Journal article
Bioreaction Engineering Leading to Efficient Synthesis of L-Glyceraldehyd-3-Phosphate
Institute of Technical Biocatalysis, Hamburg University of Technology, Hamburg, Germany.1
Department of Microbiology and Biotechnology, University of Hamburg, Hamburg, Germany.2
Sigma-Aldrich Chemie GmbH, Member of Merck Group, Buchs, Switzerland.3
Enantiopure L-glyceraldehyde-3-phosphate (L-GAP) is a useful building block in natural biological and synthetic processes. A biocatalytic process using glycerol kinase from Cellulomonas sp. (EC 2.7.1.30) catalyzed phosphorylation of L-glyceraldehyde (L-GA) by ATP is used for the synthesis of L-GAP. L-GAP has a half-life of 6.86 h under reaction conditions.
The activity of this enzyme depends on the Mg2+ to ATP molar ratio showing maximum activity at the optimum molar ratio of 0.7. A kinetic model is developed and validated showing a 2D correlation of 99.9% between experimental and numerical data matrices. The enzyme exhibits inhibition by ADP, AMP, methylglyoxal and Ca2+ , but not by L-GAP and inorganic orthophosphate.
Moreover, equal amount of Ca2+ exerts a different degree of inhibition relative to the activity without the addition of Ca2+ depending on the Mg2+ to ATP molar ratio. If the Mg2+ to ATP molar ratio is set to be at the optimum value or less, inorganic hexametaphosphate (PPi6) suppresses the enzyme activity; otherwise PPi6 enhances the enzyme activity.
Based on reaction engineering parameters such as conversion, selectivity and specific productivity, evaluation of different reactor types reveals that batchwise operation via stirred-tank reactor is the most efficient process for the synthesis of L-GAP.
Language: | English |
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Year: | 2017 |
Pages: | 1600625 |
ISSN: | 18607314 and 18606768 |
Types: | Journal article |
DOI: | 10.1002/biot.201600625 |
Bioengineering Cellulomonas Escherichia coli Glyceraldehyde 3-Phosphate Glycerol Kinase Glycerol kinase Half-Life L-Glyceraldehyde-3-phosphate instability Mg2+ to ATP ratio Phosphates Reaction kinetics Reactor simulation Reproducibility of Results Streptomyces Thermodynamics hexametaphosphate