title: [Co-expression of Rhodococcus sp. DS-3 dszABC and dszD gene with incompatible plasmids in Escherichia coli].
abstract: Biological dusulfurizaion of petroleum feedstocks and products may offer an attractive alternative to reduce sulfur oxide emissions that cause serious environmental pollution. Dibenzothiophene (DBT), a model of organic sulfur compound in petroleum, can be microbially desulfurized without degradation of the organic structure by 4S pathway. Three desulfurization enzymes (DszA, DszB and DszC) and flavin reductase (DszD) are involved in sulfur-specific DBT desulfurization. DszA and DszC are FMNH2-dependent monoxygenases, FMNH2 is provided from the freely diffusible FMNH2 pool in the cell, and is replenished by DszD. So, co-expression of the desulfurization enzymes and flavin reductase can enhance the rate of sulfur removal. In the present work two incompatible plasmids: pBADD and paN2 were constructed. The paN2 allows Escherichia coli to liberate the sulfur of DBT and DBTs and pBADD produces a flavin reductase. They were co-expressed in Escherichia coli B121 (DE3). The soluble products of DszA, DszB, DszC and DszD accounted for 7.6%, 3.5%, 3.1% and 18% of the total proteins in co-expressed system. The desulfurization rate of lysate of E. coli BL21- pBADD + paN2 is 12.03 micromol/(h x mg) Dsz protein and about 5.4-fold of that of E. coli BL2-paN2. Experiment were also conducted using resting cell with the 0.6 wt% DBT in n-hexadecane as model diesel oil. After 24 hours reaction, 0.42 mmol/L (about 84%) DBT was converted to 2-HBP by E. coli BL21- pBADD + paN2, however, there was only 0.08 mmol/L (about 16%) DBT was desulfurized by E. coli BL2-paN2. The maximum desulfurization rate of E.coli BL21-pBADD + paN2 is about 67 micromol/h. The result shows that DszD enhances the rate of 2-HBP production when co-expressed in vivo with the desulfurization enzymes.
authors.affiliation: College of Life Sciences, Nankai University, Tianjin 300071, China. coolrain@mail.nankai.edu.cn
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Li
authors.forename: Ting
authors.initials: T
authors.lastname: Ma
authors.forename: Jing-Hao
authors.initials: JH
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Ru-Lin
authors.initials: RL
authors.lastname: Liu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001673:Biodegradation, Environmental; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010105:Oxygenases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D012240:Rhodococcus / Q000201:enzymology*; D013876:Thiophenes / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D013876:Thiophenes; D010088:Oxidoreductases; D010105:Oxygenases; C421210:dibenzothiophene sulfone monooxygenase; C016366:dibenzothiophene