A family of native amine dehydrogenases for the asymmetric reductive amination of ketones

Mayol, O.; Bastard, Karine; Beloti, Lilian L.; Frese, Amina; Turkenburg, J.P.; Petit, Jean-Louis; Mariage, Aline; Debard, Adrien; Pellouin, Virginie; Perret, Alain; Berardinis, Véronique de; Zaparucha, Anne; Grogan, Gideon; Vergne-Vaxelaire, Carine

doi:10.1038/s41929-019-0249-z

Cited by 93 publications

(143 citation statements)

References 43 publications

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“…In comparison, methylamine results in 0.8% ( Sgor AmDH) to 40% ( Chat AmDH) of the original activity with ammonia at 100 equivalents of amine (see Experimental Section and Table S7 for details). This behavior is similar to that previously reported for nat‐AmDHs . In the case of MATOUAmDH2, significant activity was found also with ethylamine (6% and 16% of ammonia and methylamine activity, respectively), which again positions this enzyme as a very promising target for development.…”

Section: Resultssupporting

confidence: 89%

“…Isobutyraldehyde ( 2 a ) was the top tested substrate, with specific activities above 3.30 U mg −1 for Sgor AmDH, and up to 11.07 U mg −1 for MATOUAmDH2. Notably, activity was also detected with pentaldehyde and benzaldehyde, unlike for the reference enzymes . The dialdehyde glutaraldehyde was not able to be tested, due to crosslinking of the enzymes .…”

Section: Resultsmentioning

confidence: 96%

“…Environmental sequences that matched experimentally validated nat‐AmDHs ( Msme AmDH, Mvac AmDH, Myco AmDH, Micro AmDH, Apau AmDH and Cfus AmDH) via a Hidden Markov Model (HMM) genome mining approach, were retrieved from the OM‐RGC (12 proteins), MATOUv1 (9 proteins) and IGC (251 proteins) databases (see Experimental Section for details). Some representatives were selected for experimental assays.…”

Section: Resultsmentioning

confidence: 99%

“…No other reliable information could be extracted from the analysis of the genomic context and/or annotation of the other seven candidates with close homologs in UniprotKB database (Table S1). Many of them lack a predicted function or are annotated as putative dihydrodipicolinate reductases, as were the reference enzymes …”

Section: Resultsmentioning

confidence: 99%

“…The goal of the presented work was to access unexplored members of the recently identified natural AmDH family, and their abundance in the biosphere by an unprecedented metagenomics screen which includes eukaryotic members. This approach aims to rapidly enrich the sequence space and known properties of AmDHs for wider green chemical applications .…”

Section: Introductionmentioning

confidence: 99%

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Metagenomic Mining for Amine Dehydrogenase Discovery

et al. 2020

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Amine dehydrogenases (AmDHs) catalyze the enzymatic reduction of ketones to amines, serving as a suitable biocatalytic route for amine synthesis. A limited number of experimentally validated native AmDHs (nat‐AmDHs) have been reported recently, expanding the sequences with this function to complement the small set of engineered enzymes. Since researchers can now probe into the vast diversity of enzymes within niche environments by a metagenomics approach, a tandem metagenomic and bioinformatic approach is a powerful tool to identify new members of limited enzyme families to access new features in an iterative fashion. The previously untapped biocatalytic reservoirs of the ocean environment and human microbiome were screened for potential AmDHs using a hidden Markov model. Among the hundreds of hits, a subset of 18 enzymes was selected for further characterization and were confirmed to display AmDH activity. Additional analysis on six enzymes confirmed altered cofactor specificities and variation in substrate scopes, catalytic efficiencies, and active site residues compared to the reference nat‐AmDHs previously described. Particularly, MATOUAmDH2 from an eukaryotic organism demonstrated specific activity of 11.07 and 0.88 U mg−1 toward isobutyraldehyde and 1,2‐cyclohexadione respectively. Their abundance among the screened environments was also described. The protein sequence diversity of validated AmDHs reached by this metagenomics mining strategy highlights the success of such an approach. Metagenomically mined proteins, including eukaryotic ones, stand to increase the reach of biocatalysis towards enviromentally benign processes.

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Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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