Insights into the substrate selectivity of Bambusa oldhamii phenylalanine ammonia-lyase 1 and 2 through mutational analysis

Production of Trans-Cinnamic Acid by Immobilization of the Bambusa oldhamii BoPAL1 and BoPAL2 Phenylalanine Ammonia-Lyases on Electrospun Nanofibers

“…On SDS-gels, major monomeric PAL bands were with a molecular mass of about 75 kDa (Figure 2A-D). The purification results were corresponded with previous reports [29][30][31]33]. The eBoPAL2 exhibited the best catalytic properties and used to measure the optimum conditions of protein immobilization.…”

“…IPTG was added to a final concentration of 1 mM for protein induction. When an OD 600 of 0.6-1.0 was reached, the cells were further incubated at 30 • C with vigorous shaking for 6 h [29][30][31]33].…”

NLIP and HAD-like Domains of Pah1 and Lipin 1 Phosphatidate Phosphatases Are Essential for Their Catalytic Activities

“…E. coli DH5α was used for the proliferation of plasmids, and BL21(DE3) was transformed for the expression of the yeast Pah1 and human Lipin 1-α truncations as described previously [ 20 , 49 , 51 ]. E. coli DE3 strains carrying the plasmids ( Table 1 ) were grown at 37 °C in 250 mL of Luria–Bertani (LB, 1% tryptone, 0.5% yeast extract, 1% NaCl) medium containing 100 μg/mL ampicillin.…”

“…E. coli cells were resuspended in lysis buffer (50 mM Tris-HCl, 150 mM NaCl, 10 mM imidazole, 1 mM phenylmethanesulfonyl fluoride (PMSF), pH 7.4) and disrupted by sonication (Ultrasonic Processors, Sonicator 3000, Misonix, Farmingdale, NY, USA) [ 49 , 51 ]. Cell lysates were added into an open column containing an appropriate amount of Ni-charged resin (Bio-Rad, USA).…”

“…To determine the kinetic parameters of Pah1 truncations, the concentrations of PA were varied from 0 to 9.09 Mol %. A substrate saturation curve was obtained after a 10 min incubation [ 7 , 51 ]. A double reciprocal plot was adapted for calculation of the K m and k cat values [ 52 ].…”