Thermolabile Serratia Marcescens nuclease
AssigneeAbClonal Science, Inc.
InventorsZhenyu Zhu, Dapeng Sun, Michaela Shottes
Life of this patent
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Abstract
Disclosed is an engineered thermolabile mutant of Serratia Marcescens Nuclease. In the same buffer as used for optimal enzyme activity, after 60° C. for 20 min, while the wild type retained 12.5% activity, the thermolabile mutant R146D/D156R/D229R/D245R (SEQ ID NO: 12, without the first 21 amino acids, which are a signal peptide) retained only 0.39% activity. Heat inactivation of the mutant, when it is used for protein purification, can be used after a period when substantial DNA in a protein has been degraded, but before unwanted degradation takes place.
Claims5
A thermolabile Serratia Marcescens Nuclease mutant, comprising:
- the amino acid sequence of SEQ ID NO:
- 12 not including the first 21 amino acids from the N-terminus.
A DNA sequence encoding the amino acid sequence of SEQ ID NO:
- 12 not including the first 21 amino acids from the N-terminus.
The DNA sequence of SEQ ID NO:
- 11.
A method of removing nucleic acids from a solution following protein production in said solution, comprising:
- adding to the solution, after protein production, a thermolabile Serratia Marcescens Nuclease mutant having the amino acid sequence of SEQ ID NO:
- 12 not including the first 21 amino acids from the N-terminus, under conditions and times to permit the thermolabile Serratia Marcescens Nuclease mutant to degrade nucleic acid in the solution;
- and
- terminating the Serratia Marcescens Nuclease mutant activity after a period sufficient to degrade nucleic acid in the solution, by heating the solution.
Dependencies are inferred from the claim text.