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A method for improving the activity of nuclease p1 produced by Penicillium citrinum

A technology for producing and nuclease from Penicillium citrinum, which is applied in the field of microbial fermentation, can solve the problems of long time consumption and low enzyme activity, achieve obvious effects, improve fermentation enzyme activity, and add less amount

Active Publication Date: 2020-07-03
NANJING TECH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The production of nuclease P1 at home and abroad is mainly through liquid submerged fermentation of Penicillium citrinum, and the existing method takes a long time and has low enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Preparation of fermentation medium: glucose 20g / L, peptone 2g / L, potassium dihydrogen phosphate 0.5g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.2g / L, calcium chloride 0.2g / L, plant Calcium Magnesium 0.2g / L, Zinc Phytate 0.1g / L, Tween-80 0.5g / L, Span-80 0.5g / L, RNA 0.5g / L, pH 6, sterilized at 121°C for 15min, cooled to 30°C ℃ for later use.

[0022] Semi-continuous fermentation: Put 6L culture medium in a 10L stirring tank, insert the Penicillium citrinum strain cultured on the wort slant medium after sterilization, and ferment for 56 hours at a temperature of 30°C in the fermenter The enzyme activity was stable at about 7300U / mL, and the volume of the fermented liquid released was 4.8L. The fermented liquid retained in the fermenter was used as the next batch of fermented seeds, and 4.8L of fermentation medium was prepared and put into the fermenter for further cultivation. After 20 hours, the enzyme activity stabilized to 7300U / mL, release 4.8L of ...

Embodiment 2

[0027] Preparation of fermentation medium: glucose 20g / L, peptone 2g / L, potassium dihydrogen phosphate 0.5g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.2g / L, calcium chloride 0.2g / L, plant Calcium Magnesium Oxide 0.2g / L, Zinc Phytate 0.1g / L, Tween-80 0.5g / L, Span-80 0.5g / L, RNA 0.5g / L, pH 6, sterilized at 121°C for 15min, cooled to 30°C ℃ for later use.

[0028] Semi-continuous fermentation: Put 60L culture medium in a 100L stirring tank, insert the Penicillium citrinum cultured on the wort slant culture medium after sterilization, and ferment for 56 hours at a temperature of 30°C in the fermentation tank The enzyme activity is stable at about 7400U / mL, and the volume of the fermented liquid released is 48L. The fermented liquid retained in the fermenter is used as the next batch of fermented seeds, and 48L of fermentation medium is prepared and put into the fermenter for further cultivation. After 20 hours, the enzyme activity was stabilized to 7480U / mL, an...

Embodiment 3

[0033] Preparation of fermentation medium: glucose 20g / L, peptone 2g / L, potassium dihydrogen phosphate 0.5g / L, dipotassium hydrogen phosphate 0.5g / L, magnesium sulfate 0.2g / L, calcium chloride 0.2g / L, plant Calcium Magnesium 0.2g / L, Zinc Phytate 0.1g / L, Tween-80 0.5g / L, Span-80 0.5g / L, RNA 0.5g / L, pH 6, sterilized at 121°C for 15min, cooled to 30°C ℃ for later use.

[0034]Semi-continuous fermentation: Put 600L medium in a 1000L stirring tank, insert the Penicillium citrinum cultured on the wort slant medium after sterilization, and ferment for 56 hours at a temperature of 30°C in the fermenter The enzyme activity is stable at about 7500U / mL, and the volume of the fermented liquid released is 480L. The fermented liquid retained in the fermenter is used as the next batch of fermented seeds, and 480L of fermentation medium is prepared and put into the fermenter for further cultivation. After 20 hours, the enzyme activity was stabilized to 7500U / mL, and 480L of fermentation brot...

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Abstract

The invention relates to the technical field of biology, and particularly discloses a method for improving the enzyme activity of penicillium citrinum produced nuclease P1. By using a fermentation culture medium containing phytic acid compounds, a surfactant and a nucleic acid substance, semi-continuous fermentation is carried out on penicillium citrinum to prepare fermentation liquor containing the nuclease P1. According to the method, the single-batch fermentation time is shortened to 20 hours from 56 hours, and the enzyme activity is increased from 2500 U / mL to 7100-7500 U / mL. The additionquantity of enzyme production promoters used in the preparation method is small, the cost is low, the operation is simple, and the effect is obvious.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for improving the activity of the nuclease P1 produced by Penicillium citrinum. Background technique [0002] 5'-nucleotides have been widely used in food, biochemical and pharmaceutical industries. In the food industry, it has been 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 bread, biscuits and other foods, 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 materials for the production of many antiviral and anti-tumor drugs; in addition, nucleotides The preparation can also be used as an animal and plant growth regulator, and has obvious effects of increasing produc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/22C12R1/80
CPCC12N9/22C12Y301/04001
Inventor 陈晓春齐昌瑞张磊汤亦文周昌祝
Owner NANJING TECH UNIV
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