Literature summary for 3.2.1.139 extracted from

Ryabova, O.; Vrsanska, M.; Kaneko, S.; van Zyl, W.H.; Biely, P.
A novel family of hemicellulolytic alpha-glucuronidase (2009), FEBS Lett., 583, 1457-1462.

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Cloned(Commentary)

Cloned (Comment)	Organism
Pichia stipitis CBS 6054 grown in YNB medium, cells washed and suspended in YNB medium with a xylooligosaccharide misture and methyl-beta-xylopyranoside, 30°C, 24 h, shaker	Scheffersomyces stipitis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
120000	-	SDS-PAGE with 2 sets of protein markers: Fermentas no. SM0431, SERVA no. 39216	Scheffersomyces stipitis

Organism

Organism	UniProt	Comment	Textmining
Scheffersomyces stipitis	A3LY17	gene ABN67901, EMBL; CBS 6054	-

Purification (Commentary)

Purification (Comment)	Organism
supernatant concentrated on Amicon 10 kDa cut-off membranes, anion-exchange chromatography on HiTrap DEAE-FF column with NaCl gradient in 50 mM sodium-phosphate buffer, pH 7.0, fractions between 0.2 and 0.25 M NaCl pooled, concentrated, desalted, equilibrated in 50 mM acetate buffer, pH 4.0, with 2 M (NH4)2SO4, subjected to hydrophobic interaction chromatography on Butyl-FF column, eluted with (NH4)2SO4 gradient, active fractions between 1.1 and 0.61 M (NH4)2SO4 pooled, desalted, concentrated, subjected to 2 additional anion-exchange chromatography steps on Tricorn MonoQ 5/50GL column, first equilibrated with 50 mM sodium acetate buffer, pH 4.0, eluted with NaCl gradient, then equilibrated with 50 mM sodium phosphate buffer, pH 7.0 and NaCl gradient, concentration with Microcon 10 kDa cut-off membrane filter	Scheffersomyces stipitis

Purification (Comment)

Organism

supernatant concentrated on Amicon 10 kDa cut-off membranes, anion-exchange chromatography on HiTrap DEAE-FF column with NaCl gradient in 50 mM sodium-phosphate buffer, pH 7.0, fractions between 0.2 and 0.25 M NaCl pooled, concentrated, desalted, equilibrated in 50 mM acetate buffer, pH 4.0, with 2 M (NH4)2SO4, subjected to hydrophobic interaction chromatography on Butyl-FF column, eluted with (NH4)2SO4 gradient, active fractions between 1.1 and 0.61 M (NH4)2SO4 pooled, desalted, concentrated, subjected to 2 additional anion-exchange chromatography steps on Tricorn MonoQ 5/50GL column, first equilibrated with 50 mM sodium acetate buffer, pH 4.0, eluted with NaCl gradient, then equilibrated with 50 mM sodium phosphate buffer, pH 7.0 and NaCl gradient, concentration with Microcon 10 kDa cut-off membrane filter

Scheffersomyces stipitis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-O-Me-GlcAalpha(1-2)Xylbeta(1-4)Xylbeta(1-4)Xyl + H2O	from beechwood, de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase	Scheffersomyces stipitis	4-O-methyl-alpha-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)Xyl	de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase	?
4-O-Me-GlcAalpha(1-2)Xylbeta(1-4)Xylbeta(1-4)[4-O-Me-GlcAalpha(1-2)]Xylbeta(1-4)Xyl + H2O	hydrolysis product of endoxylanases on glucuronoxylan	Scheffersomyces stipitis	4-O-methyl-alpha-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)[4-O-Me-GlcAalpha(1-2)]Xylbeta(1-4)Xyl	de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase	?
GlcAalpha(1-2)Xylbeta(1-4)Xylbeta(1-4)[GlcAalpha(1-2)]Xylbeta(1-4)Xyl + H2O	generated from the action of endoxylanases on polysaccharides	Scheffersomyces stipitis	alpha-D-glucuronic acid + Xylbeta(1-4)Xylbeta(1-4)[GlcAalpha(1-2)]Xylbeta(1-4)Xyl	de-estrification of the methyl ester of MeGlcA with glucuronoyl esterase	?

Subunits

Subunits	Comment	Organism
?	x * 120000, SDS-PAGE	Scheffersomyces stipitis

Synonyms

Synonyms	Comment	Organism
alpha-glucuronidase	-	Scheffersomyces stipitis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
60	-	protein unstable at this temperature with 50% activity loss in 30 min	Scheffersomyces stipitis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
40	-	stable for at least 3 h, at optimal temperature loss of 50% activity after 30 min	Scheffersomyces stipitis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4.4	-	-	Scheffersomyces stipitis

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	5.5	at pH 4.0 25.5% and at pH 5.5 51.6% activity	Scheffersomyces stipitis

pI Value

Organism	Comment	pI Value Maximum	pI Value
Scheffersomyces stipitis	isoelectrofocusing on Multiphor II system using SEVALYT PRECOTES gels and markers from 3-10	-	4
Scheffersomyces stipitis	calculated from sequence	-	4.6