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.
Citations
More filters
Journal Article•
TL;DR: Crude enzyme fraction from stored corm of Amorphophallus paeoniifolius could clarify apple juices indicationg its potential use in fruit juice processing.
Abstract: Crude enzyme fraction from stored corm of Amorphophallus paeoniifolius could clarify apple juices indicationg its potential use in fruit juice processing. The extract showed presence of cellulase activity which was confirmed by Congo red plate diffusion on incubating the plates for 24 hours at 50°C. Optimum temperature, pH, incubation time for cellulase activity was also determined by DNS method. Zymography studies and SDS gel electrophoresis confirmed that the molecular weight of the enzyme is 66 kDa.
1 citations
Cites background from "Fermentative production and kinetic..."
...Cellulases have been commercially available for more than 30 years and these enzymes have represented a target for both academic as well as industrial research (Singh, 1999; Muthuvelayudham and Viruthagiri, 2006; Singh et al., 2007)....
[...]
...available for more than 30 years and these enzymes have represented a target for both academic as well as industrial research (Singh, 1999; Muthuvelayudham and Viruthagiri, 2006; Singh et al., 2007)....
[...]
TL;DR: Among 17 fungal isolates isolated from soil of Elhawia, El-hada, Elkaym and Karwa in Taif governorate in Saudi Arabia, two isolates showed high efficacy in producing cellulases enzymes as discussed by the authors.
Abstract: Among 17 fungal isolates isolated from soil of El-hawia, El-hada, El-kaym and Karwa in Taif governorate in Saudi Arabia, two isolates showed high efficacy in producing cellulases enzymes. They were identified to be Alternaria alternata and Aspergillus wentii. Some factors such as carbon and wheat bran as a raw material, nitrogen, pH and incubation temperature were investigated. Results indicated that glucose and cellulose were the most effective as a carbon source while, urea was the best nitrogen source for cellulases production. Initial pH 5.0 and incubation temperatures at 25 or 35oC achieved high cellulases production.
1 citations
Cites background from "Fermentative production and kinetic..."
...While Muthuvelayudham and Viruthagiri (2006) reported that the maximum cellulase activity was obtained at 28°C by Trichoderma reesei....
[...]
01 Jan 2015
TL;DR: Various combinations of physical/chemical pretreatments, fungal cultures, and incubation methods were investigated to metabolize GLS into cell mass, CO2, or other non-toxic components and enhance the nutritional value of canola meal.
Abstract: INCREASING THE NUTRITIONAL VALUE OF CANOLA MEAL VIA FUNGAL BIOPROCESSING JASON R. CROAT 2015 The main limitation of meals from canola and other Brassica spp. is the presence of glucosinolates (GLS), which are anti-nutritional and can even be toxic at high ingestion levels. Furthermore, large amounts of GLS can reduce palatability for livestock and thus reduce intake and growth rates. For this reason canola was bred to contain lower levels of GLS (< 30 μmol/g) and erucic acid (< 2%). However, feed inclusion rates are still limited to ~30%, and this reduces the value of canola meal. The goal of this thesis was to optimize a pretreatment and fungal conversion process to enhance the nutritional value of canola meal. Various combinations of physical/chemical pretreatments, fungal cultures, and incubation methods were investigated to metabolize GLS into cell mass, CO2, or other non-toxic components. These treatments also served to hydrolyze canola meal fiber into carbohydrates which were then metabolized by the fungi into single cell protein. Solid-state incubation conditions were initially tested, since filamentous fungi are well adapted to grow at lower moisture levels, and this would potentially reduce contamination issues with bacteria. Flask trials were performed using 50% moisture, hexane extracted (HE) or cold pressed (CP) canola meal, with incubation for 168 h at 30oC. On HE canola meal Trichoderma reesei (NRRL-3653) achieved the greatest increase in protein content (23%), while having the lowest residual levels of sugar (8%
1 citations
TL;DR: In this paper, microwave treatments of different powers (400-1600 W), pressures (1-5 bar), temperatures (120-180 °C), and treatment times (3-30 min) were applied to change the physical condition of vine-branch.
Abstract: Our research target was to utilise vine-branch, existing in huge amounts, for energetic purposes. During our experiments, microwave (MW) treatments of different powers (400–1600 W), pressures (1–5 bar), temperatures (120–180 °C), and treatment times (3–30 min) were applied to change the physical condition of vine-branch. After MW, enzymatic hydrolysis (EH) was used (85–100 h, 37 °C). In addition, beside MW, comparisons were made regarding various treatment methods: untreated (UTE), cooking plate (CP), and autoclave (AC), to determine to what extent they affect the final glucose yield. This yield can even further be increased by MW pre-treatment (50 W, 3–30 min, 40 °C) of the enzyme used during the hydrolysis, which reinforces the argument that enzyme activity can be increased by irradiation. A difference of 22.1% was detected among the glucose yield values in untreated and treated enzyme processes.
1 citations
01 Feb 2015
TL;DR: Results indicated that 1% carboxymethylcellulose (CMC) and 0.7% yeast extract were most effective as the carbon and nitrogen sources respectively.
Abstract: Effect of some nutritional and environmental factors on growth and cellulase production by Bacillus alcalophilus S39 and Bacillus amyloliquefaciens C23 was investigated. Results indicated that 1% carboxymethylcellulose (CMC) and 0.7% yeast extract were most effective as the carbon and nitrogen sources respectively. Initial pH 7 and 3% inoculum size (3.5-4.3 ×10 6 CFU/ml) found to be optimal for growth and cellulase production. Incubation temperature at 30oC and 45oC achieved the highest activity of cellulase for Bacillus alcalophilus S39 and Bacillus amyloliquefaciens C23 respectively, and the suitable shaking rate was 150 and 200 rpm.
1 citations
Cites result from "Fermentative production and kinetic..."
...Muthuvelayudham and Viruthagiri (2006) obtained similar results which showed that the cellulase activity was less when glucose was used as carbon source due to feedback inhibition....
[...]
References
More filters
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)....
[...]
...a number of applications including animal feed, pharmaceutical and textile industries (Aristidou and Penttilä, 2000)....
[...]
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)....
[...]
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)....
[...]
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)....
[...]
...The general trend is that more cellulose in the mixture results in higher levels of endoglucanase (Olsson et al., 2003)....
[...]
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