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.
Yu
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H
Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, P.R. China.
Yang
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S
Jiang
W
W
Yang
Y
Y
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