Literature summary for 3.4.21.64 extracted from

Yazawa, K.; Sugahara, M.; Yutani, K.; Takehira, M.; Numata, K.
Derivatization of proteinase K with heavy atoms enhances its thermal stability (2016), ACS Catal., 6, 3036-3046 .

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Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme, oil microbatch method, mixing of 0.001 ml of 40 mg/ml protein solution in 50 mM MES-NaOH, pH 6.5, with 0.001 ml of precipitant solution composed of 250 mM NaNO3, 50 mM CaCl2, and 50 mM MES-NaOH, pH 6.5, to form Pr3+-derivatized crystals, the precipitant solution containing additionally 25 mM PrCl3 is used, X-ray diffraction structure determination and analysis at 1.45 A resolution	Parengyodontium album

Crystallization (Comment)

Organism

purified enzyme, oil microbatch method, mixing of 0.001 ml of 40 mg/ml protein solution in 50 mM MES-NaOH, pH 6.5, with 0.001 ml of precipitant solution composed of 250 mM NaNO3, 50 mM CaCl2, and 50 mM MES-NaOH, pH 6.5, to form Pr3+-derivatized crystals, the precipitant solution containing additionally 25 mM PrCl3 is used, X-ray diffraction structure determination and analysis at 1.45 A resolution

Parengyodontium album

Protein Variants

Protein Variants	Comment	Organism
molecular biology	proteinase K is widely used in molecular biology for its broad substrate specificity, wide pH stability, and high hydrolysis activity. Aminolysis by proteinase K is also attractive for chemoenzymatic peptide synthesis	Parengyodontium album

Metals/Ions

Metals/Ions	Comment	Organism
Ca2+	1 mM CaCl2, the denaturation temperature of proteinase K derivatized with praseodymium (Pr) ions is 16.2°C, which is 5.9°C higher than those of metal-free and Ca2+-bound proteinase K, respectively. Isothermal titration calorimetry (ITC) measurements demonstrate that Pr-ion binding to proteinase K shows endothermic peaks, whereas Ca2+-ion binding shows exothermic peaks, indicating that the binding mode of Pr ions is different from that of Ca2+ ions, even though the crystal structures of proteinase K with Pr and Ca2+ ions are identical	Parengyodontium album
dysprosium	1 mM DyCl3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
europium	1 mM EuCl33, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
gadolinium	1 mM GdCl33, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
holmium	1 mM HoCl3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
lanthanum	1 mM La(NO3)3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
lutetium	1 mM LuCl3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
additional information	differential scanning calorimetry curves for proteinase K derivatized with heavy atoms, showing the correlation between atomic number and denaturation temperature, overview	Parengyodontium album
neodymium	1 mM NdCl3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
Pr3+	1 mM PrCl3, the denaturation temperature of proteinase K derivatized with praseodymium (Pr) ions is 16.2°C, which is 5.9°C higher than those of metal-free and Ca2+-bound proteinase K, respectively. Isothermal titration calorimetry (ITC) measurements demonstrate that Pr-ion binding to proteinase K shows endothermic peaks, whereas Ca2+-ion binding shows exothermic peaks, indicating that the binding mode of Pr ions is different from that of Ca2+ ions, even though the crystal structures of proteinase K with Pr and Ca2+ ions are identical. Hydrolytic activity of Pr-derivatized proteinase K shows that the hydrolytic activity is 46fold higher at 70°C using synthetic nitroanilide substrate and 9 and 76fold higher at 70°C and 80°C using fluorescein isothiocyanate-labeled casein, respectively, in comparison with the native proteinase K. Furthermore, based on the yield of chemoenzymatic peptide syntheses, the aminolysis activity of Pr-derivatized proteinase K is 3.5 and 9.5fold higher than that of the native proteinase K at 50°C and 60°C, respectively. Analysis of the mechanism by which Pr ions enhance the thermal stability of proteinase K, overview	Parengyodontium album
samarium	1 mM SmCl3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album
ytterbium	1 mM YbCl3, differential scanning calorimetry analysis bound to the enzyme	Parengyodontium album

Organism

Organism	UniProt	Comment	Textmining
Parengyodontium album	P06873	i.e. Tritirachium album or Engyodontium album	-

Source Tissue

Source Tissue	Comment	Organism	Textmining
commercial preparation	-	Parengyodontium album	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the enzyme has a broad substrate specificity. Evaluation of aminolytic activity by polymerization of glutamic acid diethyl ester oligo(glutamic acid ethyl ester)	Parengyodontium album	?	-	?
N-succinyl-L-Phe-4-nitroanilide + H2O	-	Parengyodontium album	N-succinyl-L-Phe + 4-nitroaniline	-	?

Synonyms

Synonyms	Comment	Organism
Proteinase K	-	Parengyodontium album

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
additional information	-	maximum degree of polymerization is achieved at 60°C using Pr-derivatized proteinase K, and at 50°C using the native proteinase K with Ca2+ ions. The yield of oligo(glutamic acid ethyl ester) is highest at 30°C using both the native and Pr-derivatized proteinase K	Parengyodontium album
70	-	hydrolytic activity	Parengyodontium album

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
30	80	activity range, profile overview	Parengyodontium album

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
16.2	80	the denaturation temperature of proteinase K derivatized with praseodymium (Pr) ions is 16.2°C, which is 5.9°C higher than those of metal-free and Ca2+-bound proteinase K, respectively. Isothermal titration calorimetry (ITC) measurements demonstrate that Pr-ion binding to proteinase K shows endothermic peaks, whereas Ca2+-ion binding shows exothermic peaks, indicating that the binding mode of Pr ions is different from that of Ca2+ ions, even though the crystal structures of proteinase K with Pr and Ca2+ ions are identical. Hydrolytic activity of Pr-derivatized proteinase K shows that the hydrolytic activity is 46fold higher at 70°C using synthetic nitroanilide substrate and 9 and 76fold higher at 70°C and 80°C using fluorescein isothiocyanate-labeled casein, respectively, in comparison with the native proteinase K. Furthermore, based on the yield of chemoenzymatic peptide syntheses, the aminolysis activity of Pr-derivatized proteinase K is 3.5 and 9.5fold higher than that of the native proteinase K at 50°C and 60°C, respectively	Parengyodontium album

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	-	assay at	Parengyodontium album