Secretive glutamate decarboxylase of Corynebacterium glutamicum
catalyzes an efficient conversion of glutamic acid to γ-aminobutyric
acid
Abstract
γ-Aminobutyric acid (GABA) is a non-protein amino acid produced from the
decarboxylation of glutamate by glutamate decarboxylase. Corynebacterium
glutamicum is the most promising host of γ-aminobutyric acid production
for its inherent glutamate precursor supply. However, the
intracellularly expressed glutamate decarboxylase in C. glutamicum
showed the weak catalysis capacity on the conversion of glutamate to
γ-aminobutyric acid. Here we designed an different catalysis scenario by
secretively overexpressing the glutamate decarboxylase in C. glutamicum
and moving the decarboxylation reaction into the extracellular space for
GABA synthesis. A signal peptide in the expression cassette directed the
successful secretion of glutamate decarboxylase in C. glutamicum. The
extracellular catalysis by secreted glutamate decarboxylase increased
the γ-aminobutyric acid generation by three-folds, comparing with that
by intracellular catalysis. Further efforts on enhancing the expression
of glutamate decarboxylase and decreasing the degradation of
γ-aminobutyric acid improved γ-aminobutyric acid generation by 39%. The
fed-batch fermentation of the engineered C. glutamicum strain reached
the record high titer (77.6g /L), overall yield (0.37 g/g glucose), and
productivity (1.21 g/L/h) of γ-aminobutyric acid production. This study
demonstrated a unique design of extracellular catalysis for efficient
γ-aminobutyric acid production by C. glutamicum.