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Characterization of glycerophosphoethanolamine ethanolaminephosphodiesterase from Streptomyces sanglieri

Paper ID Volume ID Publish Year Pages File Format Full-Text
20359 43171 2015 8 PDF Available
Title
Characterization of glycerophosphoethanolamine ethanolaminephosphodiesterase from Streptomyces sanglieri
Abstract

Streptomyces sanglieri extracellularly produces a glycerophosphoethanolamine ethanolaminephosphodiesterase (GPE-EP). The gene encoding the enzyme was found to consist of a 2124-bp ORF, which codes for an N-terminal 48 residue signal peptide required for secretion and a 660 amino acid mature protein with a calculated molecular mass of 72,918 Da. The maximum activity for sn-glycero-3-phosphoethanolamine (GPE) was found at pH 8.4 and 65°C in the presence of 0.1% (w/v) Triton X-100. The enzyme was activated in the presence of 2 mM EDTA; however, Zn2+ remarkably inhibited activity. During the hydrolysis of GPE at 65°C and pH 8.4, the apparent Vmax, turnover number (kcat) and Km were determined to be 0.430 mmol min−1 mg-protein−1, 522 s−1 and 0.785 mM, respectively. The enzyme exhibited specificity toward GPE and hydrolyzed ethanolamine-type substrates such as 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine, lysophosphatidylethanolamine and ethanolamine lysoplasmalogen, but not 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine. Moreover, the enzyme showed no activity toward other phospholipids, such as glycerophospholipids and plasmalogens, and sn-glycero-3-phosphodiesters except for sn-glycero-3-phosphoglycerol, suggesting that GPE-EP is not a phospholipase C (PLC). However, the amino acid sequence of GPE-EP shows 86% identity to that of PLC from Streptomyces sp. SirexAA-E (UniProt accession no. G2NFN1). Recombinant GPE-EP was functionally expressed in Escherichia coli using pET-24a(+). GPE hydrolysis by GPE-EP may represent a new pathway for phosphatidylethanolamine metabolism.

Keywords
Glycerophosphoethanolamine ethanolaminephosphodiesterase; Purification; Characterization; Molecular cloning; Expression; Streptomyces sanglieri
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Characterization of glycerophosphoethanolamine ethanolaminephosphodiesterase from Streptomyces sanglieri
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 119, Issue 2, February 2015, Pages 123–130
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
Online Support
Any Questions? feel free to contact us