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肺形侧耳:一种存在于木质纤维素残渣中可产生蛋白酶的白腐真菌。

Pleurotus pulmonarius: a protease-producing white rot fungus in lignocellulosic residues.

作者信息

Contato Alex Graça, Inácio Fabíola Dorneles, Bueno Paulo Sérgio Alves, Nolli Mariene Marques, Janeiro Vanderly, Peralta Rosane Marina, de Souza Cristina Giatti Marques

机构信息

Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA.

Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

出版信息

Int Microbiol. 2023 Jan;26(1):43-50. doi: 10.1007/s10123-022-00271-8. Epub 2022 Aug 8.

Abstract

The production of proteases by white rot fungi, such as those of the genus Pleurotus, is related to the degradation of wood proteins, the substrate on which these fungi grow in the environment. From the point of view of production, they are still little explored for this purpose. A selection of agro-industrial residues highlighted corn bagasse as the best substrate for solid-state protease production using the basidiomycete Pleurotus pulmonarius. The enzyme production was maximized through a factorial design, where the enzyme activity increased from 137.8 ± 1.9 to 234.1 ± 2.7 U/mL. Factors such as temperature stability, pH, and chemical reagents were evaluated. The optimum temperature was 45 °C, showing low thermal stability at higher temperatures. The enzyme inhibition occurred by Mn (50.3%) and Ba (76.4%); SDS strongly inhibited the activity (82.4%), while pepstatin A partially inhibited (56%), suggesting an aspartic protease character. Regarding pH, the highest protease activity was obtained at pH 5.5. Partial characterization resulted in apparent values of the K and V constants of 0.61 mg/mL and 1.79 mM/min, respectively.

摘要

白腐真菌(如侧耳属真菌)产生的蛋白酶与木材蛋白质的降解有关,木材蛋白质是这些真菌在环境中生长的底物。从生产的角度来看,针对此目的对它们的研究仍然很少。一系列农业工业残渣中,玉米秸秆被证明是使用肺形侧耳这种担子菌进行固态蛋白酶生产的最佳底物。通过析因设计使酶产量最大化,酶活性从137.8±1.9 U/mL增加到234.1±2.7 U/mL。对温度稳定性、pH值和化学试剂等因素进行了评估。最适温度为45℃,在较高温度下热稳定性较低。锰(50.3%)和钡(76.4%)会抑制该酶活性;十二烷基硫酸钠强烈抑制其活性(82.4%),而胃蛋白酶抑制剂A部分抑制(56%),表明该酶具有天冬氨酸蛋白酶的特性。关于pH值,在pH 5.5时获得最高蛋白酶活性。部分特性表征得出K和V常数的表观值分别为0.61 mg/mL和1.79 mM/min。

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