Recombinant Bacillus clausii Acetate kinase (ackA)

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Cat. No.

BT1162701

Source

Yeast

Synonyms

ackA; ABC2740Acetate kinase; EC 2.7.2.1; Acetokinase

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1162701

Source

E.coli

Synonyms

ackA; ABC2740Acetate kinase; EC 2.7.2.1; Acetokinase

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1162701

Source

E.coli

Synonyms

ackA; ABC2740Acetate kinase; EC 2.7.2.1; Acetokinase

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1162701

Source

Baculovirus

Synonyms

ackA; ABC2740Acetate kinase; EC 2.7.2.1; Acetokinase

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1162701

Source

Mammalian cell

Synonyms

ackA; ABC2740Acetate kinase; EC 2.7.2.1; Acetokinase

Purity

>85% (SDS-PAGE)

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Description

Molecular Characterization of ackA in Bacillus clausii

The ackA gene in B. clausii encodes acetate kinase, a key enzyme in the phosphotransacetylase-acetate kinase (Pta-AckA) pathway. Genomic sequencing of B. clausii strains (e.g., UBBC07, B106, and Enterogermina strains O/C, SIN, N/R, and T) reveals that ackA is conserved and shares >96% homology with B. subtilis ackA . Key features include:

Feature	Details
Gene Location	Chromosomal; linked to ccpA (catabolite control protein A) homologs
Protein Function	Catalyzes acetate production, ATP synthesis, and overflow metabolism
Transcriptional Start	90 bp upstream of the coding region in B. subtilis homologs

Regulatory Mechanisms

In B. subtilis, ackA expression is activated by CcpA and CodY, global regulators of carbon metabolism and nitrogen starvation, respectively . Similar regulatory elements are predicted in B. clausii due to:

CcpA-Binding Sites: Two amyO-like operator sequences upstream of ackA, essential for glucose-dependent activation .
CodY Interaction: CodY enhances ackA transcription in response to branched-chain amino acids (BCAAs), independent of GTP levels .

**Table 1: Comparative Features of AckA Across Bacillus Species**

Species	Regulators	Inducers	Role in Metabolism
B. clausii (predicted)	CcpA, CodY	Glucose, BCAAs	ATP synthesis, acetate overflow
B. subtilis	CcpA, CodY	Carbon/nitrogen excess	Pyruvate-to-acetate conversion

Biotechnological Applications

Recombinant AckA could enhance industrial processes due to its role in:

Probiotic Function: Acetate production supports gut barrier integrity by modulating tight junction proteins and mucin synthesis .
Metabolic Engineering: Optimizing acetate excretion in microbial cell factories for biofuel production .

Research Gaps and Future Directions

Expression Systems: No studies yet report B. clausii ackA expression in heterologous hosts (e.g., E. coli).
Enzyme Kinetics: Biochemical characterization of recombinant AckA is needed to quantify ATP/acetate yields.

Product Specs

Form

Lyophilized powder
Note: While we prioritize shipping the format currently in stock, we are happy to accommodate specific format requests. Please indicate your preference in the order notes and we will fulfill your requirement to the best of our ability.

Lead Time

Delivery time may vary depending on the purchasing method and location. We kindly request you to consult your local distributors for precise delivery timelines.
Note: Our standard shipping includes blue ice packs. If dry ice is required, please communicate this request in advance as additional fees may apply.

Notes

Repeated freezing and thawing is not recommended. For optimal usage, store working aliquots at 4°C for up to one week.

Reconstitution

Prior to opening, we recommend briefly centrifuging the vial to ensure the contents are settled at the bottom. Please reconstitute the protein in deionized sterile water to a concentration between 0.1-1.0 mg/mL. For long-term storage, we suggest adding 5-50% glycerol (final concentration) and aliquoting the solution at -20°C/-80°C. Our default glycerol concentration is 50% and can be used as a reference point.

Shelf Life

The shelf life of the protein is influenced by various factors such as storage conditions, buffer components, temperature, and the inherent stability of the protein itself.
Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C, while lyophilized form has a shelf life of 12 months at -20°C/-80°C.

Storage Condition

Upon receipt, store at -20°C/-80°C. Aliquoting is necessary for multiple uses. To avoid degradation, minimize repeated freeze-thaw cycles.

Tag Info

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The tag type is determined during the production process. If you have a specific tag type in mind, please inform us and we will prioritize its development.

Synonyms

ackA; ABC2740Acetate kinase; EC 2.7.2.1; Acetokinase

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

1-395

Protein Length

full length protein

Purity

>85% (SDS-PAGE)

Species

Bacillus clausii (strain KSM-K16)

Target Names

ackA

Target Protein Sequence

MAKIIAINAG SSSLKFQLLA MPEETVIAKG LVERIGLKKS IFTIEANGQQ HELTEDIPDH AAAVKLLLKD LTGRGIIASL EEIEGIGHRV VHGGEKFNDS VVITDEVLQG IEEVSELAPL HNPANITGIK AFKEVLPNVP AVAVFDTAFH QTMPESAFLY SLPYEYYEKY GIRKYGFHGT SHKYVTHRAA ELMETPIEEL RILSCHLGNG ASIAAVKGGK SIDTSMGFTP LAGVTMGTRS GNIDPALIPY IMEKTGQSVE EVVATLNKKS GLLGITGFSS DLRDIEAAAK EGDERAQVAL DVFASRIHKY VGSYAARMGG LDAIVFTAGI GENSATIRER VLKGLSFMGV DVDLEKNNVR GKEAFIEKAG APVKIIVIPT NEEVMIARDT VRLTS

Uniprot No.

Q5WED5

Target Background

Function

This enzyme catalyzes the formation of acetyl phosphate from acetate and ATP. It can also catalyze the reverse reaction.

Database Links

KEGG: bcl:ABC2740

STRING: 66692.ABC2740

Protein Families

Acetokinase family

Subcellular Location

Cytoplasm.

Q&A

**FAQs for Researchers on Recombinant Bacillus clausii Acetate Kinase (AckA)**

Advanced Research Questions

How to resolve contradictions in ackA activation mechanisms across Bacillus strains?
- Comparative genomics: Align ackA promoter regions of B. clausii, B. subtilis, and B. anthracis to identify conserved cre sites.
- Metabolite profiling: Quantify acetyl-CoA, ATP, and NADH levels in ackA mutants to assess metabolic bottlenecks (e.g., acetyl-CoA accumulation in B. anthracis ).
- Heterologous expression: Express B. clausii ackA in E. coli or B. subtilis mutants to isolate strain-specific regulatory factors .
What experimental designs optimize recombinant AckA activity in non-native hosts?
- Codon optimization: Adjust GC content and rare tRNA usage for expression in E. coli or Pichia pastoris.
- Inducible promoters: Use IPTG- or xylose-inducible systems to avoid toxicity from constitutive expression .
- Activity validation: Pair enzyme assays with in vivo complementation (e.g., restore growth in ackA mutants ).
Parameter Optimization Strategy
Soluble expression Use low-temperature induction (18–22°C) with chaperone co-expression
Protein stability Add 5% glycerol or 0.5 M NaCl to lysis buffers
How does the Pta-AckA pathway interact with other carbon excretion pathways?
- Flux balance analysis (FBA): Model carbon distribution between acetate, acetoin, and TCA cycles .
- Double mutants: Inactivate pta and ackA simultaneously to study compensatory pathways (e.g., Ptb-mediated butyrate synthesis in B. anthracis ).
- 13C metabolic flux analysis: Track glucose-derived carbon into acetate vs. polyhydroxybutyrate (PHB) .

Parameter	Optimization Strategy
Soluble expression	Use low-temperature induction (18–22°C) with chaperone co-expression
Protein stability	Add 5% glycerol or 0.5 M NaCl to lysis buffers

Methodological Challenges & Solutions

Why do ackA overexpression studies sometimes reduce bacterial fitness?
- Hypothesis: Toxicity from excessive acetate drainage or ATP depletion.
- Solutions:
  - Use dynamic regulation (e.g., pH- or growth phase-dependent promoters).
  - Co-express acs (acetyl-CoA synthetase) to recycle acetate .
How to validate ackA’s role in metabolic homeostasis?
- Multi-omics integration: Combine transcriptomics (RNA-seq), metabolomics (LC-MS), and fluxomics (13C labeling) .
- Energy charge measurement: Quantify ATP/ADP ratios in ackA mutants vs. wild-type strains .
Metric ackA Mutant vs. Wild-Type
ATP levels 40% reduction
NADH/NAD+ ratio 2.5-fold increase

Metric	ackA Mutant vs. Wild-Type
ATP levels	40% reduction
NADH/NAD+ ratio	2.5-fold increase

Data Contradiction Analysis

Conflicting reports on CcpA’s role as an activator/repressor of ackA:
- Root cause: Strain-specific cre site positioning or phosphorylation states of CcpA co-regulators (e.g., HPr-Ser46-P).
- Resolution:
  - Perform in vitro EMSA with phosphorylated CcpA .
  - Test ackA promoter variants with cre site deletions .
Discrepancies in acetate yield across fermentation conditions:
- Key variables: Oxygen availability, pH, and residual glucose concentrations.
- Troubleshooting:
  - Use continuous bioreactors with real-time metabolite monitoring.
  - Apply CRISPRi to titrate ackA expression levels .

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Recombinant Bacillus clausii Acetate kinase (ackA)

Description

Molecular Characterization of ackA in Bacillus clausii

Regulatory Mechanisms

**Table 1: Comparative Features of AckA Across Bacillus Species**

Biotechnological Applications

Research Gaps and Future Directions

Product Specs

Target Background

Q&A

**FAQs for Researchers on Recombinant Bacillus clausii Acetate Kinase (AckA)**

Advanced Research Questions

Methodological Challenges & Solutions

Data Contradiction Analysis

Quick Inquiry

Category

Service