角蛋白酶产生菌的分离鉴定及其kerC的克隆表达

以四川农业大学养鸡场堆砌废弃羽毛处土壤为样品,筛选出一株具有较强降解羽毛能力的细菌B-3菌株.经形态学、生理生化特性和16S rRNA基因序列分析,鉴定其为枯草芽孢杆菌(Bacillus subtilis),命名为枯草芽孢杆菌B-3.并成功克隆到该菌株的角蛋白酶基因kerC(GenBank No.:JN021789),在大肠杆菌BL21(Escherichia coli BL21)中获得了高效表达.该基因全长1146bp,GC含量46.5%,编码381个氨基酸,与已报道的枯草芽孢杆菌YYW-1的kerC基因(GenBank No.: EU362730)同源性达到100%.重组菌株经IPTG诱导后角蛋白酶酶活力达14.8U/mL,经His-Tag纯化和SDS-PAGE分析表明,重组角蛋白酶分子量约为60kDa(融合了硫氧还蛋白,Trx).重组角蛋白酶最适反应温度和pH值分别为65℃与7.0.     

硝基苯好氧降解菌筛选及其降解特性

针对化学试剂厂和制药厂废水中的硝基苯,经菌源筛选与长期好氧驯化,分离到1株能有效降解硝基苯的菌株Bacillus subilis(枯草芽孢杆菌)。系统研究了其生长条件及降解硝基苯的特性,适宜生长条件: pH为5.0～8.0,温度为30～40℃,NH4Cl浓度为100～200 mg/L,在降解硝基苯的过程中有苯胺生成,这为含低浓度硝基苯废水的生物处理提供了新的途径,即不需要厌氧工艺而直接用好氧技术就可将废水完全无害化。      

枯草芽孢杆菌对铜绿微囊藻抑制效果的研究

为探讨枯草芽孢杆菌(Bacillus subtilis)对铜绿微囊藻(Microcystis aeruginosa)的抑制效果,在实验室条件下,研究了枯草芽孢杆菌不同生长时期(延迟期、对数期、稳定期和衰亡期)无菌滤液对铜绿微囊藻生长的影响、枯草芽孢杆菌抑制铜绿微囊藻生长的作用方式以及无菌滤液影响下铜绿微囊藻丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性和光合色素含量的变化.结果显示:枯草芽孢杆菌对数期、稳定期和衰亡期滤液抑藻效果明显好于延迟期,作用第8d,对铜绿微囊藻的去除率分别达到81.19%、91.41%、91.82%;4个处理组铜绿微囊藻的叶绿素a含量均显著低于对照组.添加稳定期滤液后,铜绿微囊藻MDA含量显著升高,SOD活性先升高后降低;在对光合色素的影响中,类胡萝卜素受到的影响不如叶绿素a显著.结果表明,枯草芽孢杆菌对铜绿微囊藻的抑制效果是通过分泌胞外物质实现的,且分泌物具有很强的热稳定性.推测该胞外分泌物能够破坏光合色素,影响光合作用,抑制藻细胞的生长;同时抑制SOD活性,使细胞膜脂过氧化程度不断加深,进而破坏藻细胞的完整性,表现出对藻很强的抑制效果.     

Isolation and identification of a bioflocculant-producing strain and optimisation of cultural conditions via a response surface model

A bioflocculant-producing strain named MY6-2 was isolated from a mixed activated sludge by a nitrogen-free medium. Based on 16S rDNA sequence, biochemical and morphological characteristics, the strain MY6-2 was identified as Bacillus mucilaginosus. The chemical analysis indicated that the flocculant MY6-2 was mainly composed of extracellular polysaccharide. The result of the composition of the medium showed that MY6-2 was able to generate bioflocculant in a nutrient-poor medium that consisted of 5?g?L^?1 sucrose and no nitrogen source. Response surface methodology (RSM) was used to optimise the fermentation condition of MY6-2 for maximum flocculating activity by using central composite design (CCD). Based on the result, the optimum conditions were as follows: 100?mL of broth in a 250?mL Erlenmeyer flask, initial pH 8.0 and inoculum concentration 10%, respectively. The highest flocculating rate of 90% was achieved under these conditions by adding 0.5?mL fermentation supernatant to 95?mL of Kaolin suspension.     

Microbiological evaluation of 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one with a comparison to antibacterial activity of standard drugs

Nucleophilic substitution of 4-chloro-3-nitro-2-oxo-2H-chromen-2-one 3 by 6-chloro-7H-purin-2-ylamine (a), yielded the corresponding 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one 4. The chemical structure of synthesized compound was characterized using IR and NMR spectra, and elemental analyses. The purified synthesized compound 4 was tested at concentrations 1, 3, 5 mg/ml for its antibacterial activity against three bacterial cultures: Staphylococcus aureus, Escherichia coli, and Bacillus cereus. The antibacterial activity of synthesized compound was compared to antibacterial activity of standard antibiotics cephalexine and streptomycin.     

城市居家环境空气细菌群落结构特征

在北京市5个方向(东南西北中)共选取31户有小孩的家庭于2009年11月至2010年10月研究了城市居家环境空气细菌的群落结构特征.结果表明,从分离的632株空气细菌中共鉴定出43属细菌,其中革兰氏阳性菌32属,革兰氏阴性菌11属.优势菌属依次为微球菌属(Micrococcus)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)和库克氏菌属(Kocuria), 分别占25%~31%,12%~17%, 10%~15%,9%~18%,4属细菌百分比约占63%~71%.在北京市取样的31户家庭中,空气细菌浓度范围为47~12341cfu/m3,平均值为1821cfu/m3.总体上,春季和夏季空气细菌浓度分别为2967cfu/m3和1742cfu/m3,明显高于秋季和冬季的1334cfu/m3和1242cfu/m3(P<0.05).北京市居家环境空气细菌浓度男孩家庭(2123cfu/m3)明显高于女孩家庭(1511cfu/m3)(P<0.01).     

Biodegradation of Hexachlorocyclohexane by two species of Bacillus isolated from contaminated soil

Biodegradation of α, β, γ and δ hexachlorocyclohexane (HCH) isomers was studied in broth medium and soil microcosm by Bacillus circulans and Bacillus brevis isolated from contaminated soil. Degradation of α and γ isomers by both the bacterial isolates was higher than thermodynamically stable β and δ isomers. However, B. circulans was found more effective than B. brevis for β and δ isomers. Maximum rate of degradation was recorded at 150 mg/L followed by 100 and 50 mg/L. Soil microcosm study revealed maximum degradation of HCH isomers in the treatment containing natural soil, pesticide and bacterial inocula than the treatment having sterilised soil, pesticide and bacterial isolates. Chloride release was positively co-related with HCH degradation in broth medium as well as in soil microcosm, suggesting that B. circulans and B. brevis hold promising potential by having efficient enzyme(s) required for dechlorination of HCH from contaminated sites.     

Degradation of Polylactide by Commercial Proteases

Fifty-six commercially available proteases were tested for polylactide-degrading activity. Little or no activity was found in acid and neutral proteases, while some alkaline proteases formed appreciable amounts of lactic acid from polylactide. These polylactide-degrading proteases were derived from Bacillus species and had catalytic activity even under neutral, as well as alkaline, conditions. Savinase (Novo Nordisk) degraded polylactide the fastest among the enzymes tested and its specific activity corresponded to about one-half of proteinase K. Polylactide-degrading activity was not always present in the enzymes that affected keratin, while polylactide-degrading proteases commonly hydrolyzed keratin. A significant correlation was observed between degrading activities of polylactide and keratin in alkaline proteases.     

巨大芽孢杆菌LY-4对土壤中杀虫单农药的降解

为了探明杀虫单高效降解菌LY－4进入土壤后的生存状况及对杀虫单农药的降解动态,进行了一系列实验室土壤模拟研究．结果表明,没有外加菌时,杀虫单在土壤中的降解是缓慢的;加入降解菌后可使杀虫单的降解大大加快,当土壤中杀虫单含量一定时,随着菌量增加．杀虫单的降解加快;当菌剂量一定时,则存在一个最适降解浓度,对应此浓度,由外加菌所贡献的净增加降解率最高,而单位菌剂量对杀虫单的绝对去除量随杀虫单浓度的升高而增加．实验还证明该降解菌对外加营养的依赖性小,适应性较强     

蜡状芽孢杆菌Bacillus cereus吸附铅的研究

通过吸附实验和微量滴定法探讨了Pb²⁺与蜡状芽孢杆菌Bacillus cereus的吸附特性.结果表明,该菌体对Pb²⁺的吸附量随着pH升高而升高,最适pH值为6;菌体浓度的增加有利于对Pb²⁺的吸附;整个吸附过程符合Langmuir吸附模型,在试验的条件下,对Pb²⁺的饱和吸附容量为36.7mg/g干菌体.通过电位滴定实验,使用实时格氏图确定了滴定体系中的等当点(Ve)以及估算了表面活性位的总浓度(HS).