Shavandi Mahmoud, Sadeghizadeh Majid, Zomorodipour Alireza, Khajeh Khosro
Department of Genetics, Faculty of Science, Tarbiat Modares University, Tehran, Iran.
Bioresour Technol. 2009 Jan;100(1):475-9. doi: 10.1016/j.biortech.2008.06.011. Epub 2008 Jul 23.
The dszABC genes from newly reported dibenzothiophene biodesulfurizing bacterium, Gordonia alkanivorans RIPI90A were cloned and sequenced. The overall nucleotide sequence similarity between the dszABC genes of G. alkanivorans RIPI90A and those of Rhodococcus erythropolis IGTS8 and Gordonia nitida were 83.1% and 83.2%, respectively. A gene transfer system for G. alkanivorans RIPI90A was established employing the Escherichia coli-Rhodococcus shuttle vector pRSG43 as suitable cloning vector, resulting in transformation efficiencies up to 1.6 x 10(5)CFUs microg(-1) plasmid DNA. This stable vector was applied to cloning and efficient expression of the dsz genes under the control of lac promoter. The recombinant strain was able to desulfurize dibenzothiophene in the presence of inorganic sulfate and sulfur-containing amino acids. The maximum desulfurization activity by recombinant resting cells (131.8 microM2-hydroxybiphenylg(dry cell weight)(-1)h(-1)) was increased 2.67-fold in comparison to the highest desulfurization activity of native resting cells.
对新报道的二苯并噻吩生物脱硫细菌戈登氏嗜烷菌RIPI90A的dszABC基因进行了克隆和测序。戈登氏嗜烷菌RIPI90A的dszABC基因与红平红球菌IGTS8和光亮戈登氏菌的dszABC基因的总体核苷酸序列相似性分别为83.1%和83.2%。以大肠杆菌-红球菌穿梭载体pRSG43作为合适的克隆载体,建立了戈登氏嗜烷菌RIPI90A的基因转移系统,转化效率高达1.6×10⁵CFU/μg质粒DNA。该稳定载体用于在lac启动子控制下克隆和高效表达dsz基因。重组菌株能够在无机硫酸盐和含硫氨基酸存在的情况下对二苯并噻吩进行脱硫。与天然静止细胞的最高脱硫活性相比,重组静止细胞的最大脱硫活性(131.8μM 2-羟基联苯/(干细胞重量)⁻¹h⁻¹)提高了2.67倍。