Magnuson T S, Roberts M A, Crawford D L, Hertel G
Department of Bacteriology and Biochemistry, University of Idaho, Moscow 83843.
Appl Biochem Biotechnol. 1991 Spring;28-29:433-43. doi: 10.1007/BF02922623.
Four isoforms of the extracellular lignin peroxidase of the ligninolytic actinomycete Streptomyces viridosporus T7A (ALip-P1, P2, P3, and P4) were individually purified by ultrafiltration and ammonium sulfate precipitation, followed by electro-elution using polyacrylamide gel electrophoresis. Three of the purified peroxidases were compared for their immunologic relatedness by Western blot analysis using a polyclonal antibody preparation produced in rabbits against pure isoform P3. The anti-P3 antibody was also tested for its reactivity towards a lignin peroxidase from the white-rot fungus Phanerochaete chrysosporium and another ligninolytic actinomycete Streptomyces badius 252. Results showed that peroxidases ALip-P1 through ALip-P3 are immunologically related to one another. The peroxidases of S. badius, but not the peroxidase of P. chrysosporium, also reacted with the antibody, thus indicating that the lignin peroxidases of S. viridosporus and S. badius are immunologically related. Based upon its specific affinity, lignin peroxidase isoform ALip-P3 of S. viridosporus was readily purified using an anti-P3 antibody affinity column.
木质素分解放线菌绿产色链霉菌T7A的细胞外木质素过氧化物酶的四种同工型(ALip-P1、P2、P3和P4)通过超滤和硫酸铵沉淀进行单独纯化,随后使用聚丙烯酰胺凝胶电泳进行电洗脱。使用兔抗纯同工型P3制备的多克隆抗体制剂,通过蛋白质免疫印迹分析比较了三种纯化的过氧化物酶的免疫相关性。还测试了抗P3抗体对白腐真菌黄孢原毛平革菌的木质素过氧化物酶和另一种木质素分解放线菌浅灰链霉菌252的反应性。结果表明,过氧化物酶ALip-P1至ALip-P3彼此具有免疫相关性。浅灰链霉菌的过氧化物酶与该抗体发生反应,而黄孢原毛平革菌的过氧化物酶则未发生反应,这表明绿产色链霉菌和浅灰链霉菌的木质素过氧化物酶具有免疫相关性。基于其特异性亲和力,使用抗P3抗体亲和柱可轻松纯化绿产色链霉菌的木质素过氧化物酶同工型ALip-P3。