Polynucleotides and polypeptides useful for making alkaloid compounds

Inventors

FACCHINI, Peter James • Tucker, Joseph E.

Assignees

Antheia Inc

Abstract

Polynucleotides and polypeptides useful in the manufacture of a class of chemical compounds known as alkaloids are provided. The polynucleotides and polypeptides may be used to synthesize alkaloids, including reticuline, thebaine and morphine, in vivo and in vitro. The polynucleotides further may be used to examine the presence of the polynucleotides in a cell or a cell extract, and to modulate expression thereof in living cells.

Core Innovation

The disclosure provides compositions and genetic constructs for synthesis of alkaloids using engineered non-plant cells. The constructs include polynucleotides encoding polypeptides having thebaine synthase activity, including polynucleotides and polypeptides related to SEQ ID NO:29 and SEQ ID NO:900, and variants described as substantially identical, complementary, or hybridizing under stringent conditions.

The disclosure further provides expression vectors comprising the polynucleotide that encodes the thebaine synthase polypeptide activity. Additional elements described include promoters, transcriptional terminators, transcriptional enhancers, marker genes, origins of replication, and integration-related genetic elements for insertion into a host cell’s genome.

In addition, the disclosure describes use of heterologous hosts to produce alkaloids in vivo or in vitro and includes alkaloid transport across cellular membranes in heterologous hosts. Host cells discussed include plant cells and heterologous microbial hosts, with specific examples including Saccharomyces cerevisiae and Yarrowia lipolytica, and the alkaloid examples explicitly include reticuline and opiate alkaloids such as thebaine and morphine.

Claims Coverage

The patent includes two independent claims directed to a composition and an expression vector. The common inventive core is thebaine synthase activity with defined sequence identity relative to SEQ ID NO:29 or SEQ ID NO:900, with dependent claim refinements on higher identity thresholds and construct elements.

Overall, the claim coverage centers on polynucleotide constructs encoding a thebaine synthase polypeptide with at least 70% sequence identity to SEQ ID NO:29 or SEQ ID NO:900, in either an engineered non-plant cell composition or an expression vector. Dependent claims refine identity thresholds upward and specify yeast host types and vector components.

Stated Advantages

Documented Applications

No documented applications found