3.4.21.62 additional information - additional information - 29427, 29448, 29459, 29480 3.4.21.62 additional information - additional information kinetic analysis of the perhydrolytic wild-type and mutant enzyme activities, overview 732207 3.4.21.62 additional information - additional information Km of subtilisin Novo chemically attached to soluble DEAE-dextran and insoluble DEAE-Sephadex 29471 3.4.21.62 additional information - additional information Michaelis-Menten kinetics and thermodynamics 752829 3.4.21.62 additional information - additional information Michaelis-Menten kinetics using Lineweaver-Burk and Hanes plots 752524 3.4.21.62 additional information - additional information Michaelis-Menten kinetics using Lineweaver-Burk and Hanes plots at pH 8.5 and 22°C, purified recombinant enzyme 731172 3.4.21.62 additional information - additional information Michaelis-Menten kinetics, Km is 0.175% w/v 754435 3.4.21.62 additional information - additional information the temperature dependence of the kinetics and thermodynamic parameters suggest that the enzyme exists in two, i.e. cold and hot forms, at 22°C the cold form turns into the hot one possibly owing to a conformational change 29458 3.4.21.62 0.000655 - N-succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide 37°C, pH 10.5 669738 3.4.21.62 0.0027 - Suc-Ala-Ala-Pro-Phe-4-nitroanilide 1 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25°C 707805 3.4.21.62 0.0036 - Suc-Ala-Ala-Pro-Phe-4-nitroanilide 2.2 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25°C 707805 3.4.21.62 0.0037 - Suc-Ala-Ala-Pro-Phe-4-nitroanilide 3.5 mol lactose bound per 1 mol subtilisin, in 10 mM potassium phosphate buffer, pH 7.8, at 25°C 707805 3.4.21.62 0.0037 - Suc-Ala-Ala-Pro-Phe-4-nitroanilide subtilisin without bound lactose, in 10 mM potassium phosphate buffer, pH 7.8, at 25°C 707805 3.4.21.62 0.0054 - benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-4-nitroanilide - 667756 3.4.21.62 0.039 - L-Ala-L-Ala-L-Val-L-Ala wild-type, pH 9.0, 24°C 670743 3.4.21.62 0.053 - L-Ala-L-Ala-L-Val-L-Ala V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24°C 670743 3.4.21.62 0.059 - casein pH 10, 45°C, Vmax: 0.336 mg/min 717205 3.4.21.62 0.07 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.5, 5°C 649692 3.4.21.62 0.09 - Urea-denatured hemoglobin 30°C, pH 8.0 668316 3.4.21.62 0.11 - N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide subtilisin-like serine protease, at 20°C, in 50 mM Tris-HCl buffer, pH 7.5 710470 3.4.21.62 0.115 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant enzyme, 5°C, pH 8.6 752829 3.4.21.62 0.13 - Glp-Ala-Ala-Leu-p-nitroanilide in 2% DMSO 690033 3.4.21.62 0.13 - succinyl-Ala-Ala-Pro-Phe 4-nitroanilide - 29455 3.4.21.62 0.13 - succinyl-Ala-Ala-Pro-Phe 4-nitroanilide mutant subtilisin BPN' Asp to Ser99 29455 3.4.21.62 0.13 - succinyl-Ala-Ala-Pro-Phe 4-nitroanilide succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl 7-amide, subtilisin Sendai 29459 3.4.21.62 0.133 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant enzyme, 25°C, pH 9.4 752829 3.4.21.62 0.14 - Z-Ala-Ala-Leu-p-nitroanilide in 20% DMSO 690033 3.4.21.62 0.15 - MeO-succinyl-Ala-Ala-Phe 4-nitroanilide - 29455 3.4.21.62 0.15 - MeO-succinyl-Ala-Ala-Phe 4-nitroanilide subtilisin DY 29452 3.4.21.62 0.15 - MeO-succinyl-Ala-Ala-Phe 4-nitroanilide succinyl-Ala-Ala-Pro-Phe 4-nitroanilide, subtilisin BPN' 29455 3.4.21.62 0.15 - N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.4, 37°C, mutant S101W/G169A/V192A 717697 3.4.21.62 0.15 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/L216W, pH 7.5, 30°C 732207 3.4.21.62 0.15 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.5, 20°C 649692 3.4.21.62 0.178 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant enzyme, 25°C, pH 8.6 752829 3.4.21.62 0.18 - Glp-Ala-Ala-Leu-p-nitroanilide in 2% DMF 690033 3.4.21.62 0.19 - Z-Ala-Ala-Leu-p-nitroanilide in 20% DMF 690033 3.4.21.62 0.198 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant enzyme, 25°C, pH 7.4 752829 3.4.21.62 0.2 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165Y/L216W, pH 7.5, 30°C 732207 3.4.21.62 0.22 - acetyl-L-Phe ethyl ester 30°C, pH 8.0 668316 3.4.21.62 0.22 - MeO-succinyl-Ala-Ala-Phe 4-nitroanilide subtilisin Carlsberg 29452 3.4.21.62 0.223 - L-Phe-L-Ala-L-Ala-L-Phe wild-type, pH 9.0, 24°C 670743 3.4.21.62 0.227 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant N212G 731332 3.4.21.62 0.227 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant enzyme, 45°C, pH 8.6 752829 3.4.21.62 0.233 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant wild-type enzyme 731332 3.4.21.62 0.24 - L-Phe-L-Ala-L-Ala-L-Phe V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24°C 670743 3.4.21.62 0.25 - Glp-Ala-Ala-Leu-p-nitroanilide in 2% DMF 690033 3.4.21.62 0.25 - Z-Ala-Ala-Leu-p-nitroanilide in 20% DMSO 690033 3.4.21.62 0.278 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G 731332 3.4.21.62 0.279 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant wmutant T211G 731332 3.4.21.62 0.28 - Suc-Ala-Ala-Ala-p-nitroanilide pH 7.5, 5°C 649692 3.4.21.62 0.29 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G/T211G 731332 3.4.21.62 0.3 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.5, 40°C 649692 3.4.21.62 0.301 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210G/T211G/N212G 731332 3.4.21.62 0.32 - Suc-Ala-Ala-Ala-p-nitroanilide pH 7.5, 20°C 649692 3.4.21.62 0.325 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 8.0, 25°C, recombinant mutant P210A 731332 3.4.21.62 0.33 - Suc-Ala-Ala-Pro-Ala-p-nitroanilide pH 7.5, 5°C or 40°C 649692 3.4.21.62 0.33 - succinyl-L-Asp-L-Val-L-Arg-L-Ala-L-Phe-7-amido-4-methylcoumarin mutant Y217L, pH 8.3, 25°C 667655 3.4.21.62 0.35 - N-Succ-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.4, 37°C, wild-type 717697 3.4.21.62 0.36 - succinyl-L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin mutant Y217L, pH 8.3, 25°C 667655 3.4.21.62 0.37 - acetyl-L-Tyr ethyl ester 30°C, pH 8.0 668316 3.4.21.62 0.39 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S166C-S-a 649848 3.4.21.62 0.41 - N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide subtilisin-like serine protease, at 80°C, in 50 mM Tris-HCl buffer, pH 7.5 710470 3.4.21.62 0.42 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165L/L216W, pH 7.5, 30°C 732207 3.4.21.62 0.43 - Suc-Ala-Ala-Pro-Phe-4-nitroanilide recombinant enzyme, pH 8.5, 22°C 752524 3.4.21.62 0.44 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.5, 20°C, DTT 649692 3.4.21.62 0.45 - succinyl-L-Ala-L-Ala-L-Pro-L-Phe-4-nitroanilide mutant Y217L, pH 8.3, 25°C 667655 3.4.21.62 0.51 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S166C 649848 3.4.21.62 0.54 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant mutant T58A/G165I/L216W, pH 7.5, 30°C 732207 3.4.21.62 0.54 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C-S-d 649848 3.4.21.62 0.55 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C, wild-type and mutant M222C-S-e 649848 3.4.21.62 0.58 - Suc-Ala-Ala-Ala-p-nitroanilide pH 7.5, 20°C, DTT 649692 3.4.21.62 0.59 - N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide recombinant wild-type enzyme, pH 7.5, 30°C 732207 3.4.21.62 0.6 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C-S-c and S166C-S-f 649848 3.4.21.62 0.61 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant M222C-S-f 649848 3.4.21.62 0.62 - L-Tyr-L-Val-L-Ala-L-Asp wild-type, pH 9.0, 24°C 670743 3.4.21.62 0.63 - Suc-Ala-Ala-Ala-p-nitroanilide pH 7.5, 40°C 649692 3.4.21.62 0.63 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant M222C-S-b 649848 3.4.21.62 0.65 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C 649848 3.4.21.62 0.67 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant M222C-S-d 649848 3.4.21.62 0.68 - Suc-Ala-Ala-Pro-Leu-p-nitroanilide pH 7.5, 5°C 649692 3.4.21.62 0.68 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C-S-b and M222-S-a 649848 3.4.21.62 0.68 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C, mutant S166C-a 649872 3.4.21.62 0.69 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant M222C-S-c 649848 3.4.21.62 0.7 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S166C-S-e 649848 3.4.21.62 0.7 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C, mutant S166C-f 649872 3.4.21.62 0.71 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide subtilisin DY 29452 3.4.21.62 0.72 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S166C-S-c 649848 3.4.21.62 0.73 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C wild-type 649872 3.4.21.62 0.74 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S166C-S-d 649848 3.4.21.62 0.74 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C, mutant S166C-g 649872 3.4.21.62 0.75 - L-Tyr-L-Val-L-Ala-L-Asp V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24°C 670743 3.4.21.62 0.77 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant M222C 649848 3.4.21.62 0.78 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C-S-e 649848 3.4.21.62 0.79 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C-S-a 649848 3.4.21.62 0.79 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide subtilisin Carlsberg 29452 3.4.21.62 0.8 - succinyl-AAPA-p-nitroanilide pH 8.6, 25°C mutant S166C-S-c 649872 3.4.21.62 0.85 - acetyl-L-Phe 30°C, pH 8.0 668316 3.4.21.62 0.86 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S156C-S-f 649848 3.4.21.62 0.92 - Suc-Ala-Ala-Pro-Ala-p-nitroanilide pH 7.5, 75°C 649692 3.4.21.62 0.92 - succinyl-AAPF-p-nitroanilide pH 7.5, 25°C mutant S166C-S-b 649848 3.4.21.62 0.98 - L-Ala-L-Ala-L-Pro-L-Ala V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24°C 670743 3.4.21.62 1 - succinyl-Ala-Ala-Pro-Phe-4-nitroanilide pH 7.5, 75°C 649692 3.4.21.62 1.01 - Suc-Ala-Ala-Pro-Leu-p-nitroanilide pH 7.5, 40°C 649692 3.4.21.62 1.09 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-e 649872 3.4.21.62 1.15 - L-Ala-L-Ala-L-Pro-L-Phe wild-type, pH 9.0, 24°C 670743 3.4.21.62 1.17 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-c 649872 3.4.21.62 1.2 - Succinyl-Ala-Ala-Ala 4-nitroanilide subtilisin Carlsberg 29452 3.4.21.62 1.34 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-b 649872 3.4.21.62 1.5 - Succinyl-Ala-Ala-Ala 4-nitroanilide subtilisin DY 29452 3.4.21.62 1.5 2 succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-h 649872 3.4.21.62 1.6 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-d 649872 3.4.21.62 1.74 - succinyl-AAPA-p-nitroanilide pH 8.6, 25°C mutant S166C-S-g 649872 3.4.21.62 1.9 - succinyl-AAPA-p-nitroanilide pH 8.6, 25°C mutant S166C-S-e and S166S-f 649872 3.4.21.62 1.9 - succinyl-AAPE-p-nitroanilide pH 8.6, 25°C mutant S166C-S-a 649872 3.4.21.62 1.93 - L-Ala-L-Ala-L-Pro-L-Phe V104S/L124M/P129S/S130G/P131E/A133S/T134L/L135I, pH 9.0, 24°C 670743 3.4.21.62 1.99 - L-Ala-L-Ala-L-Pro-L-Ala wild-type, pH 9.0, 24°C 670743 3.4.21.62 2 - succinyl-AAPA-p-nitroanilide pH 8.6, 25°C wild-type 649872 3.4.21.62 2.1 - benzoyl-L-Arg ethyl ester 30°C, pH 8.0 668316 3.4.21.62 2.25 - Suc-Ala-Ala-Ala-p-nitroanilide pH 7.5, 75°C 649692 3.4.21.62 2.26 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-i 649872 3.4.21.62 2.4 - N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide subtilisin, at 20°C, in 50 mM Tris-HCl buffer, pH 8.0 710470 3.4.21.62 2.46 - succinyl-AAPF-p-nitroanilide pH 8.6, 25°C mutant S166C-S-j 649872 3.4.21.62 3.13 - N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide pH 8.5, 37°C 717855 3.4.21.62 3.37 - Suc-Ala-Ala-Pro-Leu-p-nitroanilide pH 7.5, 75°C 649692 3.4.21.62 3.4 - succinyl-AAPR-p-nitroanilide pH 8.6, 25°C mutant S166C-S-b 649872 3.4.21.62 3.74 - N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide pH 8.5, 37°C, mutant D182G 717855 3.4.21.62 3.84 - N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide pH 8.5, 37°C, wild-type 717855 3.4.21.62 4 - acetyl-L-Tyr 30°C, pH 8.0 668316 3.4.21.62 4 - Suc-Ala-Ala-p-nitroanilide pH 7.5, 20°C 649692 3.4.21.62 4 - Z-Ala-Ala-Leu-p-nitroanilide in 20% DMF 690033 3.4.21.62 4.02 - N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide pH 8.5, 37°C, mutant D180G/D182G 717855 3.4.21.62 4.28 - N-methoxysuccinyl-Ala-Ala-Pro-Val-4-nitroanilide pH 8.5, 37°C, mutant D180G 717855 3.4.21.62 4.4 - succinyl-AAPE-p-nitroanilide pH 8.6, 25°C wild-type 649872 3.4.21.62 5.2 - succinyl-AAPR-p-nitroanilide pH 8.6, 25°C mutant S166C-S-j 649872 3.4.21.62 5.3 - succinyl-AAPR-p-nitroanilide pH 8.6, 25°C mutant S166C-S-i 649872 3.4.21.62 5.5 - succinyl-AAPR-p-nitroanilide pH 8.6, 25°C mutant S166C-S-d 649872 3.4.21.62 6.67 - Suc-Ala-Ala-p-nitroanilide pH 7.5, 20°C, DTT 649692 3.4.21.62 7 - benzoyl-Arg ethyl ester subtilisin Novo, subtilisin Carlsberg 29425 3.4.21.62 7.2 - succinyl-AAPR-p-nitroanilide pH 8.6, 25°C wild-type 649872 3.4.21.62 7.9 - N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide subtilisin, at 80°C, in 50 mM Tris-HCl buffer, pH 8.0 710470 3.4.21.62 8.2 - succinyl-AAPR-p-nitroanilide pH 8.6, 25°C mutant S166C-S-h 649872 3.4.21.62 8.3 - benzyloxycarbonyl-Gly-4-nitroanilide pH 7.5, 20°C, in presence and absence of DTT 649692 3.4.21.62 10 - benzoyl-Arg ethyl ester subtilisn BPN' 29425 3.4.21.62 17 - N-acetyl-Phe ethyl ester subtilisin BPN' 29425 3.4.21.62 17 - Suc-Ala-Ala-p-nitroanilide pH 7.5, 40°C 649692 3.4.21.62 22 - N-Acetyl-Tyr ethyl ester subtilisin BPN' 29425 3.4.21.62 24 - N-Acetyl-Trp ethyl ester subtilisin BPN' 29425 3.4.21.62 30 - toluenesulfonyl-Arg methyl ester subtilisin Novo, N-acetyl-Phe methyl ester, subtilisin Carlsberg 29425 3.4.21.62 40 - toluenesulfonyl-Arg methyl ester subtilisin Carlsberg 29425 3.4.21.62 50 - N-Acetyl-Trp methyl ester subtilisin Carlsberg 29425 3.4.21.62 60 - N-Acetyl-Phe methyl ester subtilisin Novo 29425 3.4.21.62 66 - N-Acetyl-Leu methyl ester subtilisin BPN' 29425 3.4.21.62 70 - N-Acetyl-Tyr ethyl ester subtilisin Novo, N-acetyl-Tyr methyl ester, subtilisin Carlsberg 29425 3.4.21.62 90 - N-Acetyl-Tyr methyl ester subtilisin Novo, N-acetyl-Tyr ethyl ester, subtilisin Carlsberg, N-acetyl-Trp methyl ester, subtilisin Novo 29425 3.4.21.62 91 - N-Acetyl-Lys methyl ester subtilisin BPN' 29425 3.4.21.62 120 - N-Acetyl-Ala methyl ester subtilisin BPN' 29425 3.4.21.62 130 - sec-phenethyl alcohol after incubation in 1,4-dioxane, co-lyophilization with methyl-beta-cyclodextrin 685809 3.4.21.62 319 - sec-phenethyl alcohol after a 4-day incubation period in 1,4-dioxane, co-lyophilization with methyl-beta-cyclodextrin 685809