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Literature summary for 2.7.1.163 extracted from

  • Nakamura, A.; Takakura, Y.; Kobayashi, H.; Hoshino, T.
    In vivo directed evolution for thermostabilization of Escherichia coli hygromycin B phosphotransferase and the use of the gene as a selection marker in the host-vector system of Thermus thermophilus (2005), J. Biosci. Bioeng., 100, 158-163.
    View publication on PubMed
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Application

Application Comment Organism
molecular biology the mutant gene hph5 gene can be used as a selection marker in the host-vector system of Thermus thermophilus either on plasmid or by genome integration Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
gene hph, subcloning in strain JM109, expression of wild-type and mutant enzymes in Thermus thermophilus strain HB27, establishment of the host-vector system using the hpt gene as a selective marker Escherichia coli

Protein Variants

Protein Variants Comment Organism
A118V site-directed mutagenesis, the mutation is involved in conferring thermostability at 55°C to the mutant enzyme Escherichia coli
D20G site-directed mutagenesis, the mutation is involved in conferring thermostability at 55°C to the mutant enzyme Escherichia coli
D20G/A118V/S225P/Q226L/T246A site-directed mutagenesis, mutant gene hpt5, mutant enzyme is stable at up to 67°C in contrary to the wild-type enzyme Escherichia coli
Q226L site-directed mutagenesis, the mutation is involved in conferring thermostability at 55°C to the mutant enzyme Escherichia coli
S225P site-directed mutagenesis, the mutation is involved in conferring thermostability at 55°C to the mutant enzyme Escherichia coli
S52T site-directed mutagenesis, the mutation does not confer thermostability at 55°C to the mutant enzyme Escherichia coli
T246A site-directed mutagenesis, the mutation is involved in conferring thermostability at 55°C to the mutant enzyme Escherichia coli
W238C site-directed mutagenesis, the mutation does not confer thermostability at 55°C to the mutant enzyme Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
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-
-

Synonyms

Synonyms Comment Organism
hygromycin B phosphotransferase
-
 Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55
-
 inactivation of wild-type enzyme and mutants S52T and W238C Escherichia coli