Y. Ikenaka et al., Thermostability reinforcement through a combination of thermostability-related mutations of N-carbamyl-D-amino acid amidohydrolase, BIOS BIOT B, 63(1), 1999, pp. 91-95
For the improvement of N-carbamyl-D-amino acid amidohydrolase (DCase), whic
h can be used for the industrial production of D-amino acids, the stability
of DCase from Agrobacterium sp. KNK712 was improved through various combin
ations of thermostability-related mutations. The thermostable temperature (
defined as the temperature on heat treatment for 10 min that caused a decre
ase in the DCase activity of 50%) of the enzyme which had three amino acids
, H57Y, P203E, and V236A, replaced was increased by about 19 degrees C. The
mutant DCase, designated as 455M, was purified and its enzymatic propertie
s were studied. The enzyme had highly increased stability against not only
temperature but also pH, the optimal temperature of the enzyme being about
75 degrees C. The substrate specificity of the enzyme for various N-carbamy
l-D-amino acids was changed little in comparison with that of the native en
zyme. Enzymochemical parameters were also measured.