TABLE 1.

Product yields from assays of permeabilized cells of m-xylene-grown Azoarcus sp. strain T^a

Substrate added				Product (nmol)b			Product labeling
Aromatic compound	Fumarate	Succinyl-CoA	Nitrate	3-MeBS	E-3-MePI	3-Methylbenzoate
m-Xylene-d6 (360 nmol, total)	+	−	−	190	NDc	0.3	Deuterium labeled
m-Xylene-d6 (360 nmol, total)	+	+	+	120	7	42	Deuterium labeled
3-MeBS (150 nmol, total)	−	+	+	52	8	15d

^aEach anoxic assay mixture (total volume, 1 ml) contained titanium(III) chloride (200 μM) as a reductant and permeabilized cells (ca. 3 mg of protein). Initial concentrations of other substrates added are given as follows: fumarate, 500 μM; succinyl-CoA, 300 μM; and nitrate, 2 mM. A total of 360 nmol of m-xylene is equivalent to a liquid concentration of 170 μM. Assay mixtures were incubated for 1 h. Reaction products were extracted, derivatized to methyl esters, and analyzed by GC/MS. 

^b3-MeBS and 3-methylbenzoate were identified by GC/MS by using authentic standards. E-3-MePI was tentatively identified based on mass spectral similarity to an authentic E-PI standard. Assays were conducted in duplicate. Product yields of duplicate assays were typically within 5 to 20% of each other. Product yields shown are averages of the duplicate assays, and are representative of other experiments conducted. 

^cND, not detected. The detection limit of E-3-MePI was less than 1 nmol. 

^dApproximately 0.5 of the 15 nmol of 3-methylbenzoate detected in this assay may have been due to carryover from growth of the cells on m-xylene.