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  • Characterization of glycerophosphoethanolamine ethanolaminephosphodiesterase from Streptomyces sanglieri.

Characterization of glycerophosphoethanolamine ethanolaminephosphodiesterase from Streptomyces sanglieri.

Journal of bioscience and bioengineering (2014-08-20)
Shingo Mineta, Kazutaka Murayama, Daisuke Sugimori
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, Zn(2+) 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.

MATERIALS
Product Number
Brand
Product Description

Avanti
06:0 PE, 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine, chloroform
Pricing and availability is not currently available.
Avanti
06:0 PE, 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine, powder
Pricing and availability is not currently available.