耐辐射球菌对Cu2+的去除效率

耐辐射球菌是一种非致病菌,拥有极强的抗辐射、抗氧化特性,具有成为工程菌处理Cu2＋的潜力。采用酶标仪测定Cu2＋对耐辐射球菌的生长速度的影响,用电感耦合等离子光谱法测定菌液离心后得到的上清液Cu2＋浓度。结果表明,当Cu2＋初始质量浓度为1．36mg／L时,耐辐射球菌对其清除效率为57．3％;当-Cu2＋初始质量浓度为6．28mg／L时,对其清除效率为35．4％。此外,用激光共聚焦显微镜观察受到Cu2＋胁迫的耐辐射球菌,发现球菌具有明显的聚集效应。     

Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

Geranylgeranyl diphosphate synthase (GGPPS) plays a key role in the biosynthesis of antioxidative carotenoid from the extremely radioresistant bacterium Deinococcus radiodurans. In this work, the recombinant GGPPS expressed in Escherichia coli by cloning and transforming the gene dr1395 of D. radiodurans was isolated rapidly by an immobilized metal affinity supermacroporous cryogel, i.e., Cu²⁺-iminodiacetic acid (IDA)-cryogel. The properties of the Cu²⁺-IDA-cryogel were characterized using capillary-based mathematical model and experimental measurements. The obtained protein samples were analyzed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The porosity of the present Cu²⁺-IDA-cryogel is 90.4% and the water permeability is 5.04×10^-12 m². From the capillary-based model, this cryogel presents a slightly wide normal pore (capillary) size distribution with the mean diameter of 55.2 μm, the standard deviation of 28.0 μm and the half of skeleton wall thickness of 2.8 μm. The pore size distribute from about 10 to 141 μm and the effective tortuosity of these capillary pores increases from 2.60 to 9.05. The isolation of the GGPPS from cell homogenate can be achieved at the flow velocity of 3.40×10^-4 m·s^-1 by the Cu²⁺-IDA-cryogel bed. High-purity GGPPS (about 91.4%) is obtained according to the SDS-PAGE analysis of the elution samples, indicating that the present method is a promising, simple and effective approach to isolate GGPPS from cell homogenate of engineering strains.