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Efficient biocatalytic production of d-4-hydroxyphenylglycine by whole cells of recombinant Ralstonia pickettii

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Abstract

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.

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Abbreviations

Amp:

ampicillin

CGMCC:

China General Microbiological Culture Collection Center

DCase:

N-carbamoyl-D-amino-acid-amidohydrolase

dca :

DCase encoding gene

DHase:

d-hydantoinase

dha :

DHase encoding gene

dha-dca (casette):

gene casette encoding both DHase and DCase

d-HPG:

d-4-hydroxyphenylglycine

NC-d-HPG:

N-carbamoyl-d-4-hydroxyphenylglycine

dl-HPH:

dl-4-hydroxyphenylhydantoin

HPLC:

high-performance liquid chromatography

Km:

kanamycin

PS:

parent(al) strain (CGMCC1596)

RP:

recombinant plasmid (pYY05)

RS:

recombinant strain (MMYY01)

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Tc:

tetracycline

WT(s):

wild-type strain(s)

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Authors and Affiliations

  1. Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200 032, P.R. China

    H. Yu, S. Yang, W. Jiang & Y. Yang

  2. Graduate University of the Chinese Academy of Sciences, Beijing, 100 080, P.R. China

    H. Yu

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  1. H. Yu
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  2. S. Yang
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  3. W. Jiang
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  4. Y. Yang
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Correspondence to Y. Yang.

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Yu, H., Yang, S., Jiang, W. et al. Efficient biocatalytic production of d-4-hydroxyphenylglycine by whole cells of recombinant Ralstonia pickettii . Folia Microbiol 54, 509–515 (2009). https://doi.org/10.1007/s12223-009-0073-y

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  • DOI: https://doi.org/10.1007/s12223-009-0073-y

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