Purification and Characterization of an Extracellular and a Cellular  -Glucosidase from Bacillus licheniformis

Thirunavukkarasu, Mahesh; Priest, Fergus G.

doi:10.1099/00221287-130-12-3135

Cited by 9 publications

(20 citation statements)

References 11 publications

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“…Then, the amount of extracellular enzyme reached its maximum level at 32 h of cultivation without any decrease in the intracellular enzyme fraction. This type of enzyme production was reported in studies on α ‐glucosidases of B. licheniformis NCIM 6346, B. flavocaldarius KP1228 and G. thermodenitrificans HRO . Strain E134 continued to synthesize α ‐glucosidase in the cell, and the enzyme level increased in the culture medium simultaneously during growth; therefore, the way of its enzyme production was determined to be associated with extracellular as well as intracellular cell fractions according to the cellular localization of its α ‐glucosidases.…”

Section: Discussionsupporting

confidence: 52%

“…In substrate specificity experiments, substrates of phenyl α ‐D‐glucopyranoside, methyl α ‐D‐glucopyranoside, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose, isomaltose, isomaltotriose, panose, turanose, saccharose, maltitol, trehalose, lactose, raffinose, amylose, soluble starch (Sigma S2004) and dextrin (Sigma D4894) were used for glucose formation . The mode of enzyme action, exo‐type hydrolysis and transglycosylation activity analyses were performed using thin layer chromatography (TLC) as described previously …”

Section: Methodsmentioning

confidence: 99%

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Purification and characterization of intracellular and extracellular α‐glucosidases from Geobacillus toebii strain E134

Cihan

Benli

Çökmüş

2011

Cell Biochemistry & Function

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Two different α-glucosidase-producing thermophilic E134 strains were isolated from a hot spring in Kozakli, Turkey. Based on the phenotypic, phylogenetic and chemotaxonomic evidence, the strain was proposed to be a species of G. toebii. Its thermostable exo-α-1,4-glucosidases also were characterized and compared, which were purified from the intracellular and extracellular fractions with estimated molecular weights of 65 and 45 kDa. The intracellular and extracellular α-glucosidases showed optimal activity at 65 °C, pH 7.0, and at 70 °C, pH 6.8, with 3.65 and 0.83 K(m) values for the pNPG substrate, respectively. Both enzymes remained active over temperature and pH ranges of 35-70 °C and 4.5-11.0. They retained 82 and 84% of their activities when incubated at 60 °C for 5 h. Their relative activities were 45-75% and 45-60% at pH 4.5 and 11.0 values for 15 h at 35 °C. They could hydrolyse the α-1,3 and α-1,4 bonds on substrates in addition to a high transglycosylation activity, although the intracellular enzyme had more affinity to the substrates both in hydrolysis and transglycosylation reactions. Furthermore, although sodium dodecyl sulfate behaved as an activator for both of them at 60 °C, urea and ethanol only increased the activity of the extracellular α-glucosidase. By this study, G. toebii E134 strain was introduced, which might have a potential in biotechnological processes when the conformational stability of its enzymes to heat, pH and denaturants were considered.

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Section: Discussionsupporting

confidence: 52%

Section: Methodsmentioning

confidence: 99%

Purification and characterization of intracellular and extracellular α‐glucosidases from Geobacillus toebii strain E134

Cihan

Benli

Çökmüş

2011

Cell Biochemistry & Function

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“…Bacillus licheniformis spores also displayed notable α‐MUG hydrolysis, albeit at a lower rate than both B. anthracis and B. thuringiensis HD‐1 Cry − . Although a spore‐associated α‐glucosidase has not been described in the literature, B. licheniformis is known to produce both cytoplasmic and extracellular α‐glucosidases (Thirunavukkarasu and Priest ) so this activity is not unexpected.…”

Section: Resultsmentioning

confidence: 99%

A disclosure gel for visual detection of liveBacillus anthracisspores

Robinson

Bishop

2019

J Appl Microbiol

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AimsTo develop a gel formulation to trigger a visual signal for rapid disclosure of the location and extent of surface contamination with viable Bacillus anthracis spores.Methods and ResultsMethylumbelliferyl‐α‐d‐glucopyranoside was combined with hyaluronic acid to produce a gel that could be applied to a surface as a coating. It remained hydrated for a sufficient time for α‐glucosidase activity present in intact B. anthracis spores to cleave the substrate and release the fluorescent product, methylumbelliferone. The presence of B. anthracis spores could be disclosed at 5 × 104 CFU per reaction test well (0·32 cm2) both visually and using fluorescence detection equipment.ConclusionsThe disclosure gel provides a rapid, visual response to the presence of B. anthracis spores on a surface.Significance and Impact of the StudyThe disclosure gel demonstrates the first steps towards the development of a formulation that can provide nonspecialist users with a visual alert to the presence of B. anthracis spores on a surface. It is envisioned that such a formulation would be beneficial in scenarios where exposure to spore release is a risk, and could be used in the initial assessment of equipment to aid prioritization and localized execution of a decontamination strategy.

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“…Enzyme digest samples were taken from the reaction mixture at 0, 10, 30, 60 and 120 min (Suzuki et al 1979(Suzuki et al , 1984. The reaction was stopped by heat treatment for 3 min at 95°C and 15 ll digests were analyzed by thin layer chromatography using the method of Thirunavukkarasu and Priest (Thirunavukkarasu and Priest 1984).…”

Section: Mode Of Action Of A-glucosidases On Substratesmentioning

confidence: 99%

Characterization of thermostable α-glucosidases from newly isolated Geobacillus sp. A333 and thermophilic bacterium A343

Cihan

Özcan

Çökmüş

2009

World J Microbiol Biotechnol

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We have partially purified and characterized two new thermostable exo-a-1,4-glucosidases (E.C.3.2.1.20) isolated from Geobacillus sp. A333 and thermophilic bacterium A343 strains. A333 a-glucosidase showed optimum activity at 60°C, pH 6.8 and had a value of 1.38 K m for the pNPG substrate, whereas these results were found to be 65°C, 7.0 and 0.85, respectively for A343 enzyme. Specificity for 20 different substrates and thin layer chromatography studies demonstrated that the A333 enzyme had high transglycosylation activity, and A343 had wide substrate specificity. The substrate specificity of A333 a-glucosidase was determined as maltose, dextrin, turanose, maltotriose, maltopentaose, meltotetraose, maltohexaose and phenyla-D-glycopyranoside. On the other hand, the A343 a-glucosidase mostly hydrolyzed dextrin, turanose, maltose, phenyl-a-D-glucopyranoside, maltotriose, maltotetraose, maltopentaose, isomaltose, saccharose and kojibiose by acting a-1,2, a-1,3, a-1,4 and a-1,6 bonds of these substrates. The relative activites of A333 and A343 enzymes were determined to be 83 and 92% when incubated at 60°C for 5 h whereas, the pH of 50% inactivation at 60°C for 15 h were determined to be pH 4.5/10.0 and pH 5.0/10.0, respectively. In addition, the results not only showed that both of the a-glucosidases were stable in a wide range of pH and temperatures, but were also found to be resistant to most of the denaturing agents, inhibitors and metal ions tested. With this study, thermostable exo-a-1,4-glucosidases produced by two new thermophilic strains were characterized as having biotechnological potential in transglycosylation reactions and starch hydrolysis processes.

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Purification and Characterization of an Extracellular and a Cellular -Glucosidase from Bacillus licheniformis

Cited by 9 publications

References 11 publications

Purification and characterization of intracellular and extracellular α‐glucosidases from Geobacillus toebii strain E134

Purification and characterization of intracellular and extracellular α‐glucosidases from Geobacillus toebii strain E134

A disclosure gel for visual detection of liveBacillus anthracisspores

Characterization of thermostable α-glucosidases from newly isolated Geobacillus sp. A333 and thermophilic bacterium A343

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