We used a PCR‐based method to generate a single base pair mutation in the proB gene of Streptococcus thermophilus, which replaced an aspartic acid with a glycine residue at position 192 of the first proline biosynthetic enzyme γ‐glutamyl kinase. This was the first identified mutation in amino acid biosynthesis in S. thermophilus to our knowledge. The mutation caused an enhanced, feedback‐resistant γ‐glutamyl kinase activity and conferred an analogue‐resistant phenotype to an Escherichia coli transformant containing the mutated gene.
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