偏肿革裥菌Lg-mnp2基因在构巢曲霉中的表达

Expression of the Gene Lg-mnp2 Encoding for Manganese Peroxidase 2 (MnP2) from White-Rot Fungus Lenzites gibbosa in Aspergillus nidulans

锰过氧化物酶（MnP；EC1．11．1．13）是真菌分泌的胞外过氧化物酶，广泛存在于木材白腐菌中，通常以多种同工酶的形式存在。MnP在生物修复和生物制浆领域具有重要作用，其可以降解土壤和废水中的DDT、多环芳烃、多氯联苯、含有苯环的偶氮与叠氮化合物，以及其他木质素结构。探索MnP基因的高效表达，目的是为提高其产量。

The aim of this study was to enhance manganese peroxidase （MnP） gene express by Lenzites gibbosa MnP production for using Aspergillus nidulans. In this study, an auxotrophic strain, TN02A7, of A. nidulans was used as the host of L. gibbosa. The eonidiophores of TN02A7 were used to produce the protoplasts by different cell wall lyases, including lywallzyme, cellulase and snailase which were mixed together by 1： 1：1 for effectively break down the wall and produce protoplasts. PEG/CaCI2 were used to mediate transformation of L. gibbosa, containing a gene encoding for manganese peroxidase 2 （Lg-MnP2） , into the TN02A7 protoplast. The newly obtained transformant strain, TN02A7-Lg- mnp2, and TN02A7 were cultured in the same shaking medium containing lignin and the MnP activity was detected. The result showed that TNO2A7-Lg-mnp2 could produce MnP activity up to 17 U.L ＇ in 96 h, while TN02A7 did not produce MnP activity at any time, indicating that the gene Lg-mup2 had been successfully transformed into TNO2A7-Lg-mnp2 and expressed in lignin environment. This study provided a new method to produce MnP.