Molecular cloning and characterization of a novel bifunctional cellobiohydrolase/β-xylosidase from a metagenomic library of mangrove soil

Liu, Yi-De; Yuan, Ge; An, Yu-Ting; Zhu, Zi-Ran; Li, Gang

doi:10.1016/j.enzmictec.2022.110141

Cited by 3 publications

(1 citation statement)

References 42 publications

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“…The distances of residues to xylan were observably shorter at 1.9-2.8 Å, suggesting that a stably bound XynB-xylan complex. It was reported that heat resistant ability of the enzyme was correlated with the number of hydrogen bonds formed in the reaction ( Liu et al, 2023 ). However, compared with Xyn, which had higher heat resistant ability with fewer hydrogen bonds than that of XynB, indicating that there were still other factors might also influence enzyme thermostability ( Nezhad et al, 2022 ).…”

Section: Resultsmentioning

confidence: 99%

Improved secretory expression and characterization of thermostable xylanase and β-xylosidase from Pseudothermotoga thermarum and their application in synergistic degradation of lignocellulose

Chen,

Qin,

You

et al. 2023

Front. Bioeng. Biotechnol.

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Xylanase and β-xylosidase are the key enzymes for hemicellulose hydrolysis. To further improve hydrolysis efficacy, high temperature hydrolysis with thermostable hemicellulases showed promise. In this study, thermostable xylanase (Xyn) and β-xylosidase (XynB) genes from Pseudothermotoga thermarum were cloned and secretory expressed in Bacillu subtilis. Compared with Escherichia coli expression host, B. subtilis resulted in a 1.5 time increase of enzymatic activity for both recombinant enzymes. The optimal temperature and pH were 95°C and 6.5 for Xyn, and 95°C and 6.0 for XynB. Thermostability of both recombinant enzymes was observed between the temperature range of 75–85°C. Molecular docking analysis through AutoDock showed the involvement of Glu525, Asn526, Trp774 and Arg784 in Xyn-ligand interaction, and Val237, Lys238, Val761 and Asn76 in XynB-ligand interaction, respectively. The recombinant Xyn and XynB exhibited synergistic hydrolysis of beechwood xylan and pretreated lignocellulose, where Xyn and XynB pre-hydrolysis achieved a better improvement of pretreated lignocellulose hydrolysis by commercial cellulase. The observed stability of the enzymes at high temperature and the synergistic effect on lignocellulosic substrates suggested possible application of these enzymes in the field of saccharification process.

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Section: Resultsmentioning

confidence: 99%

Improved secretory expression and characterization of thermostable xylanase and β-xylosidase from Pseudothermotoga thermarum and their application in synergistic degradation of lignocellulose

Chen,

Qin,

You

et al. 2023

Front. Bioeng. Biotechnol.

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Effects of LDPE and PBAT plastics on soil organic carbon and carbon-enzymes: A mesocosm experiment under field conditions

Jia,

Yao,

Tan

et al. 2024

Environmental Pollution

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Characterization of Novel Multifunctional Xylanase from Rumen Metagenome and Its Effects on In Vitro Microbial Fermentation of Wheat Straw

Zhang,

Qiu,

Zhao

et al. 2024

Fermentation

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This study investigated the characterization of a novel multifunctional enzyme, RuXyn394, derived from the metagenome of beef cattle rumen, and its impact on the in vitro microbial fermentation of wheat straw. RuXyn394, a member of the glycosyl hydrolase 11 family, displayed optimal activity under diverse pH and temperature conditions: xylanase at pH 5.5 and 50 °C, acetyl esterase at pH 6.5 and 60 °C, exoglucanase at pH 7.0 and 50 °C, and endoglucanase at pH 6.0 and 50 °C. The enzyme’s xylanase, endoglucanase, and exoglucanase activities exhibited remarkable pH stability across the range of pH 3–8 and maintained a relatively stable performance at temperatures from 20 to 50 °C, 20 to 60 °C, and 20 to 70 °C, respectively. The xylanase function, with the highest kcat/Km ratio, was identified as the predominant activity of RuXyn394. The enzyme’s various functions responded uniquely to metal ions; notably, the addition of 5 mM K+ significantly boosted the activities of xylanase, exoglucanase, and endoglucanase by 55.5%, 53.5%, and 16.4%, respectively, without affecting its acetyl esterase activity. Over the course of three time points (30 min, 60 min, 120 min), the degradation products of wheat straw xylan, including xylopentaose, xylotetraose, xylotriose, xylobiose, xylose, and total xylooligosaccharides, constituted an average of 18.4%, 33.7%, 20.6%, 22.9%, 4.3%, and 95.7% of the total products, respectively. RuXyn394 effectively hydrolyzed wheat straw, resulting in augmented volatile fatty acid production and ammonia-N levels during in vitro microbial fermentation. These findings indicate the potential of RuXyn394 as a novel and highly efficient enzyme preparation, offering promising prospects for the valorization of wheat straw, an agricultural by-product, in ruminant diets.

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Molecular cloning and characterization of a novel bifunctional cellobiohydrolase/β-xylosidase from a metagenomic library of mangrove soil

Cited by 3 publications

References 42 publications

Improved secretory expression and characterization of thermostable xylanase and β-xylosidase from Pseudothermotoga thermarum and their application in synergistic degradation of lignocellulose

Improved secretory expression and characterization of thermostable xylanase and β-xylosidase from Pseudothermotoga thermarum and their application in synergistic degradation of lignocellulose

Effects of LDPE and PBAT plastics on soil organic carbon and carbon-enzymes: A mesocosm experiment under field conditions

Characterization of Novel Multifunctional Xylanase from Rumen Metagenome and Its Effects on In Vitro Microbial Fermentation of Wheat Straw

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