Cellulase-free xylanase by thermophilic fungi: a comparison of xylanase production by two Thermomyces lanuginosus strains

1995 ◽  
Vol 43 (4) ◽  
pp. 604-609 ◽  
Author(s):  
M. M. Hoq ◽  
W.-D. Deckwer
Keyword(s):  
Thermomyces Lanuginosus ◽  
Thermophilic Fungi ◽  
Xylanase Production

scholarly journals Optimization Of Culture Conditions For The Production Of Xylanase By Two Thermophilic Fungi Under Solid State Fermentation

2013 ◽  
Vol 39 (1) ◽  
pp. 43-51 ◽  
Author(s):  
Kumkum Azad ◽  
Md. Abdul Halim ◽  
Feroza Hossain
Keyword(s):  
Solid State ◽  
Solid State Fermentation ◽  
Culture Conditions ◽  
Maximum Activity ◽  
Thermomyces Lanuginosus ◽  
Thermophilic Fungi ◽  
Thermostable Xylanase ◽  
Optimum Time ◽  
Xylanase Production ◽  
Time Required

Two thermophilic fungi, Thermomyces lanuginosus BPJ-10 and Rhizomucor pusillus BPJ-2 were studied under solid state fermentation (SSF) using wheat bran for the production of thermostable xylanase. The optimum time required for the production of xylanase was found to be 4 days and 7 days for R. pusillus BPJ-2 and T. lanuginosus BPJ-10 respectively. The optimum temperatures for the production of xylanase by R. pusillus BPJ-2 and T. lanuginosus BPJ-10 were 45°C and 50°C respectively. The maximum activity of xylanase (1.685 IU/ml and 0.075 IU/ml) was exhibited by T. lanuginosus BPJ-10 and R. pusillus BPJ-2 at pH 7.0 and pH 4.0 respectively. The optimum moisture content for maximum xylanase production was 90% for both fungi. J. Asiat. Soc. Bangladesh, Sci. 39(1): 43-51, June 2013 DOI: http://dx.doi.org/10.3329/jasbs.v39i1.16032

Improvement of xylanase production by thermophilic fungus Thermomyces lanuginosus SDYKY-1 using response surface methodology

2011 ◽  
Vol 28 (1) ◽  
pp. 40-46 ◽  
Author(s):  
Yishan Su ◽  
Xiaoyuan Zhang ◽  
Zhongwen Hou ◽  
Xiqiang Zhu ◽  
Xueping Guo ◽  
...  
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Thermophilic Fungus ◽  
Thermomyces Lanuginosus ◽  
Xylanase Production

Optimizing Xylanase Production by a Newly Isolated Strain CAU44 of the Thermophile Thermomyces lanuginosus

2005 ◽  
Vol 21 (6-7) ◽  
pp. 863-867 ◽  
Author(s):  
Z. Q. Jiang ◽  
S. Q. Yang ◽  
Q. J. Yan ◽  
L. T. Li ◽  
S. S. Tan
Keyword(s):  
Thermomyces Lanuginosus ◽  
Xylanase Production

scholarly journals Cloning, characterization and directed evolution of a xylosidase from Aspergillus niger

2016 ◽  
Author(s):  
◽  
Bibi Khadija Khan
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Random Mutagenesis ◽  
Thermomyces Lanuginosus ◽  
Major Protein ◽  
Bioethanol Production ◽  
Xylanase Production ◽  
Fluorescent Band ◽  
Gap Promoter ◽  
Major Protein Band

β-xylosidases catalyse the hydrolyses of xylooligosaccharides into the monosaccharide sugar, xylose. In this study we report the production of xylose under different conditions in Pichia pastoris and Saccharomyces. cerevisiae, and its conversion to bioethanol using Pichia stipitis. The aim of this study was to change the characteristics of the A. niger 90196 β-xylosidase through random mutagenesis and increase expression under the control of different promoter systems in yeasts P. pastoris and S. cerevisiae. The recombinant library created through random mutagenesis was screened for changes in activity and subsequently pH and temperature stability. One variant showed an increase in enzyme expression, thermostability, and a change in amino acid sequence at residue 226. The enzyme was then cloned, expressed and characterized in P. pastoris GS115 and S. cerevisiae. β-xylosidase was constitutively expressed in P. pastoris using the GAP promoter and the inducible AOX promoter. In S. cerevisiae the enzyme was expressed using the constitutive PGK promoter and inducible ADH2 promoter systems. Enzyme functionality with the different expression systems was compared in both hosts. The GAP system was identified as the highest-producing system in P. pastoris, yielding 70 U/ml after 72 hours, followed by the PGK system in S. cerevisiae, with 8 U/ml. A 12% SDS-PAGE gel revealed a major protein band with an estimated molecular mass of 120 kDA, and the zymogram analysis revealed that this band is a fluorescent band under UV illumination, indicating enzyme activity. Stability characteristics was determined by expressing the enzyme at different pH and temperatures. Under the control of the GAP promoter in P. pastoris, enzyme activity peaked at pH4 while retaining 80% activity between pH 3 – 5. Highest activity of 70 U/ml xylosidase was recorded at 60ºC. Due to the high enzyme production in P. pastoris, the co-expression of this enzyme with a fungal xylanase was evaluated. The xylanase gene from Thermomyces lanuginosus was cloned with the GAP promoter system and expressed together with the β-xylosidase recombinant in P. pastoris. Enzyme activities of the co-expressed recombinant revealed a decrease in enzyme activity levels. The co-expressed xylanase production decreased by 26% from 136 U/ml to 100 U/ml while the xylosidase expression decreased 86% from 70 U/ml to 10 U/ml. The xylose produced from the hydrolysis of birchwood xylan was quantified by HPLC. The monosaccharide sugar was used in a separate saccharification and fermentation strategy by P. stipitis to produce bioethanol, quantified by gas chromatography. Bioethanol production peaked at 72 h producing 0.7% bioethanol from 10 g/l xylose. In conclusion a β-xylosidase from Aspergillus niger was successfully expressed in P. pastoris and was found to express large quantities of xylosidase, that has not been achieved in any prior research to date. The enzyme was also successfully co-expressed with a Thermomyces xylanase and is now capable of bioethanol production through xylan hydrolysis. This highlights potential use in industrial applications in an effort to reduce the world dependence on petroleum and fossil fuels. However the technical challenges associated with commercialization of bioethanol production are still significant.

scholarly journals Production of Phytase by three thermophilic fungi

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Ranjith Kumar R ◽  
Rajendar Reddy ◽  
Girisham S ◽  
Reddy S.M.
Keyword(s):  
Carbon Source ◽  
Nitrogen Sources ◽  
The Other ◽  
Thermomyces Lanuginosus ◽  
Thermophilic Fungi ◽  
Carbon And Nitrogen Sources ◽  
Phytase Production ◽  
Carbon And Nitrogen ◽  
Cultural Conditions ◽  
Other Hand

Production of phytase by three thermophilic fungi, Thermomyces lanuginosus, Talaromyces luteus and Rhizomucor pusillus under different cultural conditions was assessed. Temperature of 45°C, pH-6.0 were optimum for phytase production by the all three fungi under investigation . Carbon and nitrogen sources for production of phytases by the three thermophilic fungi varied with the fungus. When T. lanuginosus opted for D-glucose followed by D-fructose, T. luteus preferred D-glucose, D-mannose and mannitol for production a phytase. On the other hand, R. pusillus produced maximum phytase during its growth on mannitol and maltose as carbon source. L- asparagine, L- arginine and L-asparatic acid were preferred nitrogen sources for production of phytase by T. lanuginosus. On the other hand T. luteus, opted for L- asparagine, L-glutamic acid and L- glycine for the activity of phytase. R. pusillus produced maximum phytase in medium containing L-argine, L-asparagine and L- asparatic acid.

scholarly journals Characterization of Thermophilic Lignocellulolytic Microorganisms in Composting

2021 ◽  
Vol 12 ◽  
Author(s):  
María J. López ◽  
Macarena M. Jurado ◽  
Juan A. López-González ◽  
María J. Estrella-González ◽  
María R. Martínez-Gallardo ◽  
...  
Keyword(s):  
Thermophilic Bacteria ◽  
Wide Spectrum ◽  
Deep Understanding ◽  
Thermomyces Lanuginosus ◽  
Thermophilic Fungi ◽  
Hemicellulose Degradation ◽  
Mechanisms Of Adaptation ◽  
Prevalent Species ◽  
Selection Of

Composting involves the selection of a microbiota capable of resisting the high temperatures generated during the process and degrading the lignocellulose. A deep understanding of the thermophilic microbial community involved in such biotransformation is valuable to improve composting efficiency and to provide thermostable biomass-degrading enzymes for biorefinery. This study investigated the lignocellulose-degrading thermophilic microbial culturome at all the stages of plant waste composting, focusing on the dynamics, enzymes, and thermotolerance of each member of such a community. The results revealed that 58% of holocellulose (cellulose plus hemicellulose) and 7% of lignin were degraded at the end of composting. The whole fungal thermophilic population exhibited lignocellulose-degrading activity, whereas roughly 8–10% of thermophilic bacteria had this trait, although exclusively for hemicellulose degradation (xylan-degrading). Because of the prevalence of both groups, their enzymatic activity, and the wide spectrum of thermotolerance, they play a key role in the breakdown of hemicellulose during the entire process, whereas the degradation of cellulose and lignin is restricted to the activity of a few thermophilic fungi that persists at the end of the process. The xylanolytic bacterial isolates (159 strains) included mostly members of Firmicutes (96%) as well as a few representatives of Actinobacteria (2%) and Proteobacteria (2%). The most prevalent species were Bacillus licheniformis and Aeribacillus pallidus. Thermophilic fungi (27 strains) comprised only four species, namely Thermomyces lanuginosus, Talaromyces thermophilus, Aspergillus fumigatus, and Gibellulopsis nigrescens, of whom A. fumigatus and T. lanuginosus dominated. Several strains of the same species evolved distinctly at the stages of composting showing phenotypes with different thermotolerance and new enzyme expression, even not previously described for the species, as a response to the changing composting environment. Strains of Bacillus thermoamylovorans, Geobacillus thermodenitrificans, T. lanuginosus, and A. fumigatus exhibiting considerable enzyme activities were selected as potential candidates for the production of thermozymes. This study lays a foundation to further investigate the mechanisms of adaptation and acquisition of new traits among thermophilic lignocellulolytic microorganisms during composting as well as their potential utility in biotechnological processing.

scholarly journals Optimized Expression of a Thermostable Xylanase from Thermomyces lanuginosus in Pichia pastoris

2003 ◽  
Vol 69 (10) ◽  
pp. 6064-6072 ◽  
Author(s):  
Mônica C. Triches Damaso ◽  
Marcius S. Almeida ◽  
Eleonora Kurtenbach ◽  
Orlando B. Martins ◽  
Nei Pereira ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Cell Density ◽  
Thermomyces Lanuginosus ◽  
Small Scale ◽  
Efficient Production ◽  
Optimum Temperature ◽  
Nitrogen Base ◽  
Xylanase Production ◽  
Initial Cell ◽  
Initial Cell Density

ABSTRACT Highly efficient production of a Thermomyces lanuginosus IOC-4145 β-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 23 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°C, and they retained 60% of their original activity after 80 min at 70°C or 40 min at 80°C. The high level of fully active recombinant xylanase obtained in P. pastoris makes this expression system attractive for fermentor growth and industrial applications.

scholarly journals Investigations of the bioprocess parameters for the production of hemicellulases by Thermomyces lanuginosus strains

2012 ◽  
Author(s):  
◽  
Santhosh Kumar Kuttan Pillai
Keyword(s):  
Yield Enhancement ◽  
Thermomyces Lanuginosus ◽  
Xylanase Production ◽  
Medium Components ◽  
Pulp Properties ◽  
Agitation And Aeration ◽  
Bagasse Pulp

The aim of this study was to evaluate T. lanuginosus for the production of hemicellulases, its yield enhancement using mutagenesis and application of a selected xylanase on bagasse pupl to assess the improvement of pulp properties. The objectives were: To determine the localization of hemicellulases in T. lanuginosus strains, To develop high yielding strains of T. lanuginosus through mutagenensis, To investigate the synthesis of xylanase by T. lanuginosus MC134, To optimize the medium components and cultural conitions of T. lanuginosus MC134 strain, To study the influence of agitation and aeration on the production of xylanase by T. lanuginosus MC134 in a fermenter, To evaluate the bleach boosting abilities of T. lanuginosus xylanase on bagasse pulp, To evaluate simultaneous xylanase production and biobleaching potential of T. lanuginosus.

scholarly journals Identification and Characterization of Thermostable Y-Family DNA Polymerases η, ι, κ and Rev1 From a Lower Eukaryote, Thermomyces lanuginosus

2021 ◽  
Vol 8 ◽  
Author(s):  
Alexandra Vaisman ◽  
John P. McDonald ◽  
Mallory R. Smith ◽  
Sender L. Aspelund ◽  
Thomas C. Evans ◽  
...  
Keyword(s):  
Biochemical Characterization ◽  
Dna Polymerases ◽  
Biochemical Properties ◽  
Thermomyces Lanuginosus ◽  
Thermophilic Fungi ◽  
Domains Of Life ◽  
Y Family ◽  
Identification And Characterization ◽  
Lower Eukaryote

Y-family DNA polymerases (pols) consist of six phylogenetically separate subfamilies; two UmuC (polV) branches, DinB (pol IV, Dpo4, polκ), Rad30A/POLH (polη), and Rad30B/POLI (polι) and Rev1. Of these subfamilies, DinB orthologs are found in all three domains of life; eubacteria, archaea, and eukarya. UmuC orthologs are identified only in bacteria, whilst Rev1 and Rad30A/B orthologs are only detected in eukaryotes. Within eukaryotes, a wide array of evolutionary diversity exists. Humans possess all four Y-family pols (pols η, ι, κ, and Rev1), Schizosaccharomyces pombe has three Y-family pols (pols η, κ, and Rev1), and Saccharomyces cerevisiae only has polη and Rev1. Here, we report the cloning, expression, and biochemical characterization of the four Y-family pols from the lower eukaryotic thermophilic fungi, Thermomyces lanuginosus. Apart from the expected increased thermostability of the T. lanuginosus Y-family pols, their major biochemical properties are very similar to properties of their human counterparts. In particular, both Rad30B homologs (T. lanuginosus and human polɩ) exhibit remarkably low fidelity during DNA synthesis that is template sequence dependent. It was previously hypothesized that higher organisms had acquired this property during eukaryotic evolution, but these observations imply that polι originated earlier than previously known, suggesting a critical cellular function in both lower and higher eukaryotes.

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