Expression, crystallization and preliminary crystallographic data analysis of PigI, a putative L-prolyl-AMP ligase from the prodigiosin synthetic pathway in Serratia.
Expression, crystallization and preliminary crystallographic data analysis of PigI, a putative L-prolyl-AMP ligase from the prodigiosin synthetic pathway in Serratia.
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DOI：
10.1107/s2053230x14005780
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发表时间：
2014-05
期刊：
影响因子：
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通讯作者：
Ning Han;T. Ran;X. Lou;Yanyan Gao;Jianhua He;Lin Tang;Dongqing Xu;Weiwu Wang
中科院分区：
文献类型：
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作者：
Ning Han;T. Ran;X. Lou;Yanyan Gao;Jianhua He;Lin Tang;Dongqing Xu;Weiwu Wang
灵芝红素是一种由沙雷氏菌等微生物合成的三吡咯红色素，属于益生菌素。在沙雷氏菌中提出了一种灵芝菌素的分叉合成途径，其中MBC(4-甲氧基-2，2‘-联吡咯-5-甲醛)和MAP(2-甲基-3-N-戊基-吡咯)分别被PIGC缩合生成灵芝菌素。合成MBC的第一步是用PIGI活化L-脯氨酸，但其催化机理尚不清楚。为了阐明其作用机制，对重组PigI进行了纯化和结晶。坐滴法晶体属于空间群P1，晶胞参数为a=51.2，b=62.8，c=91.3，α=105.1，β=90.1，γ=92.2°。马修斯系数分析表明，不对称单元中含有两个分子，其Vm为2.6ä(3)Da(-1)，溶剂含量为52.69%。
Prodigiosin, a member of the prodiginines, is a tripyrrole red pigment synthesized by Serratia and some other microbes. A bifurcated biosynthesis pathway of prodigiosin has been proposed in Serratia in which MBC (4-methoxy-2,2'-bipyrrole-5-carbaldehyde) and MAP (2-methyl-3-N-amyl-pyrrole) are synthesized separately and then condensed by PigC to form prodigiosin. The first step for the synthesis of MBC is the activation of L-proline by PigI, but its catalytic mechanism has remained elusive. To elucidate its mechanism, recombinant PigI was purified and crystallized. Crystals obtained by the sitting-drop method belonged to space group P1 and diffracted to 2.0 Å resolution, with unit-cell parameters a = 51.2, b = 62.8, c = 91.3 Å, α = 105.1, β = 90.1, γ = 92.2°. Matthews coefficient analysis suggested two molecules in the asymmetric unit, with a VM of 2.6 Å(3) Da(-1) and a solvent content of 52.69%.