Colao M Ch, Garzillo A M, Buonocore V, Schiesser A, Ruzzi M
Department of Agrobiology and Agrochemistry, Tuscia University, Via C. de Lellis s.n.c., 01100 Viterbo, Italy.
Appl Microbiol Biotechnol. 2003 Dec;63(2):153-8. doi: 10.1007/s00253-003-1429-x. Epub 2003 Sep 16.
A cDNA coding for laccase was isolated from the white-rot fungus Trametes trogii 201. This cDNA corresponded to the lcc1 gene, which coded for a precursor protein of 517 amino acids with a 21 amino acid signal peptide. Comparison of the deduced sequence with known laccases showed that this enzyme was most closely related to Lac1 from basidiomycete PM1 and Trametes C30 (98% similarity). The expression of lcc1 was analysed under different growth conditions; transcription of this gene was enhanced by the addition of organic nitrogen to the medium. The level of lcc1 transcription was higher when T. trogii was grown on synthetic medium supplemented with yeast extract rather than mycological peptone or tryptone. The transcription data were in agreement with total laccase activity measured in the supernatant and suggested that laccase production and lcc1 transcription are coordinately regulated in this organism. The lcc1 cDNA was expressed in the methylotrophic yeast Pichia pastoris and the detection of laccase activity indicated that this cDNA encodes a laccase.
从白腐真菌糙皮侧耳201中分离出一个编码漆酶的cDNA。该cDNA对应于lcc1基因,其编码一个含有21个氨基酸信号肽的517个氨基酸的前体蛋白。将推导的序列与已知漆酶进行比较,结果表明该酶与担子菌PM1和糙皮侧耳C30中的Lac1关系最为密切(相似度为98%)。在不同生长条件下分析了lcc1的表达;向培养基中添加有机氮可增强该基因的转录。当糙皮侧耳在添加酵母提取物而非真菌蛋白胨或胰蛋白胨的合成培养基上生长时,lcc1的转录水平更高。转录数据与在上清液中测得的总漆酶活性一致,表明该生物体中漆酶的产生和lcc1的转录受到协同调控。lcc1 cDNA在甲基营养型酵母毕赤酵母中表达,漆酶活性检测表明该cDNA编码一种漆酶。