Alcohol dehydrogenase 1 identified from Artemisia annua (AaADH1) is a 40 kDa protein that predominately expressed in young leaves and buds, and catalyzes dehydrogenation of artemisinic alcohol to artemisinic aldehyde in artemisinin biosynthetic pathway. In this study, AaADH1 encoding gene was subcloned into vector pET-21a(+) and expressed in Escherichia coli. BL21(DE3), and purified by Co²⁺ affinity chromatography. Anion exchange chromatography was performed until the protein purity reached more than 90%. Crystallization of AaADH1 was conducted for further investigation of the molecular mechanism of catalysis, and hanging-drop vapour diffusion method was used in experiments. The results showed that the apo AaADH1 crystal diffracted to 2.95 Å resolution, and belongs to space group P1, with unit-cell parameters, a = 77.53 Å, b = 78.49 Å, c = 102.44 Å, α = 71.88°, β = 74.02°, γ = 59.97°. The crystallization condition consists of 0.1 M Bis-Tris pH 6.0, 13% (w/v) PEG 8000 and 5% (v/v) glycerol.

从青蒿中鉴定的酒精脱氢酶 1 (AaADH1) 是一种 40 kDa 的蛋白质，主要在幼叶和芽中表达，并在青蒿素生物合成途径中催化青蒿醇脱氢生成青蒿醛。本研究将AaADH1编码基因亚克隆到载体pET-21a(+)中并在大肠杆菌中表达。BL21(DE3)，并通过Co ²⁺亲和层析纯化。进行阴离子交换层析直到蛋白质纯度达到90%以上。为了进一步研究催化的分子机理，对AaADH1进行了结晶，实验采用了悬滴蒸气扩散法。结果表明，apoAaADH1 晶体衍射到 2.95 Å 分辨率，属于空间群 P1，晶胞参数a  = 77.53 Å，b  = 78.49 Å，c  = 102.44 Å，α  = 71.88°，β  = 74.02°，γ  = 59.97° . 结晶条件由 0.1 M Bis-Tris pH 6.0、13% (w/v) PEG 8000 和 5% (v/v) 甘油组成。