UNASSIGNED: @entity17932 ( @entity3445 ) is a plant natural product that serves as a precursor for the synthesis of many well-known chemotherapeutic drugs. The limited availability and high demand for the source plants of @entity3445 have led to the exploration of alternative sources for this compound. In this study, we utilized the endophytic fungus Phialocephala podophylli (strain PPE7) that we isolated from the rhizomes of Podophyllum peltatum and is known to produce detectable amounts of @entity3445 in broth culture. To date, the complete @entity3445 biosynthetic pathway has yet to be determined in any species. Since fungi are well known for clustering genes that belong to secondary metabolite pathways, use of a fungal system for investigation of the @entity3445 biosynthesis genes may ultimately lead to elucidation of the entire pathway. In this study, we investigated the @entity41150 dehydrogenase (SD) gene that facilitates the dehydrogenation of @entity41150 to form @entity42357 , a mid-pathway intermediate product in @entity3445 biosynthesis. We utilized PCR amplification to acquire the complete SD gene sequence in PPE7 and opted to synthesize the P. peltatum SD sequence for expression. Through western blotting, we confirmed the expression of the recombinant SD (PpSD) and verified protein functionality with a bioconversion assay followed by HPLC and LC-MS analyses. Here, we report the identification of the SD gene in PPE7; this is the first report of the SD gene in an endophytic fungus. Additionally, we established the groundwork for the future expression of the complete @entity3445 biosynthetic pathway in the heterologous host @entity10056 .

Investigation and Expression of the XXXX Dehydrogenase Gene Involved in @entity17932 Biosynthesis.

@entity17932 :: ('MESH:D011034', 'Chemical') :: ['Podophyllotoxin'] @entity10056 :: ('4922', 'Species') :: ['Pichia pastoris'] @entity41150 :: ('MESH:C060283', 'Chemical') :: ['secoisolariciresinol'] @entity3445 :: ('CHEBI:61453', 'Chemical') :: ['PPT'] @entity42357 :: ('MESH:C068935', 'Chemical') :: ['matairesinol']
@entity41150 :: (MESH:C060283,Chemical) :: ['Secoisolariciresinol']
