Part:BBa_K5313021

ndm B Q289A

NdmB belongs to the RO enzyme family and cannot act alone, but only together with ndmD. NdmD and ndmB together convert theobromine to 7-MX.

The ndmB gene from Pseudomonas CBB5 encodes an enzyme in the N3-demethylase complex, which is involved in the breakdown of caffeine. This enzyme is responsible for removing methyl from one of the nitrogen atoms in caffeine, promoting a step-by-step demethylation process that eventually degrades caffeine completely into simpler, non-toxic compounds that bacteria can use to grow. ndmB is an important component of the caffeine metabolic pathway in Pseudomonas CBB5 and is an important tool for biodegradation and metabolic engineering projects that degrade methylxanthine (such as caffeine).

We obtained a series of ndmB mutants through directed evolution. Compared with normal ndmB, ndmB Q289T mutates to Thr at Ala position 289. The biggest difference between this mutant and the ndmB wild type is that ndmB Q289A can remove both N3-methyl and N1-methyl, which can directly work with ndmD to convert caffeine to 7-MX.

Figure .Different mutated strains were detected by HPLC

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]