The Rhodobacter capsulatus hemA gene, which encodes 5-aminolevulinic acid synthase （ALAS）, was expressed in Escherichia coil Rosetta （DE3） and the enzymatic properties of the purified recombinant ALAS （RC-ALAS） ...The Rhodobacter capsulatus hemA gene, which encodes 5-aminolevulinic acid synthase （ALAS）, was expressed in Escherichia coil Rosetta （DE3） and the enzymatic properties of the purified recombinant ALAS （RC-ALAS） were studied. Compared with ALASs encoded by hemA genes from Agrobacterium radiobacter（AR-ALAS） and Rhodobacter sphaeroides （RS-ALAS）, the specific activity of RC-ALAS reached 198.2 U/mg, which was about 31.2% and 69.5% higher than those of AR-ALAS （151.1 U/mg） and RS-ALAS （116.9 U/mg）, respectively. The optimum pH values and temperatures of the three above mentioned enzymes were all pH 7.5 and 37 ℃, respectively. Moreover, RC-ALAS was more sensitive to pH, while the other two were sensitive to temperature. The effects of metals, ethylene diamine tetraacetic acid （EDTA）, and sodium dodecyl sulfate （SDS） on the three ALASs were also investigated. The results indicate that they had the same effects on the activities of the three ALASs. SDS and metal ions such as Co^2＋, Zn^2＋, and Cu^2＋ strongly inhibited the activities of the ALASs, while Mn^2＋ exerted slight inhibition, and K^＋, Ca^2＋, Ba^2＋, Mg^2＋, or EDTA had no significant effect. The specificity constant of succinyl coenzyme A [（kcatlKm）^S-CoA] of RC-ALAS was 1.4989, which was higher than those of AR-ALAS （0.7456） and RS-ALAS （1.1699）, showing its high catalytic efficiency. The fed-batch fermentation was conducted using the recombinant strain containing the R. capsulatus hemA gene, and the yield of 5-aminolevulinic acid （ALA） achieved was 8.8 g/L （67 mmol/L） under the appropriate conditions.展开更多
Organic triazophos wastewater was continuously treated with Rhodopseudomonas capsulatus and activated carbon and activated sludge system(PACT AS) in a plug bioreactor. A kinetic model of PACT AS wastewater trea...Organic triazophos wastewater was continuously treated with Rhodopseudomonas capsulatus and activated carbon and activated sludge system(PACT AS) in a plug bioreactor. A kinetic model of PACT AS wastewater treatment system was established to provide an useful basis for further simulate scale up treatment of toxic organic wastewater.展开更多
基金Project supported by the National Natural Science Foundation of China (No.20306026)the National Basic Research Program (973) of China (No.2007CB707805)
文摘The Rhodobacter capsulatus hemA gene, which encodes 5-aminolevulinic acid synthase （ALAS）, was expressed in Escherichia coil Rosetta （DE3） and the enzymatic properties of the purified recombinant ALAS （RC-ALAS） were studied. Compared with ALASs encoded by hemA genes from Agrobacterium radiobacter（AR-ALAS） and Rhodobacter sphaeroides （RS-ALAS）, the specific activity of RC-ALAS reached 198.2 U/mg, which was about 31.2% and 69.5% higher than those of AR-ALAS （151.1 U/mg） and RS-ALAS （116.9 U/mg）, respectively. The optimum pH values and temperatures of the three above mentioned enzymes were all pH 7.5 and 37 ℃, respectively. Moreover, RC-ALAS was more sensitive to pH, while the other two were sensitive to temperature. The effects of metals, ethylene diamine tetraacetic acid （EDTA）, and sodium dodecyl sulfate （SDS） on the three ALASs were also investigated. The results indicate that they had the same effects on the activities of the three ALASs. SDS and metal ions such as Co^2＋, Zn^2＋, and Cu^2＋ strongly inhibited the activities of the ALASs, while Mn^2＋ exerted slight inhibition, and K^＋, Ca^2＋, Ba^2＋, Mg^2＋, or EDTA had no significant effect. The specificity constant of succinyl coenzyme A [（kcatlKm）^S-CoA] of RC-ALAS was 1.4989, which was higher than those of AR-ALAS （0.7456） and RS-ALAS （1.1699）, showing its high catalytic efficiency. The fed-batch fermentation was conducted using the recombinant strain containing the R. capsulatus hemA gene, and the yield of 5-aminolevulinic acid （ALA） achieved was 8.8 g/L （67 mmol/L） under the appropriate conditions.
文摘Organic triazophos wastewater was continuously treated with Rhodopseudomonas capsulatus and activated carbon and activated sludge system(PACT AS) in a plug bioreactor. A kinetic model of PACT AS wastewater treatment system was established to provide an useful basis for further simulate scale up treatment of toxic organic wastewater.
