Recombinant Pseudomonas aeruginosa L-ornithine 5-monooxygenase (pvdA)

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Code CSB-EP706553EZX
MSDS
Size US$388
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  • (Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.
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Product Details

Purity
Greater than 90% as determined by SDS-PAGE.
Target Names
pvdA
Uniprot No.
Research Area
Others
Alternative Names
pvdA; pvd-1; PA2386; L-ornithine N(5)-monooxygenase; EC 1.14.13.195; L-ornithine N(5)-hydroxylase; Ornithine hydroxylase; L-ornithine N(5)-oxygenase; Pyoverdin biosynthesis protein A
Species
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Source
E.coli
Expression Region
1-443aa
Target Protein Sequence
MTQATATAVVHDLIGVGFGPSNIALAIALQERAQAQGALEVLFLDKQGDYRWHGNTLVSQSELQISFLKDLVSLRNPTSPYSFVNYLHKHDRLVDFINLGTFYPCRMEFNDYLRWVASHFQEQSRYGEEVLRIEPMLSAGQVEALRVISRNADGEELVRTTRALVVSPGGTPRIPQVFRALKGDGRVFHHSQYLEHMAKQPCSSGKPMKIAIIGGGQSAAEAFIDLNDSYPSVQADMILRASALKPADDSPFVNEVFAPKFTDLIYSREHAERERLLREYHNTNYSVVDTDLIERIYGVFYRQKVSGIPRHAFRCMTTVERATATAQGIELALRDAGSGELSVETYDAVILATGYERQLHRQLLEPLAEYLGDHEIGRDYRLQTDERCKVAIYAQGFSQASHGLSDTLLSVLPVRAEEISGSLYQHLKPGTAARALHEHALAS
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.
Mol. Weight
65.5kDa
Protein Length
Full Length
Tag Info
N-terminal 6xHis-SUMO-tagged
Form
Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.
Buffer
If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution
We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.
Troubleshooting and FAQs
Storage Condition
Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.
Shelf Life
The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Lead Time
3-7 business days
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Datasheet & COA
Please contact us to get it.
Description

Constructing a plasmid encoding the Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228) pvdA protein (1-443aa) initiates the general approach for generating the recombinant Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228) pvdA protein. Transformation of the plasmid into e.coli cells obtains the plasmid-containing e.coli cells, which are cultured and induced for protein expression. A N-terminal 6xHis-SUMO tag is fused to the protein. Subsequently, the protein is purified through affinity purification, and SDS-PAGE analysis is undertaken to confirm the presence and assess the purity of the protein. The protein's purity exceeds 90%.

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Target Background

Function
Catalyzes the conversion of L-ornithine to N(5)-hydroxyornithine, the first step in the biosynthesis of all hydroxamate-containing siderophores, such as pyoverdin. Pyoverdin is a hydroxamate siderophore composed of a 6,7-dihydroxyquinoline-containing fluorescent chromophore joined to the N-terminus of a partly cyclic octapeptide (D-Ser-L-Arg-D-Ser-L-N(5)-OH-Orn-L-Lys-L-N(5)-OH-Orn-L-Thr-L-Thr in strain PAO1). Specific for NADPH, which plays a role in stabilization of the C4a-hydroperoxyflavin intermediate.
Gene References into Functions
  1. PvdA is a membrane-associated protein located at the old cell pole PMID: 22498339
  2. The first two structures of a class B N-hydroxylating monooxygenase were determined with FAD in oxidized (1.9 A resolution) and reduced (3.03 A resolution) states PMID: 21757711
  3. overexpression and purification of active recombinant L-ornithine N5-hydroxylase, PvdA; detailed steady-state kinetic analysis to determine the substrate and coenzyme specificities PMID: 17015659
  4. The membrane topogenic determinants of PvdA were studies, and PvdA appears to have a single N-terminal domain anchored to the inner membrane. PMID: 18757814
  5. PvdA employs a kinetic mechanism that is a hybrid of those previously documented for other flavin-dependent hydroxylases PMID: 19368334

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Subcellular Location
Cell inner membrane; Peripheral membrane protein; Cytoplasmic side.
Protein Families
Lysine N(6)-hydroxylase/L-ornithine N(5)-oxygenase family
Database Links

KEGG: pae:PA2386

STRING: 208964.PA2386

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