Rophenyl decanoate; 4-NPP–4-nitrophenyl palmitate. The assay reaction was carried out having a nitrophenyl decanoate; substrate, and the reaction proceeded for 20The at 40 C.reaction was carr 1 mM concentration of every single 4-NPP–4-nitrophenyl palmitate. min assay Absorbance 1 mM concentration of each and every min intervals.and therepresents activity on each and every for 20 min 3 40 readings were taken at 410 nm at 1 substrate, Every bar reaction proceeded substrate, n = at along with the error bar corresponds to nm at 1 min intervals. Each bar readings have been taken at 410 the regular error with the imply (SEM). represents activity on each n two.3. Biochemical Propertiescorresponds towards the regular error from the mean (SEM). = 3 plus the error barThe biochemical properties of BaAXE had been investigated in assays with 4-NPA as the substrate. BaAXE showed optimal activity at pH eight and 40 C. The km, kcat, and kcat/km Figure 4. Activity assay. BaAXE was assayed utilizing different acyl chain esters (C2 16). 4-NPA–4-1 -1 s- (catalytic efficiency) values have been calculated as4-NPO–4-nitrophenyl , and 282 4-NPD–4- 1 , nitrophenyl acetate; 4-NPB–4-nitrophenyl butyrate; 0.43 mM, 122.four s octanoate; mM respectively. A thermostability assay showed that the BaAXEreaction was carried out activity nitrophenyl decanoate; 4-NPP–4-nitrophenyl palmitate. The assay retained about 40 having a just after incubating the every substrate, as well as the reactionC but showed no clear activity at acidic 1 mM concentration of enzyme for 2 h at 4000 proceeded for 20 min at 40 . Absorbance pHs. At were7 and at 410 nm at retained over 80 activity just after activity on every single substrate, for readings pH taken 9, BaAXE 1 min intervals. Every single bar represents incubating the enzyme n = (Figure error 4 h 3 and the5). bar corresponds towards the normal error of your mean (SEM).two.3. Biochemical Properties The biochemical properties of BaAXE had been investigated in assays with 4-NPA because the substrate. BaAXE showed optimal activity at pH eight and 40 .MMP-1, Human (HEK293, His) The km, kcat, and kcat/km (catalytic efficiency) values have been calculated as 0.IL-7, Human (HEK293, His) 43 mM, 122.PMID:23667820 four s-1, and 282 mM-1 s-1, respectively. A thermostability assay showed that the BaAXE retained about 40 activity following six of 16 incubating the enzyme for two h at 4000 but showed no clear activity at acidic pHs. At pH 7 and 9, BaAXE retained more than 80 activity just after incubating the enzyme for four h (Figure five).Molecules 2022, 27,Figure five. Biochemical properties. The biochemical properties of BaAXE had been determined in assays Figure 5. Biochemical properties. The biochemical properties of BaAXE had been determined in assays with 4-NPA. (A) The optimal temperature was determined in in assays performed different temperwith 4-NPA. (A) The optimal temperature was determined assays performed at at unique temperaatures: 20 30 40 50 , and 60 . (B) Thermostability assay determined after incubating , , , tures: 20 C, 30 C, 40 C, 50 C, and 60 C. (B) Thermostability assay determined right after incubating the the enzyme for two h at distinctive temperatures. (C) The optimal pH was determined in assays perenzyme for two h at pHs at temperatures. (C) The optimal pH was determined in assays performed formed at differentdifferentthe optimal temperature. (D) pH stability was determined soon after incuba- at various enzyme for four h at temperature. of pH stability was determined soon after incubation of tion of thepHs at the optimal20 . Following 2 h(D) incubation, residual activity was determined, and also the enzyme the h at 20 C. Right after 2 h determined. Absorb.
