Protocatechuic Acid → Gallic Acid

Alternatively, the 3-dehydroshikimic acid was added slowly with the glucose feed. The glucose feed solution was prepared by mixing 100 mL of a solution containing 18 g of 3-dehydroshikimic acid, with a 200 mL solution containing 120 g of glucose. An oxygen sensor controlled dissolved oxygen levels via addition of the solution containing glucose (0.4 g/mL) and 3-dehydroshikimic acid (0.06 g/mL, 0.35 mmol/mL). The rate of addition of the previous feed solution was about 8 mL/h and as a result approximately 0.5 g of 3-dehydroshikimic acid was added to the culture medium every hour. By the end of the fermentation run, the total amount of 3-dehydroshikimic acid added to the growing cells was 18 g, producing 6.3 g/L of gallic acid at 48 h in a 45% (mol/mol) yield based on added 3-dehydroshikimic acid. Significant amounts of protocatechuic acid (1.8 g/L) as well as unreacted 3-dehydroshikimic acid (4.2 g/L) were present at the end of the run (FIG. 5). A 20 mL aliquot of fermentation broth was taken at 24 h, 36 h, and 48 h for determination of PHB hydroxylase activity. Although pobA* activities were stable throughout the fermentation, the majority of gallic acid was produced in stationary phase (after 24 h). Interestingly, the concentration of protocatechuic acid showed an initial increase reaching a maximum value of 3.4 g/L at 36 h, after which the concentration of protocatechuic acid steadily decreased to 1.8 g/L at 48 h (FIG. 5).

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2. custom48 h

ORD record

ord-a14991c7779b4f319fd3fe1704ca91e2

Source dataset

uspto-grants-2002_10

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US06472190B1