Overexpression and export of Vibrio anguillarum metalloprotease in Escherichia coli

Vibrio anguillarum metalloprotease, an extracellular zinc metalloprotease involved in the virulence mechanism of Vibrio anguillarum, is synthesized from the empA gene as a 611-residue precursor and naturally secreted via Sec secretion pathway in Vibrio anguillarum. In this study, heterologous expression of the empA gene encoding metalloprotease and export of the recombinant metalloprotease in Escherichia coli were examined. The empA gene was subcloned into pBAD24 with arabinose promoter and sequenced. The sequence encoded a polypeptide (611 amino acids) consisting of four domains: a signal peptide, an N-terminal propeptide, a mature region and a C-terminal propeptide. The empA gene inserted in plasmid pBAD24 was overexpressed in TOP10 strain of E. coli after arabinose induction. The 36kDa polypeptide of the recombinant metalloprotease as the mature protease was further confirmed by SDS-PAGE and immunoblotting. It was found that recombinant metalloprotease with the EmpA activity and antigenicity was exported into the periplasm of Escherichia coli cells via Sec translocation pathway, whereas it was secreted into extracellular environments in V. anguillarum. The results imply that the expression, export and processing mechanism of the protein in E. coli are similar to those in V. anguillarum.     

Overexpression and export of Vibrio anguillarum metalloprotease in Escherichia coil

Vibrio anguillarum metalloprotease, an extracellular zinc metalloprotease involved in the virulence mechanism of Vibrio anguillarum, is synthesized from the empA gene as a 611-residue precursor and naturally secreted via Sec secretion pathway in Vibrio anguillarum. In this study, hetemlogous expression of the empA gene encoding metallopmtease and export of the recombinant metallopmtease in Escherichia coli were examined. The empA gene was subcloned into pBAD24 with arabinose promoter and sequenced. The sequence encoded a polypeptide （611 amino acids） consisting of four domains： a signal peptide, an Nterminal pmpoptide, a mature region and a C-terminal pmpoptide. The empA gene inserted in plasmid pBAD24 was overexpressed in TOP10 strain of E. coli after arabinose induction. The 36kDa polypeptide of the recombinant metallopmtease as the mature pmtease was further confirmed by SDS-PAGE and im- munoblotting. It was found that recombinant metallopmtease with the EmpA activity and antigenicity was exported into the poriplasm of Escherichia coli cells via Sec translocation pathway, whereas it was secreted into extracellular environments in V. anguillarum. The results imply that the expression, export and processing mechanism of the protein in E. coli are similar to those in V. anguillarum.     

极化雷达与光学遥感森林雪灾破坏协同监测

以贵州省扎佐林场为例,利用全极化雷达与光学遥感数据进行森林雪灾破坏状况协同监测,分析了倒伏森林与健康森林的后向散射特性及其差异,重点分析了倒伏森林与健康森林的极化响应特征、散射机制及其差异。基于雪灾破坏前后森林的散射机制变化,利用Freeman-Durden分解方法对倒伏森林进行了识别。为了进一步优化识别效果,引入了高分辨率光学数据,采用优选的融合方法, 将雷达与光学遥感数据融合,得到了较好的倒伏森林识别效果。研究结果表明,利用雷达与光学遥感数据协同监测森林雪灾破坏是可行的,而全极化雷达遥感数据在森林类型识别和森林破坏监测方面具有独特的优势,在我国西南地区的森林资源调查和灾害监测中具有广阔的应用前景。     

走进海洋微生物的世界

海洋占整个地球表面积的70％，其中蕴藏着丰富的海洋微生物资源，是21世纪亟待开发和利用的巨大宝藏。海洋中存在着许多自然条件特殊的环境，如低温、高温、高压、高盐、强酸、低营养和低浓度溶解氧等环境，一般的海洋生物无法在其中生存；只有某些具备特殊生理和遗传特性的微生物，如嗜冷微生物、嗜压微生物、嗜盐微生物、嗜酸微生物、寡营养微生物和嗜热古细菌，才可以在这类环境中生长和代谢，产生独特的代谢产物。     

一株地衣芽孢杆菌脲酶生产的发酵优化和酶学性质

对中国南海海绵Halichondria regosa中分离得到脲酶产生菌Bacillus licheniformis B81进行脲酶生产的发酵条件优化;并对其脲酶特性进行测试。通过对基础培养基筛选后,对选定基础培养基进行了Plachett-Bunman重要因素筛选,最终选定的重要因素通过单因素实验得到了最终的培养基配方为:葡萄糖25 g/L,蛋白胨12.5 g/L,牛肉膏5 g/L,酵母粉6.5 g/L,KH2PO42 g/L,NaCl 1 g/L,尿素0.625%(W/V),Ni(NO3)20.006 25%(W/V),H2O 1 L。优化后,使得脲酶的产量提高到了原培养基的170%。菌株所产生的脲酶的最适反应pH为7.0,最适温度为40℃,Pb2+、Cu2+对脲酶活性具有抑制作用。