Efficient biocatalytic production of D-4-hydroxyphenylglycine by whole cells of recombinant Ralstonia pickettii.
The first establishment of a homologous expression system in the host Ralstonia pickettii CGMCC1596 using the compatible broad-host-range plasmid pWB5 is described. When whole cells of the recombinant strain R. pickettii MMYY01 (CGMCC1596/pYY05) were used as the biocatalyst to transform DL-4-hydroxyphenylhydantoin (DL-HPH) to D-4-hydroxyphenylglycine (D-HPG), the conversion rate reached 94 % in first 9 h, at a production rate of 2.8 g L(-1) h(-1), with the rapid reduction of the intermediate [N-carbamoyl-2-(4-hydroxyphenyl)glycine], compared with 80 % in >50 h at a rate of 0.5 g L(-1) h(-1) for the CGMCC1596. The stability of the recombinant plasmid pYY05 is sufficient for its application in industrial batch fermentation. An alternative strategy for the conversion of DL-HPH to D-HPG by resting CGMCC1596 cells and heterologous DCase expressed by E. coli is discussed.
Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, P.R. China.
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D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D005285:Fermentation; D005818:Genetic Engineering; D005998:Glycine / Q000031:analogs & derivatives* / Q000378:metabolism; D006801:Humans; D010957:Plasmids; D049920:Ralstonia pickettii / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D011994:Recombinant Proteins; C023010:4-hydroxyphenylglycine; D000581:Amidohydrolases; C104561:N-carbamoyl-L-amino-acid hydrolase; D005998:Glycine