Ligation kit

EnzyTag has commercialized the peptiligase technology as an easy-to-use ligation kit. The kit will allow you to use our peptiligase technology at your facility, and thereby give you the freedom to choose the optimal ligation site(s) for your project as a result of the broad substrate specificity of the enzyme. Typical ligation reactions are done at room temperature in an aqueous buffered solution (pH 8.3) and are completed in 1 hour.

The kit provides 100 µg of Omniligase-1, a peptiligase variant that was designed for as broad specificity as possible, and 5 mg of standard ester and standard amine as a positive control for the ligation reactions. The kit will typically allow multiple small scale test ligations (100 µL) or ligations up to 5 mL at standard ligation conditions

Price standard kit

100 µg of omniligase (powder)
5 mg standard ester (Ac-DFSKL-Ocam-L) (powder)
5 mg standard amine (ALKKF)(powder)

 €850 excl. VAT, excl. shipping costs (€50 in EU, €150 to US)
Please contact us if larger quantities are desired.

Standard ligation conditions

200 mM Tricine/Phosphate buffer, pH 8.3
1 mg/ml TCEP
3 mM amine
2 mM ester
0.02 mg/ml Omniligase-1

As an additional service, EnzyTag will provide recommendations on the selection of the optimal ligation site for your project if the sequence of the peptide/protein can be shared (free of charge). If you do not want to share your sequence, a sheet will be provided which amino-acids are favoured in the ester and amine fragment for optimal ligations.

Since the peptiligase technology is patented,  a material transfer agreement (MTA) is required.  The MTA form will be sent via an email when you filled in the request form.

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References

Toplak et al. – 2016 – Peptiligase, an organic cosolvent stable and cation-independent peptide ligase for efficient chemoenzymatic peptide synthesis and cyclization in water (DOI: 10.1002/adsc.201600017)

Schmidt et al. – 2017 – Enzyme-mediated ligation technologies for peptides and proteins (DOI: 10.1016/j.cbpa.2017.01.017)

Schmidt et al. – 2017 – Omniligase-1 a powerful tool for peptide head-to-tail cyclisation (DOI: 10.1002/adsc.201700314)

Schmidt et al. – 2018 – Design of a substrate-tailored peptiligase variant for the efficient synthesis of thymosin-α 1 (DOI: 10.1039/c7ob02812a)

Pawlas, Nuijens et al. – 2019 – Sustainable, cost-efficient manufacturing of therapeutic peptides using chemo-enzymatic peptide synthesis (CEPS) (DOI: 10.1039/c9gc03600h).

Toplak et al. – 2021 -From Thiol-Subtilisin to Omniligase: Design and Structure of a Broadly Applicable Peptide Ligase (DOI:10.1016/j.csbj.202102002)

Please feel free to contact us

EnzyTag is open to further discussions on technology licensing for commercial purposes.

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