Production of
            <scp>l</scp>
            -Phenylalanine from
            <i>trans</i>
            -Cinnamic Acid with
            <i>Rhodotorula glutinis</i>
            Containing
            <scp>l</scp>
            -Phenylalanine Ammonia-Lyase Activity

Yamada, Shigeki; Nabe, Koichi; Izuo, Nobuhiko; Nakamichi, Katsühiko; Chibata, Ichiro

doi:10.1128/aem.42.5.773-778.1981

Cited by 140 publications

(47 citation statements)

References 15 publications

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“…The most notable among these is overcoming diffusion barriers that have been discussed above, resulting in increased product yield. The product yield (85% substrate conversion) obtained in this study is appreciably >70% substrate conversion reported for PAL catalyzed synthesis of L‐Phe from t ‐CA using Rhodotorula yeast cells by Evans et al7, 17 and Yamada et al27 In the case of PAL catalyzed L‐PM production obtained in this study, approximately 70% product yield is far >16% yield obtained by Nakamura et al,28 and comparable with the amount obtained in our earlier studies using R. glutinis yeast whole cells 10, 11. Using the conditions optimized in our earlier study,15 we obtained ˜1.9 nmoles of L‐Phe transformed/min/mg dry cells.…”

Section: Discussionsupporting

confidence: 71%

Enzyme activity evaluation of organic solvent‐treated phenylalanine ammonia lyase

Quinn

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D’Cunha

2011

Biotechnology Progress

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The direct one-step synthesis of L-phenylalanine methyl ester in an organic-aqueous biphasic system using phenylalanine ammonia lyase (E.C.4.3.1.5, PAL) containing Rhodotorula glutinis yeast whole cells was reported earlier. We report here further optimization of this biotransformation using isolated PAL, when the lyophilized enzyme is treated with different water miscible and water immiscible organic solvents. Use of isolated PAL enzyme is advantageous in overcoming diffusion barriers encountered when using PAL containing R.glutinis whole cells, and resulted in increased product yield due to better interaction of enzyme with the substrate. Among the water miscible solvents, ethanol treated and methanol-treated enzymes supported maximum PAL forward and reverse activities; respectively. In the water immiscible solvents category, heptane-treated enzyme exhibited maximal activity for both PAL forward and reverse reactions. PAL activity obtained with enzyme specimens treated with methanol, ethanol, and heptane varied in the range of 91–99% of that observed in aqueous buffer medium for the forward reaction; and 89–95% for the reverse reaction. n-butanol,acetone, and benzene were found to have a inhibitory effect on PAL enzyme, in that, it resulted in only 31–33% activity of that obtained with aqueous solution. Raman spectroscopy was used to monitor amide I and II bands which are sensitive to changes in the secondary structure of proteins. No changes in structure could be detected from the analyses of AI and AII bands of PAL spectra. This data obtained for PAL, a tetramer, could be significant in predicting how solvent interactions affect the structure and function of multimeric proteins and enzymes in nonaqueous media.

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Section: Discussionsupporting

confidence: 71%

Enzyme activity evaluation of organic solvent‐treated phenylalanine ammonia lyase

Quinn

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D’Cunha

2011

Biotechnology Progress

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“…The reverse catalytic reaction was demonstrated using the procedure of Yamada et al (1981). All four PAL isoenzymes formed L-phenylalanine from (Q-cinnamate and ammonia at identical rates, as did authentic PAL from the yeast R. glutinis, which was used both as a reference and to optimize the assay conditions.…”

Section: Comparison Of Enzymic Propertiesmentioning

confidence: 99%

Structural and Catalytic Properties of the Four Phenylalanine Ammonia‐Lyase Isoenzymes from Parsley (Petroselinum Crispum Nym.)

Appert¹,

Logemann²,

Hahlbrock³

et al. 1994

European Journal of Biochemistry

120

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Near-full-length cDNAs for the four phenylalanine ammonia-lyase (PAL) isoenzymes in parsley (Petruselinum crispum Nym.) were cloned and the complete amino acid sequences deduced. Fusion proteins with glutathione S-transferase were expressed in Escherichia coli, purified and cleaved. All of the resulting phenylalanine ammonia-lyase proteins, as well as the fusion proteins, were catalytically active. The turnover number of one selected isoenzyme, PAL-1, was estimated to be around 22 s-' for each active site. In contrast to a certain degree of differential expression in various parts of parsley plants, the four phenylalanine ammonia-lyase isoenzymes exhibited very similar apparent K,,, values for L-phenylalanine (15-24.5 pM) as well as identical temperature (58°C) and pH (8.5) optima. All of them were competitively inhibited by (E)-cinnamate with similar efficiency (K, values: 9

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“…Ammonia-lyases, in contrast, have been shown to catalyze this type of reaction with complete specificity. For example, histidine ammonialyase (HAL) and phenylalanine ammonia-lyase, which function physiologically in the nonoxidative deamination of their respective amino acids, can catalyze the amination of their a3,p-unsaturated carboxylic acids, albeit at a very slow rate (17,18). Previous results suggest that the equilibrium constant for these enzymes approaches 4 (17, 18; R. L. Fuchs, unpublished data).…”

mentioning

confidence: 99%

In vivo synthesis of histidine by a cloned histidine ammonia-lyase in Escherichia coli

Fuchs¹,

Kane²

1985

J Bacteriol

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Histidine ammonia.lyase catalyzes the first step in histidine catabolism, the deamination of histidine to urocanate and ammonia. In vitro experiments have shown that histidine ammonia-lyase also can catalyze the reverse (amination) reaction, histidine synthesis, relatively efficiently under extreme reaction conditions (4 M NH4OH, pH 10). An Escherichia coli hisB deletion strain was transformed with a pBR322 derivative plasmid (pCB101) containing the entire KkbsieUa aerogenes histidine utilization (hut) operon to determine whether the catabolic hi$tidine ammonia-lyase could function biosynthetically in vivo to satisfy the histidine auxotrophy. Although the initial construct did not grow on media containing urocanate and ammonia as a source of histidine, spontoweous mutanits possessing this ability were isolated. Four mutants characterized grew at doubling times of 4 h compared with 1 h when histidine was present, suggesting that histidine synthesis, although unequivocally present, remained growth limiting. Each mutant contained a plasmid-encoded mutation which eliminated urocanase activity, the second enzyme in the Hut catabolic pathway. This genetic block led to the accumulation of high intracellular levels of urocanate, which was subsequently converted to histidine via histidine ammonia-lyase, thus satisfying the histidine auxotrophic requirement.

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Production of l -Phenylalanine from trans -Cinnamic Acid with Rhodotorula glutinis Containing l -Phenylalanine Ammonia-Lyase Activity

Cited by 140 publications

References 15 publications

Enzyme activity evaluation of organic solvent‐treated phenylalanine ammonia lyase

Enzyme activity evaluation of organic solvent‐treated phenylalanine ammonia lyase

Structural and Catalytic Properties of the Four Phenylalanine Ammonia‐Lyase Isoenzymes from Parsley (Petroselinum Crispum Nym.)

In vivo synthesis of histidine by a cloned histidine ammonia-lyase in Escherichia coli

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