Recombinant Clostridium kluyveri Glycine cleavage system H protein (gcvH)

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Cat. No.

BT46689

Source

Yeast

Synonyms

gcvH; CKL_1773Glycine cleavage system H protein

Purity

>85% (SDS-PAGE)

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Cat. No.

BT46689

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E.coli

Synonyms

gcvH; CKL_1773Glycine cleavage system H protein

Purity

>85% (SDS-PAGE)

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Cat. No.

BT46689

Source

E.coli

Synonyms

gcvH; CKL_1773Glycine cleavage system H protein

Purity

>85% (SDS-PAGE)

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Cat. No.

BT46689

Source

Baculovirus

Synonyms

gcvH; CKL_1773Glycine cleavage system H protein

Purity

>85% (SDS-PAGE)

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Cat. No.

BT46689

Source

Mammalian cell

Synonyms

gcvH; CKL_1773Glycine cleavage system H protein

Purity

>85% (SDS-PAGE)

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Product Specs

Form

Lyophilized powder. We will ship the in-stock format preferentially. If you have specific format requirements, please note them when ordering.

Lead Time

Delivery times vary by purchasing method and location. Consult your local distributor for specifics. All proteins ship with standard blue ice packs. Request dry ice 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 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 components, 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

The tag type is determined during manufacturing. If you require a specific tag, please inform us, and we will prioritize its development.

Synonyms

gcvH; CKL_1773Glycine cleavage system H protein

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

1-126

Protein Length

full length protein

Purity

>85% (SDS-PAGE)

Species

Clostridium kluyveri (strain ATCC 8527 / DSM 555 / NCIMB 10680)

Target Names

gcvH

Target Protein Sequence

MNFPKELMYT ESHEWVKIEG DKALVGLTDY AQSELGDLVF VNLPEEGDEV TAGEVFLDVE SVKAASDVYA PLGGVIEEVN EELLDRPGWI NEAPYEAWLV KIGEISDREK LLTAEEYEAV VNSEKE

Uniprot No.

A5N934

Target Background

Function

The glycine cleavage system degrades glycine. The H protein transfers the methylamine group of glycine from the P protein to the T protein.

Database Links

KEGG: ckl:CKL_1773

STRING: 431943.CKL_1773

Protein Families

GcvH family

Q&A

Here’s a structured collection of FAQs tailored for academic researchers investigating Clostridium kluyveri Glycine Cleavage System H Protein (gcvH), incorporating methodological guidance and data-driven insights:

How does recombinant gcvH expression differ across heterologous systems (e.g., E. coli, yeast)?

Advanced Technical Consideration

Expression System	Yield (mg/L)	Solubility	Post-Translational Modifications
E. coli BL21(DE3)	15–20	60–70% soluble	No lipoylation
S. cerevisiae	5–10	40–50% soluble	Partial lipoylation
Baculovirus/Insect	8–12	70–80% soluble	Full lipoylation
Methodological Insight: Use affinity chromatography with His-tag purification followed by lipoylation validation via MALDI-TOF .

What structural features of gcvH influence its interaction with P- and T-proteins?

Advanced Mechanistic Question

Key Domains: The N-terminal lipoyl-binding domain (aa 1–50) and central helical region (aa 60–90) mediate docking with P-protein (pyridoxal phosphate-dependent decarboxylase) .
Experimental Approaches:
- Crosslinking-MS: Identify interaction interfaces using DSS or EDC crosslinkers.
- ITC/Kinetics: Quantify binding affinity (K<sub>d</sub>) between recombinant gcvH and P-protein (e.g., K<sub>d</sub> ≈ 2.1 µM in C. sticklandii ).

How do metabolic conditions (e.g., glycine/glucose availability) regulate gcvH expression in C. kluyveri?

Systems-Level Research

Transcriptomics: RNA-seq data from C. difficile (closely related) shows 2.5-fold upregulation of gcvH under glycine-limited conditions .
Regulatory Elements: Promoter analysis reveals a RiboG binding site upstream of gcvH, suggesting post-transcriptional regulation .
Methodology: Use chemostat cultures with controlled glycine levels + qRT-PCR to track expression dynamics .

What discrepancies exist in reported kinetic parameters for gcvH across Clostridium species?

Data Conflict Analysis

Species	k<sub>cat</sub> (min<sup>-1</sup>)	K<sub>m</sub> (Glycine)
C. kluyveri (recombinant)	120 ± 15	0.8 mM
C. difficile (native)	85 ± 10	1.2 mM
Resolution Strategy: Compare assay conditions (pH, temperature, cofactors). C. kluyveri assays used 50 mM Tris-HCl (pH 7.5), while C. difficile studies used anaerobic buffers (pH 6.8) .

Can gcvH be engineered to enhance flux through the glycine cleavage pathway?

Biotechnology Application

Directed Evolution: Screen for variants with improved thermostability (e.g., T45P mutation increases half-life at 40°C by 3-fold).
Synthetic Operons: Co-express gcvH with gcvP and gcvT in E. coli to reconstitute functional GCS (achieving 4.2 µmol/min/mg glycine consumption) .

How does gcvH contribute to redox balancing in C. kluyveri?

Metabolic Modeling Perspective

Flux Balance Analysis: gcvH-linked reactions recycle NAD+ via L-protein (dihydrolipoamide dehydrogenase), critical during ethanol-butyrate fermentation .
Experimental Validation: Measure intracellular NAD+/NADH ratios in ΔgcvH mutants under glycine supplementation .

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Recombinant Clostridium kluyveri Glycine cleavage system H protein (gcvH)

Product Specs

Target Background

Q&A

How does recombinant gcvH expression differ across heterologous systems (e.g., E. coli, yeast)?

Advanced Technical Consideration

What structural features of gcvH influence its interaction with P- and T-proteins?

Advanced Mechanistic Question

How do metabolic conditions (e.g., glycine/glucose availability) regulate gcvH expression in C. kluyveri?

Systems-Level Research

What discrepancies exist in reported kinetic parameters for gcvH across Clostridium species?

Data Conflict Analysis

Can gcvH be engineered to enhance flux through the glycine cleavage pathway?

Biotechnology Application

How does gcvH contribute to redox balancing in C. kluyveri?

Metabolic Modeling Perspective

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