Literature summary for 3.1.1.74 extracted from

Brissos, V.; Melo, E.P.; Martinho, J.M.; Cabral, J.M.
Biochemical and structural characterisation of cutinase mutants in the presence of the anionic surfactant AOT (2008), Biochim. Biophys. Acta, 1784, 1326-1334.

Activating Compound

Activating Compound	Comment	Organism	Structure
sodium dioctyl sulfosuccinate	in the absence of surfactant the S54D variant catalytic activity is similar to that of the wild type cutinase, whereas L153Q and T179C variants show a lower activity	Fusarium solani

Application

Application	Comment	Organism
industry	key for the design of biocatalysts with sufficient stability for practical applications in detergent industry	Fusarium solani

Cloned(Commentary)

Cloned (Comment)	Organism
over-expression of recombinant cutinase cloned in pMac5-8 and its variants (S54D, L153Q and T179C) are performed with Escherichia coli WK6 strain	Fusarium solani

Protein Variants

Protein Variants	Comment	Organism
L153Q	L153Q mutation also reduces the development of hydrophobic solvent accessible patches	Fusarium solani
S54D	S54D mutant of cutinase is significantly more resistant to sodium dioctyl sulfosuccinate denaturation than the wild type	Fusarium solani
T179C	T179C mutation located close to the active centre and to disulfide bond Cys171-Cys178 introduced changes in the cutinase structure that are observed even in the cutinase region around the tryptophan residue. This mutation also reduces the development of hydrophobic solvent accessible patches	Fusarium solani

Inhibitors

Inhibitors	Comment	Organism	Structure
sodium dioctyl sulfosuccinate	-	Fusarium solani

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.2	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
0.31	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
0.33	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
0.35	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
0.48	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
0.49	-	p-nitrophenyl butyrate	T179Cmutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
0.66	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
0.69	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
0.72	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
0.74	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
0.85	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
1.08	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani
1.09	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
1.12	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
1.14	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
1.31	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
1.56	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani
1.74	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani
3.57	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
4.33	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
22000	-	molar mass found for the wild type cutinase and its mutants is identical	Fusarium solani

Organism

Organism	UniProt	Comment	Textmining
Fusarium solani	-	pisi	-

Purification (Commentary)

Purification (Comment)	Organism
osmotic shock, acid precipitation, dialysis and two sequential anion exchange chromatographic steps, followed by a final dialysis and subsequently freeze-dried. Cutinase purity is confirmed by 12.5% SDS-PAGE	Fusarium solani

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
p-nitrophenyl butyrate + H2O	-	Fusarium solani	p-nitrophenol + butyrate	-	?

Synonyms

Synonyms	Comment	Organism
cutinase	-	Fusarium solani

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
7900	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
9600	-	p-nitrophenyl butyrate	T179Cmutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
11000	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
11000	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
11100	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
11500	-	p-nitrophenyl butyrate	T179C mutant, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani
13800	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani
14700	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
15200	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
15500	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
15600	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 0 mM	Fusarium solani
16000	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
17400	-	p-nitrophenyl butyrate	wild type, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
17900	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
24200	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 1 mM	Fusarium solani
24700	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM	Fusarium solani
25200	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
25800	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM	Fusarium solani
28500	-	p-nitrophenyl butyrate	L153Q mutant, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani
32100	-	p-nitrophenyl butyrate	S54D mutant, sodium dioctyl sulfosuccinate concentration = 2 mM	Fusarium solani

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.44	-	sodium dioctyl sulfosuccinate	L153Q mutant	Fusarium solani
0.46	-	sodium dioctyl sulfosuccinate	T179C mutant	Fusarium solani
0.72	-	sodium dioctyl sulfosuccinate	wild type	Fusarium solani
1.38	-	sodium dioctyl sulfosuccinate	S54D mutant	Fusarium solani