Part:BBa_K2372002
Beta-lactam-synthetase wildtype
Background
β-lactam synthetase (BLS), found in Streptomyces clavuligerus, catalyzes the formation of the β-lactam ring of clavulanic acid in an ATP and Mg2+ dependent manner by intramolecular closure. Clavulanic acid with its closed β-lactam ring is a potent inhibitor of β-lactamase. In our project we were using BLS for activating our chemical synthesis by formation of a β-lactam ring in vitro. In order to simplify purification a thrombin cleavage side and 6xhis-tag was added at the end of the enzyme sequence.
| BLS-Wt | |
|---|---|
| Function | β-Lactamsynthetase |
| Use in | Prokaryotic cells |
| RFC standard | RFC 10 |
| Backbone | pSB1C3 |
| Organism | Streptomyces clavuligerus |
| Source | IDT (DNA synthesis) |
| Submitted by | Tuebingen 2017 |
References
Bachmann, B. O., Li, R., & Townsend, C. A. (1998). beta-Lactam synthetase: a new biosynthetic enzyme. Proc Natl Acad Sci U S A, 95(16), 9082-9086.
Miller, M. T., Bachmann, B. O., Townsend, C. A., & Rosenzweig, A. C. (2001). Structure of beta-lactam synthetase reveals how to synthesize antibiotics instead of asparagine. Nat Struct Biol, 8(8), 684-689. doi:10.1038/90394
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 9
Illegal AgeI site found at 199
Illegal AgeI site found at 379
Illegal AgeI site found at 544
Illegal AgeI site found at 1036 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 631
Illegal BsaI site found at 1224
| None |