COMPOSITIONS AND METHODS OF PRODUCING (R)-RETICULIN AND ITS PRECURSORS Russian patent published in 2020 - IPC C12N15/63 C12N15/82 C12N9/02 C12P17/10 C12Q1/68 C12N15/52 A01H1/04 A01H1/06 A01H5/00 

FIELD: pharmaceutical industry.

SUBSTANCE: invention relates to a recombinant expression vector and methods of producing (R)-reticulin or its precursor of formula (II). Disclosed is a method of producing a compound of formula (II) from a corresponding derivative of formula (I):

, ,

where R1 is a methoxy group, R2 is hydroxyl, R3 is hydroxyl, R4 is a methoxy group, R5 is methyl, or R1 is a methoxy group, R2 is a hydroxyl, R3 is a hydrogen atom, R4 is a hydroxyl, R5 is methyl, or R1 is a methoxy group, R2 is hydroxyl, R3 is hydroxyl, R4 is hydroxyl, R5 is methyl, where the method involves contacting said derivative with a microorganism expressing a CYP450 and AKR fusion protein, which is SEQ ID NO: 323, growing said microorganism with producing (R)-reticulin or precursor thereof of formula II and recovering (R)-reticulin or precursor thereof of formula II; or the method comprises (a) providing a chimeric nucleic acid sequence capable of producing said CYP450-AKR fusion protein containing, as functionally related components, a first nucleic acid sequence encoding a CYP450 polypeptide, a second nucleic acid sequence encoding an AKR polypeptide, and one or more nucleic acid sequences capable of controlling expression in a host cell; (b) introducing said chimeric nucleic acid sequence into a host cell and growing said host cell to produce said fusion protein CYP450 and AKR to produce (R)-reticulin or a precursor thereof of formula II, where the host cell under natural conditions is able to produce said derivative or derivative is provided to cells as part of cell growth medium, (c) extraction of (R)-reticulin or precursor (R)-reticulin of formula II. Disclosed is a recombinant expression vector suitable for expression in a host cell, containing as functionally related components a nucleic acid sequence, capable of controlling expression in said host cell and a nucleic acid sequence encoding described in 1 CYP450 and AKR fusion proteins.

EFFECT: novel efficient methods of producing (R)-reticulin or its precursor of formula II, as well as an effective recombinant expression vector for their preparation.

10 cl, 1 tbl, 13 dwg, 8 ex