TY - JOUR
T1 - Native and Engineered Clifednamide Biosynthesis in Multiple Streptomyces spp.
AU - Qi, Yunci
AU - Ding, Edward
AU - Blodgett, Joshua A.V.
N1 - Funding Information:
We thank Prof. Arpita Bose for helpful comments and discussions and John M. D’Alessandro for plasmids pJMD1, pJMD2, and pJMD3. We acknowledge former WUSTL BIOL3493 students Kevin Lou for isolation of strain Streptomyces sp. strain KL33 and Naveen Jain for designing attBΦC31 integration check primers. We are grateful to Dr. Bradley Evans of the Donald Danforth Plant Science Center for high-resolution mass spectrometry assistance, Prof. Jon Clardy (Harvard Medical School) for Streptomyces albus strain J1074, and Dr. Dylan Alexander (Cubist Pharmaceuticals) for providing the sequence of PermE*pDA1652. We also thank Tyson Research Center (Dr. Kim Medley, Director) for access to soil samples used in the isolation of KL33. This work was supported by Washington University in St Louis New Faculty Start Up funds to Joshua Blodgett.
Publisher Copyright:
© 2017 American Chemical Society.
PY - 2018/2/16
Y1 - 2018/2/16
N2 - Polycyclic tetramate macrolactam (PTM) natural products are produced by actinomycetes and other bacteria. PTMs are often bioactive, and the simplicity of their biosynthetic clusters make them attractive for bioengineering. Clifednamide-type PTMs from Streptomyces sp. strain JV178 contain a distinctive ketone group, suggesting the existence of a novel PTM oxidizing enzyme. Here, we report the new cytochrome P450 enzyme (CftA) is required for clifednamide production. Genome mining was used to identify several new clifednamide producers, some having improved clifednamide yields. Using a parallel synthetic biology approach, CftA isozymes were used to engineer the ikarugamycin pathway of Streptomyces sp. strain NRRL F-2890 to yield clifednamides. Further, we observed that strong CftA expression leads to the production of a new PTM, clifednamide C. We demonstrate the utility of both genome mining and synthetic biology to rapidly increase clifednamide production.
AB - Polycyclic tetramate macrolactam (PTM) natural products are produced by actinomycetes and other bacteria. PTMs are often bioactive, and the simplicity of their biosynthetic clusters make them attractive for bioengineering. Clifednamide-type PTMs from Streptomyces sp. strain JV178 contain a distinctive ketone group, suggesting the existence of a novel PTM oxidizing enzyme. Here, we report the new cytochrome P450 enzyme (CftA) is required for clifednamide production. Genome mining was used to identify several new clifednamide producers, some having improved clifednamide yields. Using a parallel synthetic biology approach, CftA isozymes were used to engineer the ikarugamycin pathway of Streptomyces sp. strain NRRL F-2890 to yield clifednamides. Further, we observed that strong CftA expression leads to the production of a new PTM, clifednamide C. We demonstrate the utility of both genome mining and synthetic biology to rapidly increase clifednamide production.
KW - cytochrome P450
KW - genome mining
KW - metabolic engineering
KW - natural products
KW - polycyclic tetramate macrolactams
KW - Streptomyces
UR - http://www.scopus.com/inward/record.url?scp=85042183354&partnerID=8YFLogxK
U2 - 10.1021/acssynbio.7b00349
DO - 10.1021/acssynbio.7b00349
M3 - Article
C2 - 29249153
AN - SCOPUS:85042183354
SN - 2161-5063
VL - 7
SP - 357
EP - 362
JO - ACS synthetic biology
JF - ACS synthetic biology
IS - 2
ER -