Purification and Characterization of
            <scp>d</scp>
            -Aminoacylase from
            <i>Alcaligenes faecalis</i>
            DA1

Yang, Yi Tien; Lin, Chyuan-Sheng; Tseng, Ching‐Ping; Wang, Yng-Jiin; Tsai, Ying-Chieh

doi:10.1128/aem.57.4.1259-1260.1991

Cited by 26 publications

(3 citation statements)

References 12 publications

(12 reference statements)

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“…Many enzymes show a high degree of stereoselectivity. The L-aminoacylase could catalyze the hydrolysis of various N-acetyl-L-amino acids to free Lamino acids, 14,26 while the D-aminoacylase produces free D-amino acids. 26 The progress of the enzymatic reactions and the optical purity of the products and remaining substrates could be monitored by chiral HPLC.…”

Section: Resultsmentioning

confidence: 99%

“…DA1 was obtained from Prof. Ying-Chieh Tsai (Institute of Biochemistry, National Yang-Ming University, Taiwan). 26 The three chiral columns tested in this work were CHIRALPAK WH (0.46 × 25 cm) and CHIRALCEL OD-R (0.46 × 15 cm) (Daicel Chemical Industries, Japan) and CHIROBIOTIC T™ (0.46 × 25 cm) (Advanced Separation Technologies, Whippany, NJ, USA).…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

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Simultaneous analysis of enantiomeric composition of amino acids and N‐acetyl‐amino acids by enantioselective chromatography

2001

Chirality

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Among the three chiral columns, CHIROBIOTIC T, CHIRLPAK WH, and CHIRALCEL OD-R, tested for the separation of racemic amino acids and N-acetyl-amino acids, only CHIROBIOTIC T chiral column which is based on covalently bonded amphoteric glycopeptide, teicoplanin, as the stationary phase ligand could be successfully developed to enantiomerically separate racemic amino acids and N-acetyl amino acids simultaneously. This method can be used to determine the enantiomeric composition of amino acids and N-acetyl-amino acids in the catalysis of D-aminoacylase or L-aminoacylase and the conversion rate of N-acylamino acid racemases.

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Section: Resultsmentioning

confidence: 99%

Section: Materials and Methods Materialsmentioning

confidence: 99%

Simultaneous analysis of enantiomeric composition of amino acids and N‐acetyl‐amino acids by enantioselective chromatography

2001

Chirality

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“…To search for new d ‐aminoacylases suitable for d ‐amino acid production, microorganisms in various soils were screened. Two d ‐aminoacylases from Alcaligenes denitrificans DA181 (Tsai et al 1988; Yang et al 1992) and A. faecalis DA1 (Yang et al 1991; Tsai et al 1992) were isolated and showed higher specific activity and better stereospecificity than those from Pseudomonas and Streptomyces (Sugie and Suzuki 1978; Kubo et al 1980). Two other d ‐aminoacylase‐producing strains, A. denitrificans MI‐4 (Sakai et al 1991), and A. xylosoxydans A‐6 (Moriguchi et al 1993), were subsequently screened.…”

mentioning

confidence: 99%

Structural‐based mutational analysis of d‐aminoacylase from Alcaligenes faecalis DA1

Hsu¹,

Lai²,

Chang³

et al. 2002

Protein Science

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D-Aminoacylase is an attractive candidate for commercial production of D-amino acids through its catalysis in the zinc-assistant hydrolysis of N-acyl-D-amino acids. We report here the cloning, expression, and structural-based mutation of the D-aminoacylase from Alcaligenes faecalis DA1. A 1,007-bp PCR product amplified with degenerate primers, was used to isolate a 4-kb genomic fragment, encoding a 484-residue D-aminoacylase. The enzyme amino-terminal segment shared significant homology within a variety of enzymes including urease. The structural fold was predicted by 3D-PSSM to be similar to urease and dihydroorotase, which have grouped into a novel ␣/␤-barrel amidohydrolase superfamily with a virtually indistinguishable binuclear metal centers containing six ligands, four histidines, one aspartate, and one carboxylated lysine. Three histidines, His-67, His-69, and His-250, putative metal ligands in D-aminoacylase, have been mutated previously, the remaining histidine (His-220) and aspartate (Asp-366) Asp-65, and four cysteines were then characterized. Substitution of Asp-65, Cys-96, His-220, and Asp-366 with alanine abolished the enzyme activity. The H220A mutant bound approximately half the normal complement of zinc ion as did H250N. However, the C96A mutant showed little zinc-binding ability, revealing that Cys-96 may replace the carboxylated lysine to serve as a bridging ligand. According to the urease structure, the conserved amino-terminal segment including Asp-65 may be responsible for structural stabilization. Therefore, production of D-amino acids by D-aminoacylase has commercial importance.To search for new D-aminoacylases suitable for D-amino acid production, microorganisms in various soils were screened. Two D-aminoacylases from Alcaligenes denitrificans DA181 (Tsai et al. 1988;Yang et al. 1992) and A. faecalis DA1 (Yang et al. 1991;Tsai et al. 1992) were isolated and showed higher specific activity and better stereospecificity than those from Pseudomonas and Streptomyces (Sugie and Suzuki 1978;Kubo et al. 1980). Two other D-aminoacylase-producing strains, A. denitrificans MI-4 (Sakai et al. 1991), and A. xylosoxydans A-6 (Moriguchi et al. 1993), were subsequently screened. Three genes encoding for the A-6 D-aminoacylases have been cloned and expressed in Escherichia coli (Wakayama et al. 1995a,b,c). These A-6 D-aminoacylases share 45%-50% sequence iden-

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Recent Developments in Aminopeptidases, Racemases, and Oxidases

2016

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Purification and Characterization of d -Aminoacylase from Alcaligenes faecalis DA1

Cited by 26 publications

References 12 publications

Simultaneous analysis of enantiomeric composition of amino acids and N‐acetyl‐amino acids by enantioselective chromatography

Simultaneous analysis of enantiomeric composition of amino acids and N‐acetyl‐amino acids by enantioselective chromatography

Structural‐based mutational analysis of d‐aminoacylase from Alcaligenes faecalis DA1

Recent Developments in Aminopeptidases, Racemases, and Oxidases

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