Especially, TanLpl and TanLpe were affected to decrease the activity down to 46.1% and 25.2% by the presence of Zn2+, respectively. All three recombinant tannases were inhibited in the presence of 1 mM FeSO4 approximately

down to one fourth levels (Table 1). (%) Chemicals (1 mM) TanLpl TanLpa TanLpe Control 100 100 100 MnCl2 87.6 ± 22.5 111.3 ± 23.8 75.6 ± 13.2 CaCl2 98.3 ± 15.8 88.7 ± 11.5 92.3 ± 12.7 FeSO4 22.5 ± 12.2 24.1 ± 18.4 23.4 ± 13.1 ZnSO4 46.1 ± 7.64 95.4 ± 16.3 25.2 ± 17.5 MgSO4 AZD5363 123.7 ± 20.1 110.5 ± 11.9 96.7 ± 7.0 PMSF 83.2 ± 14.7 66.2 ± 20.3 81.2 ± 24.7 EDTA 97.6 ± 3.0 87.8 ± 4.2 103.7 ± 12.2 Urea 91.4 ± 8.8 96.9 ± 0.37 119.5 ± 18.3 aAssays were carried out in triplicate and the results represent the means ± standard deviations. Kinetic properties of TanLpl, TanLpa, and TanLpe K m values of TanLpl, TanLpa, and https://www.selleckchem.com/products/AZD6244.html TanLpe for the other catechin derivatives were approximately 10 times lower than those for MG (Table 2). Table 2 Kinetic properties of TanLpl, TanLpa, and TanLpe a Substarate TanLpl TanLpa TanLpe K m (mM) k cat(s-1) k cat/K m (s-1 · mM-1) K m (mM) k cat(s-1) k cat/K m (s-1 · mM-1) K m (mM) k cat(s-1) k cat/K m (s-1 · mM-1) Methyl gallate (MG) 0.37 ± 0.04 46.02 ± 0.87

125.02 ± 15.43 0.50 ± 0.06 72.73 ± 3.34 145.12 ± 10.65 0.87 ± 0.41 15.95 ± 3.13 18.79 ± 3.08 Epicatechin gallate (ECg) 0.03 ± 0.02 1.49 ± 0.19 52.23 ± 25.64 0.06 ± 0.01 11.08 ± 0.44 195.30 ± 21.53 0.05 ± 0.01 0.42 ± 0.03 8.63 ± 1.17 Epigallocatechin gallate (EGCg) 0.10 ± 0.01 Sirolimus 1.12 ± 0.03 11.68 ± 1.29 0.06 ± 0.01 14.29 ± 0.82 260.76 ± 46.52 0.06 ± 0.02 0.44 ± 0.02 7.25 ± 2.51 Catechin gallate (Cg) 0.05 ± 0.002 2.41 ± 0.10 53.65 ± 4.62 0.05 ± 0.005 8.1 ± 0.04 181.5 ± 27.71 0.08 ± 0.004 1.48 ± 0.11 19.22 ± 2.36 Gallocatechin gallate (GCg) 0.03 ± 0.008 0.89 ± 0.044 27.19 ± 6.28 0.06 ± 0.002 9.2 ± 0.09 154.68 ± 7.97 0.07 ± 0.002 1.12 ± 0.13 14.32 ± 1.95 Epigallocatechin-3-O-(3-O-methyl) gallate (EGCg3″Me) 0.04 ± 0.009 0.26 ± 0.04 6.04 ± 0.57 0.04 ± 0.004 0.35 ± 0.07 9.02 ± 2.28 0.005 ± 0.0009 0.06 ± 0.02 10.57 ± 1.33 aAssays were carried out in triplicate and the results represent the means ± standard deviations. pentosus 22A-1, selleck cloned to reveal their high amino acid identity to TanLpl from L.