Recombinant Vibrio vulnificus Lipoprotein signal peptidase (lspA)

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Cat. No.
BT2051500
Source
in vitro E.coli expression system
Synonyms
lspA; VV0688; Lipoprotein signal peptidase; Prolipoprotein signal peptidase; Signal peptidase II; SPase II
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Description

Recombinant Production and Applications

Recombinant LspA production involves heterologous expression in E. coli or Lactococcus lactis. For example:

  • Expression System: E. coli C43 (DE3) with a pET28a vector for high-yield soluble protein .

  • Tagging: N-terminal His-tag for purification via immobilized metal affinity chromatography (IMAC) .

ParameterDescription
Host StrainE. coli C43 (DE3) or BL21(DE3)
VectorpET28a (KanR^R)
Induction0.5 mM IPTG at 18°C for 16–20 hours
PurificationNi-NTA chromatography, eluted with 250 mM imidazole
Yield~2–5 mg/L culture (varies by construct)

Data extrapolated from homologous LspA studies .

Functional Insights from Homologs

  • Complementation Assays: Recombinant R. typhi LspA restored growth in E. coli Y815 at nonpermissive temperatures, confirming enzymatic activity .

  • Virulence Link: In S. aureus, LspA-deficient mutants showed reduced survival in human blood, highlighting its role in immune evasion .

V. vulnificus Lipoprotein Processing

  • IlpA Dependency: IlpA, a membrane-bound lipoprotein, requires signal peptide cleavage for adhesion to intestinal epithelial cells .

  • CRP Regulation: The cAMP-CRP system modulates V. vulnificus virulence factors (e.g., hemolysin, metalloprotease), suggesting potential crosstalk with LspA .

Comparative Analysis with Other Bacterial LspA

SpeciesEssentialityRole in VirulenceGlobomycin Sensitivity
E. coliEssentialCell envelope integrityHigh
S. aureusNon-essentialBlood survival, immune evasionModerate
R. typhiEssentialIntracellular survivalHigh
V. vulnificus (inferred)Likely essentialMaturation of IlpA, PlpAUnknown

Data synthesized from .

Future Directions

  1. Structural Studies: Cryo-EM or X-ray crystallography to resolve V. vulnificus LspA architecture.

  2. Genetic Knockout Models: Assess LspA’s role in virulence using lspA deletion mutants.

  3. Therapeutic Targeting: Screen LspA inhibitors (e.g., globomycin analogs) to disrupt lipoprotein processing .

Product Specs

Form
Lyophilized powder.
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order notes for customized fulfillment.
Lead Time
Delivery times vary depending on the purchasing method and location. Consult your local distributor for precise delivery estimates.
Note: All proteins are shipped with standard blue ice packs unless dry ice is specifically requested. Advance notification and additional fees apply for dry ice shipments.
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. We recommend adding 5-50% glycerol (final concentration) and aliquoting for long-term storage at -20°C/-80°C. Our standard glycerol concentration is 50% and can serve as a guideline.
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 forms maintain stability for 12 months at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquoting is essential for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
The tag type is determined during the manufacturing process.
Note: Tag type is determined during production. If a specific tag is required, please specify this in your order; we will prioritize its development.
Synonyms
lspA; VV0688; Lipoprotein signal peptidase; Prolipoprotein signal peptidase; Signal peptidase II; SPase II
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-168
Protein Length
full length protein
Species
Vibrio vulnificus (strain YJ016)
Target Names
lspA
Target Protein Sequence
MSGKAQIFKQTGVRWLWLALVVFLADIGIKFIVMENMGYGWANRIEVLPFFNLLYVHNYG AAFSFLSDQAGWQRWLFTGIAFVVTGLLTYWMSKLPAAEKWNNVAYAMIIGGAIGNVFDR MVHGFVVDYLDFYWGTYHWPAFNLADTAICLGAAMIILDGFRKKEEEK
Uniprot No.
Q7MNM7

Target Background

Function
This protein is a specific catalyst in the removal of signal peptides from prolipoproteins.
Database Links

KEGG: vvy:VV0688

Protein Families
Peptidase A8 family
Subcellular Location
Cell inner membrane; Multi-pass membrane protein.

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