scholarly journals Optimization of β-1,4-Endoxylanase Production by an Aspergillus niger Strain Growing on Wheat Straw and Application in Xylooligosaccharides Production

Molecules ◽  
2021 ◽  
Vol 26 (9) ◽  
pp. 2527
Author(s):  
Zahra Azzouz ◽  
Azzeddine Bettache ◽  
Nawel Boucherba ◽  
Alicia Prieto ◽  
Maria Jesus Martinez ◽  
...  

Plant biomass constitutes the main source of renewable carbon on the planet. Its valorization has traditionally been focused on the use of cellulose, although hemicellulose is the second most abundant group of polysaccharides on Earth. The main enzymes involved in plant biomass degradation are glycosyl hydrolases, and filamentous fungi are good producers of these enzymes. In this study, a new strain of Aspergillus niger was used for hemicellulase production under solid-state fermentation using wheat straw as single-carbon source. Physicochemical parameters for the production of an endoxylanase were optimized by using a One-Factor-at-a-Time (OFAT) approach and response surface methodology (RSM). Maximum xylanase yield after RSM optimization was increased 3-fold, and 1.41- fold purification was achieved after ultrafiltration and ion-exchange chromatography, with about 6.2% yield. The highest activity of the purified xylanase was observed at 50 °C and pH 6. The enzyme displayed high thermal and pH stability, with more than 90% residual activity between pH 3.0–9.0 and between 30–40 °C, after 24 h of incubation, with half-lives of 30 min at 50 and 60 °C. The enzyme was mostly active against wheat arabinoxylan, and its kinetic parameters were analyzed (Km = 26.06 mg·mL−1 and Vmax = 5.647 U·mg−1). Wheat straw xylan hydrolysis with the purified β-1,4 endoxylanase showed that it was able to release xylooligosaccharides, making it suitable for different applications in food technology.

Author(s):  
Zahra AZZOUZ ◽  
Azzeddine Bettache ◽  
Nawel Boucherba ◽  
Laura de Eugenio ◽  
Maria Martinez ◽  
...  

Plant biomass constitutes the main resource of renewable carbon in the planet and its valorization has traditionally been focused on the use of cellulose, although hemicellulose is the second most abundant group of polysaccharides on earth. Enzymes involved in its degradation are usually glycosyl hydrolases and filamentous fungi are good producers of these enzymes. In this study, a new strain of Aspergillus niger was utilized for hemicellulase production under solid state fermentation using wheat straw as a single carbon source. Physicochemical parameters for production of an endoxylanase were optimized by using one factor at a time approach and response surface methodology (RSM). Maximum xylanase yield after RSM optimization was increased 3-fold. The enzyme was purified by ultrafiltration and ion-exchange chromatography1.41-fold, with 6.2 % yield. Highest xylanase activity was observed at 50 °C and pH 6. A high pH and thermal stability were found, greater than 90% residual activity between pH 3.0-9.0 and between 30-40°C, after 24 h of incubation, presenting half-lives of 30 min at 50 and 60°C. Enzyme was mostly active for wheat arabinoxylan, and displayed the following kinetic parameters Km of 26.06 mg•ml-1 and Vmax of 5,647 U•mg-1min-1.Wheat straw xylan hydrolysis with the purified β-1,4 endoxylanase showed that it was able to release xylooligosaccharides, making it suitable for different applications in food technology.


2012 ◽  
Vol 79 (4) ◽  
pp. 1316-1324 ◽  
Author(s):  
Joost van den Brink ◽  
Gonny C. J. van Muiswinkel ◽  
Bart Theelen ◽  
Sandra W. A. Hinz ◽  
Ronald P. de Vries

ABSTRACTRapid and efficient enzymatic degradation of plant biomass into fermentable sugars is a major challenge for the sustainable production of biochemicals and biofuels. Enzymes that are more thermostable (up to 70°C) use shorter reaction times for the complete saccharification of plant polysaccharides compared to hydrolytic enzymes of mesophilic fungi such asTrichodermaandAspergillusspecies. The genusMyceliophthoracontains four thermophilic fungi producing industrially relevant thermostable enzymes. Within this genus, isolates belonging toM. heterothallicawere recently separated from the well-described speciesM. thermophila. We evaluate here the potential ofM. heterothallicaisolates to produce efficient enzyme mixtures for biomass degradation. Compared to the other thermophilicMyceliophthoraspecies, isolates belonging toM. heterothallicaandM. thermophilagrew faster on pretreated spruce, wheat straw, and giant reed. According to their protein profiles andin vitroassays after growth on wheat straw, (hemi-)cellulolytic activities differed strongly betweenM. thermophilaandM. heterothallicaisolates. Compared toM. thermophila,M. heterothallicaisolates were better in releasing sugars from mildly pretreated wheat straw (with 5% HCl) with a high content of xylan. The high levels of residual xylobiose revealed that enzyme mixtures ofMyceliophthoraspecies lack sufficient β-xylosidase activity. Sexual crossing of twoM. heterothallicashowed that progenies had a large genetic and physiological diversity. In the future, this will allow further improvement of the plant biomass-degrading enzyme mixtures ofM. heterothallica.


2020 ◽  
Vol 21 (21) ◽  
pp. 8402
Author(s):  
Wissal Ben Ali ◽  
Amal Ben Ayed ◽  
Annick Turbé-Doan ◽  
Emmanuel Bertrand ◽  
Yann Mathieu ◽  
...  

Only a few studies have examined how marine-derived fungi and their enzymes adapt to salinity and plant biomass degradation. This work concerns the production and characterisation of an oxidative enzyme identified from the transcriptome of marine-derived fungus Stemphylium lucomagnoense. The laccase-encoding gene SlLac2 from S. lucomagnoense was cloned for heterologous expression in Aspergillus niger D15#26 for protein production in the extracellular medium of around 30 mg L−1. The extracellular recombinant enzyme SlLac2 was successfully produced and purified in three steps protocol: ultrafiltration, anion-exchange chromatography, and size exclusion chromatography, with a final recovery yield of 24%. SlLac2 was characterised by physicochemical properties, kinetic parameters, and ability to oxidise diverse phenolic substrates. We also studied its activity in the presence and absence of sea salt. The molecular mass of SlLac2 was about 75 kDa, consistent with that of most ascomycete fungal laccases. With syringaldazine as substrate, SlLac2 showed an optimal activity at pH 6 and retained nearly 100% of its activity when incubated at 50°C for 180 min. SlLac2 exhibited more than 50% of its activity with 5% wt/vol of sea salt.


BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Claire Khosravi ◽  
Joanna E. Kowalczyk ◽  
Tania Chroumpi ◽  
Evy Battaglia ◽  
Maria-Victoria Aguilar Pontes ◽  
...  

Abstract Background Enzymatic plant biomass degradation by fungi is a highly complex process and one of the leading challenges in developing a biobased economy. Some industrial fungi (e.g. Aspergillus niger) have a long history of use with respect to plant biomass degradation and for that reason have become ‘model’ species for this topic. A. niger is a major industrial enzyme producer that has a broad ability to degrade plant based polysaccharides. A. niger wild-type, the (hemi-)cellulolytic regulator (xlnR) and xylulokinase (xkiA1) mutant strains were grown on a monocot (corn stover, CS) and dicot (soybean hulls, SBH) substrate. The xkiA1 mutant is unable to utilize the pentoses D-xylose and L-arabinose and the polysaccharide xylan, and was previously shown to accumulate inducers for the (hemi-)cellulolytic transcriptional activator XlnR and the arabinanolytic transcriptional activator AraR in the presence of pentoses, resulting in overexpression of their target genes. The xlnR mutant has reduced growth on xylan and down-regulation of its target genes. The mutants therefore have a similar phenotype on xylan, but an opposite transcriptional effect. D-xylose and L-arabinose are the most abundant monosaccharides after D-glucose in nearly all plant-derived biomass materials. In this study we evaluated the effect of the xlnR and xkiA1 mutation during growth on two pentose-rich substrates by transcriptome analysis. Results Particular attention was given to CAZymes, metabolic pathways and transcription factors related to the plant biomass degradation. Genes coding for the main enzymes involved in plant biomass degradation were down-regulated at the beginning of the growth on CS and SBH. However, at a later time point, significant differences were found in the expression profiles of both mutants on CS compared to SBH. Conclusion This study demonstrates the high complexity of the plant biomass degradation process by fungi, by showing that mutant strains with fairly straightforward phenotypes on pure mono- and polysaccharides, have much less clear-cut phenotypes and transcriptomes on crude plant biomass.


Author(s):  
Roland S. Kun ◽  
Sandra Garrigues ◽  
Marcos Di Falco ◽  
Adrian Tsang ◽  
Ronald P. Vries

Processes ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 30
Author(s):  
María González Martínez ◽  
Estéban Hélias ◽  
Gilles Ratel ◽  
Sébastien Thiéry ◽  
Thierry Melkior

Biomass preheating in torrefaction at an industrial scale is possible through a direct contact with the hot gases released. However, their high water-content implies introducing moisture (around 20% v/v) in the torrefaction atmosphere, which may impact biomass thermochemical transformation. In this work, this situation was investigated for wheat straw, beech wood and pine forest residue in torrefaction in two complementary experimental devices. Firstly, experiments in chemical regime carried out in a thermogravimetric analyzer (TGA) showed that biomass degradation started from lower temperatures and was faster under a moist atmosphere (20% v/v water content) for all biomass samples. This suggests that moisture might promote biomass components’ degradation reactions from lower temperatures than those observed under a dry atmosphere. Furthermore, biomass inorganic composition might play a role in the extent of biomass degradation in torrefaction in the presence of moisture. Secondly, torrefaction experiments on a lab-scale device made possible to assess the influence of temperature and residence time under dry and 100% moist atmosphere. In this case, the difference in solid mass loss between dry and moist torrefaction was only significant for wheat straw. Globally, an effect of water vapor on biomass transformation through torrefaction was observed (maximum 10%db), which appeared to be dependent on the biomass type and composition.


2020 ◽  
Vol 4 (1) ◽  
pp. 15 ◽  
Author(s):  
Mohamed H. Mohamed ◽  
Inimfon A. Udoetok ◽  
Lee D. Wilson

Pelletized biomaterial composites that contain chitosan (C) and torrefied wheat straw (S) at variable weight composition (C:S) were prepared using a facile blending process. The fractional content of the wheat straw was studied to elucidate the role of biomass on the pelletized product and effects of S-content on the physicochemical properties relevant to adsorption phenomena. Chitosan pellets (with and without S) were characterized by spectroscopic (FT-IR and 13C NMR) and thermal (TGA and DSC) techniques to provide support for their respective C:S composition. Confocal microscopy using fluorescein (FL) as a dye probe revealed the presence and an increase in the accessibility of the active sites for the composite pellets according to the S-content (wt %). Equilibrium and kinetic sorption studies using FL and reactive black (RB) dyes revealed an incremental adsorption affinity of the pellets with anionic dyes in variable charge states (FL and RB). The trend for dye adsorption parallels the incremental S-content (wt %) in the composite pellets. This study reports a first-example of a low-cost, facile, and sustainable approach for the valorization of straw and chitosan suitable for sorption-based applications in aqueous media.


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