Production of chorismate mutase-prephenate dehydrogenase by a strain of Escherichia coli carrying a multicopy, tyrA plasmid

Bhosale, Suresh B.; Rood, Julian I.; Sneddon, Margaret K.; Morrison, John F.

doi:10.1016/0304-4165(82)90372-5

Cited by 9 publications

(7 citation statements)

References 13 publications

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“…The specific activity for mutase was 102 units/mg of protein, and that for dehydrogenase was 80 units/mg of protein. These values are higher than those reported previously (Bhosale et al, 1982), and this is attributable, at least in part, to the increase in activity that occurs on increasing the concentration of dithiothreitol in stock enzyme solutions from 1 to 20 mM.…”

Section: Methodscontrasting

confidence: 72%

“…Purification of Hydroxyphenylpyruvate Synthase. The enzyme was obtained from a strain of E. cotí (JFM 30) which produces very much higher levels of hydroxyphenylpyruvate synthase than the parent strain (Bhosale et al, 1982). The purification procedure was essentially that developed by these same authors except that all buffer solutions were degassed and bubbled with nitrogen.…”

Section: Methodsmentioning

confidence: 99%

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Chorismate mutase-prephenate dehydrogenase from Escherichia coli: spatial relationship of the mutase and dehydrogenase sites

Christopherson¹,

Heyde²,

Morrison³

1983

Biochemistry

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The inhibition of the bifunctional enzyme chorismate mutase-prephenate dehydrogenase (4-hydroxyphenylpyruvate synthase) by substrate analogues has been investigated at pH 6.0 with the aim of elucidating the spatial relationship that exists between the sites at which each reaction occurs. Several chorismate and adamantane derivatives, as well as 2-hydroxyphenyl acetate and diethyl malonate, act as linear competitive inhibitors with respect to chorismate in the mutase reaction and with respect to chorismate in the mutase reaction and with respect to prephenate in the dehydrogenase reaction. The similarity of the dissociation constants for the interaction of these compounds with the free enzyme, as determined from the mutase and dehydrogenase reactions, indicates that the reaction of these inhibitors at a single site prevents the binding of both chorismate and prephenate. However, not all the groups on the enzyme, which are responsible for the binding of these two substrates, can be identical. At lower concentrations, citrate or malonate prevents reaction of the enzyme with prephenate, but not with chorismate. Nevertheless, the combining sites for chorismate and prephenate are in such close proximity that the diethyl derivative of malonate prevents the binding of both substrates. The results lead to the proposal that the sites at which chorismate and prephenate react on hydroxyphenylpyruvate synthase share common features and can be considered to overlap.

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Section: Methodscontrasting

confidence: 72%

Section: Methodsmentioning

confidence: 99%

Chorismate mutase-prephenate dehydrogenase from Escherichia coli: spatial relationship of the mutase and dehydrogenase sites

Christopherson¹,

Heyde²,

Morrison³

1983

Biochemistry

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“…These values are comparable to those reported by Hudson et al (1984) for the enzyme purified from the regulatory mutant JP2319. But they are almost twice the values reported for the enzyme isolated from JFM30 (Bhosale et al, 1982) and from the regulatory mutant JP2312 (SampathKumar & Morrison, 1982a). The use of a Matrex Blue A column represented the major purification step as judged by the 12-fold increase in specific activities over that of the ammonium sulfate fraction.…”

Section: Resultsmentioning

confidence: 77%

“…Bacterial Strains and Growth Conditions. E. coli strain JFM30, constructed by Bhosale et al (1982), was derived from strain AT2273 (F~tyrA352) carrying the multicopy plasmid pKB45 (Tcr pheA+ aroK+ tyrA+) (Zurawski et al, 1978). The strain yielded an amount of chorismate mutase-prephenate dehydrogenase that was 5000-fold higher than that yielded by the wild-type organism.…”

Section: Methodsmentioning

confidence: 99%

Chorismate mutase-prephenate dehydrogenase from Escherichia coli. 1. Kinetic characterization of the dehydrogenase reaction by use of alternative substrates

Turnbull¹,

Cleland²,

Morrison³

1990

Biochemistry

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The bifunctional enzyme involved in tyrosine biosynthesis, chorismate mutase-prephenate dehydrogenase, has been isolated from extracts of a plasmid-containing strain of Escherichia coli K12 and purified to homogeneity by a modified procedure that involves chromatography on both Matrex Blue A and Sepharose-AMP. Detailed studies of the dehydrogenase reaction have been undertaken with analogues of prephenate that act as substrates. The analogues, which included two of the four possible diastereoisomers of 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate (deoxodihydroprephenate) as well as D- and L-arogenate, were synthesized chemically. As judged by their V/K values, all analogues were poorer substrates than prephenate. The order of their effectiveness as substrates is prephenate greater than one isomer of 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate greater than L-arogenate greater than other isomer of 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate greater than D-arogenate. Thus the dehydrogenase activity is dependent on the degree and position of unsaturation in the ring structure of prephenate as well as on the type of substitution on the pyruvyl side chain. With prephenate as a substrate, the reaction is irreversible because it involves oxidative decarboxylation. By contrast, 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate undergoes only a simple oxidation, and thus, with this substrate, the reaction is reversible. Steady-state velocity data, obtained by varying substrates over a range of higher concentrations, suggest that the dehydrogenase reaction conforms to a rapid equilibrium, random mechanism with 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate as a substrate in the forward reaction or with the corresponding ketone derivative as a substrate in the reverse direction. The initial velocity patterns obtained by varying prephenate or 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate over a range of lower concentrations, at different fixed concentrations of NAD, were nonlinear and consistent with a unique model that is described by a velocity equation which is the ratio of quadratic polynomials. An equilibrium constant of 1.4 x 10(-7) M for the reaction in the presence of 1-carboxy-4-hydroxy-2-cyclohexene-1-propanoate indicates that the equilibrium lies very much in favor of ketone production.

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“…The activity of ARO7 was allosterically inhibited by tyrosine and the conversion from PREP to 4HPP catalyzed by TYR1, which was reported as a bottleneck of the L -tyrosine synthetic pathway in S. cerevisiae (Mao et al, 2017). TyrA from E. coli ( Ec TyrA ) was a bifunctional NAD + -dependent fused chorismate mutase/prephenate dehydrogenase (Bhosale et al, 1982) and overexpression of TyrA improved tyrosine production in E. coli (Lutke-Eversloh and Stephanopoulos, 2007). Furthermore, Ec TyrA was reported to be a homodimer consisted of CM domain and PDH domain, and was regulated by the feedback inhibition of tyrosine.…”

Section: Discussionmentioning

confidence: 99%

Rational Engineering of Chorismate-Related Pathways in Saccharomyces cerevisiae for Improving Tyrosol Production

Guo

Huang

Líu

et al. 2019

Front. Bioeng. Biotechnol.

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Tyrosol is extensively used in the pharmaceutical industry as an important natural product from plants. In this study, an exogenous pathway involved in catalyzing tyrosine to tyrosol was introduced into Saccharomyces cerevisiae . Furthermore, The pyruvate decarboxylase gene pdc1 was deleted to redirect the flux distribution at the pyruvate node, and a bifunctional NAD + -dependent fused chorismate mutase/prephenate dehydrogenase from E. coli ( Ec TyrA) and its' tyrosine inhibition resistant mutant ( Ec TyrA M53I/A354V ) were heterologously expression in S. cerevisiae to tuning up the chorismate metabolism effectively directed the metabolic flux toward tyrosol production. Finally, the tyrosol yield of the engineered strain GFT-4 was improved to 126.74 ± 6.70 mg/g DCW at 48 h, increased 440 times compared with that of the control strain GFT-0 (0.28 ± 0.01 mg/g DCW). The new synergetic engineering strategy developed in this study can be further applied to increase the production of high value-added aromatic compounds derived from aromatic amino acid or shikimate in S. cerevisiae .

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Production of chorismate mutase-prephenate dehydrogenase by a strain of Escherichia coli carrying a multicopy, tyrA plasmid

Cited by 9 publications

References 13 publications

Chorismate mutase-prephenate dehydrogenase from Escherichia coli: spatial relationship of the mutase and dehydrogenase sites

Chorismate mutase-prephenate dehydrogenase from Escherichia coli: spatial relationship of the mutase and dehydrogenase sites

Chorismate mutase-prephenate dehydrogenase from Escherichia coli. 1. Kinetic characterization of the dehydrogenase reaction by use of alternative substrates

Rational Engineering of Chorismate-Related Pathways in Saccharomyces cerevisiae for Improving Tyrosol Production

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