Antibiotic Technology

Course Description

COURSE CONTENT

1. Development of biotechnological antibiotic production and importance of continuous development of new antibiotics

P: History of biotechnological production of antibiotics. Nomenclature and classification of antibiotics. Application of „omics“ technologies in the development of novel antibiotics. Application of genetically modified microorganisms in the development of novel, modified and hybrid, antibiotics. Alternative antimicrobial strategies in combat of antibiotic resistance spreading.

S: Application of current „omics“ and engineering methods in the discovery and development of novel antibiotics efficient against multidrug-resistant microorganisms.

V: Determination of chloramphenicol residual concentrations in milk by ELISA method. Determination of bacterial sensitivity to antibiotics by diffusion method. Detection of antibiotic resistance genes by PCR method.

2. Biotechnological processes of antibiotics production

L: Antibiotic producing microorganisms. Genetics and biosynthesis of antibiotics as secondary metabolites. Regulation of antibiotics biosynthesis. Strain improvement and bioprocess development. Bioprocess parameters of biotechnological antibiotics production. Downstream processing: isolation and purification of antibiotics. Improvement of antibiotics stability and activity by current drug delivery technologies. Trends in bioprocesses development and optimization strategies for antibiotic production.

E: Colorimetric method for determination of oxytetracycline. Iodometric method for determination of penicillin.

3. Biotechnological processes of tetracycline antibiotics production

L: Structure, mechanism and catalytic activity of type I polyketide synthase (PKS). Biotechnological production of oxytetracycline: upstream and downstream processing.

S: Calculation of bioprocess parameters of oxytetracycline biosynthesis and isolation procedures on industrial scale.

E: Oxytetracycline (OTC) biosynthesis by submerged cultivation of Streptomyces rimosus and OTC isolation on laboratory scale.

4. Biotechnological processes of β-lactam antibiotics production

L: β-lactam antibiotics biosynthesis by thiotemplate mechanism of nonribosomal peptide synthetase (NRPS) activity. Biotechnological production of penicillin G. Isolation of penicillin G. Enzymatic preparation of 6-aminopenicillanic acid, 7-aminocephalosporanic acid and semisynthetic antibiotics. Biotechnological production of cephalosporin C and cephamycin. Enzymatic synthesis of new b-lactams. Production of clavulanic acid. Next-generation β-lactamase inhibitors.

S: Biotechnological processes of b-lactam antibiotics and β-lactamase inhibitors production.

5. Biotechnological processes of aminoglycoside, macrolide, peptide, glycolpeptide and aromatic antibiotics production

L: Biosynthesis and post-translational modification of ribosomal peptide antibiotics. Nonribosomal peptide antibiotics biosynthesis by modular nonribosomal peptide synthetase (NRPS). Polyketide antibiotics biosynthesis by type II and III polyketide synthase (PKS) and by PKS/NRPS hybrid enzymes. Biotechnological production of streptomycin, erythromycin, bacitracin, chloramphenicol and vancomycin: upstream and downstream processes for each of mentioned antibiotics production.

S: Biotechnological processes of aminoglycoside, macrolide, peptide, glycopeptide and aromatic antibiotics production

6. Biotechnological processes of antibiotics production with antifungal and cytostatic activity

L: Polyene and non-polyene antifungal antibiotics. Biotechnological production of natamycin (pimaricin): upstream and downstream processing. Anticancer (antineoplastic) antibiotics. Biotechnological production of bleomycin and daunorubicin: upstream and downstream processes. Circular (recycling) economy in antibiotic factory.

S: Biotechnological processes of antifungal and cytostatic antibiotics production

LEARNING OUTCOMES

• Argue the importance of continuous development of new antibiotics and application of the alternative antimicrobial strategies to combat the spread of antibiotic resistance
• Present the schematic diagram of the biotechnological process of antibiotic production and isolation
• Explain the main stages of the biotechnological process of production, isolation and purification of the antibiotics for preparation of the pharmaceutical preparations
• Analyse the bioprocess parameters of antibiotic production to assess the effectiveness of the biotechnological process
• Prepare the inoculum of the antibiotic-producing microorganism Streptomyces rimosus and carry out the biotechnological production of the antibiotic oxytetracycline on a laboratory scale
• Carry out the procedure of pretreatment of the fermented slurry after the biotechnological production of oxytetracycline, considering the physicochemical properties of the antibiotics (solubility, stability)
• Apply precipitation and extraction methods for purifying antibiotics
• Carry out the freeze-drying of the antibiotic by sublimation under a vacuum (lyophilisation)
• Determine the activity of tetracycline and penicillin antibiotics using quantitative methods for the determination of antibiotics
• Determine: the concentration of antibiotic residues by ELISA method, susceptibility/resistance of bacteria to antibiotics by diffusion method, and antibiotic resistance genes by PCR method

To enrol in this course, the following courses must be completed (undergraduate studies):

• Biotechnology 1
• Microbiology
• Biochemistry 1
• Biochemistry 2
• Transport Phenomena
• Unit Operations