Sequencing And Analysing On Internal Transcribed Spacer Of RRNA Of A Fungus Producing Perylenequinonoid Derivatives Photosensitizer And Its Liquid Ferment Technology Research

One fungus (AL18) was isolated from southen yunan province mountains by Prof. Chen Yuanteng (1996), which secondary metabolism is perylenequinonoid derivatives (PQD). Prof. Yang Fakui identified it as Ascomycetes Hypocreales Hypocreaceac Hypomyces (Fr.) Tul. sp.. The internal transcribed spacer (ITS) in rRNA of AL18 was cloned and sequenced. Blast result shows: the ITS sequence of AL18 shares 92% similarity with that of shiraiella bambusicola P. Henn.. Phylogenetic analyzing shows that AL18 has a distinct relationship with hypomyces geneus, but has a close relationship with shiraia bambusicola.PQD is a novel photosensitizer. AL18 is used to produce PQD by the liquid deep fermentation technique for the first time. The optimum fermentation condition determined by monosyllabic test is : 40ml culture medium per 250ml flask, inoculating amount 10%, rotating speed 180r/min, pH5.75, 30癈 fermenting 48h. Under these condition the optimized culture medium determined by monosyllabic and orthogonal test is photato extract 200g/L, white sugar 30g/L, silkworm pupa powder 12.5g/L, peptone 3.0g/L, KH2PO4 1.0g/L, MgS(V7H2O 0.5g/L ,CuSO4 '5H2O 0.05g/L, pH5.75, 21℃ sterilizer min. The PQD yield reached 1.21g/L under this culture medium.We did a small scale experiment test in 7 litter fermentor and studied the response of the stir speed and the rate of ventilate on PQD yield. The result idicated that the optimum stir speed is 520 rpm and the optimum rate of ventilate is 1:2. The yield of PQD can reach 820mg/L under the condition.