Table 1 Substrate screen of ThAOS wild type and eight V79 variants

From: Rational engineering of a thermostable α-oxoamine synthase biocatalyst expands the substrate scope and synthetic applicability

  1. The screen was carried out with 14 amino acid substrates (Aba, l-Ala, Gly, l-Ser, d-/ l-Alg, l-Asp, l-Cpa, l-Cpg, l-Hsr, l-Ile, α-Ile, l-Nva, l-Oas, l-Pra, l-Thr and l-Val) with acetyl-CoA. Mutants were screened using the DNTB assay at fixed, quasi-saturating substrate concentrations (16 mM amino acid, 1 mM acetyl-CoA). Substrates which were tested but either inactive for all variants or indeterminable include l-Cys, l-Glu, l-Phe, l-His, l-Lys, l-Leu, l-Met, l-Asn, l-Pro, l-Gln, l-Arg, l-Trp, l-Tyr, l-Nle, l-Orn, l-Phg and l-Trl. A scale bar describes the activity, the darkest blue shading in the heat map corresponds to a specific turnover number of 7.70 s-1 (for ThAOS V79S with Gly and acetyl-CoA), and white denotes no detectable activity (<0.1 s-1). The ThAOS V79C and V79E displayed no detectable activity and were omitted from the figure.
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