scholarly journals A Thermophilic GH5 Endoglucanase from Aspergillus fumigatus and Its Synergistic Hydrolysis of Mannan-Containing Polysaccharides

Catalysts ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 862
Author(s):  
Xinxi Gu ◽  
Haiqiang Lu ◽  
Lijuan Zhang ◽  
Xiangchen Meng

In this study, we isolated and identified a thermophilic strain of Aspergillus fumigatus from the “Daqu” samples. Transcriptomic analysis of A. fumigatus identified 239 carbohydrate-active enzymes (CAZy)-encoding genes, including 167 glycoside hydrolase (GH)-encoding genes, 58 glycosyltransferase (GT)-encoding genes, 2 polysaccharide lyase (PLs)-encoding genes and 12 carbohydrate esterase (CEs)-encoding genes, which indicates that the strain has a strong potential for application for enzyme production. Furthermore, we also identified a novel endoglucanase gene (AfCel5A), which was expressed in Pichia pastoris and characterized. The novel endoglucanase AfCel5A exhibited the highest hydrolytic activity against CMC-Na and the optimal activity at 80 °C and pH 4.0 and also showed good stability at pH 3.0–11.0 and below 70 °C. The Km and Vmax values of AfCel5 were 0.16 ± 0.05 mg·mL−1 and 7.23 ± 0.33 mol mg−1·min−1, respectively, using CMC-Na as a substrate. Further, the endoglucanase exhibited a high tolerance toward NaCl as well as glucose. In addition, the finding that the endoglucanase AfCel5A in combination with β-mannanse (ManBK) clearly increased the release of total reducing sugars of glucomannan by up to 74% is significant.

2018 ◽  
Vol 1145 ◽  
pp. 75-79 ◽  
Author(s):  
Hai Yun Ma ◽  
Zhi Ping Zhao ◽  
Peng Lu

The hydrolysis of cellulose into platform compounds and chemicals fuels has gained much attention to relieve the global energy crisis and environmental pollution. The filter paper (FP) cellulose with average degree of polymerization (DP) of 1000-1300 was dissolved in 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) firstly. And then acidic ionic liquids (ILs), ([(CH2)3SO3HVIm]HSO4) as the catalyst was applied to hydrolyze the FP cellulose by microwave heating. Compared with the oil bath heating method, microwave heating could effectively increase the total reducing sugars (TRS) yield about 10.7%. When the ratio of ILs catalyst to FP (w/w) was 0.167, and the ratio of deionized water to FP (w/w) was 0.833, the TRS yield was up to 60.8% within 20 min at 100°C.


2014 ◽  
Vol 7 ◽  
pp. 81-85
Author(s):  
Yakindra Prasad Timilsena ◽  
Nicolas Brosse

Various methods of pretreatments were investigated to their effect on cellulose to glucose conversion efficiency on enzymatic hydrolysis of a tropical agro-industrial waste residue (empty palm fruit bunch, EFB). Four different kinds of combinative pretreatments (autohydrolysis with and without naphthol, dilute acid prehydrolysis, soda prehydrolysis and enzymatic prehydrolysis) were tested for delignification during the first and the second steps of pretreatment. Each prehydrolysis step was seconded by the organosolv delignification with the same conditions of pretreatment. It was observed that all the combinative methods were far more efficient in delignification and enzymatic hydrolysis ability as compared to its one step counter parts. The combinative pretreatment method involving dilute acid prehydrolysis followed by organosolv delignification revealed the best result with respect to lignin removal and enzymatic hydrolysis. The resultant pulp contained very low Klason lignin (~5%) with high sugar conversion ratio (64% total reducing sugars). DOI: http://dx.doi.org/10.3126/jfstn.v7i0.10613   J. Food Sci. Technol. Nepal, Vol. 7 (81-85), 2012  


2019 ◽  
Vol 20 (9) ◽  
pp. 2261 ◽  
Author(s):  
Aline Vianna Bernardi ◽  
Deborah Kimie Yonamine ◽  
Sergio Akira Uyemura ◽  
Taisa Magnani Dinamarco

In the context of avoiding the use of non-renewable energy sources, employing lignocellulosic biomass for ethanol production remains a challenge. Cellulases play an important role in this scenario: they are some of the most important industrial enzymes that can hydrolyze lignocellulose. This study aims to improve on the characterization of a thermostable Aspergillus fumigatus endo-1,4-β-glucanase GH7 (Af-EGL7). To this end, Af-EGL7 was successfully expressed in Pichia pastoris X-33. The kinetic parameters Km and Vmax were estimated and suggested a robust enzyme. The recombinant protein was highly stable within an extreme pH range (3.0–8.0) and was highly thermostable at 55 °C for 72 h. Low Cu2+ concentrations (0.1–1.0 mM) stimulated Af-EGL7 activity up to 117%. Af-EGL7 was tolerant to inhibition by products, such as glucose and cellobiose. Glucose at 50 mM did not inhibit Af-EGL7 activity, whereas 50 mM cellobiose inhibited Af-EGL7 activity by just 35%. Additionally, the Celluclast® 1.5L cocktail supplemented with Af-EGL7 provided improved hydrolysis of sugarcane bagasse “in natura”, sugarcane exploded bagasse (SEB), corncob, rice straw, and bean straw. In conclusion, the novel characterization of Af-EGL7 conducted in this study highlights the extraordinary properties that make Af-EGL7 a promising candidate for industrial applications.


Energies ◽  
2019 ◽  
Vol 12 (15) ◽  
pp. 2952 ◽  
Author(s):  
Buzała ◽  
Kalinowska ◽  
Małachowska ◽  
Boruszewski ◽  
Krajewski ◽  
...  

The results of enzymatic hydrolysis of birch and beech kraft cellulosic pulps indicate that they may be promising feedstocks for fermentation processes including biofuel manufacturing. The aim of this study was to investigate whether birch and beech wood require the same degree of delignification by kraft pulping as pine wood. The differences observed in the efficiency of hydrolysis for the raw materials tested suggest that the differences in the anatomical structure of the examined wood in relation to pine wood is essential for the efficiency of the enzymatic hydrolysis process. The yields of glucose and other reducing sugars obtained from the birch and beech cellulosic pulps were similar (up to around 75% and 98.3% dry weight, and 76% and 98.6% dry weight, respectively). The highest glucose yields from cellulose contained in the birch and beech pulp were around 81.2% (at a Kappa number of 28.3) and 83.1% (at a Kappa number of 30.4), respectively. The maximum glucose yields and total reducing sugars of birch wood on a dry weight basis (39.8% and 52.1%, respectively) were derived from the pulp at a Kappa number of 28.3, while the highest yields of glucose and total reducing sugars of beech wood on a dry weight basis (around 36.9% and 48.2%, respectively) were reached from the pulp at a Kappa number of 25.3. To obtain the highest glucose yields and total reducing sugars of a wood on a dry weight basis, total lignin elimination from the birch and beech pulps was not necessary. However more in-depth delignification of birch and beech wood is required than for pine wood.


Processes ◽  
2019 ◽  
Vol 7 (2) ◽  
pp. 64 ◽  
Author(s):  
Nkosikho Dlangamandla ◽  
Seteno Ntwampe ◽  
Justine Angadam ◽  
Elie Itoba-Tombo ◽  
Boredi Chidi ◽  
...  

To sustainably operate a biorefinery with a low cost input in a commercial setting, the hydrolysis of lignocellulosic biomass must be undertaken in a manner which will impart environmental tolerance while reducing fermenter inhibitors from the delignification process. The challenge lies with the highly recalcitrant lignin structure, which limits the conversion of the holocelluloses to fermentable total reducing sugars (TRS). Due to these challenges, sustainable and innovative methods to pre-treat biomass must be developed for delignocellulolytic operations. Herein, Nepenthes mirabilis digestive fluids shown to have ligninolytic, cellulolytic and xylanolytic activities were used as an enzyme cocktail to hydrolyse mixed agro-waste constituted by Citrus sinensis (orange), Malus domestica (apple) peels, cobs from Zea mays (maize) and Quercus robur (oak) yard waste. The digestive fluids contained carboxylesterases (529.41 ± 30.50 U/L), β-glucosidases (251.94 ± 11.48 U/L) and xylanases (36.09 ± 18.04 U/L), constituting an enzymatic cocktail with significant potential for the reduction in total residual phenolic compounds (TRPCs), while being appropriate for holocellulose hydrolysis. Furthermore, the maximum TRS obtainable was 310 ± 5.19 mg/L within 168 h, while the TRPCs were reduced from 6.25 ± 0.18 to 4.26 ± 0.09 mg/L, which was lower than that observed when conventional methods were used. Overall, N. mirabilis digestive fluids demonstrated an ability to support biocatalytic processes with minimised cellulases hydrolysis interference. Therefore, the digestive enzymes in N. mirabilis pods can be used in an integrated system for feedstock hydrolysis in a second generation biorefinery.


RSC Advances ◽  
2015 ◽  
Vol 5 (125) ◽  
pp. 103265-103275 ◽  
Author(s):  
Mood Mohan ◽  
Robinson Timung ◽  
Narendra Naik Deshavath ◽  
Tamal Banerjee ◽  
Vaibhav V. Goud ◽  
...  

Subcritical water (SCW) treatment has gained enormous attention as an environmentally friendly technique for organic matter and an attractive reaction medium for a variety of applications. In the current work the process parameters were optimized by RSM model.


2021 ◽  
Author(s):  
Yuxiao Dong ◽  
Dongshen Tong ◽  
Laibin Ren ◽  
Xingtao Chen ◽  
Hao Zhang ◽  
...  

Author(s):  
Sara Toja Ortega ◽  
Mario Pronk ◽  
Merle K. de Kreuk

Abstract Complex substrates, like proteins, carbohydrates, and lipids, are major components of domestic wastewater, and yet their degradation in biofilm-based wastewater treatment technologies, such as aerobic granular sludge (AGS), is not well understood. Hydrolysis is considered the rate-limiting step in the bioconversion of complex substrates, and as such, it will impact the utilization of a large wastewater COD (chemical oxygen demand) fraction by the biofilms or granules. To study the hydrolysis of complex substrates within these types of biomass, this paper investigates the anaerobic activity of major hydrolytic enzymes in the different sludge fractions of a full-scale AGS reactor. Chromogenic substrates were used under fully mixed anaerobic conditions to determine lipase, protease, α-glucosidase, and β-glucosidase activities in large granules (>1 mm in diameter), small granules (0.2–1 mm), flocculent sludge (0.045–0.2 mm), and bulk liquid. Furthermore, composition and hydrolytic activity of influent wastewater samples were determined. Our results showed an overcapacity of the sludge to hydrolyze wastewater soluble and colloidal polymeric substrates. The highest specific hydrolytic activity was associated with the flocculent sludge fraction (1.5–7.5 times that of large and smaller granules), in agreement with its large available surface area. However, the biomass in the full-scale reactor consisted of 84% large granules, making the large granules account for 55–68% of the total hydrolytic activity potential in the reactor. These observations shine a new light on the contribution of large granules to the conversion of polymeric COD and suggest that large granules can hydrolyze a significant amount of this influent fraction. The anaerobic removal of polymeric soluble and colloidal substrates could clarify the stable granule formation that is observed in full-scale installations, even when those are fed with complex wastewaters. Key points • Large and small granules contain >70% of the hydrolysis potential in an AGS reactor. • Flocculent sludge has high hydrolytic activity but constitutes <10% VS in AGS. • AGS has an overcapacity to hydrolyze complex substrates in domestic wastewater. Graphical abstract


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