The invention relates to the field of protein engineering and biocatalysis, in particular to methods for oxygenation of aliphatic alkenes and terpenes using bacterial enzymes. Provided is a method for oxyfunctionalization of a substrate of interest, comprising contacting an aliphatic alkene or a terpene substrate with a source of hydrogen peroxide and a polypeptide having calcosin-like peroxy genase activity (EC 1.11.2.1), wherein the polypeptide is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 50% pairwise sequence identity when aligned to at least 150 consecutive amino acid residues of Seq. No. 2 shown in Table 1, and comprising at least the following heme-coordinating motifs: i) HXXFFD: ii) H(X)XD, wherein X is any amino acid; and (b) a fragment of the polypeptide of (a) that has calcosin-like peroxygenase activity.
Novel polypeptides having peroxygenase activity, and methods and uses related thereto. A method for the production of melanin or a melanin-like pigment, comprising the use of a polypeptide having pigment producing activity, wherein said polypeptide is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 50% pairwise sequence identity when aligned to at least 200 consecutive amino acid residues of Seq. no. 16 and comprising at least two of the following motifs: i) RXFWXRWXXGHQ; ii) LXXLXXCXD; iii) PRXXYH; iv) RXR[ML]ALQH; v) CXXL; vi) HXXIAXH; vii) DLXHXG; and viii) VDGXHHPV; (b) a polypeptide comprising an amino acid sequence having at least 30% pairwise sequence identity when aligned to at least 150 amino acid residues of Seq. no. 12, and comprising the motif HXXXC; and c) a fragment of the polypeptide of (a) or (b) that has peroxygenase activity.
C12N 9/08 - Oxidoreductases (1.), e.g. luciferase acting on hydrogen peroxide as acceptor (1.11)
C12N 15/70 - Vectors or expression systems specially adapted for E. coli
C12P 17/18 - Preparation of heterocyclic carbon compounds with only O, N, S, Se, or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
3.
METHODS FOR OXYFUNCTIONALIZATION OF VARIOUS SUBSTRATES USING BACTERIAL ENZYMES.
The invention relates to the field of protein engineering and biocatalysis, in particular to methods for oxygenation of aliphatic alkenes and terpenes using bacterial enzymes. Provided is a method for oxyfunctionalization of a substrate of interest, comprising contacting an aliphatic alkene or a terpene substrate with a source of hydrogen peroxide and a polypeptide having caleosin-like peroxygenase activity (EC 1.11.2.1), wherein the polypeptide is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 50% pairwise sequence identity when aligned to at least 150 consecutive amino acid residues of Seq. No. 2 shown in Table 1, and comprising at least the following heme-coordinating motifs: i) HXXFFD; ii) H(X)XD, wherein X is any amino acid; and (b) a fragment of the polypeptide of (a) that has caleosin- like peroxygenase activity.
The invention relates to the field of protein engineering and biocatalysis, more in particular to novel polypeptides having peroxygenase activity, and to methods and uses related thereto. Provided is a method for the production of melanin or a melanin-like pigment, comprising the use of a polypeptide having pigment producing activity, wherein said polypeptide is selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 50% pairwise sequence identity when aligned to at least 200 consecutive amino acid residues of Seq. no. 16 of Figure 1, and comprising at least two of the following motifs: i) RXFWXRWXXGHQ; ii) LXXLXXCXD; iii) PRXXYH; iv) RXR[ML]ALQH; v) CXXL; vi) HXXIAXH; vii) DLXHXG; and viii) VDGXHHPV; wherein X is any amino acid; (b) a polypeptide comprising an amino acid sequence having at least 30% pairwise sequence identity when aligned to at least 150 amino acid residues of Seq. no. 12 of Figure 2, and comprising the motif HXXXC, wherein X is any amino acid; and (c) a fragment of the polypeptide of (a) or (b) that has peroxygenase activity.
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