The Hydantoin Transport Protein from Microbacterium liquefaciens

ABSTRACT The gene hyuP from Microbacterium liquefaciens AJ 3912 with an added His6 tag was cloned into the expression plasmid pTTQ18 in an Escherichia coli host strain. The transformed E. coli showed transport of radioisotope-labeled 5-substituted hydantoins with apparent Km values in the micromolar range. This activity exhibited a pH optimum of 6.6 and was inhibited by dinitrophenol, indicating the requirement of energy for the transport system. 5-Indolyl methyl hydantoin and 5-benzyl hydantoin were the preferred substrates, with selectivity for a hydrophobic substituent in position 5 of hydantoin and for the l isomer over the d isomer. Hydantoins with less hydrophobic substituents, cytosine, thiamine, uracil, allantoin, adenine, and guanine, were not effective ligands. The His-tagged hydantoin transport protein was located in the inner membrane fraction, from which it was solubilized and purified and its identity was authenticated.

[1]  V. Blessie,et al.  Active membrane transport and receptor proteins from bacteria. , 2005, Biochemical Society transactions.

[2]  Y. Takenaka,et al.  Molecular Cloning and Expression of the hyu Genes from Microbacterium liquefaciens AJ 3912, Responsible for the Conversion of 5-Substituted Hydantoins to α-Amino Acids, in Escherichia coli , 2005, Bioscience, biotechnology, and biochemistry.

[3]  Shunichi Suzuki,et al.  Purification and Characterization of Hydantoin Racemase from Microbacterium liquefaciens AJ 3912 , 2005, Bioscience, biotechnology, and biochemistry.

[4]  R. Dorrington,et al.  Mutational analysis of the hydantoin hydrolysis pathway in Pseudomonas putida RU-KM3S , 2004, Applied Microbiology and Biotechnology.

[5]  S. Martínez-Rodríguez,et al.  Biochemical characterization of a novel hydantoin racemase from Agrobacterium tumefaciens C58. , 2004, Biochimie.

[6]  S. Martínez-Rodríguez,et al.  Molecular Cloning, Purification, and Biochemical Characterization of Hydantoin Racemase from the Legume Symbiont Sinorhizobium meliloti CECT 4114 , 2004, Applied and Environmental Microbiology.

[7]  Ian T. Paulsen,et al.  TransportDB: a relational database of cellular membrane transport systems , 2004, Nucleic Acids Res..

[8]  P. Henderson,et al.  Collection and characterisation of bacterial membrane proteins , 2003, FEBS letters.

[9]  S. Martínez-Rodríguez,et al.  Overexpression and characterization of hydantoin racemase from Agrobacterium tumefaciens C58. , 2003, Biochemical and biophysical research communications.

[10]  C. Syldatk,et al.  Hydantoinases and related enzymes as biocatalysts for the synthesis of unnatural chiral amino acids. , 2001, Current opinion in biotechnology.

[11]  C. Syldatk,et al.  Cloning and characterization of genes from Agrobacterium sp. IP I-671 involved in hydantoin degradation , 2001, Applied Microbiology and Biotechnology.

[12]  C. Syldatk,et al.  Organization of genes responsible for the stereospecific conversion of hydantoins to α-amino acids in Arthrobacter aurescens DSM 3747 , 2001, Archives of Microbiology.

[13]  C. Syldatk,et al.  Development of an Escherichia coli whole cell biocatalyst for the production of L-amino acids. , 2001, Journal of biotechnology.

[14]  P. Henderson,et al.  Expression, purification and properties of multidrug efflux proteins. , 2000, Biochemical Society transactions.

[15]  C. Syldatk,et al.  Hydantoin racemase from Arthrobacter aurescens DSM 3747: heterologous expression, purification and characterization. , 2000, Journal of biotechnology.

[16]  B. Poolman,et al.  The amplified expression, identification, putrification, assay and properties of his-tagged bacterial membrane transport proteins , 2000 .

[17]  S. Baldwin.  Membrane transport : a practical approach , 2000 .

[18]  M H Saier,et al.  Phylogenetic characterization of novel transport protein families revealed by genome analyses. , 1999, Biochimica et biophysica acta.

[19]  Andreas S. Bommarius,et al.  Biocatalysis to amino acid-based chiral pharmaceuticals—examples and perspectives , 1998 .

[20]  J. Ogawa,et al.  Diversity and versatility of microbial hydantoin-transforming enzymes , 1997 .

[21]  C. Tate,et al.  Proton-linked L-rhamnose transport, and its comparison with L-fucose transport in Enterobacteriaceae. , 1993, The Biochemical journal.

[22]  T. Ishikawa,et al.  Purification and characterization of the hydantoin racemase of Pseudomonas sp. strain NS671 expressed in Escherichia coli , 1992, Journal of bacteriology.

[23]  T. Cooper,et al.  The allantoin and uracil permease gene sequences of Saccharomyces cerevisiae are nearly identical , 1992, Yeast.

[24]  T. Ishikawa,et al.  Identification and sequencing of a gene encoding a hydantoin racemase from the native plasmid of Pseudomonas sp. strain NS671 , 1992, Journal of bacteriology.

[25]  T. Ishikawa,et al.  Cloning and sequencing of the genes involved in the conversion of 5-substituted hydantoins to the corresponding L-amino acids from the native plasmid of Pseudomonas sp. strain NS671 , 1992, Journal of bacteriology.

[26]  C. Syldatk,et al.  Production of optically pure d- and l-α-amino acids by Bioconversion of d,l-5-monosubstituted hydantoin derivatives , 1990 .

[27]  G. von Heijne,et al.  Topogenic signals in integral membrane proteins. , 1988, European journal of biochemistry.

[28]  K. Yokozeki,et al.  Mechanism of asymmetric production of L-aromatic amino acids from the corresponding hydantoins by Flavobacterium sp. , 1987 .

[29]  M. Stark.  Multicopy expression vectors carrying the lac repressor gene for regulated high-level expression of genes in Escherichia coli. , 1987, Gene.

[30]  P. Henderson,et al.  Assay, genetics, proteins, and reconstitution of proton-linked galactose, arabinose, and xylose transport systems of Escherichia coli. , 1986, Methods in enzymology.

[31]  T. Cooper,et al.  Allantoin transport in Saccharomyces cerevisiae , 1977, Journal of bacteriology.

[32]  H. Kaback,et al.  The relationship between the electrochemical proton gradient and active transport in Escherichia coli membrane vesicles. , 1977, Biochemistry.

[33]  W. Schaffner,et al.  A rapid, sensitive, and specific method for the determination of protein in dilute solution. , 1973, Analytical biochemistry.

[34]  Tateo Suzuki,et al.  Optical Rotatory Dispersion and Circular Dichroism of Amino Acid Hydantoins , 1973 .

[35]  I. West.  Lactose transport coupled to proton movements in Escherichia coli. , 1970, Biochemical and biophysical research communications.

[36]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.