偶氮染料酸性橙7脱色菌的分离、鉴定及其脱色性能研究

从活性污泥中筛选得到一株对偶氮染料酸性橙7(Acid Orange 7)具有高效脱色功能的菌株,根据16S rRNA基因序列分析鉴定,该菌属于嗜水气单胞菌属,将其命名为Aeromonas hydrophila strain JL-1.同时,研究了该菌株对偶氮染料酸性橙7的最佳脱色条件,结果表明,菌株JL-1在缺氧振荡培养条件下能对偶氮染料酸性橙7进行有效脱色;培养基、初始染料浓度、接种量、pH和NaCl浓度等环境条件对菌株JL-1脱色性能及其生长有明显影响.pH为5.0、盐度为10%、温度为30℃、转速为150 r·min~(-1)的LB(Luria-Bertani medium)培养基为菌株JL-1最佳的脱色及生长环境;在此最佳环境条件下,菌株JL-1对100 mg·L~(-1)的偶氮染料AO7在8 h的脱色率高达95%.另外,该菌株耐盐性良好,能在高达30%盐度下依然可以进行有效的生物脱色.     

柠檬酸对3种常见水产病原菌的抑菌作用

选用嗜水气单胞菌、副溶血弧菌、荧光假单胞菌3种常见水产病原菌,用于测定柠檬酸的抑菌效果及最小抑菌浓度,并研究了柠檬酸抑菌作用的持续性。3种菌均为革兰氏阴性菌。试验结果表明,柠檬酸对3种菌都有不同程度的抑菌作用,并得到最小抑菌浓度,依次是:0.0011g/mL,0.0008g/mL,0.00125g/mL;在最小浓度下抑菌作用可达6d之久,柠檬酸抑菌效果有较好的持续性。     

Cloning, expression in Escherichia coli, and enzymatic properties of laccase from Aeromonas hydrophila WL-11

A strain WL-11 with high laccase activity was isolated from activated sludge collected from the effluent treatment plant of a textile and dyeing industry.It was identified as Aeromonas hydrophila by physiological test and 16S rDNA sequence analysis.A gene encoding of laccase from a newly isolated Aeromonas hydrophila WL-11 was cloned and characterized.Nucleotide sequence analysis showed an open reading frame of 1605 bp encoding a polypeptide comprised of 534 amino acids.The primary structure of the enzyme predicted the structural features characteristic of other laccases,including the conserved regions of four histidine-rich copper-binding sites.The predicted amino acid sequence showed a high homology(more than 60%) with bacterial laccases in the genome and protein databases and the highest degree of similarity(61% identity) was observed with the multicopper oxidase of Klebsiella sp.601.When expressed in Escherichia coli,the recombinant enzyme was overproduced in the cytoplasm as soluble and active form.The purified enzyme had an optimum pH of 2.6 and 8.0 for ABTS(2,2'-azino-bis(3-ethylbenzthiazolinesulfonic acid) and DMP(2,6-dimethoxyphenol),respectively.The kinetic study on ABTS revealed a higher affinity of this enzyme to this substrate than DMP.     

脱色酶TpmD在大肠杆菌中的高效表达及纯化

用PCR方法从嗜水气单胞菌DN322基因组中扩增出编码三苯基甲烷类染料脱色酶TpmD的基因,与表达载体pET-22b(+)连接构建成重组质粒pET22-tpmD,转化大肠杆菌BL21(DE3)得到重组工程菌株.结果表明,经IPTG诱导,脱色酶基因可高效表达,粗酶液降解结晶紫、孔雀石绿、碱性品红、灿烂绿的比活力达到569.5,386.9,516.1,273.0U/g.表达产物经Ni-NTA亲和层析法一步纯化,蛋白纯度达94.05%.对4种染料的比活力分别达到1075.3,1042.8,903.9,484.3U/g,重组质粒稳定存在于工程菌中,便于规模化发酵生产.     

越冬期淡水白鲳发病之病原细菌的研究

从越冬期淡水白鲳病鱼的肝脏内分离纯化到一细菌菌株ＣＢＺＨＧ４,用ｎ（ｃｅｌｌ＝１０＾５～１０＾６ｍＬ＾－１的菌液浓度在腹腔注射试验鱼后４８ｈ内全部死亡,证实为病原菌。ＣＢＺＨＧ４经ＡＰＩ ２０ＮＥ系统鉴定为嗜水性气单胞菌（Ａｅｒｏｍｏｎａｓ ｈｙｄｒｏｐｈｉｌａ）,对卡那霉素多种抗生素和呋喃类药物敏感,对青霉素类、先锋霉素等抗生素、磺胺类药物和０／１２９不敏感。表２参２