Positive Effects of Isopropanol as a Co-Precipitant in Glycerol-3-Phosphate Acyltransferase Crystallization

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.     

从浓度定量分析探讨异丙醇作原油萃取剂的可行性

分别采用正己烷与异丙醇作为萃取剂获得不同浓度下的2组相同原油样品的萃取液,进行波长差为40nm的同步荧光光谱测量.通过插值拟合得到相应的两组相同原油样品的浓度三维荧光光谱图,并据此对相同原油样品萃取液的未知浓度进行计算,并与实际值比较,得出相应误差.文中通过对不同萃取剂中光谱谱形、峰位、荧光强度以及浓度计算的误差进行对比分析,得出采用异丙醇作为石油定量荧光分析仪的萃取剂比用正己烷更具优势,从而在定量分析的基础上论证了采用异丙醇取代正己烷作为原油萃取剂的可行性.