Improving the thermostability of a fungal GH11 xylanase via site-directed mutagenesis guided by sequence and structural analysis

Han, Nanyu; Miao, Huabiao; Ding, Junmei; Li, Junjun; Mu, Yunxiang; Zhou, Junpei; Huang, Zunxi

doi:10.1186/s13068-017-0824-y

Cited by 49 publications

(34 citation statements)

References 51 publications

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“…The exposure to physical and chemical mutagens has resulted in significant changes in the mutant strain as compared to the wild type. Han et al (2017) demonstrated the site-directed mutagenesis of XynCDBFV gene of ruminal fungus Neocallimastix patriciarum for improving the thermostability of XynCDBFV, a glycoside hydrolase (GH) family 11 xylanase. Similar work has also been carried out in different bacterial strains, a rifampin-resistant mutant of Cellulomonas biazotea, designated 7Rf, resulting in elevated levels of xylanases production as compared to the parental strain.…”

Section: Mutagenesis Of Microorganisms For Enhanced Xylanase Productionmentioning

confidence: 99%

A detailed overview of xylanases: an emerging biomolecule for current and future prospective

Bhardwaj

Kumar

Verma

2019

Bioresour. Bioprocess.

276

105

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Xylan is the second most abundant naturally occurring renewable polysaccharide available on earth. It is a complex heteropolysaccharide consisting of different monosaccharides such as l-arabinose, d-galactose, d-mannoses and organic acids such as acetic acid, ferulic acid, glucuronic acid interwoven together with help of glycosidic and ester bonds. The breakdown of xylan is restricted due to its heterogeneous nature and it can be overcome by xylanases which are capable of cleaving the heterogeneous β-1,4-glycoside linkage. Xylanases are abundantly present in nature (e.g., molluscs, insects and microorganisms) and several microorganisms such as bacteria, fungi, yeast, and algae are used extensively for its production. Microbial xylanases show varying substrate specificities and biochemical properties which makes it suitable for various applications in industrial and biotechnological sectors. The suitability of xylanases for its application in food and feed, paper and pulp, textile, pharmaceuticals, and lignocellulosic biorefinery has led to an increase in demand of xylanases globally. The present review gives an insight of using microbial xylanases as an "Emerging Green Tool" along with its current status and future prospective.

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Section: Mutagenesis Of Microorganisms For Enhanced Xylanase Productionmentioning

confidence: 99%

A detailed overview of xylanases: an emerging biomolecule for current and future prospective

Bhardwaj

Kumar

Verma

2019

Bioresour. Bioprocess.

276

105

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“…The development of thermostable lipases has clear and practical potential to meet with industrial demands. Furthermore, generating such stable lipases may simultaneously improve their kinetic efficiency (Rogers and Bommarius, 2010;Han et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

“…B-factor values for all Lipr27RCL Cα atoms were extracted from the PDB file, these B-factors were normalized such that they had a zero mean and unit variance distribution as follows: B = B−<B> σ(B) , where the b is the average of all Cα atoms and σ(B) is the standard deviation of the B-factors for the individual protein (Yuan et al, 2003). We have successfully used these equations in previous studies (Han et al, 2017).…”

Section: Lipase R27rcl Temperature Factor Calculationmentioning

confidence: 99%

“…Screening through the many colonies necessary to identify superior enzyme isoforms, however, is time-consuming and cannot be effectively performed in host species that grow slowly. An alternative approach to improving enzymatic thermal stability that has been implemented successfully is the B-factor (Xie et al, 2014;Han et al, 2017). This approach relies upon improving thermostability via increasing enzymatic rigidity at certain sites, with B-factors derived from X-ray data offering insight into atom fluctuations and rigidity relative to their equilibrium positions (Ringe and Petsko, 1986).…”

Section: Introductionmentioning

confidence: 99%

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Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis

et al. 2020

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In order to improve the thermostability of lipases derived from Rhizopus chinensis, we identified lipase (Lipr27RCL) mutagenesis sites that were associated with enhanced flexibility based upon B-factor analysis and multiple sequence alignment. We found that two mutated isoforms (Lipr27RCL-K64N and Lipr27RCL-K68T) exhibited enhanced thermostability and improved residual activity, with respective thermal activity retention values of 37.88% and 48.20% following a 2 h treatment at 50 • C relative to wild type Lipr27RCL. In addition, these Lipr27RCL-K64N and Lipr27RCL-K68T isoforms exhibited 2.4-and 3.0-fold increases in enzymatic half-life following a 90 min incubation at 60 • C. Together these results indicate that novel mutant lipases with enhanced thermostability useful for industrial applications can be predicted based upon B-factor analysis and constructed via site-directed mutagenesis.

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“…xylanase from Neocallimastix patriciarum was significantly increased [18], and mutant G414W that exhibited remarkable enhancement in half-life (6.5-fold) was obtained for Candida rugosa Lipase1 [19].…”

Section: Introductionmentioning

confidence: 99%

Improving the thermostability and acid resistance of Rhizopus oryzae α‐amylase by using multiple sequence alignment based site‐directed mutagenesis

Yang

Tang

2020

Biotech and App Biochem

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Higher thermostability or acid resistance for fungal α‐amylase will help to improve the sugar‐making process and cut down the production costs. Here, the thermostability or acid resistance of Rhizopus oryzae α‐amylase (ROAmy) was significantly enhanced by site‐directed evolution based on multiple sequence alignment (MSA) method. For instance, compared with the wild‐type ROAmy, the optimum temperature of mutants G136D and A144Y was increased from 50 to 55 °C, whereas for mutants V174R and I276P, the optimum temperature was increased from 50 to 60 °C. The optimum pH of mutants G136D and A144Y shifted from 5.5 to 5.0, whereas for mutants V174R and T253E, the optimum pH changed from 5.5 to 4.5. The results showed that mutant V174R had a 2.52‐fold increase in half‐life at 55 °C, a 2.55‐fold increase in half‐life at pH 4.5, and a 1.61‐fold increase in catalytic efficiency (kcat/Km) on soluble starch. The three‐dimensional model simulation revealed that changes of hydrophilicity, hydrogen bond, salt bridge, or rigidity observed in mutants might mainly account for the improvement of thermostability and acid resistance. The mutants with improved catalytic properties attained in this work may render an accessible and operable approach for directed evolution of fungal α‐amylase aimed at interesting functions.

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Improving the thermostability of a fungal GH11 xylanase via site-directed mutagenesis guided by sequence and structural analysis

Cited by 49 publications

References 51 publications

A detailed overview of xylanases: an emerging biomolecule for current and future prospective

A detailed overview of xylanases: an emerging biomolecule for current and future prospective

Improving the Thermostability of Rhizopus chinensis Lipase Through Site-Directed Mutagenesis Based on B-Factor Analysis

Improving the thermostability and acid resistance of Rhizopus oryzae α‐amylase by using multiple sequence alignment based site‐directed mutagenesis

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