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

A technology of nuclease and Penicillium citrinum, which is applied in the field of breeding of Penicillium citrinum by means of ion beam implantation mutagenesis, to achieve the effects of stable high-yielding traits, not easy to lose, and little damage to biological cells

Active Publication Date: 2009-08-26
NANJING TECH UNIV +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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  • Penicillium citrinum bacteria with high nuclease P1 yield and its selective breeding process
  • Penicillium citrinum bacteria with high nuclease P1 yield and its selective breeding process

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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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Abstract

The invention discloses a strain of Penicillium citrinum with high nuclease P1 production and a breeding method thereof. The Penicillium citrinum has been preserved in the General Microbiology Center of the China Committee for the Collection of Microorganisms, with a preservation number of CGMCC No.2014. The breeding method of the strain is to perform low-energy ion beam injection on the suspension of Penicillium citrinum spores, then transfer the mutated spores to a plate culture medium, select a single strain and transfer it to a wort slant for culture, and then pass through the liquid Fermentation culture is used to screen out strains producing nuclease P1 with high enzyme activity. The strain has been subcultured for more than 30 generations, and a high-yield and stable strain is obtained. The invention adopts the method of low-energy ion beam implantation, which has good mutagenic effect, high positive mutation rate, and safe operation. The enzyme activity of nuclease P1 produced by the strain through fermentation is increased by 3 to 5 times, and the high-yielding traits are stable and not easy to lose. The enzyme activity can still maintain a high level after cultivation in a 3-ton fermenter, which is suitable for large-scale 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'-nucleotides. 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 mate...

Claims

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Application Information

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Patent Type & Authority Patents(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 TECH UNIV
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