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BEC/CHE 463/563
Fermentation of Recombinant Microorganisms

Course Description

Students completing this course will be able to conduct small-scale fermentations of recombinant microorganisms as well as having an understanding of ways to approach optimization of such processes. One of the focus areas of this course is on protein production and factors that affect gene expression and recombinant protein yield.

Lecture Topics

  1. Fermentation Fundamentals for Production of Peptides, Enzymes, Antibodies and Biopharmaceutical Products
  2. Aeration and O2 Mass Transfer
  3. Sterilization and Inoculum Development
  4. Expression Systems for Recombinant Proteins
  5. Metabolic Engineering and Use of “Omics” Tools
  6. Biosensors and Process Control
  7. Downstream Processing

Lab Topics

  1. Introduction to 2-liter fermenter operation.
  2. Expression of recombinant protein Pfu CelB in E. coli (shaker flask). Introduction to enzymatic assay.
  3. Effect of aeration rate on microbial growth in the fermenter. Expression of recombinant protein Pfu CelB in E. coli (2-liter fermenter).
  4. Optimizing gene expression of Pfu CelB in E. coli (2-liter fermenter). SDS-PAGE analysis.
  5. Expression of recombinant antibody fragment scFv13R4 in E. coli (2-liter fermenter). Fed-batch strategies.
  6. ELISA of antibody fragment protein. SDS-PAGE analysis.

Prerequisites

BEC 363 or BIT 410 or MB 352/354