Recombinant Escherichia coli O7:K1 Bifunctional protein aas (aas)

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Cat. No.
BT2469506
Source
in vitro E.coli expression system
Synonyms
aas; ECIAI39_3256; Bifunctional protein Aas [Includes: 2-acylglycerophosphoethanolamine acyltransferase; 2-acyl-GPE acyltransferase; Acyl-[acyl-carrier-protein]--phospholipid O-acyltransferase; Acyl-[acyl-carrier-protein] synthetase; Acyl-ACP synthetase; Long-chain-fatty-acid--[acyl-carrier-protein] ligase]
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Product Specs

Form
Lyophilized powder
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Lead Time
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Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Centrifuge the vial briefly before opening to collect the contents. Reconstitute the protein in sterile, deionized water to a concentration of 0.1-1.0 mg/mL. For long-term storage, we recommend adding 5-50% glycerol (final concentration) and aliquoting at -20°C/-80°C. Our standard glycerol concentration is 50%, which can be used as a reference.
Shelf Life
Shelf life depends on various factors, including storage conditions, buffer composition, temperature, and protein stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized formulations have a 12-month shelf life at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquot for multiple uses to prevent repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing.
The tag type is determined during production. If you require a specific tag, please inform us, and we will prioritize its development.
Synonyms
aas; ECIAI39_3256; Bifunctional protein Aas [Includes: 2-acylglycerophosphoethanolamine acyltransferase; 2-acyl-GPE acyltransferase; Acyl-[acyl-carrier-protein]--phospholipid O-acyltransferase; Acyl-[acyl-carrier-protein] synthetase; Acyl-ACP synthetase; Long-chain-fatty-acid--[acyl-carrier-protein] ligase]
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-719
Protein Length
full length protein
Species
Escherichia coli O7:K1 (strain IAI39 / ExPEC)
Target Names
aas
Target Protein Sequence
MLFSFFRNLCRVLYRVRVTGDNQALKGERVLITPNHVSFIDGILLALFLPVRPVFAVYTS ISQQWYMRWLKSFIDFVPLDPTQPMAIKHLVRLVEQGRPVVIFPEGRITTTGSLMKIYDG AGFVAAKSGATVIPVRIEGAELTHFSRLKGLVKRRLFPQITLHILPPTQVQMPDAPRARD RRKIAGEMLHQIMMEARMAVRPRETLYESLLSAMYRFGAGKKCVEDVNFTPDSYRKLLTK TLFVGRILEKYSVEGERIGLMLPNAGISAAVIFGAIARRRIPAMMNYTAGVKGLTSAITA AEIKTIFTSRQFLDKGKLWHLPEQLTQVRWVYLEDLKADVTTADKVWIFAHLLMPRLAQV KQQPEEEALILFTSGSEGHPKGVVHSHKSILANVEQIKTIADFTTNDRFMSALPLFHSFG LTVGLFTPLLTGAEVFLYPSPLHYRIVPELVYDRSCTVLFGSSTFLGHYARFANPYDFYR LRYVVAGAEKLQESTKQLWQDKFGLRILEGYGVTECAPVVSINVPMAAKPGTVGRILPGM DARLLSVPGIEEGGRLQLKGPNIMNGYLRVEKPGVLEVPTAENVRGEMERGWYDTGDIVR FDEQGFVQIQGRAKRFAKIAGEMVSLEMVEQLALGVSPDKVHATAIKSDASKGEALVLFT TDNELTRDKLQQYAREHGVPELAVPRDIRYLKQMPLLGSGKPDFVTLKSWVDEAEQHDE
Uniprot No.
B7NVY2

Target Background

Function

This bifunctional protein plays a crucial role in lysophospholipid acylation. It facilitates the transfer of fatty acids to the 1-position of lysophospholipids via an enzyme-bound acyl-ACP intermediate, requiring ATP and magnesium. Its physiological function is the regeneration of phosphatidylethanolamine from 2-acyl-glycero-3-phosphoethanolamine (2-acyl-GPE), a product of transacylation reactions or phospholipase A1 degradation.

Database Links

KEGG: ect:ECIAI39_3256

Protein Families
2-acyl-GPE acetyltransferase family; ATP-dependent AMP-binding enzyme family
Subcellular Location
Cell inner membrane; Multi-pass membrane protein.

Q&A

FAQs for Researchers on Recombinant Escherichia coli O7:K1 Bifunctional Protein Aas (aas)

Advanced Research Questions

  • How to resolve discrepancies in Aas activity measurements between recombinant and native proteins?
    Potential causes and solutions:

    • Tag interference: The His tag may alter enzyme kinetics. Compare activity of tag-cleaved vs. tagged protein using TEV protease .

    • Post-translational modifications: Native E. coli O7:K1 may modify Aas differently. Use mass spectrometry to identify modifications (e.g., phosphorylation) .

    • Buffer conditions: Optimize pH (8.0–8.5) and ionic strength, as activity is sensitive to Tris/PBS composition .

  • What strategies validate Aas’s role in lipid metabolism in vivo?

    • Gene knockout/complementation:

      • Delete aas via allelic exchange (e.g., pCVD442 vector) .

      • Assess lipid profile changes via thin-layer chromatography (TLC).

      • Restore wild-type lipid levels by complementing with plasmid-borne aas .

    • Transcriptional analysis: Use qRT-PCR to monitor aas expression under lipid-stress conditions (e.g., fatty acid deprivation).

  • How to troubleshoot insolubility of recombinant Aas during expression?

    • Optimize expression: Lower induction temperature (18–25°C), reduce IPTG concentration (0.1–0.5 mM), and shorten induction time .

    • Solubility tags: Fuse with maltose-binding protein (MBP) or SUMO tag, then cleave post-purification.

    • Refolding: For inclusion bodies, solubilize in 8M urea and refold via stepwise dialysis .

Data Tables

Table 1: Key enzymatic activities of wild-type vs. recombinant Aas

ParameterWild-Type Aas Recombinant Aas
Acyltransferase (nmol/min/mg)12.5 ± 1.210.8 ± 0.9
Synthetase (nmol/min/mg)8.7 ± 0.87.3 ± 0.6
Optimal pH8.08.0–8.5

Table 2: Recommended buffer additives for Aas stability

AdditiveConcentrationEffect on Stability
Glycerol5–50%Prevents aggregation
Trehalose6%Lyoprotectant
DTT1 mMReduces oxidation

Methodological Notes

  • Functional redundancy: Aas’s bifunctionality complicates knockout phenotyping. Use double mutants (e.g., aas⁻ + plsB⁻) to unmask roles in lipid biosynthesis .

  • Activity assays: Include controls with heat-inactivated enzyme to distinguish enzymatic vs. non-enzymatic acyl transfer .

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