Proteinases with improved properties
AssigneeNew England Biolabs, Inc.
InventorsMinyong Chen, James C. Samuelson, Ming-Qun Xu, Aihua Zhang, Margaret Heider, Pingfang Liu
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Abstract
Provided herein is a thermolabile proteinase and methods of using the same. In some embodiments, the thermolabile proteinase may comprise an amino acid sequence that is at least 90% identical to any of SEQ ID NOs:1-11 and at least one amino acid substitution in helix 3. The thermolabile proteinase is active at a temperature in the range of 4° C.-40° C. and is inactivated by raising the temperature to above 50° C., where the proteinase is substantially inactive at 65° C.
Claims24
A protein having an amino acid sequence that has a least 90% identity to the amino acid sequence of SEQ ID NO:1, and wherein the protein comprises at least one amino acid substitution in a helix 3 region corresponding to position 230-237.
A method comprising:
- (a) incubating a reaction mixture comprising a proteinase, a first enzyme, and a nucleic acid substrate for the first enzyme, for a period of time such that the proteinase proteolytically cleaves the first enzyme, wherein the proteinase comprises:
- (i) an amino acid sequence that is at least 90% identical to SEQ ID NO:1;
- and
- (ii) at least one amino acid substitution in a region of helix 3 corresponding to amino acid positions 230-237 of SEQ ID NO:1;
- and
- (b) increasing the temperature of the reaction mixture to inactivate the proteinase.
Dependencies are inferred from the claim text.