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Method for producing mycophenolic acid by efficiently accumulating Penicillium brevicompactum and supplementing precursor in later period

A technology for precursors and mycophenolic acid, which is applied in the field of high-efficiency cumulative production of mycophenolic acid by supplementing precursors at a later stage and utilizing Penicillium breviscapae, can solve the problems of long cycle and no significant increase in yield, and achieves high production efficiency. , the effect of batch fermentation time is close, fermentation cycle is close

Active Publication Date: 2013-07-24
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current literature reports, the batch fermentation method is adopted. Xu Zhinan et al. studied that the spores of Penicillium brevismis were immobilized on a rotating fiber bed fermenter (RFB) for batch fermentation, but the yield was not significantly improved after a long cycle, while flow-through There is almost no feeding method

Method used

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  • Method for producing mycophenolic acid by efficiently accumulating Penicillium brevicompactum and supplementing precursor in later period

Examples

Experimental program
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Effect test

Embodiment 1

[0021] The slant was cultured in PDA medium for 10 days and placed in a 4°C refrigerator to store the slant. Get the slanted seeds and use 30% glycerol to elute the spores to form a spore suspension, and 2% are inoculated into the seed shake flask medium (g / L): peptone 2.0, yeast extract 1.0, starch 1.0, glucose (C 6 h 12 o 6 ·H 2 O) 10.0, pH 6.5. Cultivate for 72 hours under the conditions of 28° C. and a rotating speed of 220 r / min, and inoculate 10% of the inoculum size into a 7L fermenter according to the volume of the medium. In the initial stage of fermentation, an optimized fermentation medium (g / L): glucose (C 6 h 12 o 6 ·H 2 O) 93.4, glycine (C 2 h 5 NO 2 ) 13.4, Potassium dihydrogen phosphate (KH 2 PO 4 )3.0, magnesium sulfate (MgSO 4 ·7H 2 O) 1.0, Methionine (C 5 h 11 o 2 NS) 0.5, distilled water as the solvent, pH 4.6, the carbon and nitrogen sources were sterilized separately, and an appropriate amount of foam enemy was added.

[0022] Control the...

Embodiment 2

[0024] The slant was cultured in PDA medium for 10 days and placed in a 4°C refrigerator to store the slant. Get the slanted seeds and use 30% glycerol to elute the spores to form a spore suspension, and 2% is inoculated into the seed shake flask medium (g / L): peptone 2.0, yeast extract 1.0, starch 1.0, glucose (C 6 h 12 o 6 ·H 2 O) 10.0, pH 6.5. Cultivate for 72 hours under the conditions of 28° C. and a rotating speed of 220 r / min, and inoculate 10% of the inoculum size into a 7L fermenter according to the volume of the medium. In the initial stage of fermentation, an optimized fermentation medium (g / L): glucose (C 6 h 12 o 6 ·H 2 O) 150, glycine (C 2 h 5 NO 2 ) 14, potassium dihydrogen phosphate (KH 2 PO 4 )2.0, magnesium sulfate (MgSO 4 ·7H 2 O) 0.5, methionine (C 5 h 11 o 2 NS) 1.0, distilled water as the solvent, pH 4.4, the carbon and nitrogen sources were sterilized separately, and an appropriate amount of foam enemy was added.

[0025] Control the fer...

Embodiment 3

[0027] The slant was cultured in PDA medium for 10 days and placed in a 4°C refrigerator to store the slant. Get the slanted seeds and use 30% glycerol to elute the spores to form a spore suspension, and 2% is inoculated into the seed shake flask medium (g / L): peptone 2.0, yeast extract 1.0, starch 1.0, glucose (C 6h 12 o 6 ·H 2 O) 10.0, pH 6.5. Cultivate for 72 hours under the conditions of 28° C. and a rotating speed of 220 r / min, and inoculate 10% of the inoculum size into a 7L fermenter according to the volume of the medium. In the initial stage of fermentation, an optimized fermentation medium (g / L): glucose (C 6 h 12 o 6 ·H 2 O) 100.0, glycine (C 2 h 5 NO 2 ) 15, potassium dihydrogen phosphate (KH 2 PO 4 )4.0, magnesium sulfate (MgSO 4 ·7H 2 O) 0.5, methionine (C 5 h 11 o 2 NS) 0.5, distilled water as the solvent, pH 4.8, the carbon and nitrogen sources were sterilized separately, and an appropriate amount of foam enemy was added.

[0028] The fermentati...

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Abstract

The invention relates to a method for producing mycophenolic acid by efficiently accumulating Penicillium brevicompactum and supplementing a precursor in the later period, which comprises the following steps: culturing Penicillium brevicompactum on a PDA (potato dextrose agar) slope medium for 10 days, eluting with 30% glycerol to obtain a spore suspension, inoculating 2 wt% of spore suspension to the seed culture medium, carrying out activation culture at the controlled temperature of 28 DEG C for 72 hours, inoculating the activated Penicillium brevicompactum in a fermentation tank, carryingout fermentation and culture by using the optimized glucose-glycine liquid culture medium at the controlled temperature of 28 DEG C, and supplementing methionine in the later fermentation period, thereby obtaining the mycophenolic acid. Compared with the prior art, the invention has the following advantages: the fermentation period is less than 192 hours, and the maximum yield is up to 5.0g / L, which is about 30% higher than batch fermentation. The fermentation strategy of supplementing the precursor to the optimized carbon nitrogen source can be controlled easily, has the characteristics of short fermentation period and high yield, and has wide industrialization application prospects.

Description

technical field [0001] The invention relates to a method for producing mycophenolic acid, in particular to a method for producing mycophenolic acid with high-efficiency accumulation of Penicillium breviscellus by adding precursors later. Background technique [0002] Myhophenolc acid (MPA) is an antibiotic produced by Penicillium brevicompactum which has antifungal, antitumor and immunosuppressive effects. MPA is a reversible inhibitor of hypoxanthine mononucleotide dehydrogenase, which can selectively inhibit the activity of lymphocytes. Its 2-ethyl ester derivative - mycophenolate mofetil (MMF) is a new generation of immune Inhibitors have shown good application prospects in the treatment of clinical organ transplantation and autoimmune diseases. MPA is used as a precursor compound for synthesizing MMF, and it is very important for production enterprises to improve the industrialization level of MPA. [0003] Most of the large-scale industrial production of modern antibi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/04C12R1/80
Inventor 孙爱友魏东芝董玉国王丽华徐瑞张舰
Owner EAST CHINA UNIV OF SCI & TECH
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