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HLADH requires NADH as cofactor, TBADH and LBADH require NADPH as cofactor. The LBADH used in this work was overexpressed in a recombinant Escherichia coli strain and used in the form of crude cell extract, see: B. Riebel, Dissertation, Universität Düsseldorf, 1996.
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12444250929
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note
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To minimize changes in the phase volumes, the organic solvent was previously saturated with water. The reaction mixture was shaken vigorously for one minute resulting in the saturation of the aqueous phase with the organic solvent.
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32
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12444287819
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note
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Protocol for the standard assays are available as Supporting Information.
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33
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12444290737
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note
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Neither the nature (competitive, uncompetitive or noncompetitive) nor the cause of this instantaneous inhibition was further investigated; the values are available as supporting information.
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34
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12444266633
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note
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dea.
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35
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12444282550
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note
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After the equilibrium conversion had been reached, the organic phase containing the product was removed and replaced by a fresh solution of acetophenone in MTBE, so that conversion could take place anew.
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To improve the stability of the cofactor it was necessary to increase the aqueous pH-value to 7.0 (5.5 in reference [24]), consequently enhancing the rate of the side reaction. Under this condition the total turnover number of the cofactor (defined as the amount of product per amount of cofactor) was 2500.
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Chemical functionality is a pragmatic label for a set of molecular properties, such as the presence of certain heteroatoms and residues, the absence of others, molecular geometry, electronic density distribution and frontier orbitals, and consequent macroscopic properties of solvents, such as surface tension, solubility, solvatation energy.
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