Biomolecular Engineering in the European Community: by E. Magnien (auth.), E. Magnien (eds.)

By E. Magnien (auth.), E. Magnien (eds.)

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Extra resources for Biomolecular Engineering in the European Community: Achievements of the Research Programme (1982 – 1986) — Final Report

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M. itu rather than depolymerizing the molecule. The genetics of the mechanisms involved are now under investigation. Work done in Belgium (F. Brunei and J. Davison, Bruxelles) has focused on the demethylation of the lignin phenolic ring, as a mean of modifying its structure. Demethylation of the lijrnin substructure model compound, vanillate, to protocatechuate, was chosen as a model system and the genes encoding the demethylation function were isolated from Pseudomonas sp. The first part of the work involved the construction of efficient cloning systems for Pseudomonas.

Three genes were cloned, encoding the positive regulator (NPRl) of several ammonia sensitive permeases, the proline permease (PUT4) and glutamate dehydrogenase, which is implicated in the ammonia dependent catabolic repression of permeases. The overexpression of NPRI or PUT4 from the multicopy plasmid resulted in a 4 to 25 fold increase in permease activity and this, independently from the presence of ammonium ions, which normally reduce amino acid uptake. Such a result could prove essential in the selective pumping of amino acids from an ammonia containing complex industrial medium.

Host strain was built, which did not exhibit any plasmid encoded proteinase or lactose degrading activities. l). ~~. Moreover, second generation vectors have been derived from them, permitting the isolation of strong regulatory sigrials which allow the proper expression of heterologous genes. ~ Finally, the was increased 1600 fold so that direct screening of recombinant DNA may now be achieved. EJis proteinases and phospho-p-D-galactosidases Of particular interest is the cloning of the proteinase gene from genes.

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