IMMUNOLOGICAL RELATEDNESS OF EXTRACELLULAR LIGNINASES FROM THE ACTINOMYCETES STREPTOMYCES-VIRIDOSPORUS T7A AND STREPTOMYCES-BADIUS 252

被引:15
作者
MAGNUSON, TS
ROBERTS, MA
CRAWFORD, DL
HERTEL, G
机构
[1] Department of Bacteriology and Biochemistry, Institute for Molecular and Agricultural Genetic Engineering (IMAGE), University of Idaho, Moscow, 83843, ID
关键词
LIGNINASES; WESTERN BLOT ANALYSIS; AFFINITY PURIFICATION;
D O I
10.1007/BF02922623
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:433 / 443
页数:11
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