ABSTRACT
Fe(II)/2OG-dependent oxygenase γ-butyrobetaine hydroxylase (BBOX) stereoselectively hydroxylates inactive C-H bonds and produces L-carnitine. It has potential applications in the biosynthesis of L-carnitine and the synthesis of other small molecule alcohols. In this paper, we systematically explore the substrate range of Pseudomonas sp. AK1 BBOX (psBBOX), with emphasis on the quaternary ammonium portion of γ-butyrobetaine (γ-BB). The space limitation of the "aromatic cage" in psBBOX in the hydroxylation of large quaternary ammonium analogues was studied, and the role of four aromatic amino acid residues in the substrate binding mode was analyzed. Consequently, the F188A mutant was developed with the ability to hydroxylate cyclic quaternary ammonium analogues and generate new alcohol compounds by breaking the limitation of the "aromatic cage".
Subject(s)
Ammonium Compounds , Pseudomonas , Carnitine/chemistry , gamma-Butyrobetaine Dioxygenase/chemistry , gamma-Butyrobetaine Dioxygenase/metabolism , EthanolABSTRACT
Eleven new ophiobolin-type sesterterpenoids, asperophiobolins A-K (1-11), along with 12 known analogues (12-23) were isolated from the cultures of the mangrove endophytic fungus Aspergillus sp. ZJ-68. The structures of the new compounds were elucidated through spectroscopic analyses, and their absolute configurations were assigned by a combination of Mo2(AcO)4-induced electronic circular dichroism spectra and quantum chemical calculations. Asperophiobolins A-D (1-4) represent the first examples possessing a five-membered lactam unit between C-5 and C-21 in ophiobolin derivatives. In the bioactivity assays, compounds 8-10 and 14-17 exhibited inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW 264.7 macrophage cells with IC50 values ranging from 9.6 to 25 µM, and compound 8 was found to show comparable inhibition of Mycobacterium tuberculosis protein tyrosine phosphatase B with an IC50 value of 19 µM.