纳豆激酶的发酵工艺研究

以实验室选育的纳豆菌— 6号 (Bacillusnatto 6)为出发菌株 ,研究其固液两种发酵条件对纳豆激酶溶栓活力的影响。研究表明 ,纳豆菌适于固体发酵 ,37℃下 2 4h ,其活力可达 1 60 0IU/g以上。     

棉铃虫和甜菜夜蛾中肠丝氨酸蛋白酶活性测定以及活化、降解Cry1Ac毒素分析

昆虫中肠对Bt原毒素活化与对活化毒素降解的变化被认为是害虫对Bt产生的机制之一,研究比较棉铃虫Helicoverpa armigera(Hübner)与甜菜夜蛾Spodoptera exigua(Hübner)的中肠液、BBMV蛋白酶的活性,通过SDS-PAGE分析2种昆虫对原毒素的活化速度与对活化毒素的降解速度。2种昆虫的中肠液蛋白酶活性均显著高于BBMV蛋白酶活性,中肠液与BBMV均能迅速活化原毒素并继续降解活化后的毒素,与中肠液相比,BBMV对原毒素的活化与对活化毒素的降解均慢于中肠液,甜菜夜蛾对毒素的活化与降解又慢于棉铃虫。另外,还测定抑制剂对中肠液蛋白酶活性的抑制作用,结果表明,各抑制剂对棉铃虫和甜菜夜蛾相应酶活性的抑制表现出相同的趋势,TLCK对丝氨酶蛋白酶具较好的抑制作用,而PMSF对胰蛋白酶的抑制作用次之,TPCK对胰凝乳蛋白酶的抑制作用较弱。     

一株碱性淀粉酶产生菌Bacillus flexus XJU-3的分离鉴定及酶学特性分析

[目的]从新疆石河子市一处碱性工业污水(pH11.0)分离﹑鉴定高产碱性淀粉酶菌株,并对其所产酶酶学特性进行研究.[方法]在碱性淀粉酶分离培养基上对所分离菌株进行筛选,分离到一株高产碱性淀粉酶菌株,并将其编号XJU-3.应用生理生化试验,脂肪酸含量,16S rDNA序列以及(G C) mol%含量等方法对菌株进行鉴定,同时对XJU-3所产碱性淀粉酶的生物学特性进行研究.[结果] XJU-3可在pH4.0～12.5的LB培养基上生长,最适生长温度37℃.16S rDNA序列构建的系统进化树表明XJU-3与Bacillus flexus类聚在一起,且序列同源性为99%.该菌产生的淀粉酶最适pH10.0,最适温度40℃,且在pH9.0～13.0内有较高活性和稳定性.Co2 和Mg2 能明显提高酶的活性.[结论] XJU-3被鉴定为Bacillus flexus,由于XJU-3与B. flexus DSM 1320T在尿素水解和优势脂肪酸含量上有差异,且具有宽范围pH耐受性,因此XJU-3被认为是B.flexus的一个新菌株.XJU-3所产的碱性淀粉酶酶学特性良好,具有极大的工业应用潜力.     

生物素对地衣芽孢杆菌生长和乳酸形成的影响

生物素对地衣芽孢杆菌生长和乳酸形成的影响胡尚勤（重庆师范学院生物系重庆６３００４７）地衣芽孢杆菌（Ｂａｃｉｌｕｓｌｉｃｇｅｎｉｆｏｒｍｉｓ）是我国９０年代首次从正常待产妇阴道拭子分离出来的一个无毒新菌株。现在国内已经用它来生产“整肠生”活菌剂。它是一...     

产生物表面活性剂芽孢杆菌培养条件的优化

为了提高生物表面活性剂的表面活性,通过单因素及正交试验对已筛选的产生物表面活性剂芽孢杆菌的培养基及培养条件进行了优化,优化后的培养基成分为可溶性淀粉20 g/L,氯化铵2 g/L,KH2PO46 g/L,K2HPO42 g/L,MgSO4.7H2O 0.3 g/L,NaCl 2 g/L,CaCl20.08 g/L,EDTA 0.4 g/L。培养条件为4%接种量,种龄16 h,初始pH7,培养温度37℃,摇床转速160 r/min,发酵48 h。优化发酵条件后,发酵液表面张力由初始67.5 mN/m降低至24.8 mN/m,生物表面活性剂产量达到1.08 g/L。     

芽孢杆菌植酸酶分子生物学研究进展

来源于芽孢杆菌的中性植酸酶具有良好的热稳定性,适合作为消化道呈中性鲤科鱼类的饲料添加剂。该植酸酶晶体具有独特的螺旋桨样空间结构,钙离子在其稳定性和催化活性的结构功能关系中具有重要作用,并表现出特有的催化机理。该基因与已知植酸酶基因的结构有所不同,是一种新的植酸酶。目前主要采用多种原核表达系统对其进行表达。比较表明,芽孢杆菌表达系统和酵母表达系统是实现芽孢杆菌植酸酶规模化生产的有效途径。     

Role of lipopeptides produced by Bacillus subtilis GA1 in the reduction of grey mould disease caused by Botrytis cinerea on apple

AIM: Test of Bacillus subtilis strain GA1 for its potential to control grey mould disease of apple caused by Botrytis cinerea. METHODS AND RESULTS: GA1 was first tested for its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The potential of strain GA1 to reduce post-harvest infection caused by B. cinerea was tested on apples by treating artificially wounded fruits with endospore suspensions. Strain GA1 was very effective at reducing disease incidence during the first 5 days following pathogen inoculation and a 80% protection level was maintained over the next 10 days. Treatment of fruits with an extract of GA1 culture supernatant also exerted a strong preventive effect on the development of grey mould. Further analysis of this extract revealed that strain GA1 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families. A strong evidence for the involvement of such compounds in disease reduction arose from the recovery of fengycins from protected fruit sites colonized by bacterial cells. CONCLUSIONS: The results presented here demonstrate that, despite unfavourable pH, B. subtilis endospores inoculated on apple pulp can readily germinate allowing significant cell populations to establish and efficient in vivo synthesis of lipopeptides which could be related to grey mould reduction. SIGNIFICANCE AND IMPACT OF THE STUDY: This work enables for the first time to correlate the strong protective effect of a particular B. subtilis strain against grey mould with in situ production of fengycins in infected sites of apple fruits.     

Recombinant Bacillus thuringiensis strain shows high insecticidal activity against Plutella xylostella and Leptinotarsa decemlineata without affecting nontarget species in the field

Aims: To construct a recombinant Bacillus thuringiensis (Bt) strain with broad insecticidal spectrum and investigate its impact on nontarget organisms in field. Method and Results: The cry-type gene of wild Bt strain UV17 was identified and a novel cry1Ba gene was cloned. The cry3Aa7 gene, which was highly toxic to coleopteran pests, was introduced into UV17, and a recombinant strain designated as UV173A was obtained. Bioassay results showed that UV173A was not only highly toxic against Plutella xylostella (50% lethal concentration [LC₅₀] = 18·03 μg ml^–1), but also against coleopteran Leptinotarsa decernlineata (LC₅₀ = 0·19 mg ml^–1). The recombinant strain was then tested in field trials to monitor its spatial variation of population and to investigate the impact on nontarget invertebrates. Conclusions: A recombinant Bt stain UV173A with broad insecticidal spectrum was obtained, and it did not cause adverse effects on the population of nontarget organisms. Significance and Impact of the Study: The results obtained here indicated that cry1Ba3 gene may be useful for the resistance management of P. xylostella, and the recombinant stain UV173A was potential for field application against some crucifer vegetable pests as well as L. decemlineata.     

Diversity of commensal Bacillus cereus sensu lato isolated from the common sow bug (Porcellio scaber, Isopoda)

Although Bacillus cereus sensu lato are important both from an ecological and an economical point of view, little is known about their population structure, ecology, and relationships with other organisms. In the present work, the genotypic similarity of arthropod-borne B. cereus s.l. isolates, and their symbiotic relationship with the host are assessed. Bacilli of this group were recovered from the digestive tracts of sow bugs (Porcellio scaber) collected in three closely located sites. Their genotypic diversity was investigated using pulse-field gel electrophoresis (PFGE) following the whole-genome DNA digestions with NotI and AscI, and PCR amplification of virulence genes. The majority of the sow-bug Bacillus cereus sensu stricto isolates originating from the same but also from different sites displayed identical PFGE patterns, virulence gene content and enterotoxicity, indicating strong genetic and genomic relationships. The sow-bug Bacillus mycoides/Bacillus pseudomycoides strains displayed a higher diversity. The isopod-B. cereus s.l. relationship was also evaluated using antibiotic-resistant derivatives of B. cereus s.s., B. mycoides/B. pseudomycoides and Bacillus thuringiensis reintroduced into sow bugs. Both spores and vegetative cells of B. cereus s.l. were recovered from sow bugs over a 30-day period, strongly suggesting that these bacteria are natural residents of terrestrial isopods.     

枯草芽孢杆菌抗菌蛋白X98Ⅲ的纯化与性质

枯草芽孢杆菌（Bacillussubtilis）BS－98是一株能强烈抑制苹果轮纹病菌（Physalosporapiricola）等植物病原真菌的拮抗菌株。BS－98菌株培养液经硫酸铵分级盐析、SephadexG－100柱层析和DEAE－纤维素（DE32）柱层析后分离纯化出一种抗菌蛋白,命名为X98Ⅲ。蛋白电泳分析结果表明,此蛋白分子量为59000,等电点为4．50．醋酸纤维膜电泳后经特异染色证明X98Ⅲ含糖及胀。用DNS法测其含糖量为6％。此蛋白对热稳定,对蛋白酶部分敏感。氨基酸组分分析表明,该蛋白含11种不同氨基酸,尤富含谷氨酸和半胱氨酸等,而缺少天冬氨酸等。纯化后的X98Ⅲ对苹果轮纹病菌、芦笋茎枯病菌等有很强的抑制作用。X98Ⅲ的抑菌机理主要是溶解细胞壁,造成菌丝畸形、孢子不发芽或发芽异常。