Major proteins related to chlortetracycline biosynthesis in a Streptomyces aureofaciens production strain studied by quantitative proteomics

• Published in: Applied microbiology and biotechnology Vol. 57; no. 5-6; pp. 717 - 724
• Main Authors: LI, X.-M, NOVOTNA, J, VOHRADSKY, J, WEISER, J
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.12.2001; Springer Nature B.V
• Subjects: Anhydrotetracycline monooxygenase; Anti-Bacterial Agents - biosynthesis; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biosynthesis; Biotechnology; chlortetracycline; Chlortetracycline - biosynthesis; Electrophoresis, Gel, Two-Dimensional; elongation factor Tu; Enzymes; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; Kinetics; Microbiology; Mission oriented research; Oxidoreductases - genetics; Oxidoreductases - isolation & purification; Oxidoreductases - metabolism; Oxygenases - genetics; Oxygenases - isolation & purification; Oxygenases - metabolism; Physiology and metabolism; Proteins; Proteome; proteomics; Streptomyces aureofaciens; Streptomyces aureofaciens - drug effects; Streptomyces aureofaciens - genetics; Streptomyces aureofaciens - metabolism; Thiocyanates - pharmacology; Actinomycetales; Antibiotic; Streptomycetaceae; Tetracycline derivatives; Production; Proteomics; Bacteria; Actinomycetes; Biosynthesis; Protein; Quantitative analysis; Streptomyces aureofaciens
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s002530100807

Changes in synthesis and abundance of proteins associated with chlortetracycline (CTC) production in Streptomyces aureofaciens were investigated by two-dimensional polyacrylamide gel electrophoresis of proteins pulse-labelled in vivo with L-[35S]methionine. Eleven individual protein spots were selected as being related to formation of the antibiotic. Expression of these prominent proteins was not observed in the non-producing mutant; moreover, they were overexpressed in cultures grown in the presence of benzyl thiocyanate, a specific stimulator of CTC biosynthesis used in industrial fermentations. The expression kinetics of the selected proteins was assessed using the technique of computer-assisted image analysis with the EQIAS software and the elongation factor Tu as an internal standard. Interestingly, the kinetic profiles were generally not identical. including those of anhydrotetracycline monooxygenase and the 13-kDa subunit of tetracycline dehydrogenase, two enzymes involved, in the terminal sequential steps of the CTC biosynthetic pathway. The presence of more forms of these enzymes with different charge characteristics was observed. The data presented demonstrated how dramatically the industrial microorganism can change its protein repertoire during the production phase; at least five proteins were nearly comparable in level to the most prominent proteins, exemplified by elongation factor Tu.