Biodegradation of Structurally Diverse Phthalate Esters by a Newly Identified Esterase with Catalytic Activity toward Di(2-ethylhexyl) Phthalate.

Herein, we report a double enzyme system to degrade 12 phthalate esters (PAEs), particularly bulky PAEs, such as the widely used bis(2-ethylhexyl) phthalate (DEHP), in a one-pot cascade process. A PAE-degrading bacterium, Gordonia sp. strain 5F, was isolated from soil polluted with plastic waste. From this strain, a novel esterase (GoEst15) and a mono(2-ethylhexyl) phthalate hydrolase (GoEstM1) were identified by homology-based cloning. GoEst15 showed broad substrate specificity, hydrolyzing DEHP and 10 other PAEs to monoalkyl phthalates, which were further degraded by GoEstM1 to phthalic acid. GoEst15 and GoEstM1 were heterologously coexpressed in Escherichia coli BL21 (DE3), which could then completely degrade 12 PAEs (5 mM), within 1 and 24 h for small and bulky substrates, respectively. To our knowledge, GoEst15 is the first DEHP hydrolase with a known protein sequence, which will enable protein engineering to enhance its catalytic performance in the future.

Efficient Degradation of Malathion in the Presence of Detergents Using an Engineered Organophosphorus Hydrolase Highly Expressed by Pichia pastoris without Methanol Induction.

The biodegradation of pesticides by organophosphorus hydrolases (OPHs) requires an efficient enzyme production technology in industry. Herein, a Pichia pastoris strain was constructed for the extracellular expression of PoOPHM9, an engineered malathion-degrading enzyme. After optimization, the maximum titer and yield of fermentation reached 50.8 kU/L and 4.1 gprotein/L after 3 days, with the highest space-time yield (STY) reported so far, 640 U L-1 h-1. PoOPHM9 displayed its high activity and stability in the presence of 0.1% (w/w) plant-derived detergent. Only 0.04 mg/mL enzyme could completely remove 0.15 mM malathion in aqueous solution within 20 min. Furthermore, 12 µmol malathion on apples and cucumbers surfaces was completely removed by 0.05 mg/mL PoOPHM9 in tap water after 35 min washing. The efficient production of the highly active PoOPHM9 has cleared a major barrier to biodegradation of pesticide residues in food industry.

[Response of Soil Respiration and Organic Carbon to Returning of Different Agricultural Straws and Its Mechanism].

Soybean, maize and rice straws were selected as raw materials to study the response of the soil respiration (SR) and soil organic carbon (SOC) to returning of different straws in the Chongming Dongtan area. The results showed that all of SR, SOC and the plant biomass of the lands with returning of different straws were higher than those of the controls. The soil with soybean straw returning possessed the lowest SR and highest SOC among the three kinds of straws, meaning its higher soil organic carbon sequestration capability than corn and maize straws returning. Straw returning significantly enhanced soil dehydrogenase, ß-glycosidase activities and microbial biomass, and soil dehydrogenase activity was significantly correlated with soil respiration. The dehydrogenase activity of the soil with soybean straw returning was the lowest, thus, the lowest SR and highest SOC. Soybean straw had the highest cellulose and lignin contents and the lowest N content among the three kinds of straws, resulting in its lowest biodegradability. Therefore, when soybean straw was returned to soil, it was difficult to degrade completely by soil microorganisms, thus the lowest soil microbial activity, eventually leading to the lowest SR and highest SOC.

[Potential Carbon Fixation Capability of Non-photosynthetic Microbial Community at Different Depth of the South China Sea and Its Response to Different Electron Donors].

The seawater samples collected from many different areas with different depth in the South China Sea were cultivated using different electron donors respectively. And the variation in the potential carbon fixation capability ( PCFC ) of non-photosynthetic microbial community (NPMC) in seawater with different depth was determined after a cycle of cultivation through the statistic analysis. In addition, the cause for the variation was clarified through analyzing key gene abundance regarding CO2 fixation and characteristics of seawater with different depth. The result showed that the PCFCs of NPMC in seawater with different depth were generally low and had no significant difference when using NaNO2 as the electron donor. The PCFC of NPMC in surface seawater was higher than that in deep seawater when using H2 as the electron donor, on the contrary, the PCFC of NPMC in deep seawater was higher than that in surface seawater when using Na2S2O3 as the electron donor. The abundance of the main CO2 fixation gene cbbL in surface seawater was higher than that in deep seawater while the cbbM gene abundance in deep seawater was higher than that in surface seawater. Most hydrogen-oxidizing bacteria had the cbbL gene, and most sulfur bacteria had the cbbM gene. The tendency of seawater cbbL/cbbM gene abundance with the change of depth revealed that there were different kinds of bacteria accounting for the majority in NPMC fixing CO2 at different depth of ocean, which led to different response of PCFC of NPMC at different depth of the sea to different electron donors. The distributions of dissolved oxygen and inorganic carbon concentration with the change of the depth of the sea might be an important reason leading to the difference of NPMC structure and even the difference of PCFC at different depth of the sea.

[Breeding, optimization and community structure analysis of non-photosynthetic CO2 assimilation microbial flora].

Isolation and screening from sea water and sediments, and the optimization of electron donor and inorganic carbon source structure were performed for obtaining microbial flora with high efficient inorganic carbon fixation without the light and hydrogen. In addition, the structure of the microbial flora was studied through 16S rDNA sequence analysis and contrast for providing theoretical basis to improve carbon fixation efficiency through optimizing microbial flora structure. The result showed that non-photosynthetic microbial flora with the capacity of inorganic carbon fixation under the general aerobic and anaerobic conditions could be obtained from the sea by long-term domestication and isolation. Inorganic carbon fixation efficiency of the microbial flora was enhanced significantly by adding of sodium thiosulfate, sodium sulfide and hydrogen as electron donor. Under the aerobic and anaerobic conditions with sodium thiosulfate as electron donor, the efficiency of inorganic carbon assimilation was 10.44 mg/L and 12.56 mg/L respectively. The assimilation efficiency of the microbial flora with mixed inorganic carbon source was higher than that with single carbon source. When CO2, sodium bicarbonate and sodium carbonate were added as carbon sources, carbon fixation efficiency of the microbial flora under the aerobic and anaerobic condition was 110 mg x (L x d)(-1) and 72 mg x (L x d)(-1) respectively which had been closed to the efficiency of hydrogen-oxidizing bacteria. The analysis results showed that the predominant species of the microbial flora varied significantly after the adding of different electron donor. And 11 species of the 16 predominant species in the microbial flora was uncultured. It means that the microbial flora could only exist in symbiotic manner. The inorganic carbon fixation effect of the microbial flora may be the results of co-function of multi-microbial species. Therefore, the optimization of microbial flora structure and proportion is benefit for the further improvement of carbon fixation efficiency.