Cloning and expression of a family 10 xylanase gene (Aoxyn10) from Aspergillus oryzae in Pichia pastoris

Yin, Xin; Yan-Yan, Gong; Wang, Junqing; Tang, Cun-Duo; Wu, Minchen

doi:10.2323/jgam.59.405

Cited by 7 publications

(5 citation statements)

References 25 publications

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“…For the first time heterologous expression of 10th family xylanase Aoxyn10 from A. oryzae in Pichia pastoris was described in Yin et al’s work [ 16 ]. Gene AoXyn10 consisted of 2308 bp, including cDNA was 1689 bp, 5′,3′-non-translating region, and 1422 bp.…”

Section: Discussionmentioning

confidence: 99%

“…xAor gene was inserted into four tandem copies since it was shown that multicopy plasmids with recombinant DNA tandem clusters are effective for increasing production of target protein in case of K. phaffii transformation [ 17 ]. Extracellularly expressed Aoxyn10 activity was shown to be 45 IU/mL [ 16 ], while activity of xylanase xAor was 258,240 IU/mL in culture liquid.…”

Section: Discussionmentioning

confidence: 99%

“…For Aspergillus oryzae F1 endo-1,4-xylanase, it was shown that pH optimum is 5.0, and temperature optimum is 60 °C [ 20 ]. Extracellularly expressed Aoxyn10, pH optimum 5.5, and temperature optimum 60 °C, nonetheless xylanase was stable in pH range from 4.0 to 7.0, and temperatures lower than 50 °C [ 16 ]. Bhardwaj et al [ 5 ] showed that xylanase XynF1 from A. oryzae LC1 in expression system E. coli BL21 (DE3) had specific enzyme activity 1037.3 EU/mg, being 9.3 times higher, than in host organism.…”

Section: Discussionmentioning

confidence: 99%

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Heterologous Expression of Xylanase xAor from Aspergillus oryzae in Komagataella phaffii T07

Zadorozhny

Ushakov

Розанов

et al. 2022

IJMS

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Xylanases (EC 3.2.1.8) hydrolyze the hemicellulose of plant cell walls. Xylanases are used in the food and paper industries and for bioconversion of lignocellulose to biofuel. In this work, the producer-strain with four copies of the xAor xylanase gene was organized in two tandem copies for optimal expression in Komagataella phaffii T07 yeast. The secreted 35 kDa xylanase was purified from culture medium by gel filtration on Sephadex G-25 and anion exchange chromatography on DEAE-Sepharose 6HF. Tryptic peptides of the recombinant enzyme were analyzed by liquid chromatography-tandem mass spectrometry where the amino acid sequence corresponded to Protein Accession # O94163 for Endo-1,4-beta-xylanase from Aspergillus oryzae RIB40. The recombinant xylanase was produced in a bioreactor where the secreted enzyme hydrolyzed oat xylane with an activity of 258240 IU/mL. High activity in the culture medium suggested xylanase could be used for industrial applications without being purified or concentrated. The pH optimum for xylanase xAor was 7.5, though the enzyme was active from pH 2.5 to pH 10. Xylanase was active at temperatures from 35 °C to 85 °C with a maximum at 60 °C. In conclusion, this protocol yields soluble, secreted xylanase suitable for industrial scale production.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Heterologous Expression of Xylanase xAor from Aspergillus oryzae in Komagataella phaffii T07

Zadorozhny

Ushakov

Розанов

et al. 2022

IJMS

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“…Enzyme assay. Xylanase activity was assayed by estimating the amount of reducing sugars liberated from 0.5% birchwood xylan, using a 3,5-dinitrosalicylic acid method (Li et al, 2013;Sriprang et al, 2006;Yin et al, 2013aYin et al, , 2014. The reducing sugars were liberated by incubating 1 mL 0.5% birchwood xylan (in 50 mM Na 2 HPO 4 -citric acid buffer, pH 4.6) with 1.5 mL xylanase for 15 min at 40°C.…”

Section: Methodsmentioning

confidence: 99%

Exploration of a N-terminal disulfide bridge to improve the thermostability of a GH11 xylanase from <i>Aspergillus niger</i>

Zhou

Wang

et al. 2016

J. Gen. Appl. Microbiol.

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“…Aspergillus oryzae belongs to a group of filamentous fungi that has long been used for the commercial production of different industrial enzymes, such as alpha-amylases [ 1 ], proteases [ 2 ], glucoamylases [ 3 ], xylanases [ 4 ], other hydrolytic enzymes [ 5 ], and organic acids [ 6 ]. Not only does A. oryzae produce various biological compounds, but also it has beneficial features, such as acting as a robust host system with high production yields and acclimatization to environmental and nutritional duress [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

Sequence- and Structure-Based Functional Annotation and Assessment of Metabolic Transporters inAspergillus oryzae: A Representative Case Study

Raethong

Wong-ekkabut

Laoteng

et al. 2016

BioMed Research International

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Aspergillus oryzae is widely used for the industrial production of enzymes. In A. oryzae metabolism, transporters appear to play crucial roles in controlling the flux of molecules for energy generation, nutrients delivery, and waste elimination in the cell. While the A. oryzae genome sequence is available, transporter annotation remains limited and thus the connectivity of metabolic networks is incomplete. In this study, we developed a metabolic annotation strategy to understand the relationship between the sequence, structure, and function for annotation of A. oryzae metabolic transporters. Sequence-based analysis with manual curation showed that 58 genes of 12,096 total genes in the A. oryzae genome encoded metabolic transporters. Under consensus integrative databases, 55 unambiguous metabolic transporter genes were distributed into channels and pores (7 genes), electrochemical potential-driven transporters (33 genes), and primary active transporters (15 genes). To reveal the transporter functional role, a combination of homology modeling and molecular dynamics simulation was implemented to assess the relationship between sequence to structure and structure to function. As in the energy metabolism of A. oryzae, the H+-ATPase encoded by the AO090005000842 gene was selected as a representative case study of multilevel linkage annotation. Our developed strategy can be used for enhancing metabolic network reconstruction.

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Cloning and expression of a family 10 xylanase gene (Aoxyn10) from Aspergillus oryzae in Pichia pastoris

Cited by 7 publications

References 25 publications

Heterologous Expression of Xylanase xAor from Aspergillus oryzae in Komagataella phaffii T07

Heterologous Expression of Xylanase xAor from Aspergillus oryzae in Komagataella phaffii T07

Exploration of a N-terminal disulfide bridge to improve the thermostability of a GH11 xylanase from <i>Aspergillus niger</i>

Sequence- and Structure-Based Functional Annotation and Assessment of Metabolic Transporters inAspergillus oryzae: A Representative Case Study

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