Mapping the substrate scope of monoamine oxidase (MAO-N) as a synthetic tool for the enantioselective synthesis of chiral amines

Herter, Susanne; Medina, Florian; Wagschal, Simon; Benhaim, Cyril; Leipold, Friedemann; Turner, Nicholas J.

doi:10.1016/j.bmc.2017.07.023

Cited by 43 publications

(41 citation statements)

References 40 publications

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“…4 ). Consistent with the findings on substrate specificity seen in previous reports, 33 , 36 induced cells incubated with CHA had the lowest fluorescence, followed by AMBA and then AA. As predicted from the in vitro k cat values, induced cells incubated with BTA and BZA had the highest fluorescence.…”

Section: Resultssupporting

confidence: 92%

“…In particular, the monoamine oxidase from Aspergillus niger (MAO-N) 26 – 30 has been heavily engineered to oxidise a wide range of primary, secondary and even tertiary amines. 31 – 36 When coupled with a chemical reducing agent, a dynamic kinetic resolution can be achieved to yield enantiomerically pure amines, which are valuable building blocks for many fine and speciality chemicals, including many pharmaceuticals. 37 , 38 The most widely used screening assay for MAO-N and variants thereof involves a colony-based screen in which colonies form a red colour when amine-oxidising enzyme activity produces H 2 O 2 in the presence of an enzyme substrate, a chromogenic peroxidase substrate and an added (extracellular) peroxidase.…”

Section: Introductionmentioning

confidence: 99%

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Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting

Sadler

Currin

Kell

2018

Analyst

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Section: Resultssupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting

Sadler

Currin

Kell

2018

Analyst

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“…Turner et al. reported a deracemization reaction that uses an engineered S ‐stereoselective monoamine oxidase from Aspergillus niger in the presence of a reductant ,,. The oxidase‐catalyzed deracemization method has merit in that it does not need to be conjugated with other enzymes compared to the systems that use amine transaminase or amine dehydrogenase.…”

Section: Introductionmentioning

confidence: 99%

Expansion of the Substrate Specificity of Porcine Kidney D‐Amino Acid Oxidase for S‐Stereoselective Oxidation of 4‐Cl‐Benzhydrylamine

et al. 2018

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Discovery and development of enzymes for the synthesis of chiral amines have been a hot topic for basic and applied aspects of biocatalysts. Based on our X‐ray crystallographic analyses of porcine kidney D‐amino acid oxidase (pkDAO) and its variants, we rationally designed a new variant that catalyzed the oxidation of (S)‐4‐Cl‐benzhydrylamine (CBHA) from pkDAO and obtained it by functional high‐throughput screening with colorimetric assay. The variant I230A/R283G was constructed from the variant R283G which had completely lost the activity for D‐amino acids, further gaining new activity toward (S)‐chiral amines with the bulky substituents. The variant enzyme (I230A/R283G) was characterized to have a catalytic efficiency of 1.85 s−1 for (S)‐CBHA, while that for (R)‐1‐phenylethylamine was diminished 10‐fold as compared with the Y228L/R283G variant. The variant was efficiently used for the synthesis of (R)‐CBHA in 96 % ee from racemic CBHA by the deracemization reaction in the presence of reducing agent such as NaBH4 in water. Furthermore, X‐ray crystallographic analysis of the new variant complexed with (S)‐CBHA, together with modelling study clearly showed the basis of understanding the structure‐activity relationship of pkDAO.

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“…Ever since the recognition of mutation itself, it has been obvious that changes in the primary sequence of a protein are more or less tightly coupled to changes in its properties [323,324], whether they be k cat [325], thermotolerance [326], substrate specificity [327], solvent tolerance [328], or any other properties.…”

Section: Directed Evolution: Changing the Sequence Of The Target Tranmentioning

confidence: 99%

Control of metabolite efflux in microbial cell factories: current advances and future prospects

Kell¹

2018

Preprint

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The yield and ease of purification of biotechnological products are typically greatly enhanced if they can be secreted into the external medium. Although not universally appreciated, however, small molecule biotechnological products do not ‘float across’ the phospholipid bilayer portion of biological membranes, and thus they need transporters to assist their passage into the extramembrane and extracellular spaces. Some of these transporters may be reversible, equilibrative transporters that might more normally be used for uptake, while others may have an efflux directionality imposed on them via suitable energy coupling mechanisms. Despite the energetic costs of small molecule synthesis, natural evolution does in fact provide a number of mechanisms by which the secretion of such products can actually enhance fitness. Where available these provide useful starting points, and assaying for such activities is crucial. In particular, in a systems biology approach, we first need to identify such/suitable activities before we can seek to increase them. They may then be improved through promoter engineering, via manipulation of control elements, or by directed evolution of the transporter proteins themselves. Modern methods of synthetic biology provide enormous opportunities for all kinds of efflux transporter engineering; they are just beginning to be realised.

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Mapping the substrate scope of monoamine oxidase (MAO-N) as a synthetic tool for the enantioselective synthesis of chiral amines

Cited by 43 publications

References 40 publications

Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting

Ultra-high throughput functional enrichment of large monoamine oxidase (MAO-N) libraries by fluorescence activated cell sorting

Expansion of the Substrate Specificity of Porcine Kidney D‐Amino Acid Oxidase for S‐Stereoselective Oxidation of 4‐Cl‐Benzhydrylamine

Control of metabolite efflux in microbial cell factories: current advances and future prospects

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