Production of β-1,3-glucanase from Trichoderma harzianum in surface and submerged culture processes and its role on protoplast generation from Trichoderma reesei mycelium
TL;DR: It was observed that the activities of β-1,3-glucanase and carboxymethylcellulase varied during the entire fermentation cycle which had a distinct effect on protoplast generation from T. reesei mycelium.
Abstract: Surface and submerged culture studies were carried out for the production of extracellular β-1,3-glucanase and carboxymethylcellulase from Trichoderma harzianum, NCIM 1185. The different parameters which influence the enzyme production, viz., spore age in the slant growth, seed age, and pH (initial) of the production medium have been optimized. The ranges of these parameters studied were: Spore age of T. harzianum between 2 and 10 days, seed age between 12 h and 48 h, and pH (initial) between 3 and 7. The other conditions of fermentation were: Temperature 30°C, gyratory shaker speed 160 rev. min−1. It was observed that the activities of β-1,3-glucanase and carboxymethylcellulase varied during the entire fermentation cycle which had a distinct effect on protoplast generation from T. reesei mycelium. The optimum values of the above-mentioned parameters for lytic enzymes production were: Spore age of T. harzianum in the slant growth 5 days, seed age 36 h, and pH (initial) 5.0 for both surface and submerged culture processes.
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TL;DR: In this article, a central composite design was used to determine the optimal levels of microbiological parameters, viz., slant age, seed age and inoculum level, for enhanced griseofulvin production.
Abstract: Central composite design was used to determine the optimal levels of microbiological parameters, viz., slant age, seed age and inoculum level, for enhanced griseofulvin production by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004 in shake flask fermentation. The optimal levels of slant age, seed age and inoculum level for Penicillium griseofulvum MTCC 1898 were found to be 8.8772 days, 4.2093 days, 12% (v/v) (≡17.56 kg dry cell mass/m3) and for Penicillium griseofulvum MTCC 2004, 8.221 days, 3.4875 days and 9% (v/v) (≡8.09 kg dry cell mass/m3) respectively. The yield of griseofulvin under optimal conditions was found to be 1.65 times for Penicillium griseofulvum MTCC 1898 and 1.07 times for Penicillium griseofulvum MTCC 2004 higher than that obtained using unoptimized conditions. The fermentation time for maximum production of griseofulvin by Penicillium griseofulvum MTCC 1898 and Penicillium griseofulvum MTCC 2004 decreased by 4 days and 2 days respectively.
44 citations
Cites background from "Production of β-1,3-glucanase from ..."
...The effect of age of organism for optimal production of carboxymethyl cellulose and b-1, 3-glucanase have been reported by The ´odoe and Panda [ 10 ]....
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TL;DR: Results demonstrate that the modes of action of T. longibrachiatum involved against Th.
Abstract: Seventy-nine Trichoderma strains were isolated from soil taken from 28 commercial plantations of Agave tequilana cv. ‘Azul’ in the State of Jalisco, Mexico. Nine of these isolates produced nonvolatile metabolites that completely inhibited the growth of Thielaviopsis paradoxa on potato dextrose agar plates. These isolates were identified as Trichoderma longibrachiatum on the basis of their morphology and DNA sequence analysis of two genes (ITS rDNA and translation elongation factor EF-1β). Mycoparasitism of Th. paradoxa by T. longibrachiatum strains in dual cultures was examined by scanning electron microscopy. The Trichoderma hyphae grew alongside the Th. paradoxa hyphae, but penetration of Thielaviopsis hyphae by Trichoderma was no apparent. Aleurioconidia of Th. paradoxa were parasitized by Trichoderma. Both hyphae and aleurioconidia of Th. paradoxa lost turgor pressure, wrinkled, collapsed and finally disintegrated. In liquid cultures, all nine Trichoderma isolates produced proteases, β-1,3-glucanases and chitinases that would be responsible for the degradation of Thielaviopsis hyphae. These results demonstrate that the modes of action of T. longibrachiatum involved against Th. paradoxa in vitro experiments are mycoparasitism and the production of nonvolatile toxic metabolites.
44 citations
Cites background or methods from "Production of β-1,3-glucanase from ..."
...0 for the production of proteases [26] and b-1,3-glucanases [27], and to 5....
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...1 filter paper and used to assay enzyme activities for proteases and chitinases [25], and b-1,3-glucanases [27]....
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...The flasks were incubated on a rotary shaker at 180 rpm and 30 C for 36 h for b-1,3-glucanases [27], at 28 C for 36 h for proteases and at 28 C for 43 h for chitinases [25]....
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TL;DR: The use of high concentrations of chitin and ammonium sulphate and exclusion of peptone and urea from the medium resulted in the production of higher level of the enzyme, which was maximized to the level of 0.197 U.
Abstract: Statistical design was used to determine the optimal levels of medium components, the optimal initial pH of the enzyme production medium, the temperature of fermentation, age of the organism in the slant growth and the age of the inoculum for the production of chitinase in shake flask fermentations. The use of high concentrations of chitin and ammonium sulphate and exclusion of peptone and urea from the medium resulted in the production of higher level of the enzyme. The optimal concentrations of the medium components were 12.5 kg/m3 and 4.2 kg/m3 for the chitin and ammonium sulphate respectively. The effect of the addition of peptone and urea to the optimized medium was studied. The optimal values of initial pH and temperature were 5.6 and 28 °C respectively. The optimal age of the slant and the inoculum were found to be 105 h and 43 h respectively. The highest level of chitinase before optimization of the above variables was 0.054 U which was maximized to the level of 0.197 U.
38 citations
Cites background from "Production of β-1,3-glucanase from ..."
...Additionally, for the production of the enzyme, the age of the organism is reported as an important criterion [ 17 ]....
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TL;DR: A temperature of 30°C and an initial pH of 4.7 were found to be optimal for β-1,3-glucanase production from T. harzianum in both surface culture and submerged culture processes.
38 citations
TL;DR: The relative distribution of the enzyme in the cell is independent of the nature of the carbon source and its concentration, and is found in extracellular, cell wall bound and in the periplasmic fractions.
Abstract: Studies on the constitutive β-1,3-glucanase were conducted in submerged as well as in the stationary culture conditions, in the presence and in the absence of lactose and glucose as main carbon sources. In the absence of lactose or glucose, expression of β-1,3-glucanase was observed at 96 h in extracellular, periplasmic, cell wall bound and internal fractions during submerged fermentation. In shake flask culture, enzyme was found in all subcellular fractions using optimal glucose concentration. When Trichoderma harzianum was grown on media containing 55 kg lactose/m3 in submerged culture, activity was found in extracellular, cell wall bound and in the periplasmic fractions. The relative distribution of the enzyme in the cell is independent of the nature of the carbon source and its concentration.
9 citations
Cites methods or result from "Production of β-1,3-glucanase from ..."
...Production medium and culture conditions The optimized medium described by The ´odore and Panda [ 7 ] was used for the production of the b-1,3-glucanase....
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...Production of b-1,3-glucanase using glucose as the carbon source was reported by The ´odore and Panda [ 7 ]....
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...This appears to support the results obtained by The ´odore and Panda [ 7 ]....
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...Assay of b-1,3-glucanase To 0.5 cm 3 of substrate (50 lg/cm 3 ), 0.3 cm 3 of citrate buffer (50 mM, pH 4.8) and 0.2 cm 3 of enzyme prepa- ration were added [ 7 ]....
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...Localization of the enzyme in cells grown in the production medium From the previous studies on the optimization of production medium for b-1,3-glucanase production [ 7 ], it was observed that extracellular constitutive b-1,3-glucanase was found more using 8.6 kg glucose/m 3 of production...
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References
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25,389 citations
TL;DR: In this article, the importance of antibiotics and extracellular enzymes during hyphal interaction is discussed, and the hyphae of the majority of Trichoderma isolates coiled around hyphiae of different test fungi.
Abstract: When grown in dual culture, hyphae of the majority of Trichoderma isolates coiled around hyphae of different test fungi. Penetration of hyphae by Trichoderma hyphae seldom occurred. The importance of antibiotics and extracellular enzymes during hyphal interaction is discussed.
579 citations
TL;DR: Serratia marcescens was found to be the most active organism of 100 tested for the production of chitinase and Enterobacter liquefaciens produced nearly as much enzyme.
Abstract: Serratia marcescens was found to be the most active organism of 100 tested for the production of chitinase. Enterobacter liquefaciens produced nearly as much enzyme. Under optimal conditions high y...
532 citations
01 Jan 1976
TL;DR: In this article, a filter paper assay method and unit value is described for the measurement of enzyme saccharification action, which is simple, reproducible, and quantitative and predicts enzyme action under practical conditions.
Abstract: A filter paper assay method and unit value is described for the measurement of enzyme saccharification action. The method is simple, reproducible, and quantitative and predicts enzyme action under practical saccharification conditions. (JSR)
505 citations
01 Jan 2011
TL;DR: Isolates of T. harzianum were found to differ in the levels of hydrolytic enzymes produced when mycelium of S. rolfsii, Rhizoctonia solani, and Pythium aphanidermatum in soil was attacked, correlated with the ability of each of the Trichoderma isolates to control the respective soilborne pathogens.
494 citations