Abstract
The production of 5′-IMP and 5′-GMP by enzymatic conversion from RNA using a continuous two packed-bed reactor was investigated. 5′-Phosphodiesterase (5′PD) and 5′-adenylate deaminase (5′AD) were immobilized in an acrylic resin to produce derivatives with about 15 U/g of support. The kinetic properties of the enzymes were described by Michaelis-Menten models: no significant differences were found in the K m value of the free and immobilized 5′AD (60 and 20 μm, respectively), whereas for 5′PD the K m value was one order of magnitude higher for the immobilized enzyme (4.85 mg RNA/ml), probably due to diffusional limitations. Both enzymes remained stable after 8 h of use in a continuous packed-bed reactor whereas the half lives of the free enzymes were 193 min and 240 min at 40°C and 70°C for 5′AD and 5′PD, respectively. A procedure is proposed for the design of a continuous two packed-bed column process.
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F. Olmedo and F. Iturbe are with the Depto. de Alimentos y Biotecnologia, Facultad de Química, UNAM, México 04510, D.F., Mexico. J. Gomez-Hernández is with the Depto. de Biotecnología, UAM-1, Apdo. Postal 55-535, México 09340, D.F., Mexico. A. López-Munguía is with the Instituto de Biotecnología, Apartado Postal 510-3, Cuernavaca, Mor. 62271, Mexico
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Olmedo, F., Iturbe, F., Gomez-Hernández, J. et al. Continuous production of 5′-ribonucleotides from yeast RNA by hydrolysis with immobilized 5′-phosphodiesterase and 5′-adenylate deaminase. World Journal of Microbiology & Biotechnology 10, 36–40 (1994). https://doi.org/10.1007/BF00357560
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DOI: https://doi.org/10.1007/BF00357560