MeJA Strains Of Hericium 5L0433 Ergosterol Synthesis And Analysis Of Subtracted CDNA Library Of ESTs

In this study, the method of producing ergosterol with Hericium mycelium in liquid shake flask was established, and build subtractive library by use of suppression subtractive hybridization (Suppression subtractive hybridization, SSH). The major findings are as follows:1. The technology that MeJA promoted ergosterol synthesis has been establishedThe optimized condition to produce ergosterol with mycelium of Hericium was maltose 20g, potassium dihydrogen phosphate 3g, magnesium sulfate 1.5g, yeast extract 10g, water 1000ml. The best induction condition of MeJA(methyl jasmonate)elicitor was that eight-day old mycelium of Hericium was induced for 11 d under 25℃, the ergosterol production reached the maximum, namely 1.882mg/100ml.2. The effect of MeJA on ergosterol synthesis in Hericium 5L0433MeJA elicitors promoted the mycelium growth and ergosterol accumulation, the MeJA (methyl jasmonate) elicitors promote the accumulation of ergosterol study reveals:MeJA elicitor addition on the fermentation liquid Hericium mycelium growth, ergosterol content, growth reflects a certain role. Overall yield of mycelium and ergosterol content determined using 50μmol·L-1 best MeJA concentration on the mycelium of Hericium induction. MeJA induced the production of the mycelium when the first 8d up to 0.96g/100ml. MeJA in 10μmol·L-1,50μmol·L-1, 100μmol·L-1,200μmol·L-1 concentration conditions,50μmol·L-1 and 100μmol·L-1 mycelium yields were 0.31g/100ml and 0.33g/100ml, significantly higher than the control (0.27g/100ml). MeJA concentration of 50μmol·L-1, the dry mycelial ergosterol content of up to 1.603 mg·g-1, the control (1.260mg·g-1) of 1.27 times.3. Subtractive library-specific gene fragment preliminary bioinformatics analysisSuccessfully constructed MeJA induced Hericium forward subtractive library. BLAST software applications will be reduced library screening ESTs to the specific gene fragment sequences in GenBank sequence homology comparison, while functional classification using COG and GO functional classification of the sequence of a preliminary bioinformatics analysis. In the forward library MhmrA the GO functional classification of the results showed that the cell components function library contains cells, organelles, protein complexes of these three. Molecular function library contains the antioxidant activity, catalytic activity, signal transduction activity, translation regulation activity, etc., library functions include programmed cell biological pathways, physiological processes, regulation of biochemical processes, these four types of stimuli. In the forward library MhmrA the COG functional classification results show that the functional gene fragments from a variety of percentage terms, GO functional classification of the results showed that methyl jasmonate induced Ergosterol gene expression in post-translational modification involved, protein turnover metabolism, carbohydrate transport and metabolism, energy production and conversion, transport and metabolism of amino acids, inorganic ion transport and metabolism, coenzyme transport and metabolism in 15 categories.