Production Of N~6-(2-hydroxyethyl)-adenosine(HEA) From Cordyceps S.l. And HEA Biosynthesis Regulation

N6-(2-hydroxyethyl)-adenosine(HEA),a nucleoside analogue,has various biological activities,is the first calcium antagonists from biological sources.HEA combines the development potential as healthcare,cosmetics and biological control,has good clinical application prospects.However,HEA is now known has been detected in only a few species,identifying new sources of HEA is warranted.At the same time,HEA synthesis regulation is studied to provide guidance for the in-depth research and development of HEA.In this study,we detected HEA in a liquid culture and fruit body of 26 strains of Cordyceps sensu lato species collected in our laboratory.We found six new HEA-producing fungi,which was identified as Beauveria bassiana(Bals.-Criv.)Vuill.,Beauveria brongniartii(Sacc.)Petch,Cordyceps guangdongensis T.H.Li,Q.Y.Lin & B.Song,Lecanicillium psalliotae(Treschew)Zare & W.Gams,Paecilomyces hepiali Q.T.Chen & R.Q.Dai ex R.Q.Dai,Paecilomyces lilacinus(Thom)Samson.Moreover,the fungal kinetics of HEA production exhibited the maximum production of HEA(0.848 ± 0.044 mg/g mycelia dry weight)was achieved on day 7.HEA content in the synnemata cultivated on wheat medium was significantly higher than that of wild synnemata of Isaria cicadae.Therefore,the method was optimized of cultivation for beneficial to the growth of cicada flowers and increase to produce HEA: 25 °C,white light of 150 lux,no shell wheat,glucose and Peptone-A.In the growth period,there were significant differences in nucleosides of I.cicadae and Cordyceps militaris.The production of HEA and mycelia dry weight was highest on day 35 and then decreased slightly by the end of the growth period.I.cicadae have autofluorescence,and we only detected single mating type MATing 1-2.We established an Agrobacterium tumefaciens-mediated transformation system,a mutant library consisting of 33 transformants with A.tumefaciens AGL-1 and I.cicadae spores,with a transformation rate of 16.37%.The mutant strains were cultivated with obvious difference in the morphology of the plate,to screen high yield of HEA.It was found that the transformant Z4 was the highest HEA strains,and the mutant strains of I.cicadae grow to be unsound.The difference in the price of conidia powder and synnema is huge.The experimental results showed that the difference between them was analyzed.In this paper,we isolated the differential compounds by silica gel column chromatography and high performance liquid chromatography.