US 9,803,225 B2 · Means and methods for the enzymatic production of L-methionine from O-phospho-L-homoserine and methanethiol

Abstract

Provided is a method for producing L-methionine in which O-phospho-L-homoserine and methanethiol are enzymatically converted into L-methionine and H3PO4. Such a conversion is achieved by an enzyme called O-phospho-L-homoserine (OHPS) dependent methionine synthase. Also described are O-phospho-L-homoserine (OHPS) dependent methionine synthases, i.e. proteins which are able to enzymatically convert O-phospho-L-homoserine and methanethiol into L-methionine and H3PO4 as well as microorganisms which have been genetically modified so as to be able to produce L-methionine from O-phospho-L-homoserine and methanethiol. Furthermore described are methods to screen for enzymes that catalyze the conversion of O-phospho-L-homoserine and methanethiol into L-methionine and H3PO4.

Claims12

Dependent claims

A method for producing L-methionine, comprising enzymatically converting O-phospho-L-homoserine (OPHS) and methanethiol into L-methionine and H3PO4 according to the following reaction scheme:

O-phospho-L-homoserine+CH3.SH<=>L-methionine+H3PO4;
wherein
the enzymatic conversion is achieved by a OPHS dependent methionine synthase that is a protein derived from a cystathionine gamma synthase (EC 2.5.1.48) by mutation;
and
the OPHS dependent methionine synthase is a protein selected from the group consisting of:
(a) a protein comprising the amino acid sequence as shown in any one of SEQ ID NOs:
6 to 29;
and
(b) a protein having a sequence identity of at least 90% to any one of SEQ ID NOs:
6 to 29 and having the enzymatic activity of converting O-phospho-L-homoserine and methanethiol into L-methionine and H3PO4.

7 more elements

A method comprising enzymatically converting O-phospho-L-homoserine and methanethiol into L-methionine and H3PO4 via a protein that is:

(I) a cystathionine γ synthase having the amino acid sequence shown in SEQ ID NO:
3 by substitution or deletion of at least one amino acid residue in SEQ ID NO:
3 selected from the group consisting of:
(a) proline 10;
(b) asparagine 11;
(c) glutamine 15;
(d) isoleucine 27;
(e) alanine 30;
(f) leucine 45;
(g) serine 47;
(h) valine 60;
(i) alanine 68;
(j) phenylalanine 150;
(k) threonine 178;
(l) aspartate 183;
(m) isoleucine 185;
(n) threonine 220;
(o) methionine 232;
(p) valine 245;
(q) alanine 257;
(r) asparagine 259;
(s) phenylalanine 261;
(t) phenylalanine 275;
(u) isoleucine 287;
(v) histidine 289;
(w) tyrosine 324;
(x) glycine 326;
(y) proline 356;
(z) threonine 371;
(aa) valine 396;
(bb) proline 405;
(cc) aspartate 431;
(dd) isoleucine 436;
(ee) isoleucine 457;
(ff) aspartate 459;
(gg) proline 470;
(hh) glutamate 472;
(ii) alanine 506;
and
(jj) isoleucine 507;
or
(II) a cystathionine gamma synthase, the amino acid sequence of which has at least 90% sequence identity to the amino acid sequence shown in SEQ ID NO:
3, by substitution or deletion of at least one amino acid residue corresponding to any one of (a) to (jj) listed above in SEQ ID NO:
3;
or
(III) the cystathionine gamma synthase of (I) or (II) by deletion of one or more N-terminal amino acids corresponding to amino acids 1 to 103 of the amino acid sequence as shown in SEQ ID NO:
3, wherein substitution or deletion of at least one amino acid residue is present.

44 more elements

Dependencies are inferred from the claim text.

Means and methods for the enzymatic production of L-methionine from O-phospho-L-homoserine and methanethiol

Life of this patent

Means and methods for the enzymatic production of L-methionine from O-phospho-L-homoserine and methanethiol

Life of this patent

Abstract

Claims12