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Accession Number ADA581260
Title Manufacture of TATB and TNT from Biosynthesized Phloroglucinols.
Publication Date Jul 2010
Media Count 47p
Personal Author J. W. Frost
Abstract In route to the microbial synthesis of mono-O-methylphloroglucinols, phloroglucinol O-methyl transferase (POMT) from Rosa chinensis var. spontanea has been successfully de novo synthesized in codon-optimized form for expression in E. coli, which is the host currently used for microbial synthesis of phloroglucinol (PG) from glucose. The specific activity of heterologously- expressed, codon-optimized POMT is at 0.02 U/mg, which is 5-fold higher than the specific activity of POMT purified to homogeneity from Rosa chinensis var. spontanea petals. Similarly, orcinol O-methyl transferase (OOMT) protein sequence was identified from GenBank, codon optimized for E. coli expression using the DNA2.0 algorithm, and synthesized. Expressing this gene in E. coli gave a specific activity at 0.006 U/mg using crude lysate. Efforts had been made to create the first E. coli mono-O-methylphloroglucinol synthesizing construct. E. coli PG1/pKIT1.008 synthesized 0.7 g/L methoxyresorcinol in the medium under fed-batch fermentor-controlled conditions. A methionine supplementation strategy was developed to give a 40% increase in mono-O-methylphloroglucinol production under fed-batch fermentor-controlled conditions. It is believed that mono-O-methylphloroglucinol is toxic to E. coli cells and difficulties were experienced in maintaining a healthy culture in the fermentor. To circumvent this issue, resin-based extractive fermentation was carried out as an external loop throughout the run. By using a strong anion- exchange resin under optimized conditions, E. coli PG1/pKIT1.008 synthesized 2 g/L mono-O-methylphloroglucinol in 70 h after inoculation. Simultaneously, efforts had been made in both strain development and optimizing fermentation conditions for microbial phloroglucinol synthesis. Under optimized resin-based extractive fermentation, E. coli PG1/pKIT10.080 synthesized 25 g/L of phloroglucinol.
Keywords Biosynthesis
Microbial synthesis
Serdp collection
Serdp(Strategic environmental research and development progr

Source Agency Non Paid ADAS
NTIS Subject Category 57B - Biochemistry
79A - Ammunition, Explosives, & Pyrotechnics
Corporate Author Strategic Environmental Research and Development Program, Arlington, VA.
Document Type Technical report
Title Note Final rept.
NTIS Issue Number 1325
Contract Number N/A

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