Genome-guided investigation of anti-inflammatory sesterterpenoids with 5-15 trans-fused ring system from phytopathogenic fungi

Norditerpenoids biosynthesized by variediene synthase-associated P450 machinery along with modifications by the host cell Aspergillus oryzae

“…Complementary DNA (cDNA) of DvVS was used as a template of the PCR reactions for direct sequencing and subcloning each gene into the pXW55 vector using Spe I and Pml I restriction sites to afford pXW55- DvVS . The methods of cDNA synthesis and transformants SC- DvVS construction were the same as those in our previous study [ 19 ]. Yeast competent cell preparation and transformation were performed with Frozen-EZ Yeast Transformation II kit™ (Zymo Research) according to the manufacturer's protocols.…”

“…Mycelia of the transformant AO- orfABC/vndDE was inoculated into 250 mL Erlenmeyer flasks containing 100 mL of MPY medium containing 0.1% (NH 4 ) 2 SO 4 and 0.01% adenine to prepare the seed culture. Fermentation was then carried out in 125 aseptic bags (each containing 80 g rice and 120 mL water), and cultivation methods were the same as those in our previous research [ 19 ] incubated stationarily at 30 °C for 24 days.…”

“…Among fungi-derived terpene synthases, bifunctional terpene synthases (BFTSs) have been discovered as unique types of chimeric enzymes containing a prenyltransferase (PT) domain and a terpene cyclase (TC) domain, and show the possibility of catalyzing the formation of structurally diversified di/sesterterpenes [ 1 , 8 , 9 ]. Since 2007, a series of BFTS genes or BFTS-containing biosynthetic gene clusters (BGCs) have been obtained from various fungal species by using a combination of genome mining and heterologous expression in either Aspergillus oryzae (AO) or Saccharomyces cerevisiae (SC) [ [9] , [10] , [11] , [12] , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] ]. Based on bioinformatics analysis and tandem heterologous expression, an anti-inflammatory 5/15 bicyclic sesterterpenoids was characterized from Bipolaris sorokiniana , and 5/6/7/3/5 pentacyclic sesterterpene synthases (FoFS and AtAS) from Fusarium oxysporum and A. terreus [ 18 , 19 ].…”

“…Since 2007, a series of BFTS genes or BFTS-containing biosynthetic gene clusters (BGCs) have been obtained from various fungal species by using a combination of genome mining and heterologous expression in either Aspergillus oryzae (AO) or Saccharomyces cerevisiae (SC) [ [9] , [10] , [11] , [12] , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] , [22] , [23] ]. Based on bioinformatics analysis and tandem heterologous expression, an anti-inflammatory 5/15 bicyclic sesterterpenoids was characterized from Bipolaris sorokiniana , and 5/6/7/3/5 pentacyclic sesterterpene synthases (FoFS and AtAS) from Fusarium oxysporum and A. terreus [ 18 , 19 ]. The FoFS catalyzed the formation of fusoxypenes, which are enantiomeric with the precursor of the products of AtAS [ 18 , 19 ].…”

“…Based on bioinformatics analysis and tandem heterologous expression, an anti-inflammatory 5/15 bicyclic sesterterpenoids was characterized from Bipolaris sorokiniana , and 5/6/7/3/5 pentacyclic sesterterpene synthases (FoFS and AtAS) from Fusarium oxysporum and A. terreus [ 18 , 19 ]. The FoFS catalyzed the formation of fusoxypenes, which are enantiomeric with the precursor of the products of AtAS [ 18 , 19 ].…”