| Summary: | <i>Corynebacterium glutamicum</i> is regarded as an industrially important microbial cell factory and is widely used to produce various value-added chemicals. Because of the importance of <i>C</i>. <i>glutamicum</i> applications, current research is increasingly focusing on developing <i>C. glutamicum</i> synthetic biology platforms. Because of its ability to condense with adipic acid to synthesize the industrial plastic nylon-46, putrescine is an important platform compound of industrial interest. Developing a high-throughput putrescine biosensor can aid in accelerating the design–build–test cycle of cell factories (production strains) to achieve high putrescine-generating strain production in <i>C. glutamicum</i>. This study developed a putrescine-specific biosensor (pSenPuuR) in <i>C</i>. <i>glutamicum</i> using <i>Escherichia coli</i>-derived transcriptional factor PuuR. The response characteristics of the biosensor to putrescine were further improved by optimizing the genetic components of pSenPuuR, such as the response promoter, reporter protein, and promoter for controlling PuuR expression. According to the findings of the study, pSenPuuR has the potential to be used to assess putrescine production in <i>C. glutamicum</i> and is suitable for high-throughput genetic variant screening.
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