Development of enzyme-cartridge flow-injection analysis for industrial process monitoring: Part I. development and characterization

doi:10.1016/0003-2670(94)00264-9

Analytica Chimica Acta

Volume 298, Issue 2, 30 November 1994, Pages 141-149

https://doi.org/10.1016/0003-2670(94)00264-9 Get rights and content

Abstract

The preparation of cartridges with five different enzymes: glucose oxidase (GOD), pyranose oxidase (PyOD), alcohol oxidase (AOD), l-amino acid oxidase (l-ASOD) and l-lactate-2-monooxigenase (LMO) immobilized on VA-Epoxy Biosynth carrier were optimized and standardized. The enzyme cartridges were characterized, integrated in flow injection analysis (FIA) systems and the optimal application conditions and salt effects were evaluated.

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Carbohydrate oxidases are found in all kingdoms of life but are mostly found in fungi. Their natural role is not always clear. Usage of molecular oxygen as electron acceptor is not a logical choice when the enzyme is part of a catabolic pathway. This chapter provides an overview of the occurrence and properties of carbohydrate oxidases. The physiological role of the different enzymes is discussed in relation to their origin, and the catalytic and structural properties are discussed in relation to their family background. It also provides a summary of the biocatalytic applications of carbohydrate oxidases. Carbohydrate oxidases are valuable enzymes for several applications. They are relatively stable and do not need expensive coenzymes. Carbohydrate oxidases are widely used in diagnostic applications, in the food and drinks industry, and for carbohydrate synthesis. They are also used for bleaching (production of H₂O₂) and as oxygen scavenger.
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Citation Excerpt :
However, the presence of 2.0 g l−1 acetate in a sample solution caused a reduction in the relative peak height to about 30%. The inhibitory effect of acetate on the activity of the enzyme reactor may be caused by the change of pH in the sample [25]. This effect can be removed by controlling the pH value in the sample and/or employing the calibration solution for fumaric acid, which contains some acetate in known concentrations.
A flow injection system has been developed for monitoring fumaric acid in biological processes by using fumarase and malic dehydrogenase immobilized on VA-Epoxy Biosynth E3-carrier. An immobilized fumarase and malic dehydrogenase reactor had a good operational lifetime (up to 18 h) and storage stability (up to 10 days). The flow injection analysis (FIA) system with the immobilized enzymes cartridge was characterized according to the factors affecting the activity of the immobilized enzymes, such as pH, temperature, sample matrix, etc. The optimal pH value of the immobilized enzymes was slightly shifted in the alkaline range, i.e. 9.0. Some components such as 1 mM imidazole, 5 mM sodium azide and 2 g l⁻¹ semicarbazide in a carrier buffer solution had significant inhibitory or activating effects on the response of the FIA system. But many components in the sample solution had no significant effects on the activity of the immobilized enzymes, except in the case of the 2 g l⁻¹ acetate sample. The linear concentration ranges of fumaric acid were investigated by changing the volumetric packing ratio of two immobilized enzymes in a cartridge. The cartridge with a 1:1 packing ratio had a linear concentration range of 0.05–0.3 g l⁻¹ with a correlation coefficient (r²) of 0.996, while the enzyme cartridge with a 1:2 packing ratio (or 1:4 packing ratio) had 0.1–0.6 g l⁻¹ with r²=0.998 (or 0.3–1.2 g l⁻¹ with r²=0.996). The FIA system utilizing the enzyme cartridge has been applied in order to monitor on-line the concentrations of fumaric acid in a few biological processes, i.e. a continuously stirred reactor and a cultivation process of Rhizopus oryzae Korean Collection for Type Cultures 6946.
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Development of enzyme-cartridge flow-injection analysis for industrial process monitoring: Part I. development and characterization

Abstract

J. Biotechnol.

J. Biol. Chem.

Anal. Chim. Acta