Temperature and solvent exposure response of three fatty acid peroxygenase enzymes for application in industrial enzyme processes

Plasma‐driven biocatalysis: In situ hydrogen peroxide production with an atmospheric pressure plasma jet increases the performance of OleTJE when compared to adding the same molar amount of hydrogen peroxide in bolus

“…The OleT JE pET28a‐based construct was expressed in Escherichia coli Arctic Express (DE3) (Agilent) as previously established. [ 44 ] Briefly, overnight cultures (60 ml) were grown at 30°C in Luria‐Bertani medium (LB) with 50 µg/ml kanamycin and 20 µg/ml gentamycin and used (50 ml) to inoculate cultures (1 L) in the same media the next day for expression. The inoculated cultures were then grown at 30°C in the same media until an OD of 0.6–0.8 was achieved.…”

“…The use of Triton‐X‐100 has been demonstrated previously to increase the myristic acid‐binding and catalytic activity of OleT JE . [ 3,11,44 ] Reactions were incubated at 27°C for 20 min while rotating at 150 rpm before being quenched with 25 µl of 37% hydrochloric acid, spiked with 1 mM 1‐heptadecene as an internal standard, and extracted using 400 µM ethyl acetate. Sample tubes were then centrifuged at 13 300 rpm for 20 min to separate the layers.…”

“…The top layer was extracted, dried with anhydrous MgSO 4 , and then derivatized 1:1 with BSTFA at 70°C for 20 min. [ 44 ]…”

“…The top layer was extracted, dried with anhydrous MgSO 4 , and then derivatized 1:1 with BSTFA at 70°C for 20 min. [44] Multiple concentrations of H 2 O 2 were tested, including 238, 500, 700, and 1140 µM, corresponding to the H 2 O 2 concentrations measured after 1, 2, 3, and 5 min He/H 2 O plasma treatment, respectively.…”

Microbial P450 repertoire (P450ome) and its application feasibility in pharmaceutical industry, chemical industry, and environmental protection

“…CYP152K6 was stable up to 50°C and showed great stability in 3% acetone and dimethylformamide. Stability in solvents helps the enzyme's substrates remain soluble in solution for more efficient catalysis, and heat stability allows enzymes to remain active longer during industrial processes (Wapshott‐Stehli & Grunden, 2021).…”