Release of Cell-associated Tannase of Serratia ficaria DTC by Sonication, Surfactants and Solvents

Belur, Prasanna D.; Mugeraya, Gopal; Nainegali, Basavaraj S.

doi:10.3923/ajbkr.2011.91.97

Cited by 5 publications

(4 citation statements)

References 16 publications

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“…Ta n n a s e s a re e n zy m e s u s e d i n different industrial applications as in food and pharmaceutical. In the last decade, there have been efforts looking for new sources for tannase enzyme production 27 . Many researches on tannase synthesis showed clearly that almost all species of Aspergillus are the predominant source of tannase for industrial production and application 28 .…”

Section: Results and Discussion Partial Purification Of Tannasementioning

confidence: 99%

Purification, Characterization and Application of Tannase Enzyme Isolated from Marine Aspergillus nomius GWA5

Farag¹,

Hassan²,

El-Says³

et al. 2018

J Pure Appl Microbiol

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Tannase enzyme (EC 3.1.1.20) is an enzyme used in many biotechnological applications as in chemical, beverage, pharmaceutical and food industries. was isolated and purified from marine Aspergillus nomius GWA5 by 75% acetone fractional precipitation, followed by gel filtration in Sephadex G-100 and ion exchange chromatography on DEAE-Sephadex A-50 yielding 4.48-fold purification. Estimation of tannase molecular weight was carried out using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showing a molecular weight of 30 kDa. The highest activity (291 U/mg protein) were at pH 6.0 and 50 °C, respectively. Tannase stability was observed in acidic range (4-6) and was stable to heat treatment. In absence of its substrate it retained about 84.5% of its activity at 80 °C for 15 min. Effect of some metal ions and chelator on tannase activity was investigated. Mg 2+ activated as activator of the pure enzyme while EDTA, Cd 2+ , Pb 2+ and Hg 2+ inhibited its activity and retained about 40.78, 51.55, 30.24 and 24.55% of its activity, respectively. Promising activity of Tannase was shown in removing tannin stains of tea from clothes.

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Section: Results and Discussion Partial Purification Of Tannasementioning

confidence: 99%

Purification, Characterization and Application of Tannase Enzyme Isolated from Marine Aspergillus nomius GWA5

Farag¹,

Hassan²,

El-Says³

et al. 2018

J Pure Appl Microbiol

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“…Similar observation was also reported by Saptarshi and Lele [ 17 ] whom demonstrated that the recovery of intracellular L-asparaginase is directly proportional to the cell concentration but reduction of enzyme yield occurred beyond the optimum cell concentration. This phenomenon could be due to the distortion of frequency on sonic wave’s passages and severe attenuation on sound intensity under highly saturated condition [ 16 , 29 ]. Both effects cause the reduction in the cavitation zone and consequently the disruption of mycelia cell becomes less effective, which can be observed from Figure 2 a where GAD activity and soluble proteins were started to decline at 12% of cell concentration.…”

Section: Resultsmentioning

confidence: 99%

A comparative study of extraction techniques for maximum recovery of glutamate decarboxylase (GAD) from Aspergillus oryzae NSK

et al. 2013

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Backgroundγ-Amino butyric acid (GABA) is a major inhibitory neurotransmitter of the mammalian central nervous system that plays a vital role in regulating vital neurological functions. The enzyme responsible for producing GABA is glutamate decarboxylase (GAD), an intracellular enzyme that both food and pharmaceutical industries are currently using as the major catalyst in trial biotransformation process of GABA. We have successfully isolated a novel strain of Aspergillus oryzae NSK that possesses a relatively high GABA biosynthesizing capability compared to other reported GABA-producing fungal strains, indicating the presence of an active GAD. This finding has prompted us to explore an effective method to recover maximum amount of GAD for further studies on the GAD’s biochemical and kinetic properties. The extraction techniques examined were enzymatic lysis, chemical permeabilization, and mechanical disruption. Under the GAD activity assay used, one unit of GAD activity is expressed as 1 μmol of GABA produced per min per ml enzyme extract (U/ml) while the specific activity was expressed as U/mg protein.ResultsMechanical disruption by sonication, which yielded 1.99 U/mg of GAD, was by far the most effective cell disintegration method compared with the other extraction procedures examined. In contrast, the second most effective method, freeze grinding followed by 10% v/v toluene permeabilization at 25°C for 120 min, yielded only 1.17 U/mg of GAD, which is 170% lower than the sonication method. Optimized enzymatic lysis with 3 mg/ml Yatalase® at 60°C for 30 min was the least effective. It yielded only 0.70 U/mg of GAD. Extraction using sonication was further optimized using a one-variable-at-a-time approach (OVAT). Results obtained show that the yield of GAD increased 176% from 1.99 U/mg to 3.50 U/mg.ConclusionOf the techniques used to extract GAD from A. oryzae NSK, sonication was found to be the best. Under optimized conditions, about 176% of GAD was recovered compared to recovery under non optimized conditions. The high production level of GAD in this strain offers an opportunity to conduct further studies on GABA production at a larger scale.

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“…Lactobacillus brevis has a relatively weak proteolytic system and thus fewer activities are reported 15 whereas Pediococcus acidilactici is poorly studied with respect to its proteolytic system. This protocol has also been assessed and compared with previously described methods for DPP-III and aminopeptidase B [8][9][10][11][12][13] and the results are shown in Fig. 3 This newly developed method resulted in even better extraction.…”

Section: Resultsmentioning

confidence: 99%

“…To date, several methods for extracting membrane proteins have been described. [8][9][10][11][12][13] All these have their own inherent advantages and disadvantages and none benefit from the existence of internationally recognized standards for universally applicable extraction of membrane proteins.…”

Section: Introductionmentioning

confidence: 99%

An improved protocol for rapid extraction of membrane enzymes from Gram positive bacteria

et al. 2012

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Release of Cell-associated Tannase of Serratia ficaria DTC by Sonication, Surfactants and Solvents

Cited by 5 publications

References 16 publications

Purification, Characterization and Application of Tannase Enzyme Isolated from Marine Aspergillus nomius GWA5

Purification, Characterization and Application of Tannase Enzyme Isolated from Marine Aspergillus nomius GWA5

A comparative study of extraction techniques for maximum recovery of glutamate decarboxylase (GAD) from Aspergillus oryzae NSK

An improved protocol for rapid extraction of membrane enzymes from Gram positive bacteria

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