Green Synthesis of Phloroglucinol: Exploiting Pseudomonas fluorescens and Scale-Up
Abstract
Through a combination of metabolic engineering and reaction engineering, a new generation of phloroglucinol synthesizing microbe has been developed and evaluated using resin-based extractive fermentation process. Phloroglucinol synthase, phlD, from P. fluorescens PF-5 had been cloned and expressed in E. Coli PG1/pBC2.274. The isolated gene's product has been unambiguously identified as phloroglucinol synthase using electrospray ionization tandem mass spectroscopy. The expression of phlD in E. Coli PG1/pBC2.274 under fermentor-control conditions generated inclusion bodies of inactive protein. By attenuating the expression level of phlD in E. coli, a more stable phloroglucinol production E. coli strain, PG1/pKIT10.080, has been developed and capable of producing 25 g/L of phloroglucinol under fermentor-controlled resin-based extractive cultivation conditions. The use of resin-based extractive technique facilitates the removal of toxic phloroglucinol product from the fermentation process as it is being formed allowing for maximum production of phloroglucinol in the E. coli host. In contrast, microbial synthesis of phloroglucinol by a wild-type P. fluorescens species under similar culturing condition produced only 0.1g/L phloroglucinol.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 14, 2009
- Accession Number
- ADA548825
Entities
People
- John W. Frost