| Cloning, expression and characterization of d-aminoacylase from Achromobacter xylosoxidans subsp. denitrificans ATCC 15173. | |
| | |
MedLine Citation:
|
PMID: 23369306 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
|
d-Aminoacylase catalyzes the conversion of N-acyl-d-amino acids to d-amino acids and fatty acids. The aim of this study was to identify the d-aminoacylase gene from Achromobacter xylosoxidans subsp. denitrificans ATCC 15173 and investigate the biochemical characterization of the enzyme. A previously uncharacterized d-aminoacylase gene (ADdan) from this organism was cloned and sequenced. The open reading frame (ORF) of ADdan was 1467bp in size encoding a 488-amino acid polypeptide. ADdan, with a high amino acid similarity to N-acyl-d-aspartate amidohydrolase from Alcaligenes A6, showed relatively low sequence similarities to other characterized d-aminoacylases. The recombinant ADdan protein was expressed in Escherichia coli BL21 (DE3) using pET-28a with a T7 promoter. The enzyme was purified in a single chromatographic step using nickel affinity gel column. The molecular mass of the expressed protein, calculated by SDS-PAGE, was about 52kDa. The purified ADdan showed optimal activity at pH 8.0 and 50°C, and was stable at pH 6.0-8.0 and up to 45°C. Its activity was inhibited by Cu(2+), Fe(2+), Ca(2+), Mn(2+), Ni(2+), Zn(2+) and Hg(2+), whereas Mg(2+) had no significant influence on this recombinant d-aminoacylase. This is the first report on the characterization of d-aminoacylase with activity towards both N-acyl derivatives of neutral d-amino acids and N-acyl-d-aspartate. The characteristics of ADdan could prove to be of interest in industrial production of d-amino acids. |
| | |
Authors:
|
Wei Wang; Huange Xi; Qirui Bi; Ying Hu; Yang Zhang; Mengxiang Ni |
Related Documents
:
|
8726286 - Short-chain fatty acids in the human colon: relation to gastrointestinal health and dis... 20156686 - Pretreatment of wheat straw by nonionic surfactant-assisted dilute acid for enhancing e... 23131896 - Suppression of ccr impacts metabolite profile and cell wall composition in pinus radiat... 23354716 - A hydrolase from lactobacillus sakei moonlights as transaminase. 1883086 - Colonization control of lactose-fermenting salmonella typhimurium in young broiler chic... 20408596 - Self-ordered anodic alumina with continuously tunable pore intervals from 410 to 530 nm. |
Publication Detail:
|
Type: JOURNAL ARTICLE Date: 2013-1-28 |
Journal Detail:
|
Title: Microbiological research Volume: - ISSN: 1618-0623 ISO Abbreviation: Microbiol. Res. Publication Date: 2013 Jan |
Date Detail:
|
Created Date: 2013-2-1 Completed Date: - Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 9437794 Medline TA: Microbiol Res Country: - |
Other Details:
|
Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
|
Copyright © 2013 Elsevier GmbH. All rights reserved. |
Affiliation:
|
School of Life Science & Technology, China Pharmaceutical University, No. 24, Tong Jia Xiang, Nanjing 210009, PR China. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Overexpression of polyphosphate kinase gene (ppk) increases bioinsecticide production by Bacillus th...
Next Document: The role of axillary ultrasound in the detection of metastases from primary breast cancers.