Purification and properties of d-aminoacylase from Alcaligenes denitrificans subsp. xylosoxydans MI-4

Kenji Sakai; Taketeru Obata; Kohtaro Ideta; Mitsuaki Moriguchi

doi:10.1016/0922-338X(91)90227-8

Purification and properties of d-aminoacylase from Alcaligenes denitrificans subsp. xylosoxydans MI-4

Kenji Sakai, Taketeru Obata, Kohtaro Ideta, Mitsuaki Moriguchi

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Abstract

d-Aminoacylase has been purified 144-fold to electrophoretic homogeneity by ammonium sulfate fractionation, DEAE-Toyopearl and affinity column chromatographies, and Sephadex G-100 gel filtration from the crude extracts of Alcaligenes denitrificans subsp. xylosoxydans MI-4. The enzyme was composed of a single polypeptide of about 51,000. The enzyme catalyzed hydrolysis of N-acyl-derivatives of neutral d-amino acids. Optimal pH and temperature were 7.8 and 50°C. The apparent K_m and the V_max for N-acetyl-d-phenylalanine were 14.1 mM and 1331 units/mg protein, respectively. The activity of the enzyme was inhibited by N-acetyl-d-valine (K_i=2.15 mM) and N-acetyl-d-alloisoleucine (K_i=1.47 mM), but not by its products (i.e., amino acids and acetate). The enzyme also had dipeptidase activity. Activation by metal ions was not observed.

Original language	English
Pages (from-to)	79-82
Number of pages	4
Journal	Journal of Fermentation and Bioengineering
Volume	71
Issue number	2
DOIs	https://doi.org/10.1016/0922-338X(91)90227-8
Publication status	Published - 1991
Externally published	Yes

All Science Journal Classification (ASJC) codes

Biotechnology
Applied Microbiology and Biotechnology

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10.1016/0922-338X(91)90227-8

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title = "Purification and properties of d-aminoacylase from Alcaligenes denitrificans subsp. xylosoxydans MI-4",

abstract = "d-Aminoacylase has been purified 144-fold to electrophoretic homogeneity by ammonium sulfate fractionation, DEAE-Toyopearl and affinity column chromatographies, and Sephadex G-100 gel filtration from the crude extracts of Alcaligenes denitrificans subsp. xylosoxydans MI-4. The enzyme was composed of a single polypeptide of about 51,000. The enzyme catalyzed hydrolysis of N-acyl-derivatives of neutral d-amino acids. Optimal pH and temperature were 7.8 and 50°C. The apparent Km and the Vmax for N-acetyl-d-phenylalanine were 14.1 mM and 1331 units/mg protein, respectively. The activity of the enzyme was inhibited by N-acetyl-d-valine (Ki=2.15 mM) and N-acetyl-d-alloisoleucine (Ki=1.47 mM), but not by its products (i.e., amino acids and acetate). The enzyme also had dipeptidase activity. Activation by metal ions was not observed.",

author = "Kenji Sakai and Taketeru Obata and Kohtaro Ideta and Mitsuaki Moriguchi",

year = "1991",

doi = "10.1016/0922-338X(91)90227-8",

language = "English",

volume = "71",

pages = "79--82",

journal = "Journal of Bioscience and Bioengineering",

issn = "1389-1723",

publisher = "The Society for Biotechnology, Japan",

number = "2",

}

TY - JOUR

T1 - Purification and properties of d-aminoacylase from Alcaligenes denitrificans subsp. xylosoxydans MI-4

AU - Sakai, Kenji

AU - Obata, Taketeru

AU - Ideta, Kohtaro

AU - Moriguchi, Mitsuaki

PY - 1991

Y1 - 1991

N2 - d-Aminoacylase has been purified 144-fold to electrophoretic homogeneity by ammonium sulfate fractionation, DEAE-Toyopearl and affinity column chromatographies, and Sephadex G-100 gel filtration from the crude extracts of Alcaligenes denitrificans subsp. xylosoxydans MI-4. The enzyme was composed of a single polypeptide of about 51,000. The enzyme catalyzed hydrolysis of N-acyl-derivatives of neutral d-amino acids. Optimal pH and temperature were 7.8 and 50°C. The apparent Km and the Vmax for N-acetyl-d-phenylalanine were 14.1 mM and 1331 units/mg protein, respectively. The activity of the enzyme was inhibited by N-acetyl-d-valine (Ki=2.15 mM) and N-acetyl-d-alloisoleucine (Ki=1.47 mM), but not by its products (i.e., amino acids and acetate). The enzyme also had dipeptidase activity. Activation by metal ions was not observed.

AB - d-Aminoacylase has been purified 144-fold to electrophoretic homogeneity by ammonium sulfate fractionation, DEAE-Toyopearl and affinity column chromatographies, and Sephadex G-100 gel filtration from the crude extracts of Alcaligenes denitrificans subsp. xylosoxydans MI-4. The enzyme was composed of a single polypeptide of about 51,000. The enzyme catalyzed hydrolysis of N-acyl-derivatives of neutral d-amino acids. Optimal pH and temperature were 7.8 and 50°C. The apparent Km and the Vmax for N-acetyl-d-phenylalanine were 14.1 mM and 1331 units/mg protein, respectively. The activity of the enzyme was inhibited by N-acetyl-d-valine (Ki=2.15 mM) and N-acetyl-d-alloisoleucine (Ki=1.47 mM), but not by its products (i.e., amino acids and acetate). The enzyme also had dipeptidase activity. Activation by metal ions was not observed.

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U2 - 10.1016/0922-338X(91)90227-8

DO - 10.1016/0922-338X(91)90227-8

M3 - Article

AN - SCOPUS:0025974624

SN - 1389-1723

VL - 71

SP - 79

EP - 82

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

IS - 2

ER -

Purification and properties of d-aminoacylase from Alcaligenes denitrificans subsp. xylosoxydans MI-4

Abstract

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