Characterization of the cytochrome P450 monooxygenase genes (P450ome) from the carotenogenic yeast Xanthophyllomyces dendrorhous

Abstract

BackgroundThe cytochromes P450 (P450s) are a large superfamily of heme-containing monooxygenases involved in the oxidative metabolism of an enormous diversity of substrates. These enzymes require electrons for their activity, and the electrons are supplied by NAD(P)H through a P450 electron donor system, which is generally a cytochrome P450 reductase (CPR). The yeast Xanthophyllomyces dendrorhous has evolved an exclusive P450-CPR system that specializes in the synthesis of astaxanthin, a carotenoid with commercial potential. For this reason, the aim of this work was to identify and characterize other potential P450 genes in the genome of this yeast using a bioinformatic approach.ResultsThirteen potential P450-encoding genes were identified, and the analysis of their deduced proteins allowed them to be classified in ten different families: CYP51, CYP61, CYP5139 (with three members), CYP549A, CYP5491, CYP5492 (with two members), CYP5493, CYP53, CYP5494 and CYP5495. Structural analyses of the X. dendrorhous P450 proteins showed that all of them have a predicted transmembrane region at their N-terminus and have the conserved domains characteristic of the P450s, including the heme-binding region (FxxGxRxCxG); the PER domain, with the characteristic signature for fungi (PxRW); the ExxR motif in the K-helix region and the oxygen-binding domain (OBD) (AGxDTT); also, the characteristic secondary structure elements of all the P450 proteins were identified. The possible functions of these P450s include primary, secondary and xenobiotic metabolism reactions such as sterol biosynthesis, carotenoid synthesis and aromatic compound degradation.ConclusionsThe carotenogenic yeast X. dendrorhous has thirteen P450-encoding genes having potential functions in primary, secondary and xenobiotic metabolism reactions, including some genes of great interest for fatty acid hydroxylation and aromatic compound degradation. These findings established a basis for future studies about the role of P450s in the carotenogenic yeast X. dendrorhous and their potential biotechnological applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3942-9) contains supplementary material, which is available to authorized users.

Sex steroids and steroidogenesis-related genes in the sea cucumber, Holothuria scabra and their potential role in gonad maturation

“…The well-known steroidogenesis CYP, CYP11 (or P450 side-chain cleavage enzyme), was not identified in H. scabra ; however we identified a partial transcript that encodes the related HscCYP10 precursor. HscCYP10 contains the cytochrome P450 domain, as well as a proline-rich motif (PPGTPITP) and PERF/W motif, which is involved in heme incorporation and/or stability/catalytic activity of cytochrome P450 39 , 40 (Fig. 4 A).…”

“…4 B). The heme-binding region (PFxxGxxxCxG) of CYP17 shows high similarity in all species, which contain the highly conserved cysteine residue that holds the heme-group in place 40 (Fig. S1 B).…”

Sterol Regulatory Element-Binding Protein (Sre1) Promotes the Synthesis of Carotenoids and Sterols in Xanthophyllomyces dendrorhous

“…In our previous studies, potential SRE sequences, which are conserved SREBP DNA binding sequences, were identified in the promoter regions of X. dendrorhous genes of the MVA pathway (Werner et al, 2016) and genes encoding P450s enzymes (Córdova et al, 2017). Considering that HMGS (hydroxymethylglutaryl-CoA synthase) of the MVA pathway is a well-known target of SREBPs in organisms like S. pombe (Hughes et al, 2005), C. neoformans (Chang et al, 2007) and mammals (Smith et al, 1988; Goldstein and Brown, 1990; Wang et al, 1994; Horton et al, 2003), we analyzed the promoter region of this gene and identified two potential SRE sequences (SRE1: CGTCTCCTGAC and SRE2: TGTAACACCAC, located at positions -584 to -574 and -232 to -220 from the translation start codon, respectively).…”

Changes in the Biotransformation of Green Tea Catechins Induced by Different Carbon and Nitrogen Sources in Aspergillus niger RAF106

“…However, the dramatic declines of tannase activities in yeast extract and peptone samples at the late stage of fermentation might be due to the dramatic changes in pH which was not affected significantly by different carbon sources (Supplementary Figures S3B,C) and/or the non-correlation between tannase and the degradation of non-ester-catechins and GA at the late stage of fermentation. Moreover, P450 monooxygenases are involved in aromatic compound degradation, and dioxygenases including extradiol and intradiol dioxygenases catalyze the cleavage of catechol/phenolic/aromatic rings (Contreras-Domínguez et al, 2006; Roopesh et al, 2010; Wadke et al, 2016; Córdova et al, 2017; Kawabata et al, 2019). Meanwhile, non-ester-catechins from the degradation of ester-catechins could be metabolized by gut microbiota to produce the ring fission metabolites (γ-valerolactones), which further degrade to form lower molecular weight phenolic acids (Takagaki and Nanjo, 2010; Chen and Sang, 2014).…”