Activity enhancement of a Bacillus circulans xylanase by introducing ion-pair interactions into an α-helix

Pokhrel, Subarna; Joo, Jeong Chan; Lee, Hoe-Suk; Yoo, Young Je

doi:10.1016/j.procbio.2013.07.008

Cited by 5 publications

(1 citation statement)

References 36 publications

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“…The locations of glycosylated residues Asn37 and Asn88 in the structure of XynCDBFV were indicated in the left panel. b The enzyme activities of the wild-type and deglycosylated mutants were determined at the indicated temperatures, and the relative activity of each sample was presented as a percentage of the value of wild type at 55°C reactivity at higher temperature and alkaline conditions to meet the demands of bio-bleaching (Paes and O'Donohue 2006;Song et al 2012;Stephens et al 2009;Pokhrela et al 2013;Umemoto et al 2009). Notwithstanding, the needs for enzymes with higher reactivity at milder conditions remain.…”

Section: Discussionmentioning

confidence: 99%

Improving the catalytic performance of a GH11 xylanase by rational protein engineering

Cheng

Chen

Huang

et al. 2015

Appl Microbiol Biotechnol

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XynCDBFV from Neocallimastix patriciarum is among the most effective xylanases and holds great potentials in a wide variety of industrial applications. In the present study, several active site residues were modified referring to the instrumental information of the complex structure of XynCDBFV and xylooligosaccharides. Among the 12 single active site mutants, W125F and F163W show increased activity comparing to the wild-type protein. The double mutant W125F/F163W was then generated which displayed nearly 20 % increase in the enzyme activity. Although W125F/F163W showed 5 °C reduction in the optimal temperature, it still preserves similar thermostability and is more active than the wild-type enzyme at temperatures lower than 60 °C. These properties make the double mutant a suitable candidate for commercial applications that involve lower operating temperatures. Furthermore, we investigated the effect of N-glycosylation on the thermostability of XynCDBFV when expressed in the yeast strain Pichia pastoris for industrial use. Two potential glycosylation sites (Asn-37 and Asn-88) were examined, and their roles in enzyme performance were validated. We found that the N-glycosylations of XynCDBFV are related to both catalytic activity and heat stability, with Asn-37 motif playing a dominant role. Collectively, the enzymatic properties of XynCDBFV were improved by molecular engineering, and the influences of N-glycosylations on the enzyme have been clearly elucidated herein.

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Section: Discussionmentioning

confidence: 99%

Improving the catalytic performance of a GH11 xylanase by rational protein engineering

Cheng

Chen

Huang

et al. 2015

Appl Microbiol Biotechnol

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Engineering improved thermostability of the GH11 xylanase from Neocallimastix patriciarum via computational library design

Cui

Peng

et al. 2018

Appl Microbiol Biotechnol

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Xylanases, which cleave the β-1,4-glycosidic bond between xylose residues to release xylooligosaccharides (XOS), are widely used as food additives, animal feeds, and pulp bleaching agents. However, the thermally unstable nature of xylanases would hamper their industrial application. In this study, we used in silico design in a glycoside hydrolase family (GH) 11 xylanase to stabilize the enzyme. A combination of the best mutations increased the apparent melting temperature by 14 °C and significantly enhanced thermostability and thermoactivation. The variant also showed an upward-shifted optimal temperature for catalysis without compromising its activity at low temperatures. Moreover, a 10-fold higher XOS production yield was obtained at 70 °C, which compensated the low yield obtained with the wild-type enzyme. Collectively, the variant constructed by the computational strategy can be used as an efficient biocatalyst for XOS production at industrially viable conditions.

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IgE binding to peanut allergens is inhibited by combined d-aspartic and d-glutamic acids

Chung

Reed

2015

Food Chemistry

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Activity enhancement of a Bacillus circulans xylanase by introducing ion-pair interactions into an α-helix

Cited by 5 publications

References 36 publications

Improving the catalytic performance of a GH11 xylanase by rational protein engineering

Improving the catalytic performance of a GH11 xylanase by rational protein engineering

Engineering improved thermostability of the GH11 xylanase from Neocallimastix patriciarum via computational library design

IgE binding to peanut allergens is inhibited by combined d-aspartic and d-glutamic acids

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