Hemicellulolytic and cellulolytic functions of the domains of a β-mannanase cloned from Caldicellosiruptor saccharolyticus

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dc.contributor.authorFrangos, Tracy
dc.contributor.authorBullen, Denise
dc.contributor.authorBergquist, Peter L.
dc.contributor.authorDaniel, Roy M.
dc.date.accessioned2010-08-30T02:00:09Z
dc.date.available2010-08-30T02:00:09Z
dc.date.issued1999-08
dc.description.abstractVarious combinations of the four domains of the multifunctional mannanase from Caldicellosiruptor saccharolyticus have been cloned and expressed in Escherichia coli. The four domains comprise two catalytic domains (1 and 4), and two putative cellulose binding domains (2 and 3). Each of the six gene products (Man1, Man123, Man1234, Man23, Man234 and Man4) was partially purified by heat treatment. The enzymes Man1234, Man123 and Man1 exhibited activity on mannans, and Man1234, Man234 and Man4 exhibited activity on xylan and carboxymethylcellulose (CMC). For the complete enzyme (Man1234) all activities were of the same order of magnitude. Activities were additive against a mixture of mannan and xylan or mannan and CMC (but not xylan and CMC). The expression product Man23 exhibited activity on none of the substrates tested, nor did its presence influence thermostability or significantly reduce the Km value for any of the substrates. However, when expressed in combination with domains 1 or 4 it greatly increased their activity. We conclude that domain 1 catalyses mannan hydrolysis and domain 4 catalyses xylan and CMC hydrolysis at the same active site: domains 2 and 3 have no obvious function, since they do not reduce substrate Km nor affect thermostability. However, their effect on rates of substrate hydrolysis may indicate a role influencing the conformation of the adjacent catalytic domains.en_NZ
dc.identifier.citationFrangos, T., Bullen, D., Bergquist, P. & Daniel, R.M. (1999). Hemicellulolytic and cellulolytic functions of the domains of a β-mannanase cloned from Caldicellosiruptor saccharolyticus. The International Journal of Biochemistry & Cell Biology, 31(8), 853-859.en_NZ
dc.identifier.doi10.1016/S1357-2725(99)00036-9en_NZ
dc.identifier.urihttps://hdl.handle.net/10289/4462
dc.language.isoen
dc.publisherElsevieren_NZ
dc.relation.isPartOfThe International Journal of Biochemistry and Cell Biologyen_NZ
dc.subjectmannanaseen_NZ
dc.subjectcelluloseen_NZ
dc.subjectXylanaseen_NZ
dc.subjectenzyme domainsen_NZ
dc.subjectcaldicellosiruptor saccharolyticusen_NZ
dc.titleHemicellulolytic and cellulolytic functions of the domains of a β-mannanase cloned from Caldicellosiruptor saccharolyticusen_NZ
dc.typeJournal Articleen_NZ
pubs.begin-page853en_NZ
pubs.elements-id40878
pubs.end-page859en_NZ
pubs.volume31en_NZ
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