- Widely applicable background depletion step enables transaminase evolution through solid-phase screening
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Directed evolution of transaminases is a widespread technique in the development of highly sought-after biocatalysts for industrial applications. This process, however, is challenged by the limited availability of effective high-throughput protocols to evaluate mutant libraries. Here we report a rapid, reliable, and widely applicable background depletion method for solid-phase screening of transaminase variants, which was successfully applied to a transaminase from Halomonas elongata (HEWT), evolved through rounds of random mutagenesis towards a series of diverse prochiral ketones. This approach enabled the identification of transaminase variants in viable cells with significantly improved activity towards para-substituted acetophenones (up to 60-fold), as well as tetrahydrothiophen-3-one and related substrates. Rationalisation of the mutants was assisted by determination of the high-resolution wild-type HEWT crystal structure presented herein.
- Planchestainer, Matteo,Hegarty, Eimear,Heckmann, Christian M.,Gourlay, Louise J.,Paradisi, Francesca
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- Implementation of biocatalysis in continuous flow for the synthesis of small cyclic amines
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Significant progress has been made in establishing transaminases as robust biocatalysts for the green and scalable synthesis of a diverse range of chiral amines.[1] However, very few examples on the amination of small cyclic ketones have been reported.[2] Cyclic ketones are particularly challenging for transaminase enzymes because they do not display the well-defined small and large substituent areas that are characteristic for the biocatalytic mechanism. In this work, we exploited the broad substrate scope of the (S)-selective transaminase from Halomonas elongata (HeWT) to develop an efficient biocatalytic system in continuous flow to generate a range of small cyclic amines which feature very often in pharmaceuticals and agrochemicals.[3] Tetrahydrofuran-3-one and other challenging prochiral ketones were rapidly (5-45 min) transformed to their corresponding amines with excellent molar conversion (94-99%) and moderate to excellent ee.
- Hegarty, Eimear,Paradisi, Francesca
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p. 890 - 894
(2020/12/25)
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- PURINE DERIVATIVES FOR USE IN THE TREATMENT OF ALLERGIC, INFLAMMATORY AND INFECTIOUS DISEASES
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Compounds of formula (I): wherein R1 is C1-6alkylamino, or C1-6alkoxy; m is an integer having a value of 3, 4, or 5; n is an integer having a value of 0 to 3; p is an integer having a value of 1 or 2 and salts thereof are inducers of human interferon. Compounds which induce human interferon may be useful in the treatment of various disorders, for example the treatment of allergic diseases and other inflammatory conditions for example allergic rhinitis and asthma, the treatment of infectious diseases and cancer, and may also be useful as vaccine adjuvants.
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Page/Page column 48
(2010/04/03)
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- Method of treating arrhythmias
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A method for treating arrhythmias is provided comprising administering a low dose of an adenosine receptor agonist, in particular CVT-510, to a mammal in need of such treatment.
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