Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA

Liew, Kok Jun; Ngooi, Chen Yi; Shamsir, Mohd Shahir; Sani, Rajesh K.; Chong, Chun Shiong; Goh, Kian Mau

doi:10.1016/j.pep.2019.105464

Cited by 22 publications

(25 citation statements)

References 55 publications

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“…Bifunctional CtCel5E was fused with a β-glucosidase from Clostridium cellulovorans (CcBglA), the fusion expression improved the enzyme stability and glucose yield [25]. The analysis of the enzymatic hydrolysis products of heterologous expression xylanase is also an important research focus [26,27]; The other part focuses on the synergistic effect of recombinant expressed xylanase and commercial cellulase, such as two different sources of xylanase (rXYN10A MALCI and rXYN11A MALCI) act synergistically with commercial cellulases and resulted in 1.54 and 1.58 folds improved hydrolysis of acid treated rice straw and alkali treated rice straw [28]. In another research, two different sources of xylanase (rXyn162 and rXylCD2) exhibited synergistic hydrolysis of oat spelts xylan and sodium hydroxide pretreated corn stover (SHPC).…”

Section: Discussionmentioning

confidence: 99%

Co-expression of cellulase and xylanase genes in Saccharomyces cerevisiae toward enhanced bioethanol production from corn stover

Xiao

Xia

et al. 2019

Bioengineered

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Lignocellulose is considered as a good resource for producing renewable energy. Previous in vitro studies have shown the synergistic action between cellulase and xylanase during lignocellulose biohydrolysis. In order to achieve the same effect in S. cerevisiae to enhance the practical biotransformation, two recombinant Saccharomyces cerevisiae strains (INVSc1-CBH-CA and INVSc1-CBH-TS) with co-expressed cellulase and xylanase were constructed. The cellulase and xylanase activities in INVSc1-CBH-CA and INVSc1-CBH-TS were 716.43 U/mL and 205.13 U/mL, 931.27 U/mL and 413.70 U/ mL, respectively. The recombinant S. cerevisiae can use the partly delignified corn stover (PDCS) more efficiently and more ethanol producted than S. cerevisiae only expressing cellulase. Fermentation with INVSc1-CBH-CA and INVSc1-CBH-TS using PDCS ethanol yields increased by 1.7 and 2.1 folds higher than INVSc1-CBH, 2.8 and 3.4 folds higher than the wild type S. cerevisiae. The strategy of co-expression cellulase and xylanase in saccharomyces cerevisiae is effective and can be a foundation to research the mechanism of synergy effect of cellulose and xylanase.

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Section: Discussionmentioning

confidence: 99%

Co-expression of cellulase and xylanase genes in Saccharomyces cerevisiae toward enhanced bioethanol production from corn stover

Xiao

Xia

et al. 2019

Bioengineered

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“…Strain RA has 20 genes related to hemicellulose degradation (Table 5, Figure 6). These hemicellulose-acting enzymes belong to GH 2,10,16,29,31,43,51,53,67,78,92,106,113,130,and 154. These enzymes also shared low similarity with the existing sequences deposited in databases (57%-72% identity).…”

Section: Cazymes In R Sacchariphilus Strain Ramentioning

confidence: 99%

“…The majority of strain RA's proteins are likely to be thermostable and salt-tolerant to ensure all biological pathways are functional in high salt. So far, only two xylanases from this bacterium have been characterized [31,32].…”

Section: Gh3mentioning

confidence: 99%

“…Interestingly, xylanase XynRA1 (AWN76_008205, upregulated in MB+xylan) functions better in low salinity, while XynRA2 (AWN76_003690, non-DEG xylanase) exhibits a higher salt tolerance up to 5.0 M NaCl. Both enzymes displayed different characteristics, including product specificity, kinetic performance, and responded differently to temperature, pH, substrate, and metal ions [31,32].…”

Section: Gh3mentioning

confidence: 99%

“…Based on the genome annotation, strain RA harbors two non-homologous xylanases, XynRA1 and XynRA2. Both proteins exhibit slightly different in vitro biochemical characteristics [31,32]. Additionally, strain RA expresses up to four non-homologous cellulases, two β-glucosidases, and four α-l-rhamnosidases [38].…”

Section: Introductionmentioning

confidence: 99%

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Global Transcriptomic Responses of Roseithermus sacchariphilus Strain RA in Media Supplemented with Beechwood Xylan

et al. 2020

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The majority of the members in order Rhodothermales are underexplored prokaryotic extremophiles. Roseithermus, a new genus within Rhodothermales, was first described in 2019. Roseithermus sacchariphilus is the only species in this genus. The current report aims to evaluate the transcriptomic responses of R. sacchariphilus strain RA when cultivated on beechwood xylan. Strain RA doubled its growth in Marine Broth (MB) containing xylan compared to Marine Broth (MB) alone. Strain RA harbors 54 potential glycosyl hydrolases (GHs) that are affiliated with 30 families, including cellulases (families GH 3, 5, 9, and 44) and hemicellulases (GH 2, 10, 16, 29, 31,43, 51, 53, 67, 78, 92, 106, 113, 130, and 154). The majority of these GHs were upregulated when the cells were grown in MB containing xylan medium and enzymatic activities for xylanase, endoglucanase, β-xylosidase, and β-glucosidase were elevated. Interestingly, with the introduction of xylan, five out of six cellulolytic genes were upregulated. Furthermore, approximately 1122 genes equivalent to one-third of the total genes for strain RA were upregulated. These upregulated genes were mostly involved in transportation, chemotaxis, and membrane components synthesis.

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An insight into microbial sources, classification, and industrial applications of xylanases: A rapid review

Kaur

Joshi

Sharma

et al. 2023

Biotech and App Biochem

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Endo 1,4‐β‐d‐xylanases (EC3.2.1.8) are one of the key lignocellulose hydrolyzing enzymes. Xylan, which is present in copious amounts on earth, forms the primary substrate of endo‐xylanases, which can unchain the constituent monosaccharides linked via β‐1,4‐glycosidic bonds from the xylan backbone. Researchers have shown keen interest in the xylanases belonging to glycoside hydrolase families 10 and 11, whereas those placed in other glycoside hydrolase families are yet to be investigated. Various microbes such as bacteria and fungi harbor these enzymes for the metabolism of their lignocellulose fibers. These microbes can be used as miniature biofactories of xylanase enzymes for a plethora of environmentally benign applications in pulp and paper industry, biofuel production, and for improving the quality of food in bread baking and fruit juice industry. This review highlights the potential of microbes in production of xylanase for industrial biotechnology.

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Heterologous expression, purification and biochemical characterization of a new endo-1,4-β-xylanase from Rhodothermaceae bacterium RA

Cited by 22 publications

References 55 publications

Co-expression of cellulase and xylanase genes in Saccharomyces cerevisiae toward enhanced bioethanol production from corn stover

Co-expression of cellulase and xylanase genes in Saccharomyces cerevisiae toward enhanced bioethanol production from corn stover

Global Transcriptomic Responses of Roseithermus sacchariphilus Strain RA in Media Supplemented with Beechwood Xylan

An insight into microbial sources, classification, and industrial applications of xylanases: A rapid review

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