A thermophilic, pH-tolerant, and highly active GH10 xylanase from Aspergillus fumigatus boosted pre-treated sugarcane bagasse saccharification by cellulases

Bernardi, Aline Vianna; Gerolamo, Luís Eduardo; Uyemura, Sérgio Akira; Dinamarco, Taísa Magnani

doi:10.1016/j.indcrop.2021.113697

Cited by 16 publications

(9 citation statements)

References 117 publications

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“…4,5 To date, D-mannose isomerase has not, to our knowledge, been produced by K. phaffii, which is an excellent candidate for secretory expression of numerous enzymes. 9,10 In this study, XpMIaseA was successfully produced by K. phaffii with the production levels of 282.0 U mL −1 (as D-fructose forming) and 106.6 U mL −1 (as D-mannose forming) (Figure 2A), which is 2.1 times higher than that of the D-mannose isomerase produced by B. subtilis (51.2 U mL −1 as mannose forming). 4 This is the highest production level of D-mannose isomerases reported to date.…”

Section: Production and Purification Ofmentioning

confidence: 73%

“…coli shows a production level of 51.2 U mL –1 Komagataella phaffii (Pichia pastoris) is another excellent host for secretory expression, which is widely used to produce glycoside hydrolases such as β-mannanase and β-xylanase. , This host provides an effective alternative for the production and application of d -mannose isomerases. However, none of the d -mannose isomerases is produced by K.…”

Section: Introductionmentioning

confidence: 99%

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One-Pot Three-Enzyme System for Production of a Novel Prebiotic Mannosyl-β-(1 → 4)-Fructose Using a d-Mannose Isomerase from Xanthomonas phaseoli

Xiaohan

Yan

et al. 2022

J. Agric. Food Chem.

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The present supply of prebiotics is entirely inadequate to meet their demand. To produce novel prebiotics, a d-mannose isomerase (XpMIaseA) from Xanthomonas phaseoli was first produced in Komagataella phaffii (Pichia pastoris). XpMIaseA shared the highest amino acid sequence identity (58.0%) with the enzyme from Marinomonas mediterranea. Efficient secretory production of XpMIaseA (282.0 U mL–1) was achieved using high cell density fermentation. The optimal conditions of XpMIaseA were pH 7.5 and 55 °C. It showed a broad substrate specificity, which isomerized d-mannose, d-talose, mannobiose, epilactose, and mannotriose. XpMIaseA was employed to construct a one-pot three-enzyme system for the production of mannosyl-β-(1 → 4)-fructose (MF) using mannan (5%, w/v) as the substrate. The equilibrium yield of MF was 58.2%. In in vitro fermentations, MF significantly stimulated (≤3.2-fold) the growth of 12 among 15 tested Bifidobacterium and Lactobacillus strains compared with fructo-oligosaccharides. Thus, the novel d-mannose isomerase provides a one-pot bioconversion strategy for efficiently producing novel prebiotics.

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Section: Production and Purification Ofmentioning

confidence: 73%

Section: Introductionmentioning

confidence: 99%

One-Pot Three-Enzyme System for Production of a Novel Prebiotic Mannosyl-β-(1 → 4)-Fructose Using a d-Mannose Isomerase from Xanthomonas phaseoli

Xiaohan

Yan

et al. 2022

J. Agric. Food Chem.

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“…2 b), which was slightly higher than the theoretical mass of 50 kDa. The difference in sizes suggested that post-translational modifications occurred in Pf GH5 during heterologous expression in P. pastoris and these post-translational modifications had effects on the migration rate of the Pf GH5 on SDS-PAGE 23 . To gain insights into the action of this glycoside hydrolase in biomass deconstruction, the enzyme was screened on various polysaccharides, for its specificity on glycosidic bonds (Table 1 ).…”

Section: Resultsmentioning

confidence: 99%

Characterization of a GH5 endoxylanase from Penicillium funiculosum and its synergism with GH16 endo-1,3(4)-glucanase in saccharification of sugarcane bagasse

Ogunyewo

Okereke

Kumar

et al. 2022

Sci Rep

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The production of second-generation fuels from lignocellulosic residues such as sugarcane bagasse (SCB) requires the synergistic interaction of key cellulose-degrading enzymes and accessory proteins for their complete deconstruction to useful monomeric sugars. Here, we recombinantly expressed and characterized unknown GH5 xylanase from P. funiculosum (PfXyn5) in Pichia pastoris, which was earlier found in our study to be highly implicated in SCB saccharification. The PfXyn5 has a molecular mass of ~ 55 kDa and showed broad activity against a range of substrates like xylan, xyloglucan, laminarin and p-nitrophenyl-β-d-xylopyranoside, with the highest specific activity of 0.7 U/mg against xylan at pH 4.5 and 50 °C. Analysis of the degradation products of xylan and SCB by PfXyn5 showed significant production of xylooligosaccharides (XOS) with a degree of polymerization (DP) ranging from two (DP2) to six (DP6), thus, suggesting that the PfXyn5 is an endo-acting enzyme. The enzyme synergistically improved the saccharification of SCB when combined with the crude cellulase cocktail of P. funiculosum with a degree of synergism up to 1.32. The PfXyn5 was further expressed individually and simultaneously with a notable GH16 endoglucanase (PfEgl16) in a catabolite-derepressed strain of P. funiculosum, PfMig188, and the saccharification efficiency of the secretomes from the resulting transformants were investigated on SCB. The secretome of PfMig188 overexpressing Xyn5 or Egl16 increased the saccharification of SCB by 9% or 7%, respectively, over the secretome of PfMig188, while the secretome of dual transformant increased SCB saccharification by ~ 15% at the same minimal protein concentration.

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“…It was reported that bivalent cations such as Mg 2+ and Ca 2+ could act as activators of some enzymes by helping to stabilize the enzyme−substrate complex. 38,39 The inhibitory effect of metal ions and EDTA on the activity of BaCDA might be related to the formation of metalloprotein. 29,35 3.6.…”

Section: Deacetylation Activity and Kineticmentioning

confidence: 99%

Chitin Deacetylase from Bacillus aryabhattai TCI-16: Heterologous Expression, Characterization, and Deacetylation Performance

Li,

Liang,

Fang

et al. 2024

J. Agric. Food Chem.

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Chitin deacetylase (CDA) removes the acetyl group from the chitin molecule to generate chitosan in a uniform, highquality deacetylation pattern. Herein, BaCDA was a novel CDA discovered from our previously isolated Bacillus aryabhattai strain TCI-16, which was excavated from mangrove soil. The gene BaCDA was cloned and overexpressed in Escherichia coli BL21 (DE3) to facilitate its subsequent purification. The purified recombinant protein BaCDA was obtained at a concentration of about 1.2 mg/mL after Ni 2+ affinity chromatography. The molecular weight of BaCDA was around 28 kDa according to the sodium dodecyl sulfate− polyacrylamide gel electrophoresis (SDS−PAGE) analysis. In addition, BaCDA exhibited a significant deacetylation effect on colloidal chitin, and the deacetylation degree was measured from the initial 25.69 to 69.23% by Fourier transform infrared (FT-IR) spectroscopy. Scanning electron microscopy (SEM) observation showed that the surface of colloidal chitin after enzymatic digestion was rough, the crystal fibers disappeared, and the chitin structure was loose and porous with grooves. The results of electrospray ionization mass spectrometry (ESI-MS) showed that BaCDA had full-deacetylation activity against (GlcNAc) 4 . Molecular docking revealed that BaCDA had an open active pocket capable of binding to the GlcNAc unit. This study not only provides a novel enzymatic resource for the green and efficient application of chitin but also helps to deepen the understanding of the catalytic mechanism of CDA.

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A thermophilic, pH-tolerant, and highly active GH10 xylanase from Aspergillus fumigatus boosted pre-treated sugarcane bagasse saccharification by cellulases

Cited by 16 publications

References 117 publications

One-Pot Three-Enzyme System for Production of a Novel Prebiotic Mannosyl-β-(1 → 4)-Fructose Using a d-Mannose Isomerase from Xanthomonas phaseoli

One-Pot Three-Enzyme System for Production of a Novel Prebiotic Mannosyl-β-(1 → 4)-Fructose Using a d-Mannose Isomerase from Xanthomonas phaseoli

Characterization of a GH5 endoxylanase from Penicillium funiculosum and its synergism with GH16 endo-1,3(4)-glucanase in saccharification of sugarcane bagasse

Chitin Deacetylase from Bacillus aryabhattai TCI-16: Heterologous Expression, Characterization, and Deacetylation Performance

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