Fermentation process of ocean penicillium and secondary metabolite Flufuran thereof
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A technology of secondary metabolites and fermentation process, which is applied in the field of fermentation process of marine Penicillium and its secondary metabolite Flufuran, which can solve the problems of low yield and achieve the effects of increasing yield, avoiding sample waste, and shortening fermentation time
Active Publication Date: 2014-09-10
NINGBO XINUOYA MARINE BIOTECH CO LTD
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Therefore, the low yield is the limiting factor for the comprehensive activity evaluation and development application of this compound
Method used
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specific Embodiment 1
[0032] A marine Penicillium fungus named after the taxonomy Penicillium dipodomyicola , was deposited in China Center for Type Culture Collection in March 2014 with the accession number CCTCC M 2014087. The colony was pale yellow at the initial stage of culture, and the color of the colony gradually deepened to brown with the prolongation of culture time. The specific steps of using the marine Penicillium to ferment and produce the secondary metabolite Flufuran are as follows:
[0033] (1) Activation of strains
[0034] The marine Penicillium with the preservation number CCTCC M 2014087 stored in the refrigerator at -80°C was inoculated on the potato dextrose broth solid medium (PDA), cultured in the incubator at 25°C for 2 days, and a single colony was picked to inoculate again Put it on the PDA medium, and culture it in the incubator at 25 ℃ for 2 days before use;
[0035] (2) Seed cultivation
[0036] The Penicillium maritima obtained from activation in step (1) was in...
specific Embodiment 2
[0048] 1. Optimization of medium formula
[0049] The potato extract powder in the medium was taken as a whole, and based on this, the carbon source was changed according to the ratio of 1:2, including soluble starch, glucose, maltose, sucrose and fructose, and six different formulations of the medium were prepared as follows:
[0050] Recipe 1: 10 g of potato extract, 1 L of sea water (add 35 g of sea crystal to 1 L of pure water);
[0051] Formula 2: 10 g of potato extract, 20 g of soluble starch, 1 L of sea water (add 35 g of sea crystal to 1 L of pure water);
[0052] Recipe 3: 10 g of potato extract, 20 g of glucose, 1 L of sea water (35 g of sea crystals added to 1 L of pure water);
[0053] Formula 4: 10 g of potato extract, 20 g of maltose, 1 L of sea water (add 35 g of sea crystal to 1 L of pure water);
[0054] Formula 5: 10 g of potato extract, 20 g of sucrose, 1 L of sea water (add 35 g of sea crystal to 1 L of pure water);
[0055] Formula 6: 10 g of potato ext...
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Abstract
The invention discloses a fermentation process of ocean penicillium and secondary metabolite Flufuran thereof. The ocean penicillium is characterized in that the preservation number of penicillium is CCTCCM2014087, and the fermentation process of the penicillium comprises the following steps: inoculating PDA (Potato Dextrose Agar) with the ocean penicillium, culturing for 2 days in an incubator with the temperature of 25 DEG C, picking a single bacterial flora and then inoculating on a PDA culture medium, and culturing for 2 days at the temperature of 25 DEG C for later use; (2) inoculating PDB with the activated strain, culturing for 1 day at the temperature of 25 DEG C and at a rotating speed of 150rpm to obtain a seed solution; (3) inoculating the seed solution in a fermentation culture medium and culturing for 12 days at the temperature of 25 DEG C and at the rotating speed of 150rpm, wherein the inoculated amount of the seed solution is 5vol%,; (4) filtering a fermentation solution to obtain a supernatant liquor, extracting for three times by use of ethyl acetate, removing an organic phase, and dissolving again by use of methanol to obtain a sample introduction solution; (5) separating and purifying the sample introduction solution by adopting a high-speed counter-current chromatographic method to obtain a product, wherein the fermentation process has the advantages of high preparation speed and high product purity.
Description
technical field [0001] The invention relates to the technical field of microbial fermentation, in particular to a fermentation process of marine Penicillium and its secondary metabolite Flufuran. Background technique [0002] Flufuran was first obtained from Gabriela M. Cabrera et al. in 2002 Polyporus ciliatus isolated from (Gabriela M.; Robertia M.; Jorge E.; Alicia M. Phytochem. 2002, 61, 189-193.). Antonio Evidente et al. 2009 from Aspergillus flavus and found that the compound had significant activity against Phytophthora (Antonio E.; Gennaro C.; Biancavaleria P.; Anna A.; Antonino T.; Dominique M. Chem. Biodivers. 2009, 6, 328-334.). In addition, analogues of this compound, such as 2-tetrahydrofurancarboxylic acid, are widely used as pharmaceutical intermediates. [0003] In the literature mentioned above, the fermentation yields of Flufuran are 0.625 mg / L and 43.7 mg / L, and they need to be cultured for 15 days and 14 days, respectively. Therefore, low yield i...
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