Zhang Yu, Dong Zhongqi, Luo Yuan, Yang En, Xu Huini, Chagan Irbis, Yan Jinping
Laboratory of Bioconversion, Life Science and Technology College, Kunming University of Science and Technology, No. 272 Jingming Road, Kunming 650500, China.
Microorganisms. 2021 Dec 16;9(12):2595. doi: 10.3390/microorganisms9122595.
Manganese peroxidases (MnPs), gene family members of white-rot fungi, are necessary extracellular enzymes that degrade lignocellulose and xenobiotic aromatic pollutants. However, very little is known about the diversity and expression patterns of the MnP gene family in white-rot fungi, especially in contrast to laccases. Here, the gene and protein sequences of eight unique MnP genes of S0301 were characterized. Based on the characteristics of gene sequence, all TtMnPs here belong to short-type hybrid MnP (type I) with an average protein length of 363 amino acids, 5-6 introns, and the presence of conserved cysteine residues. Furthermore, analysis of MnP activity showed that metal ions (Mn and Cu) and static liquid culture significantly influenced MnP activity. A maximum MnP activity (>14.0 U/mL) toward 2,6-DMP was observed in static liquid culture after the addition of Mn (1 mM) or Cu (0.2 or 2 mM). Moreover, qPCR analysis showed that Mn obviously upregulated the Group I MnP subfamily (_09901, 09904, 09903, and 09906), while Cu and HO, along with changing temperatures, mainly induced the Group II MnP subfamily (_11984, 11971, 11985, and 11983), suggesting diverse functions of fungal MnPs in growth and development, stress response, etc. Our studies here systematically analyzed the gene structure, expression, and regulation of the TtMnP gene family in , one of the important lignocellulose-degrading fungi, and these results extended our understanding of the diversity of the MnP gene family and helped to improve MnP production and appilications of strains and other white-rot fungi.
锰过氧化物酶（MnPs）是白腐真菌的基因家族成员，是降解木质纤维素和外源芳香族污染物所必需的胞外酶。然而，人们对白腐真菌中MnP基因家族的多样性和表达模式了解甚少，尤其是与漆酶相比。在此，对S0301的八个独特MnP基因的基因和蛋白质序列进行了表征。基于基因序列特征，这里所有的TtMnPs都属于短型杂合MnP（I型），平均蛋白质长度为363个氨基酸，有5 - 6个内含子，且存在保守的半胱氨酸残基。此外，MnP活性分析表明，金属离子（Mn和Cu）和静置液体培养显著影响MnP活性。在添加Mn（1 mM）或Cu（0.2或2 mM）后的静置液体培养中，观察到对2,6 - DMP的最大MnP活性（>14.0 U/mL）。此外，qPCR分析表明，Mn明显上调了I组MnP亚家族（_09901、09904、09903和09906），而Cu和H₂O₂以及温度变化主要诱导了II组MnP亚家族（_11984、11971、11985和11983），这表明真菌MnPs在生长发育、应激反应等方面具有多种功能。我们在此的研究系统地分析了重要的木质纤维素降解真菌之一中TtMnP基因家族的基因结构、表达和调控，这些结果扩展了我们对MnP基因家族多样性的理解，并有助于提高菌株和其他白腐真菌的MnP产量及应用。
