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【目的】筛选能抗营养阻遏产漆酶的黄孢原毛平革菌,论证其产漆酶的确定性及抗营养阻遏产木质素酶的可行性,为白腐菌产酶代谢调控、木质素降解机理的研究奠定基础。【方法】利用重复紫外诱变法,以愈创木酚富氮鉴别培养基筛选目标菌株;比较不同营养条件下菌体生长与产酶动力学差异研究产酶营养调控机理;通过热处理、排除锰离子和加入过氧化氢酶等不同措施论证黄孢原平毛平革菌能否产生漆酶。【结果】3种不同方法均证实选育到的pcR5305和pcR5324菌株在限氮与富氮条件下均能产生漆酶,pcR5305和pcR5324在限氮条件下产漆酶分别达到203.5、187.6 U/L;在富氮条件下为220.6、183.9 U/L,而原菌株pc530在两种条件下都基本不产生漆酶。二菌株产漆酶调控方式不同,pcR5305漆酶产生与菌体生长同步,而pcR5324漆酶产生却受营养氮阻遏。二菌株同时具有抗营养阻遏高产木质素过氧化物酶(LiP)和锰过氧化物酶(MnP)(分别为LiP 1343.2、MnP 252.2 U/L;LiP 1169.5、MnP 172.4 U/L)的能力。【结论】筛选到的黄孢原毛平革菌变异菌株能产漆酶,同时表现了抗营养阻遏产漆酶、木质素过氧化物酶和锰过氧化物酶的能力,具有重要的生产应用与理论研究价值,为白腐菌产酶代谢调控机理研究提供了原始菌株并奠定了良好的基础。
【Objective】 The objective of this study was to screen Phanerochaete chrysosporium, an antifeedant production-resistant laccase, to demonstrate the certainty of its laccase production and the feasibility of anti-nutritional deterrent production of ligninase. Degradation mechanism of research laid the foundation. 【Method】 The target strains were screened by repeated UV mutagenesis with the guaiacol-rich nitrogen-enrichment medium. The growth and enzyme kinetics differences were studied under different nutrient conditions to study the mechanism of enzyme production and nutrition regulation. Manganese Ions and adding catalase and other measures to demonstrate whether the Phanerochaete chrysosporium can produce laccase. 【Result】 Three different methods confirmed that all the strains of pcR5305 and pcR5324 were capable of producing laccase under nitrogen-limited and nitrogen-rich conditions. The laccase yield of pcR5305 and pcR5324 reached 203.5 and 187.6 U / L ; 220.6,183.9 U / L in the condition of rich nitrogen, while the original strain pc530 basically did not produce laccase under both conditions. Two strains of laccase production in different ways, pcR5305 laccase production and cell growth synchronization, and pcR5324 laccase production by nutrient nitrogen repression. The two strains also exhibited the ability of anti-nutrition to suppress the production of high-yielding lignin peroxidase (LiP) and manganese peroxidase (MnP) (LiP 1343.2, MnP 252.2 U / L; LiP 1169.5, MnP 172.4 U / L), respectively. 【Conclusion】 The selected mutants of Phanerochaete chrysosporium could produce laccase and showed the ability of anti-nutrition to suppress the production of laccase, lignin peroxidase and manganese peroxidase, and had important production and application The value of theoretical research provided the original strain for the study of the regulation mechanism of the production of white rot fungi and laid a good foundation.