Recombinant Enzyme Product Specification Sheet

Prozomix Limited, Station Court, Haltwhistle, Northumberland, NE49 9HN, UK. Tel: +44 (0) 1434 400455 Fax: +44 (0) 1434 322822 [email protected] [email protected] www.prozomix.com [full product listing at www.prozomix.com/products/listing]

Cat. No.: PRO-E0018

LOT: 2008-0018

Activity: Licheninase and cellulase

Synonyms: Lichenase; β-(1→4)-D-glucan 4-glucanohydrolase; 1,3;1,4-β-glucan endohydrolase; 1,3;1,4-β-glucan 4-glucanohydrolase; 1,3-1,4-β-D- glucan 4-glucanohydrolase; (1→3)-(1→4)-β-D-glucan 4- glucanohydrolase; beta-(1→4)-D-glucan 4-glucanohydrolase; 1,3;1,4-beta-glucan endohydrolase; 1,3;1,4-beta-glucan 4- glucanohydrolase; 1,3-1,4-beta-D-glucan 4-glucanohydrolase; (1→3)-(1→4)-beta-D-glucan 4-glucanohydrolase, and, endo-1,4-β-D- glucanase; β-1,4-glucanase; β-1,4-endoglucan hydrolase; celluase A; cellulosin AP; endoglucanase D; alkali cellulase; cellulase A 3; celludextrinase; 9.5 cellulase; avicelase; pancellase SS; 1,4- (1,3;1,4)-β-D-glucan 4-glucanohydrolase; 4-(1,3;1,4)-β-D-glucan 4- glucanohydrolase; endo-1,4-beta-D-glucanase; beta-1,4-glucanase; beta-1,4-endoglucan hydrolase; 1,4-(1,3;1,4)-beta-D-glucan 4- glucanohydrolase; 4-(1,3;1,4)-beta-D-glucan 4-glucanohydrolase, respectively

Nomenclature: CAZy [GH5 and GH26, glycoside hydrolase families 5 and 26, both members of clan GH-A], CtGH26-GH5 has an N-terminal GH26 licheninase catalytic module and a C-terminal GH5 cellulase catalytic module

Source organism: Clostridium thermocellum NCIB 10682

Enzyme Commission No.: 3.2.1.73 and 3.2.1.4, respectively

Activity: 270 U/mL (60oC; pH 7; barley β-glucan) Specific activity: 450 U/mg

Purity: > 95 % as judged by SDS-PAGE

Form and storage: Supplied in 3.2 M ammonium sulphate, store at 4oC (shipped at room temperature) pH optimum: 7 (stable from 6 - 8)

Temperature optimum: 60oC (stable up to 65oC)

[Protein]: 0.6 mg/mL

Sequence length: 601 amino acids (view sequence)

Accession No.: AAA23225

Molecular weight: 70487.6 Da (theoretical)

~ 69400 Da (observed by SDS-PAGE)

- (observed by mass spectrometry)

Biological function: The licheninase catalytic module hydrolyses (1→4)-β-D-glucosidic linkages in β-D-glucans containing (1→3)- and (1→4)-bonds. It acts on lichenin and cereal β-D-glucans, but not on β-D-glucans containing only 1,3- or 1,4-bonds. The cellulase catalytic module endohydrolyses (1→4)-β-D-glucosidic linkages in cellulose, lichenin and cereal β-D-glucans

Potential application(s): Biomass conversion, carbohydrate research

Comments: CtGH26-GH5 contains an N-terminal GH26 licheninase catalytic module and a C-terminal GH5 cellulase catalytic module

Usage: Agitate bottle sufficiently to fully homogenise enzyme precipitate before use

Assay: One unit is defined as the amount of enzyme required to release 1 μmol of glucose-reducing-sugar equivalents per minute from barley β-glucan in 50 mM phosphate buffer, pH 7.0, at 60°C, where reducing sugars are measured by the method of Miller (1959; Anal. Chem. 31, 426-428)

Primary sequence:

MLKIGAWVGTQPSESAIKSFQELQGRKLDIVHQFINWSTDFSWVRPYADAVYNNGSILMITWEPWEYNTVDIKNG KADAYITRMAQDMKAYGKEIWLRPLHEANGDWYPWAIGYSSRVNTNETYIAAFRHIVDIFRANGATNVKWVFNVN CDNVGNGTSYLGHYPGDNYVDYTSIDGYNWGTTQSWGSQWQSFDQVFSRAYQALASINKPIIIAEFASAEIGGNK ARWITEAYNSIRTSYNKVIAAVWFHENKETDWRINSSPEALAAYREAIGAGSSNPTPTPTWTSTPPSSSPKAVDP FEMVRKMGMGTNLGNTLEAPYEGSWSKSAMEYYFDDFKAAGYKNVRIPVRWDNHTMRTYPYTIDKAFLDRVEQVV DWSLSRGFVTIINSHHDDWIKEDYNGNIERFEKIWEQIAERFKNKSENLLFEIMNEPFGNITDEQIDDMNSRILK IIRKTNPTRIVIIGGGYWNSYNTLVNIKIPDDPYLIGTFHYYDPYEFTHKWRGTWGTQEDMDTVVRVFDFVKSWS DRNNIPVYFGEFAVMAYADRTSRVKWYDFISDAALERGFACSVWDNGVFGSLDNDMAIYNRDTRTFDTEILNALF N

Literature: 1. Taylor et al. (2005) J. Biol. Chem. 280, 32761-32767