Glycerol-3-phosphate acyltransferase(GPAT) is considered as the rate-limiting enzyme of glycerolipid synthesis pathway and the core element in lysophosphatidic acid(LPA) synthesis. For understanding its catalytic mech...Glycerol-3-phosphate acyltransferase(GPAT) is considered as the rate-limiting enzyme of glycerolipid synthesis pathway and the core element in lysophosphatidic acid(LPA) synthesis. For understanding its catalytic mechanism, the structural biology study is expected, but is always hindered by obtaining crystals for X-ray diffraction analysis. In this study, a progressive strategy to optimize the crystal of microalgae plastidial GPAT was presented. After the expression and purification of GPAT, the crystals were screened by hanging-drop and only clusters were obtained. The crystals were optimized by adjusting temperature, pH, protein concentration, or precipitant, but little improvement. To improve the interaction between protein and precipitant, the isopropanol was applied as co-precipitant. The qualified crystals formed. It's suggested that isopropanol is critical to affect protein crystallization by altering polyethylene glycol(PEG)-water-protein interaction when PEG serves as precipitant. The resulting crystal diffracted to a resolution of 2.75 ? and belonged to space group P1, with unit-cell parameters a = 50.79, b = 80.09, c = 88.21 ?, and α = 62.85, β = 73.04, γ = 80.53?. This work introduced a new strategy to optimize the crystal of the heterogeneous catalysis enzymes like GPAT and provided the fundamental structural information for the oriented synthesis of marine microalgae glycerolipid.展开更多
The mechanical force borne by the lintel and“brackets”in the inclined channel area of CDQ and the thermal stress generated by the top-down temperature gradient were simulated by ANSYS software.The following conclusi...The mechanical force borne by the lintel and“brackets”in the inclined channel area of CDQ and the thermal stress generated by the top-down temperature gradient were simulated by ANSYS software.The following conclusions are drawn.(1)The mechanical force has very mild effect on the damage of the“brackets”.(2)The temperature gradient caused by the uneven temperature distribution of CDQ oven is the key factor affecting the“bracket”damage.The different expansion of refractory materials in various parts results in the thermal stress concentration which tears the“brackets”.(3)The structure adjustment of the“brackets”has very little effect on the thermal stress distribution.(4)The multi-phase nitride bonded silicon carbide for CDQ can effectively alleviate the stress concentration at the“brackets”,realizing the long-life and stable operation.展开更多
Multi-phase nitrides bonded silicon carbide lintel blocks were prepared using industrial SiC(SiC≥98 mass%,3-0.5,≤0.5 and≤0.044 mm),Si powder(Si≥98 mass%,≤0.044 mm),and SiO2 micropowder(SiO2≥96 mass%,d50=0.15 pm)...Multi-phase nitrides bonded silicon carbide lintel blocks were prepared using industrial SiC(SiC≥98 mass%,3-0.5,≤0.5 and≤0.044 mm),Si powder(Si≥98 mass%,≤0.044 mm),and SiO2 micropowder(SiO2≥96 mass%,d50=0.15 pm)as raw materials,and calcium lignosulfonate as the additive,batching,mixing,and molding on a vibration pressure molding machine,drying and then firing at 1420℃for 10 h in high-purity N2.The apparent porosity,the bulk density,the cold modulus of rupture,the hot modulus of rupture,and the linear expansion coefficient of the samples were tested.The phase composition and the microstructure of the samples at different nitriding depths(50,100,and 150 mm)were analyzed by XRD and SEM.The field application effects of the blocks were studied.The results show that:(1)the multi-phase nitrides bonded silicon carbide refractories can dynamically adjust their own phase composition and minimize structural and thermal stresses,improving the service life of key parts of dry quenching furnaces;(2)calcium lignosulfonate can improve the nitriding micro-environment of multi-phase nitrides bonded silicon carbide lintel blocks,successfully increasing the effective nitriding thickness of the blocks to 300 mm;(3)Sinosteel LI RR provides a unique concept in the design of materials and block types as well as the stable and scientific overall structure,promoting the industrialization process of dry quenching furnaces with long service life in China.展开更多
基金financially supported by the National Natural Science Foundation of China (Nos. 21576253, 31500294 and 31470432)
文摘Glycerol-3-phosphate acyltransferase(GPAT) is considered as the rate-limiting enzyme of glycerolipid synthesis pathway and the core element in lysophosphatidic acid(LPA) synthesis. For understanding its catalytic mechanism, the structural biology study is expected, but is always hindered by obtaining crystals for X-ray diffraction analysis. In this study, a progressive strategy to optimize the crystal of microalgae plastidial GPAT was presented. After the expression and purification of GPAT, the crystals were screened by hanging-drop and only clusters were obtained. The crystals were optimized by adjusting temperature, pH, protein concentration, or precipitant, but little improvement. To improve the interaction between protein and precipitant, the isopropanol was applied as co-precipitant. The qualified crystals formed. It's suggested that isopropanol is critical to affect protein crystallization by altering polyethylene glycol(PEG)-water-protein interaction when PEG serves as precipitant. The resulting crystal diffracted to a resolution of 2.75 ? and belonged to space group P1, with unit-cell parameters a = 50.79, b = 80.09, c = 88.21 ?, and α = 62.85, β = 73.04, γ = 80.53?. This work introduced a new strategy to optimize the crystal of the heterogeneous catalysis enzymes like GPAT and provided the fundamental structural information for the oriented synthesis of marine microalgae glycerolipid.
文摘The mechanical force borne by the lintel and“brackets”in the inclined channel area of CDQ and the thermal stress generated by the top-down temperature gradient were simulated by ANSYS software.The following conclusions are drawn.(1)The mechanical force has very mild effect on the damage of the“brackets”.(2)The temperature gradient caused by the uneven temperature distribution of CDQ oven is the key factor affecting the“bracket”damage.The different expansion of refractory materials in various parts results in the thermal stress concentration which tears the“brackets”.(3)The structure adjustment of the“brackets”has very little effect on the thermal stress distribution.(4)The multi-phase nitride bonded silicon carbide for CDQ can effectively alleviate the stress concentration at the“brackets”,realizing the long-life and stable operation.
文摘Multi-phase nitrides bonded silicon carbide lintel blocks were prepared using industrial SiC(SiC≥98 mass%,3-0.5,≤0.5 and≤0.044 mm),Si powder(Si≥98 mass%,≤0.044 mm),and SiO2 micropowder(SiO2≥96 mass%,d50=0.15 pm)as raw materials,and calcium lignosulfonate as the additive,batching,mixing,and molding on a vibration pressure molding machine,drying and then firing at 1420℃for 10 h in high-purity N2.The apparent porosity,the bulk density,the cold modulus of rupture,the hot modulus of rupture,and the linear expansion coefficient of the samples were tested.The phase composition and the microstructure of the samples at different nitriding depths(50,100,and 150 mm)were analyzed by XRD and SEM.The field application effects of the blocks were studied.The results show that:(1)the multi-phase nitrides bonded silicon carbide refractories can dynamically adjust their own phase composition and minimize structural and thermal stresses,improving the service life of key parts of dry quenching furnaces;(2)calcium lignosulfonate can improve the nitriding micro-environment of multi-phase nitrides bonded silicon carbide lintel blocks,successfully increasing the effective nitriding thickness of the blocks to 300 mm;(3)Sinosteel LI RR provides a unique concept in the design of materials and block types as well as the stable and scientific overall structure,promoting the industrialization process of dry quenching furnaces with long service life in China.
