BEC 363
Foundations of Recombinant Microorganisms for Biomanufacturing

Course Description

Introduction to basic biomanufacturing techniques with particular focus on the most commonly used recombinant microbes in industry. It includes microbial identification, metabolism, and growth kinetics; recombinant production and analysis of low molecular weight molecules, alcohols, recombinant enzymes and others. The laboratory portion of this half-semester course provides students with practical experience on basic biomanufacturing techniques carried out in small scale.

Lecture Topics

  1. Survey of microorganisms used in industry. Rationale for strain selection, development, process optimization. Overview of recombinant protein therapeutic and vaccine products.
  2. Microbial identification by morphology, physiology and genome sequencing (MicroSeq). Inoculum preparation and storage of microbial cultures in cell banks.
  3. Introduction to microbial metabolism. Stoichiometry of microbial growth. Types of growth media (defined, semi-defined, complex) and media optimization. Quantification of cell growth, growth yields and batch growth kinetics. Measurement of growth of aerobic or anaerobic microbes, animal cells in micro wells, T-flasks, shaker flasks and spinner flasks.
  4. Microbial production and detection of alcohols, organic acids, and antibiotics.
  5. Basic genetic engineering to alter microbes to produce recombinant peptides and proteins useful as therapeutics or diagnostics: plasmids, promoters and gene expression.
  6. Microbial production and detection of recombinant products: peptides, proteins, hormones and antibodies.
  7. Microbial production and detection of enzymes. Use of enzymes and cells as biocatalysts.

Lab Topics

Note: Some topics for slow-growing microbes may extend more than one laboratory period.

  1. Biological safety basics, aseptic techniques for avoiding contamination in microbial cultures and cell lines. Preparing liquid culture media and solid agar media. Environmental monitoring.
  2. Microbial identification by microscopy/plate morphology and metabolism tests. Environmental monitoring analysis: culture contamination by microscopy and plating methods. Generation of cell banks: cell counting/freezing of E. coli and CHO cells,preservation of S. cerevisiae. Cultivation of cell lines used in industry: thawing and cultivating E. coli in shaker flasks and on agar plates, thawing CHO and MDCK cells and seeding shaker flasks and T flasks, respectively.
  3. Measuring microbial growth rate and cell yields: aerobic growth of E. coli, anaerobic growth of E. coli. E. coli cell banking post freezing analysis.
  4. Microbial production of alcohols & antibiotics: anaerobic growth and ethanol production from starch, corn or hydrolyzed cellulose by S. cerevisiae plus glucoamylase, measuring antibiotic production by B. licheniformis using bioassays.
  5. Genetic transformation to alter microbial characteristics: plasmid isolation and transformation of E. coli, selection for transformants expressing GFP.
  6. Microbial production of recombinant products: transformation efficiency, quantification of growth and intracellular product by E. coli expressing GFP, quantification of intracellular and extracellular GFP in recombinant CHO cell culture.
  7. Enzymes or microbial cells as biocatalysts: CelB production by recombinant S. cerevisiae, glucoamylase production by Aspergillus sps and purified glucoamylase enzyme kinetics.
  8. Final Practical Exam

Prerequisites & Requirements


BIO 183

Text Requirements

All lectures, lab modules, and reading assignments will be available on the Moodle site for the course or will be provided in hard copy by the instructor.