Suggested that the HpaBC complicated has in vivo activity towards p-CA, and Af, with high catalytic efficiency. Our Our final results showed activity towards p-CA, N, N, and Af, with higher catalytic efficiency. outcomes showed that that these substrates (except anthocyanin) could be converted to the corresponding orthothose substrates (except anthocyanin) might be converted to the corresponding ortho-prodproducts the optimaloptimal circumstances. Similarly, the conversion of these by P2 3ucts below below the circumstances. Similarly, the conversion of those substrates substrates by P2 3-carrying also drastically higher than greater than that of P2-carrying strains. carrying strains was strains was also considerably that of P2-carrying strains. The converThe conversion to E was 58 , followed by that for Af by that three.14), p-CA (32.93 2.01), sion price from N price from N to E was 58 , followed (35.two for Af (35.2 3.14), p-CA (32.93 two.01), DHK (23.74 1.75) and K (23.84 0.88), though substrate PEL was not PEL DHK (23.74 1.75) and K (23.84 0.88), even though activity on the activity on the substratedewas not TrkA Purity & Documentation detected (Table 2). These results provided proof that the HpaBC complex had tected (Table 2). These final results provided evidence that the HpaBC complex had in vivo in vivo activity towards monohydroxylated flavanol and flavonol. activity towards monohydroxylated flavanol and flavonol.Molecules 2021, 26, 2919 Molecules 2021, 26, x FOR PEER REVIEW9 of 13 9 ofFigure 6. HPLC analysis of the enzymatic items of the HpaBC complicated, when feeding with diverse substrates. HPLC Figure 6. HPLC analysis in the enzymatic goods of the HpaBC complex, when feeding with diverse substrates. HPLC chromatogram chromatogram (left) and regular compound (correct) analyses of your enzymatic reaction. N, E,E, K, Q, DHK, DHQ, and Af typical compound (appropriate) analyses with the enzymatic reaction. N, K, Q, DHK, DHQ, C C and have been monitored at at nm, and p-CA and CA had been monitored at 340 340 The substrates and and corresponding solutions Af had been monitored280280 nm, and p-CA and CA have been monitored at nm. nm. The substrates corresponding goods had been were detected by HPLCLC-MS. The ortho-hydroxylation activities of (a): p-CAp-CA to (b): N to N to E; (c): Af to C;K to Q; detected by HPLC and and 2021, 26, x FOR PEER Review Molecules 2021, 26,Molecules LC-MS. The ortho-hydroxylation activities of (a): to CA; CA; (b): E; (c): Af to C; (d): (d): K x FOR PEER Review to Q;(e): DHKDHK to DHQ. Final substrate concentration of 80 mg-1, 3. = 3. L and and (e): to DHQ. Final substrate concentration of 80 mg -1 , n = n10 ofTable two. The yield and conversion rate of ortho-hydroxylated flavonoids for the HpaBC complicated Table two. The yield and2. The yield and conversion price of ortho-hydroxylated flavonoidscomplex when feeding with difTable conversion price of ortho-hydroxylated flavonoids for the HpaBC for the HpaBC complex when feedin when feeding with various substrates. The horizontal lines 12-LOX Inhibitor web inside the table indicate that no activities has ferent substrates. The horizontal lines horizontal lines inside the table indicate that no activities has Information are shownData are sho ferent substrates. The within the table indicate that no activities has been detected. been detected. as the been implies s.d.s (n suggests s.d.s (n =Data are shown because the signifies s.d.s (n = 3). = 3). detected. three).SubstratesSubstrates SubstratesProducts ProductsYield Yield Conversion Price ( ) Conver L Conversion Items (mg )(mg-1) Yield (mg-1) Rate ( ) L-2.
