Oesterhelt C, Vogelbein S, Shrestha R P, Stanke M, Weber A P M
Institut für Biochemie und Biologie, Universität Potsdam, Karl-Liebknecht-Str. 24-25, Haus 20, 14476, Potsdam-Golm, Germany.
Planta. 2008 Jan;227(2):353-62. doi: 10.1007/s00425-007-0622-z. Epub 2007 Sep 25.
We report the identification of a small family of secreted class III plant peroxidases (Prx) from the genome of the unicellular thermoacidophilic red alga Galdieria sulphuraria (Cyanidiaceae). Apart from two class I ascorbate peroxidases and one cytochrome c peroxidase, the red algal genome encodes four class III plant peroxidases, thus complementing the short list of algal cell wall peroxidases (Passardi et al. in Genomics 89:567-579, 2007). We have characterized the family gene structure, analyzed the extracellular space and cell wall fraction of G. sulphuraria for the presence of peroxidase activity and used shotgun proteomics to identify candidate extracellular peroxidases. For a detailed enzymatic characterization, we have purified a secreted peroxidase (GsPrx04) from the cell-free medium using hydrophobic interaction chromatography. The enzyme proved heat and acid-stable and exhibited an apparent molecular mass of 40 kDa. Comparative genomics between endolithically growing G. sulphuraria and a close relative, the obligatory aquatic, cell wall-less Cyanidioschyzon merolae, revealed that class III peroxidases only occur in the terrestrial microalga, thus supporting the key function of these enzymes in the process of land colonization.
我们报告了从单细胞嗜热嗜酸红藻硫黄加尔迪藻(蓝藻科)的基因组中鉴定出一小类分泌型III类植物过氧化物酶(Prx)。除了两种I类抗坏血酸过氧化物酶和一种细胞色素c过氧化物酶外,该红藻基因组还编码四种III类植物过氧化物酶,从而补充了藻类细胞壁过氧化物酶的简短清单(Passardi等人,《基因组学》89:567 - 579,2007年)。我们对该家族基因结构进行了表征,分析了硫黄加尔迪藻的细胞外空间和细胞壁部分是否存在过氧化物酶活性,并使用鸟枪法蛋白质组学来鉴定候选细胞外过氧化物酶。为了进行详细的酶学表征,我们使用疏水相互作用色谱法从无细胞培养基中纯化了一种分泌型过氧化物酶(GsPrx04)。该酶被证明具有热稳定性和酸稳定性,表观分子量为40 kDa。对在岩石内生长的硫黄加尔迪藻与其近缘种、 obligatory aquatic、无细胞壁的梅氏蓝纤维藻进行比较基因组学分析,结果表明III类过氧化物酶仅存在于陆生微藻中,从而支持了这些酶在陆地定殖过程中的关键作用。