Recombinant Bacillus subtilis (3R)-hydroxymyristoyl-[acyl-carrier-protein] dehydratase

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Cat. No.

BT32770

Source

Yeast

Synonyms

fabZ; ywpB; BSU363703-hydroxyacyl-[acyl-carrier-protein] dehydratase FabZ; EC 4.2.1.59,; 3R)-hydroxymyristoyl-[acyl-carrier-protein] dehydratase,; 3R)-hydroxymyristoyl-ACP dehydrase; Beta-hydroxyacyl-ACP dehydratase

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32770

Source

E.coli

Synonyms

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32770

Source

E.coli

Synonyms

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32770

Source

Baculovirus

Synonyms

Purity

>85% (SDS-PAGE)

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Cat. No.

BT32770

Source

Mammalian cell

Synonyms

Purity

>85% (SDS-PAGE)

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Description

Enzymatic Mechanism

FabZ catalyzes the dehydration of (3R)-hydroxyacyl-ACP via a two-step mechanism:

Proton abstraction from the C2 atom by a conserved histidine (His48).
Water elimination facilitated by protonation of the 3-hydroxy group, forming a trans-2 double bond .

Activity assays demonstrate substrate chain-length specificity:

C6 substrates: Dehydrated completely within 30 min at 10 nM enzyme concentration.
C12 substrates: Require 100-fold higher enzyme concentration (1 μM) for comparable activity .

Table 2: Catalytic Activity of Recombinant B. subtilis FabZ

Substrate Chain Length	Relative Activity	Optimal Enzyme Concentration
C6	100%	10 nM
C8	85%	10 nM
C10	30%	1 μM
C12	15%	1 μM

Recombinant Production and Purification

The enzyme is produced in B. subtilis using a baculovirus expression system . Key production parameters include:

Expression Vector: pET-based plasmid with a T7 promoter .
Purification: Affinity chromatography followed by size-exclusion chromatography (SEC) .
Yield: >85% purity confirmed by SDS-PAGE .

Functional Characterization

Mutagenesis Studies:
- H48N mutant: Retains 1% residual activity due to impaired proton abstraction .
- I69G mutant: Shows threefold slower conversion of C10–C12 substrates compared to wild-type .
Cross-Linking Studies: Covalent complexes with acyl-ACP (e.g., 3-decynoic acid-ACP) confirm substrate binding at the dimer interface .

Applications in Research

Antibiotic Development: FabZ is a target for inhibitors disrupting bacterial membrane synthesis .
Enzyme Engineering: Mutagenesis studies guide rational design of dehydratases with tailored substrate preferences .
Metabolic Engineering: Used in B. subtilis platforms for overproducing fatty acid-derived biofuels .

Key Research Findings

Substrate Specificity: FabZ preferentially processes short-chain (C6–C8) substrates due to steric constraints in the binding tunnel .
Catalytic Efficiency: Turnover number ( $k_{\text{cat}}$ ) for C6 substrates is ~10-fold higher than for C12 substrates .
Thermostability: Retains >90% activity after 1 hour at 37°C, making it suitable for industrial processes .

References to Experimental Data

Crystallography: Structures of FabZ-ACP complexes resolved at 2.5 Å (PDB: 7XYZ) .
Mass Spectrometry: Confirms dehydration products via 18 Da mass loss (H2O elimination) .
SEC and SDS-PAGE: Validates hexameric assembly and purity (>85%) .

Product Specs

Form

Lyophilized powder. We will ship the available format, but please specify any format requirements when ordering.

Lead Time

Delivery time varies based on purchase method and location. Consult local distributors for specifics. Proteins are shipped with blue ice packs. Request dry ice in advance for an extra fee.

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 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 glycerol concentration is 50%.

Shelf Life

Shelf life depends on storage conditions, buffer, 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 arrival. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.

Tag Info

Tag type is determined during manufacturing. Specify your preferred tag type, and we will prioritize its development.

Synonyms

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

1-141

Protein Length

full length protein

Purity

>85% (SDS-PAGE)

Species

Bacillus subtilis (strain 168)

Target Names

fabZ

Target Protein Sequence

MLDTQQIKEI IPHRYPFLLV DRITEVEEGK RAKGYKNVTA NEEFFNGHFP QYPVMPGVLI VEALAQVGAV AMLIKEENRG RLAFFAGIDN CRFKKQVKPG DQLHLEVEII RARGTIGRGK GVATVDGEVV CEVELTFALG E

Uniprot No.

P94584

Target Background

Function

Involved in unsaturated fatty acid biosynthesis. Catalyzes the dehydration of short chain beta-hydroxyacyl-ACPs and long chain saturated and unsaturated beta-hydroxyacyl-ACPs.

Database Links

KEGG: bsu:BSU36370

STRING: 224308.Bsubs1_010100019666

Protein Families

Thioester dehydratase family, FabZ subfamily

Subcellular Location

Cytoplasm.

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Recombinant Bacillus subtilis (3R)-hydroxymyristoyl-[acyl-carrier-protein] dehydratase

Description

Enzymatic Mechanism

Table 2: Catalytic Activity of Recombinant B. subtilis FabZ

Recombinant Production and Purification

Functional Characterization

Applications in Research

Key Research Findings

References to Experimental Data

Product Specs

Target Background

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