Methods and systems for sulfimidation or sulfoximidation of organic molecules
The disclosure generally relates to the fields of synthetic organic chemistry. In particular, the present disclosure relates to methods and systems for the imidation of sulfides.
1. A reaction mixture comprising a nitrene source, an organosulfur substrate and a heme enzyme or an engineered heme enzyme for producing a product having a new S—N bond.
2. The reaction mixture of claim 1 , wherein the nitrene source is an azide.
3. The reaction mixture of claim 2 , wherein the azide has the general formula R 1 —N 3 ,
wherein R 1 is:
(i) a substituted or unsubstituted aryl, a substitute or unsubstituted alkyl, —OR 2 , or —NR 2 , wherein R 2 is a substituted or unsubstituted aryl, a substitute or unsubstituted alkyl;
(ii) —SO 2 R 3 , wherein R 3 a substituted or unsubstituted aryl, a substitute or unsubstituted alkyl, —OR 2 or NR 2 , wherein R 2 are any alkyl or aryl;
(iii) —COR 4 wherein R 4 is a substituted or unsubstituted aryl, a substitute or unsubstituted alkyl, —OR 2 or NR 2 , wherein R 2 are any alkyl or aryl; or
(iv) —P(O)(OR 5 )(OR 6 ), wherein R 5 and R 6 are independently H, a substituted or unsubstituted aryl, a substitute or unsubstituted alkyl.
4. The reaction mixture of claim 3 , wherein the azide has a structure selected from the group consisting of:
wherein R 1 is any alkyl, aryl, —OR, NR 2 , wherein R, R 2 and R 3 are any alkyl, or aryl.
5. The reaction mixture of claim 1 , wherein the nitrene source is selected from the group consisting of:
wherein R 1 is any alkyl, aryl, —OR, NR 2 , wherein R, R 2 and R 3 are any alkyl, or aryl.
6. The reaction mixture of claim 1 , wherein the S—N containing product is an aliphatic amine.
7. The reaction mixture of claim 1 , wherein the product is generated through a nitrenoid intermediate.
8. The reaction mixture of claim 1 , wherein the engineered heme enzyme is a cytochrome P450 enzyme, P450BM3 enzyme or a variant thereof.
9. The reaction mixture of claim 1 , wherein the cytochrome P450 enzyme is expressed in a bacterial, archaeal or fungal host organism.
10. The reaction mixture of claim 8 , wherein the cytochrome P450 BM3 enzyme comprises the amino acid sequence set forth in SEQ ID NO: 1 or a variant thereof.
11. The reaction mixture of claim 8 , wherein the cytochrome P450 enzyme variant comprises a mutation at the axial position of the heme coordination site.
12. The reaction mixture of claim 11 , wherein the mutation is an amino acid substitution of Cys with a member selected from the group consisting of Ala, Asp, Arg, Asn, Glu, Gin, Gly, His, He, Lys, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr and Val at the axial position.
13. The reaction mixture of claim 11 , wherein the mutation is an amino acid substitution of Cys with Asp or Ser at the axial position.
14. The reaction mixture of claim 8 , wherein the P450 BM3 enzyme variant comprises at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, or all thirteen of the following amino acid substitutions in SEQ ID NO: 1: V78A, F87V, P142S, T175I, A184V, S226R, H236Q, E252G,T268A, A290V, L353V, I366V, and E442K.
15. The reaction mixture of claim 8 , wherein the cytochrome P450 enzyme variant comprises a T268A mutation and/or a C400X mutation in SEQ ID NO: 1, wherein X is any amino acid other than Cys.
16. The reaction mixture of claim 8 , wherein the cytochrome P450 enzyme variant comprises a T438S mutation and/or a C400X mutation in SEQ ID NO: 1, wherein X is any amino acid other than Cys.
17. The reaction mixture of claim 8 , wherein the cytochrome P450 enzyme variant comprises a T268A mutation, a C400X mutation and a T438S mutation in SEQ ID NO:1, wherein X is any amino acid other than Cys.
18. The reaction mixture of claim 8 , wherein the engineered heme enzyme comprises a fragment of the cytochrome P450 enzyme or variant thereof.
19. The reaction mixture of claim 8 , wherein the engineered heme enzyme is a cytochrome P450 BM3 enzyme variant selected from Table 4, Table 5 and Table 6.
20. The reaction mixture of claim 1 , wherein the product is a compound of Formula Ia:
wherein R 1 is a sulfoxide, a carbonyl or a phosphonate; wherein R 2 is H or any alkyl or aryl; and wherein R 3 is H, O or an optionally substituted aryl group.