80（TW-80）对苯并[a]芘的增溶特性及对苯并[a]芘高效降解菌Bacillus pumilus strain Bap9生长的影响,结果表明,2种表面活性剂对苯并[a]芘均有良好的增溶效果,均能作为碳源和能源被菌株Bap9所利用,TX-100增溶能力和增殖能力相对更强;不同浓度的TX-100对菌株降解苯并[a]芘的影响不同,当浓度为1 000 mg/L时,对降解的促进作用最强,可将苯并[a]芘降解率提高20.8%;在苯并[a]芘降解过程中,TX-100亦能作为碳源被菌株Bap9利用,不产生二次污染,因此可用于苯并[a]芘污染环境的生物修复。     

南京城市污泥发酵苏云金杆菌培养基成分优化

研究南京江心州污水处理厂污泥作为原料发酵生产苏云金杆菌(Bacillus thringiensis)的可行性,并与常规的工业发酵黄豆粉培养基发酵B.thringiensis做比较,结果表明,以固形物含量3%的污泥为基质发酵B.thringiensis,其菌体数及芽孢数只有108个/mL,而工业发酵黄豆粉培养基发酵B.thringiensis菌体数及芽孢数为109个/mL。优化污泥发酵生产苏云金杆菌培养基成分,将污泥与不同量的黄豆粉复配作基质发酵B.thringiensis,结果发现,在污泥中添加15 g/L黄豆粉可基本满足工业发酵生产的需要,发酵72 h获得的B.thringiensis细胞总数及芽孢数均在109个/mL以上。以黄粉虫作试虫的生物毒性实验表明,复配后的基质与工业发酵黄豆粉培养基生产的B.thringiensis孢晶混合液的毒性相当。     

Detection, isolation, and identification of cadmium-resistant bacteria based on PCR-DGGE

This study focused on the screening of cadmium-resistant bacterial strains from Pb-Zn tailing. We investigated the diversity of microbial community inhabiting Dong-san-cha Pb-Zn tailing in Beijing, China, by polymerase chain reaction-denaturing gradient gel electrophoresis （DGGE） of 16S rRNA gene of bacterial strain, and found two dominant strains in the DGGE profile. Using special culture media, we isolated two strong cadmium-resistant bacterial strains. On the basis of morphological, physiological, and biochemical characteristics, BIOLOG, and 16S rDNA sequencing, the two strains were identified as Bacillus cereus and Enterobacter cloacae. Minimal inhibitory concentrations （MICs） of heavy metals for the bacteria were determined. E. cloacae showed higher MIC values for heavy metals and a larger range of antibiotic resistance than B. cereus.     

苏云金芽孢杆菌无质粒突变株BMB171的转化和表达性能

研究了苏云金芽孢杆菌无质粒突变株ＢＭＢ１７１的转化性能和表达ｃｒｙ１Ａｃ和ｃｒｙ１Ｃａ基因的性能．用ｐＨＴ３１０１、ｐＢＭＢ３０５、ｐＢＭＢ１７３６、ｐＢＭＢ６７１、ｐＢＴＬ１和ｐＨＶ１２４９等６种外源质粒电转化无质粒突变株ＢＭＢ１７１,其转化频率分别是Ｂｔ４Ｑ７、Ｂｔ４Ｄ１０和Ｂｔｉ．ＩＰＳ．７８／１１等３种常用受体菌相应最高转化频率的１０００、６．７、１２．５、６６．７、３５００和２倍,其每μｇＤＮＡ的最高转化子数达１０７．导入ＢＭＢ１７１的外源质粒的稳定性与其复制子类型和质粒大小有关．无质粒突变株ＢＭＢ１７１表达ｃｒｙ１Ａｃ基因的表达量高于对照受体菌Ｂｔ４Ｑ７,略低于Ｂｔ４Ｄ１０和Ｂｔｉ．ＩＰＳ．７８／１１,而ＢＭＢ１７１表达ｃｒｙ１Ｃａ基因的表达量高于这３种受体菌;对小菜蛾３龄幼虫和甜菜夜蛾初孵幼虫的毒力测定结果表明,ＢＭＢ１７１表达ｃｒｙ１Ａｃ基因的杀虫毒力高于Ｂｔ４Ｑ７和Ｂｔ４Ｄ１０,略低于Ｂｔｉ．ＩＰＳ．７８／１;而表达ｃｒｙ１Ｃａ基因的毒力高于这３种受体菌     

BACILLUS SP. BT-7木聚糖酶基因克隆与鉴定

报道了从一株有较高半纤维素酶活性的Ｂａｃｉｌｕｓｓｐ．ＢＴ７克隆内切木聚糖酶基因的结果．以大肠杆菌ＸＬ１ｂｌｕｅ为宿主菌,采用水解圈检测法,在含有ＲＢＢ木聚糖（４氧甲基Ｄ葡萄糖苷Ｄ木聚糖Ｒｅｍａｚｏｌ亮蓝Ｒ）的平板上分离到能水解ＲＢＢ木聚糖的阳性克隆３个,对它们进行的限制酶作图和Ｓｏｕｔｈｅｒｎ杂交分析表明,它们属于３种不同的内切木聚糖酶基因     

土壤分离菌株去除水溶液中铅离子研究

金属及微生物种类、pH和接触时间是决定生物吸附效果的关键因素.从受铅污染的土壤中分离出5株有生物吸附铅离子能力的菌株,并鉴定吸附能力最强的菌株1为芽孢杆菌;试验了pH、接触时间和初始铅离子浓度对该芽孢杆菌生物吸附铅的影响,在温度为25℃时,pH值3.0、初始铅离子浓度250 mg/L、吸附时间不超过15 min有最大吸附量92.27 mg/g,此时去除率为73.82%,且25℃吸附平衡基本符合Langmuir等温模型.因此,用该芽孢杆菌生物吸附去除水溶液中铅是可行的.     

一株1，2-二氯苯降解菌的分离鉴定及其降解特性

采用富集驯化方法,从盐城芦苇湿地根际土壤中分离得到一株可高效降解1,2-二氯苯的菌株,命名为DL-1。该菌株可以在以1,2-二氯苯为惟一碳源的无机培养基上生长,能够耐受最高浓度为200 mg/L的1,2-二氯苯。根据形态特征观察、生理生化鉴定和16S rDNA序列同源性分析,该目标菌株被鉴定为蜡质芽孢杆菌（Bacillus cereus）。菌株DL-1对1,2-二氯苯降解性能研究表明,该菌株为一株兼性厌氧菌,其适宜降解浓度、适宜温度、适宜pH值和适宜接种量分别为120mg/L、32℃、7和10%,在适宜降解条件下降解12,-二氯苯4 d其降解率达到80.3%。本实验为利用该菌株降解12,-二氯苯污水的应用提供了理论基础。     

Influence of extracellular polymeric substances (EPS) on Cd adsorption by bacteria

The role of extracellular polymeric substances (EPS) in Cd adsorption by Bacillus subtilis and Pseudomonas putida was investigated using a combination of batch adsorption experiments, potentiometric titrations, Fourier transform infrared spectroscopy (FTIR). An increased adsorption capacity of Cd was observed for untreated bacteria relative to that for EPS-free bacteria. Surface complexation modeling of titration data showed the similar pK_a values of functional groups (carboxyl, phosphate and hydroxyl) between untreated and EPS-free bacteria. However, site concentrations on the untreated bacteria were found to be higher than those on the EPS-free bacteria. FTIR spectra also showed that no significant difference in peak positions was observed between untreated and EPS-free bacteria and carboxyl and phosphate groups were responsible for Cd adsorption on bacterial cells. The information obtained in this study is of fundamental significance for understanding the interaction mechanisms between heavy metals and biofilms in natural environments.     

Effect of ultrasonication and Fenton oxidation on biodegradation of bis(2-ethylhexyl) phthalate (DEHP) in wastewater sludge

The presence of bis(2-ethylhexyl) phthalate (DEHP) and its metabolites, i.e. 2-ethylhexanol, 2-ethylhexanal, and 2-ethylhexanoic acid in wastewater sludge (WWS) were investigated during aerobic digestion and Bacillus thuringiensis (Bt)-based fermentation of WWS. Ultrasonication and Fenton oxidation pre-treatment was applied to improve biodegradability of WWS and bioavailability of the target compounds for digestion and fermentation. DEHP and 2-ethylhexanoic acid were observed at higher concentration, meanwhile 2-ethylhexanol and 2-ethylhexanal were observed at lower concentration in WWS. After 20-day aerobic digestion, DEHP removal was 72%, 89%, and 85%, and 2-ethylhexanoic acid removal was 71%, 84%, 79%, respectively for raw, ultrasonicated, and Fenton-oxidized sludges. Bt was found to degrade DEHP, leading to DEHP removal of 21%, 40%, and 30%, respectively for raw, ultrasonicated, and Fenton-oxidized sludges in the fermentation. The results suggested that aerobic stabilization and Bt-based fermentation can remove the phthalates, and pre-treatment of WWS was also effective in improvement of DEHP biodegradation. Hence, Bt-based biopesticide production from WWS can be applied safely when taking into consideration the phthalate contaminants.