ABSTRACT
A method for the immobilization of chloroperoxidase from the bacterial strain Serratia marcescens in semi-permeable membranes was developed based on the following proteins: bovine serum albumin, gelatin, ribonuclease, cytochrom C, and the protein of the covering layer of Bacillus sphaericus cells. Estimation of the activity and stability of immobilized preparations in a batch reactor was carried out.
Subject(s)
Chloride Peroxidase/chemistry , Enzymes, Immobilized/chemistry , Serratia marcescens/enzymology , Enzyme Stability , Membranes/chemistry , Permeability , Proteins/chemistryABSTRACT
The gene of tryptophan 7-halogenase was isolated from the Pseudomonas aureofaciens ACN strain producing pyrrolnitrin, a chlorocontaining antibiotic, and sequenced. A high homology degree (over 95%) was established for the genes and the corresponding halogenases from P. aureofaciens ACN and P. fluorescens BL915. The tryptophan 7-halogenase gene was amplified by PCR, and the corresponding enzyme was expressed in Escherichia coli cells using the pBSII SK+ vector.
Subject(s)
Genes, Bacterial , Oxidoreductases/genetics , Pseudomonas/genetics , Amino Acid Sequence , Cloning, Molecular , Escherichia coli/enzymology , Molecular Sequence Data , Oxidoreductases/biosynthesis , Polymerase Chain Reaction , Pseudomonas/enzymologyABSTRACT
The gene of tryptophan-7-halogenase from the Pseudomonas fluorescens strain CHA0, a producer of the halogenated antibiotic pyrrolnitrin, has been cloned and sequenced.
Subject(s)
Oxidoreductases/chemistry , Oxidoreductases/genetics , Pseudomonas fluorescens/enzymology , Chromosomes, Bacterial/genetics , Cloning, Molecular , Escherichia coli/genetics , Molecular Sequence Data , Molecular Structure , Oxidoreductases/biosynthesis , Pseudomonas fluorescens/classification , Pseudomonas fluorescens/genetics , Pyrrolnitrin/biosynthesis , Pyrrolnitrin/chemistry , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Sequence Analysis, Protein , Sequence Homology, Amino AcidABSTRACT
Using blot hybridization, it has been shown that microorganisms producing halogen-containing antibiotics--Pseudomonas pyrrocinia, P. aureofaciens ACN, P. aureofaciens Pa1, P. fluorescens CHA0, Actinoplanes sp., Kitasatasporia sp., Sacharothrix aerocolonigenes, Actinomadura melliaura, and Streptomyces albogriseolus--contain the genes of the halogenating enzymes related to tryptophan-7-halogenase and monodechloroaminopyrrolnitrin halogenase from P. fluorescens BL 915.
Subject(s)
Anti-Bacterial Agents/biosynthesis , Bacteria/enzymology , Bacteria/genetics , Halogens/metabolism , Anti-Bacterial Agents/chemistry , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Nucleic Acid Hybridization , Oxidoreductases/genetics , Oxidoreductases/metabolismABSTRACT
Heme- and metal-independent chloroperoxidase from Serratia marcescens W 250 is shown to be capable of catalyzing the p-nitrophenyl phosphate hydrolysis. The parameters of the phosphatase reaction are determined and inhibitors and activators of the process are found. A hypothetical mechanism of the hydrolysis of phosphoesters by heme- and metal-independent haloperoxidases is suggested. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru.
Subject(s)
Chloride Peroxidase/metabolism , Esters/metabolism , Organophosphorus Compounds/metabolism , Serratia marcescens/enzymology , Electrophoresis, Polyacrylamide Gel , Heme/metabolism , HydrolysisABSTRACT
A bacterial non-heme chloroperoxidase from Serratia marcescens W 250 was immobilized in calcium-alginate gel. Methods for stabilization of the immobilized enzyme were developed, and some kinetic parameters of the immobilized preparations were determined. The enzyme encapsulated into the gel granules in the presence of potassium ferricyanide followed by treatment with glutaraldehyde demonstrated the highest stability under the reaction conditions.
Subject(s)
Chloride Peroxidase/metabolism , Enzymes, Immobilized/metabolism , Serratia marcescens/enzymology , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Chloride Peroxidase/chemistry , Enzyme Stability , Enzymes, Immobilized/chemistryABSTRACT
Indole, indolylacetic acid, and tryptophan were oxidized by cloroperoxidases isolated from strains of Streptomyces lividans and Pseudomonas pyrrocinia. Indigo (indoxyl), isatin, and anthranilic acid (intermediate products of oxidative degradation of indole and indole derivatives) were extracted from the reaction medium.
Subject(s)
Chloride Peroxidase/chemistry , Indoleacetic Acids/chemistry , Indoles/chemistry , Tryptophan/chemistry , Heme , Oxidation-Reduction , Pseudomonas , StreptomycesABSTRACT
The influence of various factors on the activity of chloroperoxidase from Serratia marcescens was investigated. The enzyme is active only in acetate-containing buffers within the pH range 4.2-5.8. F-, Cu2+, [Fe(CN)6]4+, and [Fe(CN)6]3+ inhibit the enzyme. The chloroperoxidase is thermostable and resistant to the effect of lower alcohols.
