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KMID : 1160619970020040354
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Preventive Nutrition and Food Science
1997 Volume.2 No. 4 p.354 ~ p.359
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Production and Characterization of Phenylalanine Ammonia-lyase from Rhodotorula aurantiaca K-505
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Cho Dae-Haeng
Chae Hee-Jeong
Kim Eui-Yong
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Abstract
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Optimal cultivation conditions for the production of phenylalanine ammonia-lyase(PAL) from Rhodotorula aurantiaca K-505 were selected, and the kinetic parameters of the produced PAL were determined. The most suitable carbon and nitrogen sources were glucose and tryptone, respectively. The strain expressed PAL constitutively when using the optimized semi-complex media. High cell density culture could be critical for maximal production of PAL since the PAL biosynthesis was growth associated. Maximum PAL activity was observed at initial pH 6.0. Although the cell growth was not markedly affected by temperature between 22 and 28℃, the cells yielded the maximum PAL activity when cultivated at 22℃. The maximum activity for the deamination of L-phenylalanine to trans-cinnamic acid was observed around pH 8.8. The PAL activity gave the maximum at 45℃, and greatly decreased at higher than 50℃. Activation energy(E_a) calculated from Arrhenius equation was 6.28㎉/mol in the range of 22℃ to 40℃. A Hanes-Woolf plot showed that the enzyme reaction follows Michaelis- Menten equation, whose K_M and V_(max) values were 4.65×10^(-3)M and 0.89μ mol/mg-min respectively.
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KEYWORD
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Phenylalanine ammonia-lyase(PAL), Rhodotorula aurantiaca, phenylalanine, culture conditions, kinetic parameters
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