YALI0C22088g from Yarrowia lipolytica catalyses the conversion of l‐methionine into volatile organic sulfur‐containing compounds

Abstract

The enzymatic conversion of L-methionine (L-Met) into volatile organic sulfur-containing compounds (VOSCs) plays an important role in developing the characteristic aroma of foods. However, the mechanism for the direct conversion of L-Met into VOSCs is still unclear in yeast cells used to make food products. Here, we show that the transcription profile of YALI0C22088g from Yarrowia lipolytica correlates positively with L-Met addition. YALI0C22088g catalyses the c-elimination of L-Met, directly converting L-Met into VOSCs. YALI0C22088g also exhibits strong C-S lysis activities towards L-cystathionine and the other sulfur-containing compounds and forms a distinct cystathionine-c-lyase subgroup. We identified eight key amino acid residues in YALI0C22088g, and we inferred that the size of the tunnel and the charges carried by the entrance amino acid residue are the determinants for the enzymatic conversion of L-Met into VOSCs. These findings reveal the formation mechanism of VOSCs produced directly from L-Met via the demethiolation pathway in Yarrowia lipolytica, which provides a rationale for engineering the enzymatic conversion of L-Met into VOSCs and thus stimulates the enzymatic production of aroma compounds.

Depletion of L-Methionine in Foods with an Engineered Thermophilic Methionine γ-lyase Efficiently Inhibits Tumor Growth

“…PDB IDs and KEGG entries of MGLs are provided. YHR112C and YALI0C22088g from yeast are generally recognized as safe and have low immunogenic potential, but their ability to degrade l -Met is weaker than that of hMGL and pMGL. (b, c) Hydrophobic analysis of hCGL and pMGL.…”

“…In both mutants, the loop region comprising amino acids 50–60 blocks the entrance (predicted by CAVER software) of the active sites existing at the interface of the dimer (Figure S3a and Figure S3b). , Hydrophobic interactions and charge interactions convey the substrate flux to the active site through the entrance, and entrance properties such as polarity, shape and size may also impact substrate entry and product departure. ,,, Mutant hMGL (E59N/E339V) has better hydrophobic and electronic properties for the entry of l -Met than mutant hMGL (F58Y/E339V) (Figure S3c and Figure S 3d). At pH 8.0 and 65 °C, mutant hMGL (F58Y/E339V) exhibited higher catalytic efficiency toward l -Met and l -cysteine ( l -Cys) than mutant hMGL (E59N/E339V) (Table and Figures S4–S7).…”

“…There is an urgent need to develop powerful mutants to deplete l -Met in foods. Methionine γ-lyase (MGL) catalyzes the cleavage of the C–S bond in l -Met, producing α-ketobutyric acid, ammonia and methanethiol . Methanethiol and its derivatives, termed volatile organic sulfur-containing compounds (VOSCs), can be extracted from liquid fermentation medium at 60 °C for 10 min .…”

Enzymatic Synthesis of l‐Methionine Analogues and Application in a Methyltransferase Catalysed Alkylation Cascade**

“…HPLC analysis : For HPLC method A [35] (analysis of l ‐methionine analogues) and HPLC method C (analysis of SAM analogues) an Agilent G1311A System equipped with a quaternary pump in combination with an ISAspher 100‐5‐C18 Aq column (250×4.0 mm) was used. For HPLC method A, a gradient with water (pH 3, H 3 PO 4 ) and acetonitrile; and for HPLC method C, a gradient with ammonium formate (10 m m, pH 3.5) and acetonitrile was used (s. Table S4).…”

Engineering the Entrance of a Flavonoid Glycosyltransferase Promotes the Glycosylation of Etoposide Aglycone

“…We used YASARA software to dock DMEP to the entrance of UGT78D2. A molecular dynamics (MD) simulation of UGT78D2 complexed with DMEP was performed following the methods described by Zhao et al 26 ■ ASSOCIATED CONTENT * sı Supporting Information…”