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Penicillium citrinum strain with high nuclease P1 yield and its selective breeding process

A technology of nuclease and Penicillium citrinum, applied in the field of ion beam implantation mutagenesis to breed the Penicillium citrinum, to achieve the effects of high yield, stable traits, little damage to biological cells, and not easy to lose

Active Publication Date: 2007-11-07
NANJING UNIV OF TECH +1
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Problems solved by technology

There are no relevant patents and literature reports on the application of ion implantation mutagenesis technology to the production of nuclease P1 by Penicillium citrinum

Method used

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Experimental program
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Embodiment 1

[0025] Get the spores of Penicillium citrinum cultivated on the inclined plane, and make 10 5 For spore suspension, divide 0.2ml of spore suspension into glass surface dishes, dry aseptically for 1 hour, put the culture dish containing bacteria to be treated into the ion implanter for continuous N + Ion implantation, the vacuum degree in the target chamber is 5×10 -5 Pa, the injection energy is 30kev, the dose is 50~500×10 13 ions cm -2 ·s -1 . Then dilute the mutated spore suspension 100 times with sterile water, take 0.05ml and spread it on the PDA plate medium and culture it at 28~32℃ for 7~10d, from which the pure strains are selected and cultivated on the wort slant for 5~ After 10 days, transfer to 25ml liquid fermentation medium (components: glucose 40g, peptone 4g, potassium dihydrogen phosphate 0.06g, dipotassium hydrogen phosphate 0.06g, magnesium sulfate 0.06g, calcium chloride 0.04g, dilute to 1L, pH 5-7) in a 250ml shaker flask, cultured at 28-32°C, 250rpm sh...

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PUM

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Abstract

The present invention discloses one Penicillium citrinum strain with high nuclease P1 yield and its selective breeding process. The Penicillium citrinum strain is preserved in the preservation number of CGMCC No. 2014. Its selective breeding process includes low energy ion beam implantation on Penicillium citrinum spore suspension, transferring the mutagenized spore to plate culture medium for culturing, selecting single strain and transferring to malt wort and slant cultivating, liquid fermenting culturing to screen out strain with high nuclease P1 yield, and passage culturing for over 30 generations to obtain strain with stable high yield character. The Penicillium citrinum strain has high enzyme activity and stable high yield character, and is suitable for industrial production.

Description

technical field [0001] The invention belongs to the technical field of microbial mutagenesis, and relates to a Penicillium citrinum high-yield nuclease P1 and a method for breeding the Penicillium citrinum by means of ion beam implantation mutagenesis. Background technique [0002] Nuclease P1 obtained from the fermentation broth of Penicillium citrinum is a phosphodiesterase that can hydrolyze RNA to obtain four 5' mononucleotides. 5'-nucleotides have been widely used in food, biochemical and pharmaceutical industries. In the food industry, it has expanded from the initial food freshness aid to a functional food additive that can improve the immune function of organisms. It can be added to foods such as bread and biscuits, especially in infant food. It can effectively enhance the ability of infants and young children to resist bacillary dysentery and reduce the occurrence of diarrhea; in the field of medicine, 5'-nucleotides can not only be used as drugs, but also the raw ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/15C12N15/55C12N9/22
Inventor 应汉杰杨蕴毅贺沁婷张磊阮文辉
Owner NANJING UNIV OF TECH
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