Improving the Thermostability of a Fungal GH11 Xylanase via Fusion of a Submodule (C2) from Hyperthermophilic CBM9_1-2

Miao, Huabiao; Ying-jie, MA; Zhe, Yuanyuan; Tang, Xianghua; Wu, Qian; Huang, Zunxi; Han, Nanyu

doi:10.3390/ijms23010463

Cited by 16 publications

(5 citation statements)

References 42 publications

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“…Table 2 showed that the EcsXyl4 has 227 hydrogen bonds, while CFxyl3 and EcsXyl1-3 only have 224, 216, 220, and 226, respectively, which results in a more thermal stability for EcsXyl4 than for other xylanases. In a previous study, the thermal stability of a xylanase from N. patriciarum increased after insertion of a CBM9_1-2, which attributed to four additional hydrogen bonds (S42–S462, T59–E277, S41–K463, and S44–G371) ( Miao et al, 2022 ). Figure 2E shows the 3D structure of EcsXyl4.…”

Section: Resultsmentioning

confidence: 95%

Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution

Tian

Zhang²,

Yang³

et al. 2022

Front. Bioeng. Biotechnol.

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Xylanase, a glycoside hydrolase, is widely used in the food, papermaking, and textile industries; however, most xylanases are inactive at high temperatures. In this study, a xylanase gene, CFXyl3, was cloned from Cellulomonas flavigena and expressed in Escherichia coli BL21 (DE3). To improve the thermostability of xylanase, four hybrid xylanases with enhanced thermostability (designated EcsXyl1–4) were engineered from CFXyl3, guided by primary and 3D structure analyses. The optimal temperature of CFXyl3 was improved by replacing its N-terminus with the corresponding area of SyXyn11P, a xylanase that belongs to the hyperthermostable GH11 family. The optimal temperatures of the hybrid xylanases EcsXyl1–4 were 60, 60, 65, and 85°C, respectively. The optimal temperature of EcsXyl4 was 30 C higher than that of CFXyl3 (55°C) and its melting temperature was 34.5°C higher than that of CFXyl3. After the hydrolysis of beechwood xylan, the main hydrolysates were xylotetraose, xylotriose, and xylobiose; thus, these hybrid xylanases could be applied to prebiotic xylooligosaccharide manufacturing.

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Section: Resultsmentioning

confidence: 95%

Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution

Tian

Zhang²,

Yang³

et al. 2022

Front. Bioeng. Biotechnol.

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“…The C2 submodule had a considerable influence on improving the CDBFV's thermostability, according to enzymatic property study. The half-lives of the three chimaeras C2-CDBFV, CDBFV-C2, and C2-CDBFV-C2 are 1.5 times (37.5 min), 4.9 times (122.2 min), and 3.8 times (93.1 min) higher than wild-type CDBFV (24.8 min) at the ideal temperature (60.0°C) [55].…”

Section: Genetic Engineeringmentioning

confidence: 90%

Engineering to enhance thermostability of xylanase: For the new era of biotechnology

Pandey¹,

Sharma²,

Gupta³

2022

J App Biol Biotech

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“…For example, the catalytic efficiency of the Paenibacillus xylanase, using birchwood xylan as substrate, was increased by 1.5 fold after the introduction of a xylan-binding domain [ 34 ]. Furthermore, inserting the submodule C2 from Carbohydrate-binding modules 9_1-2 into the N- and/or C-terminal regions of Neocallimastix patriciarum xylanase increased (18–28%) its catalytic activity toward corncob xylanase [ 35 ]. In the present study, the proline-rich C-terminal oligopeptide enhanced the catalytic activity of xylanase by 2.48 fold, which is a remarkable effect in this type of studies.…”

Section: Discussionmentioning

confidence: 99%

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

et al. 2022

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Xylanases are widely used in the degradation of lignocellulose and are important industrial enzymes. Therefore, increasing the catalytic activity of xylanases can improve their efficiency and performance. In this study, we introduced the C-terminal proline-rich oligopeptide of the rumen-derived XynA into XylR, a GH10 family xylanase. The optimum temperature and pH of the fused enzyme (XylR-Fu) were consistent with those of XylR; however, its catalytic efficiency was 2.48-fold higher than that of XylR. Although the proline-rich oligopeptide did not change the enzyme hydrolysis mode, the amount of oligosaccharides released from beechwood xylan by XylR-Fu was 17% higher than that released by XylR. This increase may be due to the abundance of proline in the oligopeptide, which plays an important role in substrate binding. Furthermore, circular dichroism analysis indicated that the proline-rich oligopeptide might increase the rigidity of the overall structure, thereby enhancing the affinity to the substrate and catalytic activity of the enzyme. Our study shows that the proline-rich oligopeptide enhances the catalytic efficiency of GH10 xylanases and provides a better understanding of the C-terminal oligopeptide-function relationships. This knowledge can guide the rational design of GH10 xylanases to improve their catalytic activity and provides clues for further applications of xylanases in industry.

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Improving the Thermostability of a Fungal GH11 Xylanase via Fusion of a Submodule (C2) from Hyperthermophilic CBM9_1-2

Cited by 16 publications

References 42 publications

Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution

Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution

Engineering to enhance thermostability of xylanase: For the new era of biotechnology

Fusion of a proline-rich oligopeptide to the C-terminus of a ruminal xylanase improves catalytic efficiency

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