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. Author manuscript; available in PMC: 2017 Apr 15.
Published in final edited form as: ACS Chem Biol. 2016 Feb 10;11(4):1137–1147. doi: 10.1021/acschembio.5b00913

Figure 3.

Figure 3

(A) Possible substrates tested for BexE activity. (B) HPLC analysis of reactions using the Streptomyces galilaeus (ATCC 31615) lysate with BexE and BexF. The main products in the lysate are aklaviketone and dehydro-aklaviketone, shown at 16.20 and 24.61 min, respectively. The addition of BexE to the Streptomyces galilaeus (ATCC 31615) lysate results in the appearance of two new peaks at 19.11 and 24.98 min. (C) The fractionation of the Streptomyces galilaeus (ATCC 31615) lysate to identify the BexE substrate. HPLC was used to fractionate areas with increased peak sizes in the lysate after 5 h of incubation to identify the BexE substrate (shown in dotted boxes). Fraction 2 contained a peak at 10.49 min, which contains the true BexE substrate. Fraction 2 was assayed with BexE and BexF in the presence of NADPH or NADH. Assays with the purified fraction 2 confirmed that the BexE reaction is NADPH dependent.