Protecting group chemistry for clean, reductant-free dyeing

A method of producing a compound of Formula (A), the method comprising:

• culturing a host cell in vitro in a growth medium,
• wherein the host cell is genetically modified with one or more heterologous nucleic acids comprising nucleotide sequences encoding:
• i) a heterologous oxygenase polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:1;
• and
• ii) a heterologous glucosyltransferase polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to SEQ ID NO:3,
• wherein the oxygenase polypeptide modifies an indole compound present in the host cell to produce a compound of Formula (II), and
• wherein the glucosyltransferase polypeptide modifies the compound of Formula (II) to produce the compound of Formula (A),
• wherein the compound of Formula (A) is:
• wherein:
• R1, R2, R3, and R4 are independently selected from the group consisting of H, halo, nitro, sulfate, phosphate, hydroxyl, C(O)ORb, R2C(O)ORb,
• —RaOC(O)Rb, —ORa, —NRbRc, and alkyl;
• wherein alkyl is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halo, nitro, sulfate, phosphate, hydroxyl, —C(O)ORb, —RaC(O)ORb, —RaOC(O)Rb, —ORa, —NRbRc, and —RaORd;
• R5 is H or alkyl, wherein alkyl is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halo, hydroxyl, ═O,
• C(O)ORb, —RaC(O)ORb, —RaOC(O)Rb, —ORa, and —NRbRc;
• Ra and Rd are independently alkyl;
• Rb and Rc are independently H or alkyl;
• Z is a glycone;
• and
• wherein the compound of Formula (II) is:
• wherein R1, R2, R3, R4, and R5 are defined as for Formula (A).