ArticleApplied and environmental microbiology2003
Optimized expression of a thermostable xylanase from Thermomyces lanuginosus in Pichia pastoris.
Article in Applied and environmental microbiology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 121 citations in OpenAlex.
- Isolation, expression, and in silico profiling of a thermostable xylanase from Geobacillus stearothermophilus strain NASA267: insights into structural features and agro-waste valorization.Microbial cell factories · 2025Article
- A Lipase Gene ofInternational journal of molecular sciences · 2024Article
- Aspergillus oryzae Rutinosidase: Biochemical and Structural Investigation.Applied and environmental microbiology · 2021Article
- Insights Into the Role of Exposed Surface Charged Residues in the Alkali-Tolerance of GH11 Xylanase.Frontiers in microbiology · 2020Article
- Exploiting xylan as sugar donor for the synthesis of an antiproliferative xyloside using an enzyme cascade.Microbial cell factories · 2019Article
- Engineered Pichia pastoris production of fusaruside, a selective immunomodulator.BMC biotechnology · 2019Article
- Aspergillus oryzae S2 AmyA amylase expression in Pichia pastoris: production, purification and novel properties.Molecular biology reports · 2019Article
- Single-Cell Approach to Monitor the Unfolded Protein Response During Biotechnological Processes WithFrontiers in microbiology · 2019Article
- Differential expression, molecular cloning, and characterization of porcine beta defensin 114.Journal of animal science and biotechnology · 2019Article
- Biobleaching of paper pulp with xylanase produced by3 Biotech · 2017Article
- Perspectives for clinical use of engineered human host defense antimicrobial peptides.FEMS microbiology reviews · 2017Review
- Expression, Docking, and Molecular Dynamics of Endo-BioMed research international · 2017Article
- Cloning and heterologous expression of cellulose free thermostable xylanase from Bacillus brevis.SpringerPlus · 2014Article
- Identification of a novel fungus, Leptosphaerulina chartarum SJTU59 and characterization of its xylanolytic enzymes.PloS one · 2013Article
- Characterization and pH-dependent substrate specificity of alkalophilic xylanase from Bacillus alcalophilus.Journal of industrial microbiology & biotechnology · 2012Article
- Recombinant sterol esterase from Ophiostoma piceae: an improved biocatalyst expressed in Pichia pastoris.Microbial cell factories · 2012Article
- Expression of lignocellulolytic enzymes in Pichia pastoris.Microbial cell factories · 2012Article
- High expression of recombinant Streptomyces sp. S38 xylanase in Pichia pastoris by codon optimization and analysis of its biochemical properties.Molecular biology reports · 2011Article
- Heterologous expression of a gene for thermostable xylanase from Chaetomium thermophilum in Pichia pastoris GS115.Molecular biology reports · 2011Article
- Thermostable enzymes as biocatalysts in the biofuel industry.Advances in applied microbiology · 2010Review
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Highly efficient production of a Thermomyces lanuginosus IOC-4145 beta-1,4-xylanase was achieved in Pichia pastoris under the control of the AOX1 promoter. P. pastoris colonies expressing recombinant xylanase were selected by enzymatic activity plate assay, and their ability to secrete high levels of the enzyme was evaluated in small-scale cultures. Furthermore, an optimization of enzyme production was carried out with a 2(3) factorial design. The influence of initial cell density, methanol, and yeast nitrogen base concentration was evaluated, and initial cell density was found to be the most important parameter. A time course profile of recombinant xylanase production in 1-liter flasks with the optimized conditions was performed and 148 mg of xylanase per liter was achieved. Native and recombinant xylanases were purified by gel filtration and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, circular dichroism spectroscopy, matrix-assisted laser desorption ionization-time of flight-mass spectrometry and physicochemical behavior. Three recombinant protein species of 21.9, 22.1, and 22.3 kDa were detected in the mass spectrum due to variability in the amino terminus. The optimum temperature, thermostability, and circular dichroic spectra of the recombinant and native xylanases were identical. For both enzymes, the optimum temperature was 75 degrees C, and they retained 60% of their original activity after 80 min at 70 degrees C or 40 min at 80 degrees C. The high level of fully active recombinant xylanase obtained in P. pastoris makes this expression system attractive for fermentor growth and industrial applications.
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