Real-time assay of immobilized tannase with a stopped-flow conductometric device

Chang, Fu-Shiang; Chen, Pochung; Chen, Richie L.C.; Lu, Faming; Cheng, Tzong‐Jih

doi:10.1016/j.bioelechem.2005.12.004

Cited by 10 publications

(5 citation statements)

References 10 publications

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“…12 The kinetics of solutes uptake by colloidal particles of synthetic resins was studied by the means of the technique equipped with a conductivity detection. 13 Previous work 14 had shown that reaction between 1,1,1-trichloro-3-methyl-3-phospholene (TCMP) and methanol for generation of 1-methoxy-3-methyl-2-phospholene oxide (MMPO) is the two step reaction with an initial fast step. The fast reaction between TCMP and methanol could not be followed successfully by stopped-flow spectrophotometric detection because the optical absorptions of TCMP (reactant) and MMPO (product) are very close to each other and may be difficult to recognize by this method.…”

Section: Introductionmentioning

confidence: 99%

Establishing a new conductance stopped-flow apparatus to investigate the initial fast step of reaction between 1,1,1-trichloro-3-methyl-3-phospholene and methanol under a dry inert atmosphere

Habibi‐Khorassani

Ebrahimi

Maghsoodlou

et al. 2011

Analyst

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In the present work, for the first time, investigation of the initial fast step of reaction between 1,1,1-trichloro-3-methyl-3-phospholene (TCMP) and methanol was studied under a dry inert atmosphere by a newly constructed CSF apparatus by means of a further development in the configuration of the previous stopped-flow spectrophotometer (SFS). Hence, it was necessary to make many changes to the stopped-flow apparatus: replacement of the spectrophotometer amplifier with a conductance amplifier and the use of a conductivity cell to replace the optical one. The conductivity cell was made of polyethylene capillary tube (1 mm internal diameter; i.d.) and its inside dimensions were 1 mm i.d. × 3 mm long. Two tube electrodes which are made of stainless steel (0.8 mm i.d. × 14 mm long) were fixed at opposite ends of the observation cell and their outer surfaces were connected to the conductivity bridge amplifier by the two lead wires. The mixer was a 3-way Teflon valve and the distance between the mixer and the end of the observation cell was 30 mm. For each run, at least 24 µL of solution was required for a typical trace with a dead time of about 5 ms. Because of the extreme sensitivity of TCMP to moisture, the stopped-flow (CSF) apparatus was used inside a glove bag under a dry nitrogen atmosphere. Kinetic parameters for pseudo first-order reaction involving k(obs) = 30 s(-1) at 22 °C and activation energy E(a) = 13.55 KJ mol(-1) were successfully calculated for the initial fast step of the reaction between TCMP and methanol at 22 °C.

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Section: Introductionmentioning

confidence: 99%

Establishing a new conductance stopped-flow apparatus to investigate the initial fast step of reaction between 1,1,1-trichloro-3-methyl-3-phospholene and methanol under a dry inert atmosphere

Habibi‐Khorassani

Ebrahimi

Maghsoodlou

et al. 2011

Analyst

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“…Glass Bead Activated With GA Glass beads (5 g) were activated in 4 ml of GA 9% (v/v) (GA), previously prepared in sodium acetate buffer (10 mmol L −1 pH 5.0) (Chang et al, 2006). The suspension was kept under agitation for 2 h at room temperature.…”

Section: Amberlite Activated With Gamentioning

confidence: 99%

Immobilization of the Tannase From Aspergillus fumigatus CAS21: Screening the Best Derivative for the Treatment of Tannery Effluent Using a Packed Bed Reactor

Cavalcanti

Maestrello

Guimarães³

2021

Front. Bioeng. Biotechnol.

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Enzyme immobilization is an important alternative to stabilize enzyme properties favoring the efficiency of derivatives (enzyme + support/matrix) for different purposes. According to this, the current study aimed to immobilize the Aspergillus fumigatus CAS21 tannase and the use of the derivatives in the treatment of the effluent produced by the tannery industry. The tannase was immobilized on sodium alginate, DEAE-Sephadex, amberlite, and glass pearls as supports. Calcium alginate was the most adequate support for tannase immobilization with 100% yield and 94.3% for both efficiency and activity. The best tannase activity for the calcium alginate derivative was obtained at 50°C–60°C and pH 5.0. Thermal and pH stabilities evaluated for 24 h at 30°C–60°C and pH 4–7, respectively, were improved if compared to the stability of the free enzyme. Considering the reuse of the calcium alginate derivative, 78% of the initial activity was preserved after 10 catalytic cycles, and after the 9-month storage at 4°C, the activity was maintained in 70%. This derivative was applied in a packed bed reactor (PBR) for the treatment of tannin-rich effluents from the tannery industry. The reduction of the tannin content was effective reaching degradation of 74–78% after 48 h of PBR operation. The concentration of total phenolic compounds was also reduced, and the color and clarity of the effluent improved. In conclusion, the calcium alginate derivative is an attractive alternative as biocatalyst for large-scale treatment of the effluents from the tannery industry.

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“…In many industries, enzymes are immobilized on a solid surface for repetitive use. The application of the stopped flow system for study of the kinetics of the immobilized enzyme was reported by stopping the substrate solution at the reactor packed with enzyme coated beads coupled with oncolumn detector, that is, conductometer, for real-time detection [67]. Arrangement of the manifold to reduce operation steps while yielding more kinetic data points was successfully attempted.…”

Section: Applications Of Flow-based Analysis Systems In Enzyme Kinmentioning

confidence: 99%

Flow-Based Systems for Rapid and High-Precision Enzyme Kinetics Studies

Hartwell

Grudpan

2012

Journal of Analytical Methods in Chemistry

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Enzyme kinetics studies normally focus on the initial rate of enzymatic reaction. However, the manual operation of steps of the conventional enzyme kinetics method has some drawbacks. Errors can result from the imprecise time control and time necessary for manual changing the reaction cuvettes into and out of the detector. By using the automatic flow-based analytical systems, enzyme kinetics studies can be carried out at real-time initial rate avoiding the potential errors inherent in manual operation. Flow-based systems have been developed to provide rapid, low-volume, and high-precision analyses that effectively replace the many tedious and high volume requirements of conventional wet chemistry analyses. This article presents various arrangements of flow-based techniques and their potential use in future enzyme kinetics applications.

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Real-time assay of immobilized tannase with a stopped-flow conductometric device

Cited by 10 publications

References 10 publications

Establishing a new conductance stopped-flow apparatus to investigate the initial fast step of reaction between 1,1,1-trichloro-3-methyl-3-phospholene and methanol under a dry inert atmosphere

Establishing a new conductance stopped-flow apparatus to investigate the initial fast step of reaction between 1,1,1-trichloro-3-methyl-3-phospholene and methanol under a dry inert atmosphere

Immobilization of the Tannase From Aspergillus fumigatus CAS21: Screening the Best Derivative for the Treatment of Tannery Effluent Using a Packed Bed Reactor

Flow-Based Systems for Rapid and High-Precision Enzyme Kinetics Studies

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