Fermentative production and kinetics of cellulase protein on Trichoderma reesei using sugarcane bagasse and rice straw
TL;DR: This is the first attempt of combining the synthetic substrate (xylose, lactose) with natural substrate (sugarcane bagasse, rice straw) and the mixture of substrates produced the highest maximal enzyme activity on cellulose with xylose.
Abstract: Cellulase a multienzyme made up of several proteins finds extensive applications in food, fermentation and textile industries. Trichoderma reesei is an efficient producer of cellulase protein. The comparative study was made on various carbon sources on the production of cellulase using strains of T. reesei QM 9414, 97.177 and Tm3. Pretreatment of sugarcane bagasse and rice straw offers very digestible cellulose and potentially less inhibition. Cellulase production was enhanced by multiple carbon sources because of diauxic pattern of utilization of substrates. This is the first attempt of combining the synthetic substrate (xylose, lactose) with natural substrate (sugarcane bagasse, rice straw). The mixture of substrates produced the highest maximal enzyme activity on cellulose with xylose, cellulose with lactose, bagasse with xylose, bagasse with lactose, rice straw with xylose and rice straw with lactose. In addition Monod growth kinetics and Leudeking piret product formation kinetics were studied using T. reesei with optimized medium under optimized conditions of inoculum concentration, D.O. level, agitator speed, temperature and pH.
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TL;DR: Of the two developed models from shake-flasks, xylanase model validated successfully the fermenter results giving 18.7 IU/mL of experimental xylan enzyme in 1.5-L working volume bioreactor.
Abstract: Most of the corn-ethanol plants use dry-mill process and generate large volume of distillers dried grains with solubles (DDGS) as the co-product. The DDGS is a promising feedstock with rich content of cellulose and xylan. The goal of this study was to optimize cellulase and xylanase production by Trichoderma reesei (NRRL 6156) using DDGS as the carbon source at shake-flask and validate the results at benchtop bioreactors. The fermentation medium containing varied amounts of dilute-acid pretreated DDGS, yeast extract, and peptone were optimized for maximal enzyme production at 30 °C, pH 5.0 and 180 rpm with 3% (v/v) inoculum using Box-Behnken response surface methodology (RSM). Co-production of xylanase and cellulase enzymes was not achieved in the same medium; higher DDGS hydrolysate favored cellulase enzyme only while lower amount induced the xylanase enzyme. Based on the RSM results, maximal xylanase of 18.5 IU/mL was predicted with 5% DDGS hydrolysate, 0.1 g/L yeast extract, and 2 g/L peptone, while maximal cellulase of 1.1 IU/mL was predicted with 20% DDGS hydrolysate, 1 g/L yeast extract, and 0.5 g/L peptone. Of the two developed models from shake-flasks, xylanase model validated successfully the fermenter results giving 18.7 IU/mL of experimental xylanase in 1.5-L working volume bioreactor.
14 citations
TL;DR: Production of CMCase from newly isolated strain of Aspergillus versicolor was studied by optimizing different physico-chemical parameters and could easily be used for the production of cellulase using renewable biomass.
13 citations
Cites background from "Fermentative production and kinetic..."
...…reported that high concentration of nitrogenous compounds alters the hydrophobicity of cell wall which ultimately decreases the production of CMCase (Muthuvelayudham & Viruthagiri, 2006). a b c d e f d 0 200 400 600 800 1000 0 0.25 0.5 1 1.5 2 2.5 E n zy m e a ct iv it y (U /m l/ m in ) NaNO3 (gm...
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02 Sep 2010
TL;DR: In this paper, a 0.16 kW/L ultrasonic treatment was applied to treat rice straw during enzymatic hydrolysis and fermentation process, which showed a breaking of lignin and exposition of cellulose surface.
Abstract: Pretreatment is one of the approaches to yield the higher saccharification process leading to optimization of ethanol yield. The effect of ultrasonic pretreatment on ethanol production from rice straw was studied. A 0.16 kW/L ultrasonic treatment was applied to treat rice straw during enzymatic hydrolysis and fermentation process. The enzymatic hydrolysis process was carried out at 50°C and pH 4.8 by using a commercial enzyme and followed by fermentation with S.cereviceae. Morphological structure of rice straw was analyzed by a scanning electron microscopy (SEM) and the glucose concentration was detected by biochemistry analyzer. The result showed that the structure of rice straw after ultrasonic treatment showed a breaking of lignin and exposition of cellulose surface. The glucose concentration of the pretreated rice straw increased significantly by 57.65% (p = 0.003) when compared to the untreated rice straw. The total ethanol concentration of the untreated was not obviously different from the pretreated rice straw. Results revealed that ultrasonic treatment could enhance the ethanol production from rice straw.
12 citations
Cites background from "Fermentative production and kinetic..."
...Muthuvelayudham and Viruthagiri (2006) [11] also reported that removal of lignin is crucial for increasing cellulase activity which can be observed in sugarcane bagasse and rice straw whereby the cellulose provided is more digestible....
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TL;DR: In this article, the optimal conditions for biogas generation from anaerobic digestion of rice straw using Response Surface Methodology (RSM) were obtained using a 1m3 floating drum aaerobic digester.
Abstract: The experimental and analytical study is to obtain the optimal conditions for biogas generation from anaerobic digestion of rice straw using Response Surface Methodology (RSM). This experiment is carried out using 1m3 floating drum anaerobic digester. The parameters temperature, pH, and substrate concentration and agitation time on biogas production from rice straw were investigated. The experimental results showed that the linear model terms of temperature, substrate concentration and the pH , agitation time had significant of interactive effects (P<0.05) were studied on biogas. The highest level of biogas 0.72m3 is produced at optimum temperature 50°c, pH 7.5, substrate concentration 110.70 kg, and agitation time of 5sec respectively.
12 citations
Cites background from "Fermentative production and kinetic..."
...Mainly used to study the effect of factors and estimating optimum levels of the experiments [12]....
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TL;DR: In this article, Aspergillus niger under solid state fermentation (SSF) on a laboratory scale was investigated, and different fermentation parameters, such as moisture content, initial pH, temperature, depth of the substrate, and inoculum size were optimized.
Abstract: β-Exoglucanase production on the lignocellulosic material, wheat bran, by Aspergillus niger under solid state fermentation (SSF) on a laboratory scale was investigated. Different fermentation parameters, such as moisture content, initial pH, temperature, depth of the substrate, and inoculum size on exoglucanase production were optimized. Moisture content of 40 %, pH of 7.0, substrate depth of 1.0 cm, inoculum size of 2 × 106 spores/g of wheat bran, and temperature at 30 °C were optimal for maximum production of exoglucanase. Maximum yields of exoglucanase with 28.60 FPU/g of wheat bran were obtained within 3 days of incubation under optimal conditions.
12 citations
Cites background from "Fermentative production and kinetic..."
...The importance of the depth of the bed of medium on enzyme production is discussed by Muthuvelayudham and Viruthagiri (2006)....
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References
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TL;DR: The conversion of both cellulose and hemicellulose for the production of fuel ethanol is being studied intensively, with a view to developing a technically and economically viable bioprocess.
492 citations
"Fermentative production and kinetic..." refers background in this paper
...…growth rate (day)-1; µ, specific growth rate (day)–1; Ks, Monod constant; P, product concentration (g/l); α and β, Leudeking Piret constants; and X, substrate concentration (g/l). a number of applications including animal feed, pharmaceutical and textile industries (Aristidou and Penttilä, 2000)....
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...a number of applications including animal feed, pharmaceutical and textile industries (Aristidou and Penttilä, 2000)....
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TL;DR: RNA slot blot experiments showed that no expression could be observed on glucose-containing medium and that high glucose levels abolish the inducing effect of sophorose, showing that distinct and clear-cut mechanisms of induction and glucose repression regulate cellulase expression in an actively growing fungus.
Abstract: Basic features of regulation of expression of the genes encoding the cellulases of the filamentous fungus Trichoderma reesei QM9414, the genes cbh1 and cbh2 encoding cellobiohydrolases and the genes egl1, egl2 and egl5 encoding endoglucanases, were studied at the mRNA level. The cellulase genes were coordinately expressed under all conditions studied, with the steady-state mRNA levels of cbh1 being the highest. Solka floc cellulose and the disaccharide sophorose induced expression to almost the same level. Moderate expression was observed when cellobiose or lactose was used as the carbon source. It was found that glycerol and sorbitol do not promote expression but, unlike glucose, do not inhibit it either, because the addition of 1 to 2 mM sophorose to glycerol or sorbitol cultures provokes high cellulase expression levels. These carbon sources thus provide a useful means to study cellulase regulation without significantly affecting the growth of the fungus. RNA slot blot experiments showed that no expression could be observed on glucose-containing medium and that high glucose levels abolish the inducing effect of sophorose. The results clearly show that distinct and clear-cut mechanisms of induction and glucose repression regulate cellulase expression in an actively growing fungus. However, derepression of cellulase expression occurs without apparent addition of an inducer once glucose has been depleted from the medium. This expression seems not to arise simply from starvation, since the lack of carbon or nitrogen as such is not sufficient to trigger significant expression.
483 citations
"Fermentative production and kinetic..." refers background in this paper
...Hypercelluloytic mutant strains secrete large amounts of cellulases (Ilmen et al., 1997)....
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TL;DR: This mini-review summarises the considerable amount of data accumulated over the past three decades regarding promoters of genes encoding hydrolytic enzymes, inducer molecules, and models for the recognition of the insoluble substrates cellulose and xylan.
Abstract: The genus Trichoderma comprises a group of filamentous ascomycetes that are now widely used in industrial applications because of their ability to produce extracellular hydrolases in large amounts. In addition, strong inducible promoters together with high secretory capacity have made Trichoderma an attractive host for heterologous protein production. Several promoters of genes encoding hydrolytic enzymes have been investigated in detail regarding their cis-acting elements and trans-acting factors. Potent inducer molecules, for both xylanolytic and cellulolytic enzyme systems, have been identified and characterized. Furthermore, models for the recognition of the insoluble substrates cellulose and xylan have been developed based on a large set of experiments. This mini-review summarises the considerable amount of data accumulated over the past three decades.
207 citations
"Fermentative production and kinetic..." refers background in this paper
...The genus Trichoderma, filamentous ascomycetes are widely used in industrial applications because of high secretory capacity and inducible promoting characteristics (Mach and Zeilinger, 2003)....
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TL;DR: The growth and enzyme production by Trichoderma reesei Rut C-30 using different lignocellulosic materials as carbon source were investigated and it was found that endoglucanase and endoxylanse activities were produced throughout the cultivations, whereas α-arabinosidase was induced late during the cultivation.
157 citations
"Fermentative production and kinetic..." refers background in this paper
...Biosynthesis of cellulase was also made on lignocellulo-sic materials such as sugar beet pulp and alkaline extracted sugar beet pulp and cellulose (Olsson et al., 2003)....
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...The general trend is that more cellulose in the mixture results in higher levels of endoglucanase (Olsson et al., 2003)....
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TL;DR: The filamentous fungus Penicillium brasilianum IBT 20888 was cultivated on a mixture of 30 g l−1 cellulose and 10 g xylan for 111 h and the resulting culture filtrate was used for protein purification and revealed similarity in molecular mass, pI and hydrolytic properties suggested that these two enzymes were identical, but the smaller was lacking the cellulose-binding domain or an essential part of it.
134 citations