Papain with nitrile hydratase activity from Carica papaya

Enzyme Description

Species

Extremophile

EC Number

4.2.1.84 ↗ (standard : 3.4.22.2)

Sequence

Length: 212 amino acids

IPEYVDWRQKGAVTPVKNQGSCGSCWAFSAVVTIEGIIKIRTGNLNEYSEQELLDCDRRSYGCNGGYPWSALQLVAQYGIHYRNTYPYEGVQRYCRSREKGPYAAKTDGVRQVQPYNEGALLYSIANQPVSVVLEAAGKDFQLYRGGIFVGPCGNKVDHAVAAVGYGPNYILIKNSWGTGWGENGYIRIKRGTGNSYGVCGLYTSSFYPVKN

Andrea Versari et al. (2002) — Enzymatic hydrolysis of nitrides by an engineered nitrile hydratase (Papain Gln19Glu) in aqueous-organic media

Biotechnology and Bioengineering · doi:10.1002/bit.10267 ↗ · Activity - Michaelis-Menten

5 measurements ▼

Database ID

UniProt: P00784 ↗

Sequence Annotation

Inferred - from protein name (132 first residues from UniProt sequence removed from mature protein, presence of 1 mutation compared to UniProt)

Protein Source

Recombinant, host yeast Pichia pastoris

Experimental Data (5 measurements)

5 measurements

Mutation	Property	Assay	Solvent	Solvent Volume	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	10%	—	6.8	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in microM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	20%	—	6.7	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in microM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	30%	—	4.9	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in microM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	40%	—	3.6	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in microM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	50%	—	2.7	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in microM/hr) \| Insoluble substrate in aqueous phase

Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	10%	—	6.8	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in µM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	20%	—	6.7	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in µM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	30%	—	4.9	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in µM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	40%	—	3.6	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in µM/hr) \| Insoluble substrate in aqueous phase
Q19E	Activity - Michaelis-Menten	Vmax (in µM/hr), measured in the presence of organic solvent by HPLC	Acetone	50%	—	2.7	µM/hr	50 mM citrate buffer, 2mM DTT, 1mM EDTA	5	22°C	N-acetyl-L-phenylalanyl-L-alanine nitrile	N-acetyl-phenylalanyl-alanine amide	—	—	Classical aqueous control (in µM/hr) \| Insoluble substrate in aqueous phase

Mutations in this dataset (1)

Q19E

Visualization : Activity — Michaelis-Menten

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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