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b Proposed modified hydantoinase process for the synthesis of enantiopure β-amino acids starting from racemic 6-monosubstituted dihydrouracils.
In a previous study we tested the potential of using a modified hydantoinase process for the production of optically pure β-amino acids (see Figure 1B).
Since kinetic resolutions merely enable a maximum yield of 50%%, lately the application of a modified hydantoinase process was proposed (Fig. 1b) (Engel et al. 2014).
Furthermore the hydrolysis of dihydrouracils was discovered, making this enzyme a potential tool toward enantiopure β-amino acids applying a modified hydantoinase process (Engel et al. 2012b).
Even the second step of the modified hydantoinase process was realized by Martínez-Goméz et al.: The synthesis of α-methyl-β-alanine from 5-methyl-5,6-dihydrouracil was accomplished using the dihydropyrimidinase from Sinorhizobium meliloti CECT4114 and the β-ureidopropionase from Agrobacterium tumefaciens C58 (Martínez-Gómez et al. 2012).
The viability of the first step in this proposed modified hydantoinase process, the hydrolysis of dihydrouracil as well as the hydrolysis of differently substituted dihydrouracils was shown in previous works (May et al. 1998; Servi et al. 2005; O'Neill et al. 2011).
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Fig. 1 Hydantoinase process for the synthesis of optically pure α-amino acids starting from racemic 5-monosubstituted hydantoins [Modified after Syldatk et al. (1999)].
This total conversion demonstrates that the "double-racemase hydantoinase process" upgrades the classical "hydantoinase process" for natural and non-natural l-amino acid production.
L159V variant was used to convert HPAH to l-homophenylalanine (l-HPA) in the hydantoinase process.
In this work the hydantoinase and carbamoylase from Arthrobacter crystallopoietes DSM 20117 were investigated with respect to their applicability in a cell-free hydantoinase process.
The "hydantoinase process" is a well-established method for the industrial production of optically pure d-amino acids.
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