Genenase I

  • This product was discontinued on 01/01/2013
recombinant
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Description

Genenase™ I is a variant of subtilisin BPN´ that has been engineered to have increased specificity by substituting amino acids in its active site (1,2). When designing fusion proteins for cleavage with Genenase I, we recommend the site Pro-Gly-Ala-Ala-His-Tyr. Genenase I will cleave at other histidine residues depending on the surrounding amino acids and the 3-dimensional conformation of the protein. Genenase I cleaves His-Tyr-Glu and His-Tyr-Asp slowly, but will not cleave His-Tyr-Pro or His-Tyr-Ile (2).

Product Source

Isolated from a Bacillus subtilis strain carrying the protease gene derived from Bacillus amyloliquefaciens.

Specificity

Genenase I cleaves His-Tyr-Glu and His-Tyr-Asp slowly, but will not cleave His-Tyr-Pro or His-Tyr-Ile (2).

Properties and Usage

Unit Definition

0.5 μg of Genenase I will cleave 50 μg of test substrate to 95% completion in a total reaction volume of 50 μl in 8 hours or less at 23°C in 20 mM Tris-HCl (pH 8.0 @ 25°C) with 200 mM NaCl.

Storage Temperature

-20°C

Molecular Weights

Apparent: 28 kDa
Theoretical: 27.4 kDa

Notes

  1. The rate of Genenase™ I cleavage has been shown to depend on the amino acid following the cleavage site (position P1')(3). Rates of cleavage when P1' is Asp or Glu are slowest, 40-fold slower than when P1' is Arg or Cys. The rate when P1'= Glu or Asp is enhanced 10- and 14-fold, respectively, by including 2 M KCl in the reaction.

References

  1. Carter, P. and Wells, J.A. Science. 237
  2. Carter, P. et al. Biochemistry. 30
  3. Carter, P. et al. Proteins: Structure, Function, and Genetics. 6

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

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Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.