Streptomyces thermoviolaceus is a thermophilic actinomycete that was found to produce relatively large amounts of extracellular peroxidase activity when grown on xylan as primary carbon source. The activity was due to multiple isoforms of peroxidase, of which two, designated P-3 and P-5, were predominant. The two proteins were purified to homogeneity by a combination of ultrafiltration, ammonium sulphate precipitation, anion-exchange chromatography, gel filtration and preparative gel electrophoresis. The peroxidases were found to be haemoproteins that catalysed the oxidation of a range of substrates in the presence of hydrogen peroxide. Both are monomeric acidic proteins (P-3: 82 kDa, pl 5·0; P-5: 60 kDa, pl 4·75) but with some differences in substrate specificity, P-3 exhibiting the broader substrate range. Peroxidase activity was optimal at pH values close to neutrality, and both enzymes were robust, exhibiting activity at elevated temperatures in the presence of denaturing agents such as SDS or 8 M urea. Peroxidase P-3 was stable at 50° for more than 24 h and had a half-life of 70 min at 70°. Polyclonal antibodies prepared against each isoform cross-reacted, indicating that the proteins were antigenically related. No cross-reactions were detected against horseradish peroxidase or crude peroxidase preparations from two other thermophilic streptomycetes.
AdhiT. P., KorusR. A., CrawfordD. L.1989; Production of major extracellular enzymes during lignocellulose degradation by two streptomycetes in agitated submerged culture. Appl Environ Microbiol 55:1165–1168
BradfordM.M.1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
DattaA., BettermannA., KirkT. K.1991; Identification of a specific manganese peroxidase among ligninolytic enzymes secreted by Phanerochaete chrysosporium during wood decay. Appl Environ Microbiol 57:1453–1460
FarrellR. L., MurtaghK. E., TienM., MozuchM. D., KirkT. K.1989; Physical and enzymatic properties of lignin peroxidase isoenzymes from Phanerochaete chrysosporium. Enzyme Microb Technol 11:322–328
GoddenB., BallA. S., HelvensteinP., McCarthyA. J., PenninckxM. J.1992; Towards elucidation of the lignin degradation pathway in actinomycetes. J Gen Microbiol 138:2441–2448
GoldM. H., WariishiH., ValliK.1989; Extracellular peroxidases involved in lignin degradation by the white-rot basidiomycete Phanerochaete chrysosporium. In Biocatalysis in Agricultural Biotechnology, (ACS Symposium Series 389) pp. 127–140 Edited by WhitakerJ. R., SonnetP. E. . Washington, DC: American Chemical Society;
GoldM. H., WariishiH., MayfieldM. B., KishiK.1993; Recent studies on lignin and manganese peroxidase from the basidiomycete Phanerochaete chrysosporium. In Plant Peroxidases: Biochemistry and Physiology pp. 87–95 Edited by WelinderK. G., RasmussenS. K. , PenelC., GreppinH. . Geneva: University of Geneva;
HochmanA.1993; The diversity of bacterial hydroperoxidases. In Plant Peroxidases: Biochemistry and Physiology pp. 103–112 Edited by WelinderK. G., RasmussenS. K. , PenelC., GreppinH. . Geneva: University of Geneva;
IshidaA., FutamauraA., MatsusakaT.1987; Detection of peroxidase activity and its localisation in the forespore envelopes of Bacillus cereus. Appl Environ Microbiol 33:27–32
JamesP.D.A., EdwardsC.1989; The effects of temperature on growth and production of the antibiotic granaticin by a thermotolerant streptomycete. J Gen Microbiol 135:1997–2003
LodhaS. J., KorusR. A., CrawfordD. L.1991; Synthesis and properties of lignin peroxidase from Streptomyces viridosporus T7A. Appl Biochem Biotechnol 28:29411–420
MlikiA., ZimmermannW.1992; Purification and characterisation of an intracellular peroxidase from Streptomyces cyaneus. Appl Environ Microbiol 58:916–919
OuchterlonyO., NilssonL. A.1978; Immunodiffusion and immunoelectrophoresis. In Handbook of Experimental Immunology pp. 191–19 Edited by WeirD. W. Oxford: Blackwell Scientific;
RamachandraM., CrawfordD. L., HertelG.1988; Characterisation of an extracellular lignin peroxidase of the lignocellulolytic actinomycete Streptomyces viridosporus. Appl Environ Microbiol 54:3057–3063
SpikerJ. K., CrawfordD. L., ThielE. C.1992; Oxidation of phenolic and non-phenolic substrates by the lignin peroxidase of Streptomyces viridosporus T7A. Appl Microbiol Biotechnol 37:518–523
TuiselH., SinclairR., BumpusJ. A., AshbaugjhW., BrockB. J., AustS. D.1990; Lignin peroxidase H2 from Phanerochaete chrysosporium: purification, characterisation and stability to temperature and pH. Arch Biochem Biophys 279:158–166
WelinderK.G., GajhedeM.1993; Structure and evolution of peroxidases. In Plant Peroxidases: Biochemistry and Physiology pp. 35–42 Edited by WelinderK. G., RasmussenS. K. , PenelC., GreppinH. . Geneva: University of Geneva;
WinterB., FiechterA., ZimmermannW.1991; Degradation of organochlorine compounds in spent sulfite bleach plant effluents by actinomycetes. Appl Environ Microbiol 57:2858–2863