1 / 7

Introduction: Polyketide and Nonribosomal Peptide Natural Products

Introduction: Polyketide and Nonribosomal Peptide Natural Products. Doxorubicin -Antitumor agent. Erythromycin -Antibiotic. Cyclosporin -Immunosuppressant. Enniatin -Antibiotic. Yersiniabactin -Siderophore. Epothilone -Antitumor agent. OH. -Gene Expression.

dakota
Download Presentation

Introduction: Polyketide and Nonribosomal Peptide Natural Products

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Introduction: Polyketide and Nonribosomal Peptide Natural Products Doxorubicin -Antitumor agent Erythromycin -Antibiotic Cyclosporin -Immunosuppressant Enniatin -Antibiotic Yersiniabactin -Siderophore Epothilone -Antitumor agent OH

  2. -Gene Expression -Enzyme Activity (Temperature Dependent) KDa 212 158 97.2 66.4 DEBS1+TE DEBS2 DEBS3 DEBS1+TE DEBS2 DEBS3 Standard Gene Expression and Active Enzyme Requirement DEBS1+TE, DEBS2, or DEBS3 BL21(DE3) E. coli strain sfp SDS-PAGE -T7 promoter -lac operator In vitro reaction Analytical TLC

  3. AT ACP KS AT KR ACP KS AT KR ACP TE O H O O Standard Complete Negative Culture Control In vivo Polyketide Production -DEBS1+TE-Triketide Lactone Example DEBS1+TE DEBS1+TE BAP1 PCC PrpE PCC Propionyl-CoA (2S)-Methylmalonyl-CoA Triketide Lactone 14C-Propionate Induce/Feed 37°C 22°C

  4. In vivo Polyketide Production -DEBS-6dEB Example PCC DEBS1 BAP1 Full DEBS DEBS2 DEBS3 PrpE PCC Propionyl-CoA (2S)-Methylmalonyl-CoA Propionate 6dEB

  5. S S O O H H O O S S N N S S OH O O H H O O S S N N O O H H S S N N S S S S N N S S N N S S O O N N S S O O H H O O S S N N H H O O S H - O H N C O O H O H O 2 YbtE HMWP1 BAPint - C O O + H N 3 HMWP2 YbtU C H 2 C H 2 + S e A d e n o s i n C H 3 Salicylate Extending Production Capabilities -Using the yersiniabactin synthetase (YBTS) system HMWP2 (230 kDa) HMWP1 (350 kDa) NRPS NRPS PKS YbtE (60 kDa) NRPS YbtU (41 kDa) Salicylate Cysteine Yersiniabactin, Ybt S-adenosylmethionine Malonyl-CoA

  6. YbtE YbtU HMWP1 22C HMWP2 Extending Production Capabilities -Gene Expression -LC-MS Analysis Fe3+

  7. Summary -Complex polyketide biosynthesis +E. coli polyketide production -Gene transfer -Gene expression -Post-translational modification and protein folding -Substrate requirement +First complex natural product generated through E. coli -Process development/improvement +Fed-batch bioreactor process +Production improved ~200 fold -New production capabilities +Both polyketide and nonribosomal peptide products

More Related