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
TL;DR: The results suggested that NU-6 should be an attractive producer for both cellulose and xylanase production.
Abstract: The strain of Trichoderma reesei Rut C-30 was subjected to mutation after treatment with N-methyl-N′-nitro-N-nitrosoguanidine (NG) for 6 h followed by UV irradiation for 15 min. Successive mutants showed enhanced cellulase production, clear hydrolysis zone and rapid growth on Avicel-containing plate. Particularly, the mutant NU-6 showed approximately two-fold increases in activity of both FPA and CMCase in shake flask culture when grown on basal medium containing peptone (1%) and wheat bran (1%). The enzyme production was further optimized using eight different media. When a mixture of lactose and yeast cream was used as cellulase inducer, the mutant NU-6 yielded the highest enzyme and cell production with a FPase activity of 6.2 U ml−1, a CMCase activity of 54.2 U ml−1, a β-glucosidase activity of 0.39 U ml−1, and a fungal biomass of 12.6 mg ml−1. It deserved noting that the mutant NU-6 also secreted large amounts of xylanases (291.3 U ml−1). These results suggested that NU-6 should be an attractive producer for both cellulose and xylanase production.
33 citations
Cites background from "Fermentative production and kinetic..."
...21), all of which act synergistically in the conversion of cellulose into glucose [7, 11]....
[...]
TL;DR: In this paper, the authors evaluated the effect of some variables on β-glucosidase production by Penicillium verruculosum using passion fruit peel as substrate, and on hydrolysis of this process residue with P. verruculusosum crude extract, by applying a full factorial central composite design.
Abstract: β-Glucosidases show great potential as catalysts for various biotechnology processes including biomass hydrolysis for bioethanol production. In this study, response surface methodology was used to evaluate the effect of some variables on β-glucosidase production by Penicillium verruculosum using passion fruit peel as substrate, and on hydrolysis of this process residue with P. verruculosum crude extract, by applying a full factorial central composite design. Process optimization resulted in a 5.7 fold increase in β-glucosidase activity. The enzymes were more active at 65 °C, pH 4.5, remaining stable at 55 and 60 °C and over a broad pH range. P. verruculosum crude extract hydrolyzed passion fruit peel with glucose yield of 45.54%. This article provides, for the first time, the production of remarkable yields of β-glucosidase and the achievement of expressive levels of glucose through the use of passion fruit peel, an abundant and inexpensive agro-industrial residue.
32 citations
TL;DR: During application in saccharification tests, the isolated halotolerant endoglucanase was able to hydrolyse sugarcane bagasse, rice husk, and wheat bran, with the highest production of reducers/fermentable sugars within 24 h of saccharization for wheat Bran.
31 citations
TL;DR: Different bioreactors have been reviewed for cellulase production with the aim to investigate main factors for designing a bioreactor, including efficient mixing, sufficient mass transfer, low shear stress, low foaming problems and low consumption of water and energy.
Abstract: Cellulases are costly, a principal challenge of enzymatic hydrolysis of cellulosic materials for bioethanol production For an efficient cellulase production, fungi are preferred over bacteria due to their permeation capability and versatile substrate consumption Some limitations in the enzymatic hydrolysis step prevent the process to be economically feasible Different strategies have been investigated to overcome these limitations, including genetic engineering, enzyme recycling, high solid loadings, pretreatment technologies, supplementation of cellulases with additives and application of nanomaterials for improving the thermal and pH stability of cellulases Several studies have been performed in various bioreactors with the target to procure higher yields of glucose in the enzymatic hydrolysis step The key factors for designing a bioreactor include efficient mixing, sufficient mass transfer, low shear stress, low foaming problems and low consumption of water and energy In this scenario, different bioreactor configurations, including stirred tank bioreactor, horizontal rotating tubular bioreactor, airlift bioreactor, membrane bioreactor, reciprocating plate bioreactor, solid-state fermentation bioreactors have been reviewed for cellulase production with the aim to investigate main factors for designing a bioreactor
29 citations
Cites background from "Fermentative production and kinetic..."
...Effective parameters on cellulases complex Researchers have indicated that the cellulolytic system of T. reesei includes 60–80% exo-glucanase, 20–36% Endo-Glucanase and 1% b-glucosidase (Gritzali and Brown Jr. 1979; Zaldı́var et al. 2001; Muthuvelayudham and Viruthagiri 2006)....
[...]
...Muthuvelayudham and Viruthagiri (2006) investigated the effects of various carbon sources on the production of cellulases using strains of T. reesei....
[...]
TL;DR: The results indicate that the production of cellulase from green pea hulls may provide a novel and economical solution for industrial waste disposal.
Abstract: Pea hulls have a significant contribution to agricultural waste and are left unattended. It has a rich nutrient media and may have application in waste utilization. In this investigation, the potential of pea hulls for the production of cellulase has been explored. The effect of H2O2 concentration, agitation speed and harvesting time on cellulase production by Trichoderma reesei QM9414 under submerged fermentation at constant pH of 5.0 and temperature 30 ± 0.1 °C were studied. The maximum filter paper (FP) cellulase activity of 0.372 ± 0.019 U/mL was obtained at 91 h incubation period and 120 rpm agitation speed. Based on the optimized results of fermentation parameters, 120 rpm agitation speed, 5% H2O2 concentration and 91 h harvesting time was recommended for efficient cellulase production. The effect of harvesting time on protein, reducing sugar and cellulase activity was pre-dominant. The purified cellulase enzyme specific activity was recorded as 13.8 U/mL. The results indicate that the production of cellulase from green pea hulls may provide a novel and economical solution for industrial waste disposal.
27 citations
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