Huang S T, Tzean S S, Tsai B Y, Hsieh H J
Department of Plant Pathology and Microbiology, National Taiwan University, Taipei 10617, Taiwan, ROC.
Microbiology (Reading). 2009 Feb;155(Pt 2):424-433. doi: 10.1099/mic.0.022459-0.
A novel ligninolytic peroxidase gene (ACLnP) was cloned and characterized from a poroid brown-rot fungus, Antrodia cinnamomea. The genomic DNA of the fungus harboured two copies of ACLnP, with a length of 2111 bp, interlaced with 12 introns, while the full-length cDNA was 1183 bp, with a 66 bp signal peptide and an ORF of 990 bp. The three-dimensional molecular structure model was comparable to that of the versatile peroxidase of Pleurotus eryngii. ACLnP was cloned into vector pQE31, successfully expressed in Escherichia coli strain M15 under the control of the T5 promoter and produced a non-glycosylated protein of about 38 kDa, pI 5.42. The native and recombinant ACLnP was capable of oxidizing the redox mediator veratryl alcohol, and also decolorized bromophenol blue and 2,6-dimethoxyphenol dyes, implicating a functional extracellular peroxidase activity. The significance of discovering a functional ACLnP gene in A. cinnamomea in terms of wood degradation and colonization capacity in its unique niche is discussed.
从一种多孔状褐腐真菌——肉桂拟层孔菌中克隆并鉴定了一个新的木质素分解过氧化物酶基因（ACLnP）。该真菌的基因组DNA含有两个ACLnP拷贝，长度为2111 bp，间隔有12个内含子，而全长cDNA为1183 bp，带有一个66 bp的信号肽和一个990 bp的开放阅读框。三维分子结构模型与刺芹侧耳的多功能过氧化物酶的模型相当。ACLnP被克隆到载体pQE31中，在T5启动子的控制下于大肠杆菌菌株M15中成功表达，并产生了一种约38 kDa、pI为5.42的非糖基化蛋白。天然和重组的ACLnP能够氧化氧化还原介质藜芦醇，还能使溴酚蓝和2,6 - 二甲氧基苯酚染料脱色，这表明其具有功能性细胞外过氧化物酶活性。本文讨论了在肉桂拟层孔菌中发现功能性ACLnP基因在其独特生态位中的木材降解和定殖能力方面的意义。
