Application ob hydrogen high pressure bioreactor using immobilized microorganisms

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Immobilized microorganisms have been used for NADH regeneration under hydrogen high pressure. This NADH regeneration system has been applied to production of L-phenylalanine. We attempt to produce L-phenlalanine from phenylpyruvate by immobilized cells of Nocardis opaca. N.opace was cultivated in nutrient broth. The cultured cells were immobilized in 2% Ca-clginate. Continual production of phenylalanine was carried out in the medium containing phenylpyruvate, NH_4Cl and immobilized whole cells at 37゜C for 10h under 100atm hydrogen. Production rate of phenylalanine was 0.24 umol/min-cm^3 gel with a conversion of 82%. Immobilized cells retained 76% of phenylalanine production rate after 10h reactions were repeated 11 times with two intervening reactivation.The immobilized N.opaca was applied to phenylalanine methyl ester production. NADH regeneration system was also cooperated in this system. production. of phenylalanine methyl ester was carrid out in water-organic solvent system containing phenlpyruvate methy ester, NH_4 Cl, NAD^+ and immobilized cells in the presence of hydrogen at 100atm. Immobilized cells of N. opara produced phenylalanine methy ester at the rate of 0.02 <micrn>mol/ min/cm^3 gel by 36h activated immobilized cells with a conversion of 80%. The phenylalanine methyl ester production was followed by synthesis of zaspartame from phenylpyruvate methyl ester, NH_4Cl, NAD^+, Z-aspartic acid by immobilized N.opara - thermolysin. production rate of Z-aspartame was 22.4 <micrn>mol/min/g thermolysin. The conversion was 86%.Hydrogenation of fatty acid was carried out by metal catalysts and immobilized tumen bacteria. Linoleic acid was hydrogenated with high selectivity to oleic acid by palladium catalysts in the presence og added phenylacetaldehyde. Oleic acid was hydrogenated to steatic acid by immobilized Fusocillus sp. in water-ethyl acetate system.