Kim Se-Kwon, Lee Baek-Suk, Wilson David B, Kim Eun-Ki
Department of Biological Engineering, Inha University, Incheon 402-751, Korea.
J Biosci Bioeng. 2005 Feb;99(2):109-14. doi: 10.1263/jbb.99.109.
Recombinant Escherichia coli JM109 (pZH3-5/pMT), harboring a manganese transport gene (mntA) and a metal-sequestering protein (metallothionein [MT]) gene, was cultivated to accumulate cadmium (Cd) in an aqueous phase. Isopropyl beta-D-thiogalactoside (IPTG)-induced cells showed rapid Cd(2+) ion accumulation (90% of maximum accumulation in 15 min) and had an accumulation six times higher than that of the control. Under optimum conditions, i.e., pH 7, 37 degrees C and 0.5 (OD600), 1.5 mM IPTG induction resulted in the accumulation of 21.5 micromol Cd/g dry cell. Storage at 37 degrees C for 24 h had no effect on the accumulation. Significantly, Cd was selectively accumulated in a solution containing an equal concentration of three other metals, resulting in more than 90% of the total accumulated metals being Cd. The accumulation of Cd was reduced by the presence of Mn2+ ion whereas no significant effect was observed with Cu2+, Zn2+ and Pb2+ ions. A chelator, EDTA, had no effect on the accumulation up to 100 mM. The bioaccumulation rate followed Michaelis-Menten kinetics (Vm=2.7 micromol Cd2+/min.g dry cell, Km=0.67 microM). The equilibrium isotherm showed a Langmuir isotherm. In the membrane reactor experiment, 1 mg/l Cd in an inlet solution decreased to 0.2 mg/l in the effluent, removing 80% of Cd, continuously. These results indicated the potentials of a genetically modified microorganism for the highly selective accumulation of Cd at a low concentration and the future application to the removal and recovery of Cd.
携带锰转运基因(mntA)和金属螯合蛋白(金属硫蛋白[MT])基因的重组大肠杆菌JM109(pZH3 - 5/pMT)在水相中培养以积累镉(Cd)。异丙基β - D - 硫代半乳糖苷(IPTG)诱导的细胞显示出快速的Cd(2+)离子积累(15分钟内达到最大积累量的90%),并且积累量比对照高六倍。在最佳条件下,即pH 7、37℃和0.5(OD600),1.5 mM IPTG诱导导致每克干细胞积累21.5微摩尔Cd。在37℃储存24小时对积累没有影响。值得注意的是,在含有等浓度的其他三种金属的溶液中,Cd被选择性积累,导致积累的总金属中超过90%是Cd。Mn2+离子的存在会降低Cd的积累,而Cu2+、Zn2+和Pb2+离子未观察到显著影响。螯合剂EDTA在高达100 mM时对积累没有影响。生物积累速率遵循米氏动力学(Vm = 2.7微摩尔Cd2+/分钟·克干细胞,Km = 0.67微摩尔)。平衡等温线显示为朗缪尔等温线。在膜反应器实验中,进水溶液中1毫克/升的Cd在流出物中降至0.2毫克/升,连续去除80%的Cd。这些结果表明了转基因微生物在低浓度下对Cd进行高度选择性积累的潜力以及未来在Cd去除和回收方面的应用。