Fermentation media and production of exotoxin by three varieties of Bacillus thuringiensis

The insecticidal activities of the exotoxin produced by three varieties of Bacillus thuringiensis grown in six fermentation media were determined by testing the supernatants against larvae of the house fly, Musca domestica, and the black cutworm, Agrotis ipsilon. The activities of the exotoxins from the isolates varied when they were grown in the same medium and also when they were grown in different media. When an isolate of B. thuringiensis var. thuringiensis, and of var. tolworthi were grown in proflo broth, the supernatants produced were more toxic to house fly than to black cutworm larvae, indicating the presence of more than one exotoxin. Autoclaving the supernatants for 15 and 30 min further demonstrated the presence of several exotoxins.     

Fermentation broth of Bacillus thuringiensis as a source of precursors for production of nikkomycins

Production of nikkomycins (nucleoside antibiotics inhibiting chitin synthesis) by Streptomyces tendae (ATCC 31160) was considerably enhanced by addition of fermentation wastes of Bacillus thuringiensis var. kurstaki (HD 263) to the basal soymannitol medium. Analysis of this fermentation broth for nucleic acid derivatives showed that they contained about 0·8 g/l of various nucleosides and bases (uracil 0·43 g/l; hypoxanthine 0·21 g/l; as well as uridine, cytosine and adenine).     

产耐热蛋白酶苏云金芽胞杆菌的发酵动力学

对苏云金芽胞杆菌FZ62耐热蛋白酶的代谢过程进行初步研究,酶生成与菌体生长为耦联过程。以Logistic方程和Luedeking—Piret方程为基础,建立该菌发酵过程中菌体生长、产物生成和基质消耗的动力学模型,求得模型参数,该动力学模型与试验结果拟和程度较好。     

苏云金芽孢杆菌蛋白酶发酵动力学模型的构建

对苏云金芽孢杆菌FS140蛋白酶分枇发酵的代谢特性进行了研究.首先描述了FS140分枇发酵过程中细胞生长、产物积累、糖消耗的变化规律.基于Logistic方程和Luedeking-Piret方程,建立了苏云金芽孢杆菌蛋白酶发酵过程细胞生长、产物合成及基质消耗随时间变化的数学模型.动力学模型计算值结果与实验值拟合良好,较好反映了苏云金芽孢杆菌分批发酵过程的动力学特征.     

苏云金杆菌发酵上清液中生物活性物质的研究

对苏云金芽胞杆菌MP-342发酵上清液中生物活性物质的抑菌作用及化学结构进行初步研究。经717树脂脱色和732树脂吸附、氨水洗脱、真空浓缩及冷冻干燥,得到了纯度较高的精制品。然后进行了抑菌活性试验,结果表明该物质对苹果轮纹菌(Dothiorella gregaria)和苹果褐斑病菌(Marssonin mali)等植物病害微生物有明显的抑菌作用。最后通过质谱分析,表明该活性物质的分子量为396,推测该物质与国外文献报道的zwittermicin A可能为同一物质。     

苏云金杆菌“79007”菌株发酵工艺的研究

对苏云金杆菌79007菌进行了生长条件的研究,获得了该菌最佳发酵培养基配方和最优发酵工艺。最佳培养基配方(g/100 mL):豆饼粉2.5,淀粉2.0,酵母粉2.0,玉米浆2.0,添加剂Ⅰ号0.1,葡萄糖1.0,KH2PO40.3,CaCO30.1,CaCl20.05,MgCl20.01,CuCl20.005,ZnSO40.002,CoCO30.005;15 m3发酵罐最优工艺:通气量1∶0.5~1.1,培养温度28~33℃,搅拌转速0~200 r/min。40 m3发酵罐连续4批试验表明,平均发酵效价6 053.5 IU/μL、平均晶体蛋白含量达0.5%,产品质量达到原粉优质品。     

苏云金芽孢杆菌培养基优化及间歇发酵

对苏云金芽孢杆菌的培养基配方进行室内摇瓶优化筛选,首先用摇瓶培养筛选到Ⅱ号培养基,在此配方的基础上,将培养基组分划分为氮源、碳源及无机盐三因素,采用三因素二水平正交旋转组合设计的方法进行培养基优化组合研究,建立其芽孢产量依氮源、碳源、无机盐的响应面方程。借助此方程获得响应面最佳点即培养基各组分的最佳配比。实验结果表明,该方法是苏云金芽孢杆菌培养基优化中十分简便、实用、快速的途径。此外,对其间歇发酵过程也进行了初步考察。     

A purification and some properties of an insecticidal exotoxin from Bacillus thuringiensis Berliner

An insecticidal exotoxin from Bacillus thuringiensis var. thuringiensis (Berliner) has been purified. The efficiency of each stage of the purification has been ascertained and the yield of toxic material estimated by means of a quantitative bioassay. It is shown that the exotoxin is an adenine derivative substituted at position 9 and having a molecular weight of approximately 825. It can be dephosphorylated enzymically or chemically under conditions that define the exotoxin as a phosphomonoester. This results in loss of toxicity, both to insects and to mice. Spectroscopic and kinetic data are presented which suggest that a beta-ribofuranosyl moiety may be attached to the adenine. Glucose and allomucic acid have been positively identified as hydrolysis fragments from the exotoxin. These results are discussed and compared with the results of others on similar (or possibly identical) compounds.     

苏云金芽胞杆菌Bt 0601发酵条件的优化研究

应用正交实验和单因子实验,结合杀虫毒力测定,对携带杀虫基因cry1Ac和虎纹捕鸟蛛毒素基因Hwtx-I的苏云金芽胞杆菌工程菌Bt 0601的发酵工艺条件进行研究。实验获得的苏云金芽胞杆菌Bt 0601工程菌的最佳发酵条件为:发酵初始酸碱度控制在pH 7.5、发酵温度(30±0.5)℃、转速200 r/min、通气量为30 mL/300 mL,接种量为4%,培养44 h。此外,本文还对Bt 0601菌株在30 L发酵罐中的发酵特性进行了研究,摸索了通气量对其大罐发酵的影响,确定大罐发酵的最佳通气量为1.75 L/min。     

Fermentation during growth and sporulation of Bacillus thuringiensis HD-1

During growth in a glucose-tryptone-mineral salts broth, B. thuringiensis HD-1 Produced lactate, pyruvated, acetate, acetoin, and poly-beta-hydroxybutyrate (PHB). Based on the type and sequence of products found, all or part of the acetate appears to have been produced via the 2,3-butanediol cycle. PHB was produced and utilized during spore formation and may have provided energy for that process.     

Effect of mineral salts on exotoxin formation and productivity of a Bacillus thuringiensis culture

The effect of ten mineral salts on the productivity and toxin synthesis was studied in Bacillus thuringiensis IPM-1140. Exotoxin synthesis was stimulated by Zn2+, Mn2+ and NH4+ ions as well as by potassium phosphates. The direct correlation between the number of viable spores and the exotoxin accumulation was disordered at extreme salt concentrations. Optimal salt concentrations in the yeast-polysaccharide medium were found using the method of a fractional factor experiment, which made it possible to increase the productivity of the culture to 5 X 10(9) spores/ml and the yield of the exotoxin to 730 micrograms/ml. The thermoresistance and the entomopathogenic activity of crystals increased when B. thuringiensis IPM-1140 was grown in this medium.     

苏云金杆菌Bt0601菌株发酵培养基的优化

运用正交试验L18(37)设计对携带杀虫基因cry1Ac和虎纹捕鸟蛛毒素基因hwtx-Ⅰ的苏云金杆菌工程菌Bt0601的发酵培养基进行了优化研究.试验获得的最佳优化复合培养基配方为(%)玉米粉1.0,黄豆饼粉0.50,酵母膏0.15,蛋白胨0.05,磷酸二氢钾0.75,碳酸钙0.05,硫酸镁0.035.Bt0601在该优化培养基下发酵,48 h每mL发酵液产芽孢11.4×109个,伴孢晶体23.0×107个,对3龄小菜蛾(Plutella xylostella)48 h的致死率达96.7%.普通发酵培养基下相应的芽孢产量为9.33×109个,伴孢晶体9.57×107个,毒力为68.5%.     

Cost-effective production of Bacillus thuringiensis by solid-state fermentation

Production of Bacillus thuringiensis (Bt) was standardized on wheat bran based media in 250 ml Erlenmeyer flasks. Scale-up of Bt production on the best medium in plastic tubs with aeration at 8 h intervals starting 16 h after incubation yielded a significant increase in spore count and toxin content of the product. Maximum lysis of Bt cells was obtained by 60 h of incubation at 30 degrees C. This protocol was suitable for production of Bt strains and local isolates. The Bt produced proved highly effective at 0.1% concentration against larvae of castor semilooper, Achaea janata L, resulting in complete mortality by three days in laboratory bioassays. In field trials, the population of castor semilooper larvae on the castor bean crop was reduced significantly by three days after application. The cost for material production of 1 kg of Bt was approximately US dollars 0.70.     

A pangenomic study of Bacillus thuringiensis

Bacillus thuringiensis (B.thuringiensis) is a soil-dwelling Gram-positive bacterium and its plasmid-encoded toxins (Cry) are commonly used as biological alternatives to pesticides.In a pangenomic study,we sequenced seven B.thuringiensis isolates in both high coverage and base quality using the next-generation sequencing platform.The B.thuringiensis pangenome was extrapolated to have 4196 core genes and an asymptotic value of 558 unique genes when a new genome is added.Compared to the pangenomes of its closely related species of the same genus,B.thuringiensis pangenome shows an open characteristic,similar to B.cereus but not to B.anthracis; the latter has a closed pangenome.We also found extensive divergence among the seven B.thuringiensis genome assemblies,which harbor ample repeats and single nucleotide polymorphisms (SNPs).The identities among orthologous genes are greater than 84.5％ and the hotspots for the genome variations were discovered in genomic regions of 2.3-2.8 Mb and 5.0-5.6 Mb.We concluded that high-coverage sequence assemblies from multiple strains,before all the gaps are closed,are very useful for pangenomic studies.     

Novel fermentation media for production of Bacillus thuringiensis subsp. israelensis

The production of Bacillus thuringiensis subsp. israelensis (deBarjac) (Bti) as a biopesticide is not cost-effective using existing fermentation technology. In this study, we explored the use of several less expensive alternative culture media (potato, common sugar, and Bengal gram) for the growth and production of Bti. Growth was obtained in all tested media and was comparable to that obtained in conventional medium (Luria-Bertani). Toxicity assays showed that the toxin produced from the novel growth media were effective in killing larvae of Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti and toxicity was comparable to that produced from Luria-Bertani medium. These observations suggest that potato can be used as a cheap source of culture medium for the production of Bti toxin in mosquito control programs.     

Identification of beta-exotoxin production, plasmids encoding beta-exotoxin, and a new exotoxin in Bacillus thuringiensis by using high-performance liquid chromatography.

An improved high-performance liquid chromatography separation was developed to detect and quantify beta-exotoxin production in Bacillus thuringiensis culture supernatants. Exotoxin production was assigned to a plasmid in five strains, from three subspecies (B. thuringiensis subsp. thuringiensis serotype 1, B. thuringiensis subsp. tolworthi serotype 9, and B. thuringiensis subsp. darmstadiensis serotype 10). A new exotoxin, called type II beta-exotoxin in this report, was discovered in B. thuringiensis subsp. morrisoni serotype 8ab, purified, and partially characterized. This material is more specific than type I beta-exotoxin and is very active against the Colorado potato beetle, Leptinotarsa decemlineata.