Compositions and methods for making (S)-norcoclaurine and (S)-norlaudanosoline, and synthesis intermediates thereof
Methods that may be used for the manufacture of the chemical compound (S)-norcoclaurine, (S)-norlaudanosoline, and (S)-norcoclaurine or [S]-norlaudanosoline synthesis intermediates are provided. (S)-Norcoclaurine, (S)-norlaudanosoline, and (S)-norcoclaurine or (S)-norlaudanosoline synthesis intermediates are useful as precursor products in the manufacture of certain medicinal agents.
1. The method of making (S)-norcoclaurine or (S)-norlaudanosoline, comprising:
(a) providing at least one (S)-norcoclaurine or (S)-norlaudanosoline pathway precursor selected from a first L-tyrosine derivative; and
(b) contacting the (S)-norcoclaurine or (S)-norlaudanosoline pathway precursor with at least one of the enzymes selected from the group of enzymes consisting of (i) monoamine oxidase (MAO); and (ii) norcoclaurine synthase (NCS) under reaction conditions permitting the catalysis of the (S)-norcoclaurine or (S)-norlaudanosoline pathway precursor to form (S)-norcoclaurine or (S)-norlaudanosoline; and
wherein the first L-tyrosine derivative has the chemical formula (1):
wherein R 1 represents hydrogen or hydroxyl;
wherein R 2 represents hydrogen; and
wherein R 3 represents an amino group —(NH 2 ); and
wherein R 3 ′ represents a hydrogen atom, provided R 1 represents hydroxyl; or
wherein R 3 and R 3 ′ taken together, form a carbonyl group;
and wherein monoamine oxidase (MAO) is a polypeptide having the sequence set forth in SEQ.ID. NO. 96; and
and wherein norcoclaurine synthase (NCS) is a polypeptide having a sequence selected from the group of sequences set forth in SEQ.ID NO: 13; SEQ.ID NO: 14; SEQ.ID NO: 17; SEQ.ID NO: 28; SEQ.ID NO: 34; and SEQ.ID NO: 42; or is encoded by SEQ.ID NO: 109 or SEQ.ID NO: 113.
2. The method of making (S)-norcoclaurine according to claim 1 wherein the (S)-norcoclaurine pathway precursor is dopamine and 4-hydroxyphenylacetaldehyde; and the enzyme is norcoclaurine synthase (NCS).
3. The method of making (S)-norlaudanosoline according to claim 1 wherein the (S)-norlaudanosoline pathway precursor is dopamine and the enzymes are (i) monoamine oxidase (MAO) and (ii) norcoclaurine synthase (NCS).
4. The method of making (S)-norlaudanosoline according to claim 1 wherein the (S)-norlaudanosoline pathway precursors are dopamine and 3,4-dihydroxyphenylacetaldehyde and the enzyme is norcoclaurine synthase (NCS).
5. The method according to claim 1 wherein the reaction conditions are in vitro reaction conditions.
6. The method according to claim 1 wherein the reaction conditions are in vivo reaction conditions.
7. The method according to claim 6 , wherein the in vivo reaction conditions are attained by expressing the monoamine oxidase (MAO); or norcoclaurine synthase (NCS) in yeast or in E. coli.
8. A method for preparing (S)-norcoclaurine or (S)-norlaudanosoline; comprising:
(a) providing a chimeric nucleic acid sequence comprising as operably linked components:
(i) one or more nucleic acid sequences encoding one or more of the D polypeptides selected from the group of polypeptides consisting of monoamine oxidase (MAO); and NCS norcoclaurine synthase (NCS);
and
(ii) one or more nucleic acid sequences capable of controlling expression in a host cell;
(b) introducing the chimeric nucleic acid sequence into a host cell and growing the host cell to produce the polypeptide selected from the group of polypeptides consisting of monoamine oxidase (MAO); and norcoclaurine synthase (NCS) and to produce one or more of (S)-norcoclaurine or (S)-norlaudanosoline;
(c) recovering (S)-norcoclaurine, or (S)-norlaudanosoline;
wherein monoamine oxidase (MAO) is a polypeptide having the sequence set forth in SEQ.ID. NO. 96; and wherein NCS norcoclaurine synthase (NCS) is a polypeptide having a sequence selected from the group of sequences set forth in SEQ.ID NO: 13; SEQ.ID NO: 14; SEQ.ID NO: 17; SEQ.ID NO: 28; SEQ.ID NO: 34; and SEQ.ID NO: 42; or is encoded by SEQ.ID NO: 109 or SEQ.ID NO: 113.
9. The method according to claim 8 wherein the host cell is a yeast cell or an E. coli cell.
10. A method for preparing an (S)-norcoclaurine or (S)-norlaudanosoline pathway precursor selected from the group consisting of: dopamine; 4-hydroxyphenylacetaldehyde; and 3,4-dihydroxyphenylacetaldehyde; and comprising:
(a) providing a chimeric nucleic acid sequence comprising (i) one or more nucleic acid sequences complementary all or a portion of the mRNA synthesized by the nucleic acid sequence encoding the polypeptides selected from the group of polypeptides consisting of monoamine oxidase (MAO); and norcoclaurine synthase (NCS); and (ii) one or more elements capable of controlling transcription of the complementary nucleic acid sequence, wherein the chimeric nucleic acid sequence is capable of producing an antisense RNA complementary all or a portion of the mRNA of the nucleic acid sequence encoding the polypeptides selected from the group of polypeptides consisting of MAO; and norcoclaurine synthase (NCS);
(b) introducing the chimeric nucleic acid sequence into a host cell;
(c) growing the host cell to produce the antisense RNA and inhibit synthesis of the polypeptide selected from the group of polypeptides consisting of monoamine oxidase (MAO); and norcoclaurine synthase (NCS) and to produce one or more of an norcoclaurine or (S)-norlaudanosoline pathway precursors selected from the group consisting of: dopamine; and 4HPAA and 3,4 HPAA; and
(d) recovering a (S)-norcoclaurine pathway precursor or (S)-norlaudanosoline pathway precursor selected from the group consisting of dopamine; 4-hydroxyphenylacetaldehyde; and 3,4-dihydroxyphenylacetaldehyde; wherein monoamine oxidase (MAO) is a polypeptide having the sequence set forth in SEQ.ID. NO. 96; and wherein norcoclaurine synthase (NCS) is a polypeptide having a sequence selected from the group of sequences set forth in SEQ.ID NO: 13; SEQ.ID NO: 14; SEQ.ID NO: 17; SEQ.ID NO: 28; SEQ.ID NO: 34; and SEQ.ID NO: 42; or is encoded by SEQ.ID NO: 109 or SEQ.ID NO: 113.
11. The method according to claim 10 wherein the host cell is a yeast cell or an E. coli cell.
12. The method according to claim 8 wherein the nucleic acid sequence encoding the norcoclaurine synthase (NCS) polypeptide is selected from the group consisting of SEQ.ID NO 51; SEQ.ID NO: 52; SEQ.ID NO: 55; SEQ.ID NO: 66; SEQ.ID NO: 72; SEQ.ID NO: 80; SEQ.ID NO: 109; and SEQ.ID NO: 113 and wherein the nucleic acid sequence encoding the monoamine oxidase (MAO) polypeptide is SEQ.ID NO: 95.