Fusion of family VI cellulose binding domains to Bacillus halodurans xylanase increases its catalytic activity and substrate-binding capacity to insoluble xylan

Mangala, Selanere L.; Kittur, Farooqahmed S.; Nishimoto, Mamoru; Sakka, Kazuo; Ohmiya, Kunio; Kitaoka, Motomitsu; Hayashi, Kiyoshi

doi:10.1016/s1381-1177(02)00226-6

Cited by 41 publications

(21 citation statements)

References 29 publications

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“…In addition, the presence of CBM32 in rGH5-CBM32 may decrease resistance of the substrate to rGH5 attack. Activity-stimulating effects of CBMs toward insoluble substrates have been observed in cellulases and other glycoside hydrolases (20,21,25). For example, the family 3 CBM of Paenibacillus curdlanolyticus B-6 Xyn10D has been recently shown to be important for hydrolysis of insoluble arabinoxylan and natural biomass by comparison of the parental and truncated enzymes (25), while the addition of family 6 CBMs of Clostridium stercorarium XynA to Bacillus halodurans xylanase XylA increased hydrolytic activity of the chimeric enzyme toward insoluble oat spelt xylan but not soluble birchwood xylan (21).…”

Section: Discussionmentioning

confidence: 99%

Influence of a Mannan Binding Family 32 Carbohydrate Binding Module on the Activity of the Appended Mannanase

Mizutani

Fernandes²,

Karita

et al. 2012

Appl Environ Microbiol

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eIn general, cellulases and hemicellulases are modular enzymes in which the catalytic domain is appended to one or more noncatalytic carbohydrate binding modules (CBMs). CBMs, by concentrating the parental enzyme at their target polysaccharide, increase the capacity of the catalytic module to bind the substrate, leading to a potentiation in catalysis. Clostridium thermocellum hypothetical protein Cthe_0821, defined here as C. thermocellum Man5A, is a modular protein comprising an N-terminal signal peptide, a family 5 glycoside hydrolase (GH5) catalytic module, a family 32 CBM (CBM32), and a C-terminal type I dockerin module. Recent proteomic studies revealed that Cthe_0821 is one of the major cellulosomal enzymes when C. thermocellum is cultured on cellulose. Here we show that the GH5 catalytic module of Cthe_0821 displays endomannanase activity. C. thermocellum Man5A hydrolyzes soluble konjac glucomannan, soluble carob galactomannan, and insoluble ivory nut mannan but does not attack the highly galactosylated mannan from guar gum, suggesting that the enzyme prefers unsubstituted ␤-1,4-mannoside linkages. The CBM32 of C. thermocellum Man5A displays a preference for the nonreducing ends of mannooligosaccharides, although the protein module exhibits measurable affinity for the termini of ␤-1,4-linked glucooligosaccharides such as cellobiose. CBM32 potentiates the activity of C. thermocellum Man5A against insoluble mannans but has no significant effect on the capacity of the enzyme to hydrolyze soluble galactomannans and glucomannans. The product profile of C. thermocellum Man5A is affected by the presence of CBM32.

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Section: Discussionmentioning

confidence: 99%

Influence of a Mannan Binding Family 32 Carbohydrate Binding Module on the Activity of the Appended Mannanase

Mizutani

Fernandes²,

Karita

et al. 2012

Appl Environ Microbiol

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“…Most information on the role of CBDs has been obtained from the removal [1], domain exchange [13] and site-directed mutagenesis of the CBDs [15], or by artiWcial addition of the CBD [9]. It therefore seems that the CBDs are interchangeable to a certain degree, but much more data are needed on diVerent catalytic domain-CBD combinations to elucidate the exact functional role of the CBDs.…”

Section: Introductionmentioning

confidence: 99%

Effect of the cellulose-binding domain on the catalytic activity of a β-glucosidase from Saccharomycopsis fibuligera

Gundllapalli

Pretorius

Otero

2007

J Ind Microbiol Biotechnol

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Enzyme engineering was performed to link the beta-glucosidase enzyme (BGL1) from Saccharomycopsis fibuligera to the cellulose-binding domain (CBD2) of Trichoderma reesei cellobiohydrolase (CBHII) to investigate the effect of a fungal CBD on the enzymatic characteristics of this non-cellulolytic yeast enzyme. Recombinant enzymes were constructed with single and double copies of CBD2 fused at the N-terminus of BGL1 to mimic the two-domain organization displayed by cellulolytic enzymes in nature. The engineered S. fibuligera beta-glucosidases were expressed in Saccharomyces cerevisiae under the control of phosphoglycerate-kinase-1 promoter (PGK1 ( P )) and terminator (PGK1 ( T )) and yeast mating pheromone alpha-factor secretion signal (MFalpha1 ( S )). The secreted enzymes were purified and characterized using a range of cellulosic and non-cellulosic substrates to illustrate the effect of the CBD on their enzymatic activity. The results indicated that the recombinant enzymes of BGL1 displayed a 2-4-fold increase in their hydrolytic activity toward cellulosic substrates like avicel, amorphous cellulose, bacterial microcrystalline cellulose, and carboxy methyl cellulose in comparison with the native enzyme. The organization of the CBD in these recombinant enzymes also resulted in enhanced substrate affinity, molecular flexibility and synergistic activity, thereby improving the ability of the enzymes to act on and hydrolyze cellulosic substrates, as characterized by adsorption, kinetics, thermal stability, and scanning electron microscopic analyses.

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“…Many xylanases have an additional CBM, which can be a cellulose binding domain (CBD) or a xylan binding domain (XBD) (1,5,7,22,25,28). XBD typically increases activity against insoluble xylan (1,5,24), although some XBDs also bind soluble xylans (21,25).…”

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confidence: 99%

Effect of Glycosylation and Additional Domains on the Thermostability of a Family 10 Xylanase Produced by Thermopolyspora flexuosa

Anbarasan

Jänis

Paloheimo

et al. 2010

Appl Environ Microbiol

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The effects of different structural features on the thermostability of Thermopolyspora flexuosa xylanase XYN10A were investigated. A C-terminal carbohydrate binding module had only a slight effect, whereas a polyhistidine tag increased the thermostability of XYN10A xylanase. In contrast, glycosylation at Asn26, located in an exposed loop, decreased the thermostability of the xylanase. The presence of a substrate increased stability mainly at low pH.

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Fusion of family VI cellulose binding domains to Bacillus halodurans xylanase increases its catalytic activity and substrate-binding capacity to insoluble xylan

Cited by 41 publications

References 29 publications

Influence of a Mannan Binding Family 32 Carbohydrate Binding Module on the Activity of the Appended Mannanase

Influence of a Mannan Binding Family 32 Carbohydrate Binding Module on the Activity of the Appended Mannanase

Effect of the cellulose-binding domain on the catalytic activity of a β-glucosidase from Saccharomycopsis fibuligera

Effect of Glycosylation and Additional Domains on the Thermostability of a Family 10 Xylanase Produced by Thermopolyspora flexuosa

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