Stability and kinetics of β-1,3-glucanse from Trichoderma harzianum

Rana, Deb Sankar; Théodore, K.; Naidu, G. S. N.; Panda, T.

doi:10.1016/s0032-9592(02)00323-0

Cited by 19 publications

(11 citation statements)

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“…The enzyme was sensitive to copper and fairly sensitive to zinc and manganese, but insensitive to magnesium, calcium and aluminum (Table 3). Glucanases produced by Rhizopus oryzae [29], Bacillus clausii [30], Bacillus halodurans [32] and Trichoderma harzianum [31] show similar sensitivity to the divalent metal ion copper.…”

Section: Resultsmentioning

confidence: 99%

Characterization of a β-glucanase produced by Rhizopus microsporus var. microsporus, and its potential for application in the brewing industry

2006

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BackgroundIn the barley malting process, partial hydrolysis of β-glucans begins with seed germination. However, the endogenous 1,3-1,4-β-glucanases are heat inactivated, and the remaining high molecular weight β-glucans may cause severe problems such as increased brewer mash viscosity and turbidity. Increased viscosity impairs pumping and filtration, resulting in lower efficiency, reduced yields of extracts, and lower filtration rates, as well as the appearance of gelatinous precipitates in the finished beer. Therefore, the use of exogenous β-glucanases to reduce the β-glucans already present in the malt barley is highly desirable.ResultsThe zygomycete microfungus Rhizopus microsporus var. microsporus secreted substantial amounts of β-glucanase in liquid culture medium containing 0.5% chitin. An active protein was isolated by gel filtration and ion exchange chromatographies of the β-glucanase activity-containing culture supernatant. This isolated protein hydrolyzed 1,3-1,4-β-glucan (barley β-glucan), but showed only residual activity against 1,3-β-glucan (laminarin), or no activity at all against 1,4-β-glucan (cellulose), indicating that the R. microsporus var. microsporus enzyme is a member of the EC 3.2.1.73 category. The purified protein had a molecular mass of 33.7 kDa, as determined by mass spectrometry. The optimal pH and temperature for hydrolysis of 1,3-1,4-β-glucan were in the ranges of 4–5, and 50–60°C, respectively. The Km and Vmax values for hydrolysis of β-glucan at pH 5.0 and 50°C were 22.39 mg.mL-1 and 16.46 mg.min-1, respectively. The purified enzyme was highly sensitive to Cu+2, but showed less or no sensitivity to other divalent ions, and was able to reduce both the viscosity and the filtration time of a sample of brewer mash. In comparison to the values determined for the mash treated with two commercial glucanases, the relative viscosity value for the mash treated with the 1,3-1,4-β-glucanase produced by R. microsporus var. microsporus. was determined to be consistently lower.ConclusionThe zygomycete microfungus R. microsporus var. microsporus produced a 1,3-1,4-β-D-glucan 4-glucanhydrolase (EC 3.2.1.73) which is able to hydrolyze β-D-glucan that contains both the 1,3- and 1,4-bonds (barley β-glucans). Its molecular mass was 33.7 kDa. Maximum activity was detected at pH values in the range of 4–5, and temperatures in the range of 50–60°C. The enzyme was able to reduce both the viscosity of the brewer mash and the filtration time, indicating its potential value for the brewing industry.

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Section: Resultsmentioning

confidence: 99%

Characterization of a β-glucanase produced by Rhizopus microsporus var. microsporus, and its potential for application in the brewing industry

2006

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“…There is no relation between these values. Rana et al (2003) studied the stability and kinetics of b-1,3-glucanase from Trichoderma harzianum. A central composite design with two independent variables, pH and temperature, was applied.…”

Section: Other Applications Of Rsmmentioning

confidence: 99%

“…Stability and kinetics of b-1,3-glucanase from Trichoderma harzianum were investigated using RSM. Also simple enzyme kinetics of alcohol dehydrogenase was evaluated using RSM (Rana, Theodore, Naidu, & Panda, 2003).…”

Section: Introductionmentioning

confidence: 99%

Modeling and optimization I: Usability of response surface methodology

Baş

Boyaci

2007

Journal of Food Engineering

1,728

517

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“…(CCMB 300 and CCMB 306) cells. Enzyme activity depends mainly on pH and temperature, and a novel approach was used to control the combined effect of pH and temperature on β-1,3-glucanase and chitinase using Response Surface Methodology (RANA et al, 2003;NAWANI;KAPADNIS, 2005).…”

Section: Resultsmentioning

confidence: 99%

Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis

et al. 2012

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Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis Aplicação do modelo Doehlert na otimização das variáveis experimentais para a lise de Pseudozyma sp. (CCMB 306) e Pseudozyma sp. (CCMB 300)Amanda Reges de SENA 1,3 , Gildomar Lima VALASQUES JÚNIOR 1,2 , Ingara Keisle São Paulo BARRETTO 1 , Sandra Aparecida ASSIS 1 * IntroductionThe cell wall of yeast is basically made of glucan and mannan-protein. Lytic enzymes such as chitinases, proteases and β-1,3-glucanases are usually necessary. There are numerous applications for biotechnologies: preparation of protoplasts, extraction of pigments, mass treatment of residual yeast cell fermentation industries for preparation of animal feed, in studies of the mechanism of cell wall synthesis for control of pathogenic yeast, etc. (HUNTER;ASENJO, 1988;SATO, 2005;SALAZAR; ASENJO, 2007).These enzymes are capable of lysing the cell wall of Saccharomyces cereviseae, Candida sp. and other genera of yeasts, which extends their use allowing the selective manufacture of products, regardless of scale, and can be performed under temperature and pH conditions that do not involve the denaturation of cellular products of interest (FLEURI; SATO, 2005). Mathematical models have been increasingly used to help explain responses of biochemical reactions. This technique is often used for optimization and/or verification of the influence of medium components and cultivation parameters for enzyme production (FLEURI; SATO, 2008).The production and optimization of the parameters that affect the enzymatic synthesis should always be investigated because the optimal conditions vary among different organisms and different enzymes (BRAVO et al., 2000). In many cases, the interaction of parameters that influence fermentation processes can be evaluated with a reduced number of tests using an experimental design (THÉODORE; PANDA, 1995). Several experimental design models could be used to reduce the number of experiments under different conditions. Among them are the Plackett-Burman, Doehlert, Central Composite, and Box-Behnken (LI et al., 2007).The choice of Doehlert design is justified by a number of advantages such as (1) its spherical experimental domain with an uniformity in space filling, (2) its ability to explore the whole of the domain, and (3) its potential for sequentially where the experiments can be reused when the boundaries have not been well chosen at first (BENSALAH et al., 2010). Doehlert designs are easily applied to optimized variables and offer advantages over the Central Composite (CCD) and Box-Behnken designs, used in response surface analysis. They need fewer experiments, ResumoO presente trabalho visou verificar a influência do pH e temperatura na lise de leveduras utilizando planejamento experimental. No estudo, foi utilizado o extrato enzimático, contendo β-1,3-glucanase e quitinase líticas, obtidas do micro-organismo Moniliophthora perniciosa. O delineamento experiment...

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Stability and kinetics of β-1,3-glucanse from Trichoderma harzianum

Cited by 19 publications

References 11 publications

Characterization of a β-glucanase produced by Rhizopus microsporus var. microsporus, and its potential for application in the brewing industry

Characterization of a β-glucanase produced by Rhizopus microsporus var. microsporus, and its potential for application in the brewing industry

Modeling and optimization I: Usability of response surface methodology

Application of Doehlert experimental design in the optimization of experimental variables for the Pseudozyma sp. (CCMB 306) and Pseudozyma sp. (CCMB 300) cell lysis

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