Fig. 3: Kinetic characterization of pMT7 and pMT10 enzymes.
From: Total enzymatic synthesis of cis-α-irone from a simple carbon source
a Fold change in pMT7 activity when SAH (0-200 μM), homocysteine (0-200 μM), adenine (0–200 μM), or adenosine (0–200 μM) was supplemented into purified pMT7 reaction. It was calculated by dividing the pMT7 activity where no additive was added. b Fold change in pMT7 and pMT10 activities when SAH (0–40 μM) was supplemented into the purified enzyme reactions. It was calculated by dividing the pMT7 and pMT10 activity where no additive was added. c Effect of introducing auxiliary enzymes, Mtn and MtaD, to hydrolyse SAH on cis-α-irone production by purifying pMT7 and pMT10 with or without initial 20 μM SAH. pMT10 increased production of cis-α-irone by ~4.5 and ~6.5-fold without and with MtaD enzyme, respectively compared to pMT7 reaction alone. a to c, The average and s.d. of three biologically independent experiments are shown. Significance (p-value) was evaluated by a two-sided Student’s t-test, n.s. represents p > 0.05. d In vitro biotransformation of psi-ionone to cis-α-irone by using cell lysates containing overexpressed pMT10. The bar chart represents the mean value of irone concentration. The orange diamonds represent the percentage conversion calculated based on the starting substrate concentration. The average and the s.d. of two biologically independent experiments are shown in the graph. The abbreviations are as follows. Mtn, S-adenosylhomocysteine nucleosidase. MtaD, S-adenosylhomocysteine deaminase. SAH, S-adenosylhomocysteine. Source data are provided as a Source Data file.
