Recombinant Methanocaldococcus jannaschii Isopropylmalate/citramalate isomerase large subunit (leuC)

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Cat. No.
BT43158
Source
Yeast
Synonyms
leuC; MJ0499Isopropylmalate/citramalate isomerase large subunit; EC 4.2.1.33; EC 4.2.1.35,; R)-2-methylmalate dehydratase,; R)-citramalate dehydratase; 3-isopropylmalate dehydratase; Alpha-isopropylmalate dehydratase; Citraconate hydratase; Isopropylmalate isomerase; IPMI; Maleate hydratase; Malease; EC 4.2.1.31
Purity
>85% (SDS-PAGE)
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Cat. No.
BT43158
Source
E.coli
Synonyms
leuC; MJ0499Isopropylmalate/citramalate isomerase large subunit; EC 4.2.1.33; EC 4.2.1.35,; R)-2-methylmalate dehydratase,; R)-citramalate dehydratase; 3-isopropylmalate dehydratase; Alpha-isopropylmalate dehydratase; Citraconate hydratase; Isopropylmalate isomerase; IPMI; Maleate hydratase; Malease; EC 4.2.1.31
Purity
>85% (SDS-PAGE)
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Cat. No.
BT43158
Source
E.coli
Synonyms
leuC; MJ0499Isopropylmalate/citramalate isomerase large subunit; EC 4.2.1.33; EC 4.2.1.35,; R)-2-methylmalate dehydratase,; R)-citramalate dehydratase; 3-isopropylmalate dehydratase; Alpha-isopropylmalate dehydratase; Citraconate hydratase; Isopropylmalate isomerase; IPMI; Maleate hydratase; Malease; EC 4.2.1.31
Purity
>85% (SDS-PAGE)
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Cat. No.
BT43158
Source
Baculovirus
Synonyms
leuC; MJ0499Isopropylmalate/citramalate isomerase large subunit; EC 4.2.1.33; EC 4.2.1.35,; R)-2-methylmalate dehydratase,; R)-citramalate dehydratase; 3-isopropylmalate dehydratase; Alpha-isopropylmalate dehydratase; Citraconate hydratase; Isopropylmalate isomerase; IPMI; Maleate hydratase; Malease; EC 4.2.1.31
Purity
>85% (SDS-PAGE)
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Cat. No.
BT43158
Source
Mammalian cell
Synonyms
leuC; MJ0499Isopropylmalate/citramalate isomerase large subunit; EC 4.2.1.33; EC 4.2.1.35,; R)-2-methylmalate dehydratase,; R)-citramalate dehydratase; 3-isopropylmalate dehydratase; Alpha-isopropylmalate dehydratase; Citraconate hydratase; Isopropylmalate isomerase; IPMI; Maleate hydratase; Malease; EC 4.2.1.31
Purity
>85% (SDS-PAGE)
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Description

Introduction to Recombinant Methanocaldococcus jannaschii Isopropylmalate/Citramalate Isomerase Large Subunit (leuC)

The leuC gene encodes the large subunit of a bifunctional enzyme, 3-isopropylmalate dehydratase, critical for isoleucine and leucine biosynthesis in Methanocaldococcus jannaschii. This heterodimeric enzyme (leuC + leuD) catalyzes reversible dehydration/hydration reactions in branched-chain amino acid pathways. Recombinant leuC is engineered for heterologous expression in microbial systems, enabling studies of its catalytic mechanisms and metabolic engineering applications .

Enzyme Architecture

FeatureDescription
Subunit CompositionHeterodimer: leuC (large subunit) + leuD (small subunit)
Catalytic DomainsBroad-spectrum dehydration/hydration activity for hydroxyacids
Key InteractionsForms complexes with leuD via conserved motifs for substrate binding

Catalytic Reactions

SubstrateProductPathway Role
2-Isopropylmalate3-Isopropylmalate (via 2-isopropylmaleate)Leucine biosynthesis
2-Methylmalate3-Methylmalate (via citraconate)Isoleucine biosynthesis
Maleate(R)-MalateMalease activity (side reaction)

Core Metabolic Functions

  • Isoleucine Biosynthesis: Converts 2-methylmalate to 3-methylmalate (citraconate intermediate), a precursor for isoleucine .

  • Leucine Biosynthesis: Isomerizes 2-isopropylmalate to 3-isopropylmalate, a key step in leucine production .

  • Redox Flexibility: Functions under both aerobic and anaerobic conditions, critical for archaeal metabolism .

Industrial Applications in Metabolic Engineering

ApplicationImpactSource
Citramalate ProductionE. coli strains lacking leuC show elevated citramalate yields (up to 46.5 g/L) due to reduced acetate formation .
Amino Acid Flux ControlDisruption of leuC redirects metabolic flux toward citramalate, a precursor for methacrylic acid polymers .

Pathway Interactions

The leuC-leuD complex interacts with enzymes in:

  1. Glyoxylate Bypass: Competes with malate synthase (aceB) for acetyl-CoA utilization .

  2. Acetate Metabolism: Knockouts reduce acetate yield, enhancing carbon flux toward target metabolites .

Recombinant Engineering Challenges

  • Thermal Stability: Native M. jannaschii enzymes require high-temperature conditions (optimal activity at 80°C), complicating heterologous expression .

  • pH Dependence: Activity peaks at alkaline pH (8.5–9.0), necessitating buffer optimization in E. coli .

Future Directions and Unresolved Questions

  1. Catalytic Mechanism: Detailed kinetic studies of substrate specificity (e.g., maleate vs. citraconate) are lacking .

  2. Structural Biology: Crystallographic data for the leuC-leuD complex remain unpublished, limiting rational engineering .

  3. Industrial Scalability: Fed-batch fermentation strategies for leuC-engineered strains require optimization to achieve >60 g/L citramalate yields .

Product Specs

Form
Lyophilized powder. We will preferentially ship the format we have in stock. If you have special format requirements, please note them when ordering.
Lead Time
Delivery times vary by purchase method and location. Consult your local distributor for specific delivery times. All proteins are shipped with normal blue ice packs by default. For dry ice shipment, contact us in advance (extra fees apply).
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Briefly centrifuge the vial before opening. Reconstitute protein in sterile deionized water to 0.1-1.0 mg/mL. Add 5-50% glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final glycerol concentration is 50%.
Shelf Life
Shelf life depends on storage conditions, buffer ingredients, storage temperature, and protein stability. Liquid form: 6 months at -20°C/-80°C. Lyophilized form: 12 months at -20°C/-80°C.
Storage Condition
Store at -20°C/-80°C upon receipt. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing. If you have a specific tag type requirement, please inform us, and we will prioritize developing it.
Synonyms
leuC; MJ0499Isopropylmalate/citramalate isomerase large subunit; EC 4.2.1.33; EC 4.2.1.35,; R)-2-methylmalate dehydratase,; R)-citramalate dehydratase; 3-isopropylmalate dehydratase; Alpha-isopropylmalate dehydratase; Citraconate hydratase; Isopropylmalate isomerase; IPMI; Maleate hydratase; Malease; EC 4.2.1.31
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-424
Protein Length
full length protein
Purity
>85% (SDS-PAGE)
Species
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440) (Methanococcus jannaschii)
Target Names
leuC
Target Protein Sequence
MGMTIVEKIL AKASGKKEVS PGDIVMANID VAMVHDITGP LTVNTLKEYG IEKVWNPEKI VILFDHQVPA DSIKAAENHI LMRKFVKEQG IKYFYDIREG VCHQVLPEKG HVAPGEVVVG ADSHTCTHGA FGAFATGIGS TDMAHVFATG KLWFKVPETI YFNITGDLQP YVTSKDVILS IIGEVGVDGA TYKACQFGGE TVKKMSIASR MTMTNMAIEM GGKTGIIEPD EKTIQYVKEA MKKHGTERPF EVIKGDEDAE FAEVYEIEAD KIEPVFACPH NVDNVKQARE VAGKPIDQVF IGSCTNGRLE DLRMAIKIIE KHGGIADDVR VVVTPASREE YLKALKEGII EKFLKYGCVV TNPSCSACMG SLYGVLGPGE VCVSTSNRNF RGRQGSLEAE IYLASPITAA ACAVKGELVD PRDL
Uniprot No.
P81291

Target Background

Function
This enzyme has broad specificity and catalyzes reversible hydroxyacid isomerizations through dehydration/hydration reactions. It isomerizes 2-isopropylmalate and 3-isopropylmalate (via 2-isopropylmaleate) in leucine biosynthesis, and 2-methylmalate and 3-methylmalate (via 2-methylmaleate/citraconate) in isoleucine biosynthesis. It also exhibits malease activity, hydrating maleate to (R)-malate.
Gene References Into Functions
1. The first structures of oxidized (ox-MJ0499) and reduced (red-MJ0499) forms of the large subunit of IPM isomerase from M. jannaschii are reported at 1.8 Å and 2.7 Å resolution, respectively (PMID: 24699638).
Database Links

KEGG: mja:MJ_0499

STRING: 243232.MJ_0499

Protein Families
Aconitase/IPM isomerase family, LeuC type 2 subfamily

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