Related Experiment Video
Updated: Jul 4, 2026

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Conversion of alpha-ketoglutarate into L-glutamic acid with urea as ammonium source using multienzyme systems and
1Artificial Cells and Organs Research Centre, Faculty of Medicine, McGill University, 3655 Drummond Street, Montreal, Quebec, Canada.
Abstract:
Urea could be effectively converted into L-glutamic acid with semipermeable nylon-polyethylenimine artificial cells containing L-glutamic dehydrogenase (EC 1.4.1. 3), yeast alcohol dehydrogenase (EC 1.1.1.1), urease (EC 3.5.1. 5) and soluble dextran-NAD(+). For batch conversion, the artificial cell suspension to total reaction volume ratios ranged from 1 in 5 to 1 in 60. From 22.6 to 53.4 micromol of L-glutamic acid could be produced by 0.4 mL artificial cell suspension within 2 h. The corresponding conversion ratios were 56.5-11. 1%. The L-glutamic dehydrogenase multienzyme system showed a good storage stability: 66.0% of the original activity was retained after 1 month of storage at 4 degrees C. A small bioreactor was prepared to contain 4.0 mL artificial cells. At a flow rate of SV = 1.5 h(-1), the maximum conversion rate was 49.6 micromol L-glutamic acid/p h. Thirty-eight percent of the maximum activity was retained when continuously used for four days at 22 degrees C. A kinetic analysis for the L-glutamic dehydrogenase multienzyme system was studied. The Michaelis constants are as follows: alpha-ketoglutarate is 0.838 mM; urea is 1.90 mM; dextran- NAD(+) is 0.345 mM; and ethanol is 5.31 mM.
More Related Videos
12:47Workflow Based on the Combination of Isotopic Tracer Experiments to Investigate Microbial Metabolism of Multiple Nutrient Sources
Published on: January 22, 2018
11:47Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System
Published on: August 1, 2016
Related Concept Videos
Amino Acid Catabolism
Inorganic Nitrogen Assimilation
Urea Cycle
Production of Organic Acids
Phase II Reactions: Glucuronidation
Production of Pharmaceuticals