Structural highlights
FunctionPublication Abstract from PubMedAdenylation enzymes catalyze the selective incorporation of aminoacyl building blocks in the biosynthesis of nonribosomal peptides and related natural products. Although beta-amino acid units are one of the important aminoacyl building blocks in natural product biosynthesis, very little is known about the engineering of beta-amino acid adenylation enzymes. In this study, we engineered the substrate specificity of the (S)-beta-phenylalanine adenylation enzyme, HitB, involved in the biosynthesis of macrolactam polyketide hitachimycin. Based on the previously determined structure of HitB wild-type, we mutated Phe328 and Ser293, which are located near the meta and ortho position of the (S)-beta-phenylalanine moiety, respectively. As a result, the HitB F328V and F328L mutants efficiently activated meta-substituted (S)-beta-phenylalanine analogs, and the HitB T293G and T293S mutants efficiently activated ortho-substituted (S)-beta-phenylalanine analogs. Structural analysis of the HitB F328L and T293G mutants with the corresponding nonhydrolyzable intermediate analogs revealed an enlarged substrate binding pocket for (S)-beta-phenylalanine analogs, providing detailed insights into the structural basis for creating enzyme substrate promiscuity. Our findings may be useful for production of various beta-amino acid-containing natural product analogs. Engineering the Substrate Specificity of (S)-beta-Phenylalanine Adenylation Enzyme HitB.,Wang D, Miyanaga A, Chisuga T, Kudo F, Eguchi T Chembiochem. 2024 May 28:e202400383. doi: 10.1002/cbic.202400383. PMID:38805007[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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