Difference between revisions of "Part:BBa K4621004"

Latest revision as of 18:06, 5 October 2023

A whole functional sequence in front of the monooxygenase gene, including its promoter.

Usage and Biology

LPMO hypothetical promoter is a whole functional sequence in front of the Lytic polysaccharide monooxygenase LPMO gene, which was cloned from SCUT-3 gene. In our project, through transcriptome analysis, qPCR and other means, we found that the gene expression of monooxygenase LPMO was up-regulated when SCUT-3 was in chitin medium, and we speculated that its promoter Pro lpmo may have properties that can be induced by chitin.

Clonal electrophoresis images of the fragments are shown in Figure. 1, with lane 4 representing LPMO hypothetical promoter

Fig.1 Gel electrophoresis results

LPMO hypothetical promoter and genes encoding a series of enzymes responsible for 5-Hydroxyectoine synthesis (ectoperon) were overexpressed using an integrated plasmid in Streptomyces SCUT-3. The modified strain was inoculated in 40 mL LB medium and fermented at 40℃ and 220 rpm for 120h. The production of Ectoine and 5-Hydroxyectoin in the presence or without chitin (the first day of fermentation) was detected to evaluate the chitin inducability of the promoter. The results are shown in Figure. 2.

Fig.2 Fermentation of products with or without the addition of an inducer（chitin) (a).Ectoine (b).5-Hydroxyectoin(120h)

By adding chitin at different days after fermentation, we also determined that the optimal time for chitin induction of LPMO hypothetical promoter was the second day of fermentation, and the specific experimental data are shown in Figure 3:

Fig.3 Fermentation of the product of SCUT-3-EctP at different induction times

Sequence and Features