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Method for producing nuclease P1 through fermentation of penicillium citrinum

A production method and nuclease technology, which are applied in the production field of nuclease P1 produced by Penicillium citrinum fermentation, can solve the problems of low nuclease P1 enzyme activity, high production cost, low yield and the like, and achieve industrialized production, increase yield, The effect of changing the market landscape

Active Publication Date: 2017-01-11
ANGELYEAST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem solved by the present invention is that the existing method for producing nuclease P1 by fermentation of Penicillium citrinum has the defects of high production cost, low enzyme activity of nuclease P1, and low yield

Method used

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  • Method for producing nuclease P1 through fermentation of penicillium citrinum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] 1. Strain activation: Take a glycerol tube of Penicillium citrinum spore suspension, pick a loop with a 10ul inoculation loop, insert it into 100g of PDA medium in a sterile environment, and culture it at 28°C for 3 days. And eluted with 50ml sterile water to obtain spore suspension.

[0073] 2. Seed preparation: inoculate the spore suspension into 450ml seed shake flask culture medium according to the inoculum size of 1% by weight, and place it at 28°C to cultivate for 22 hours to obtain the seed bacteria liquid, wherein the formula of the seed shake flask culture medium Same as the recipe for the seed medium in step 3.

[0074] 3. Seed tank cultivation: according to the inoculum amount of 0.1% by weight, the seed bacteria solution is inserted into a seed tank (0.5 tons of storage capacity) equipped with 450L seed culture medium, and cultivated with aeration and stirring at a temperature of 30°C for 24 hours Prepare the seed culture solution, wherein the stirring spee...

Embodiment 2

[0081] 1. Strain activation: Take a glycerol tube of Penicillium citrinum spore suspension, pick a loop with a 10ul inoculation loop, insert it into 100g of PDA medium in a sterile environment, and culture it at 29°C for 2 days. And eluted with 50ml sterile water to obtain spore suspension.

[0082] 2. Seed preparation: inoculate the spore suspension into the culture medium containing 6L seed shake flasks according to the inoculum size of 2.5% by weight, and cultivate them at 29°C for 20 hours to obtain the seed bacterial liquid, wherein the seed shake flasks The formula of medium is the same as the formula of seed medium in step 3.

[0083] 3. Seed tank cultivation: according to the inoculum amount of 0.5% by weight, the seed bacteria solution is inserted into a seed tank (2 tons of storage capacity) equipped with 1200L seed medium, and cultivated with aeration and stirring at a temperature of 29°C for 28 hours Obtain the seed culture solution, wherein the stirring speed is ...

Embodiment 3

[0090] 1. Strain activation: Take a glycerin tube of Penicillium citrinum spore suspension, pick a loop with a 10ul inoculation loop, put it into 100g of PDA medium in a sterile environment, culture it at 28°C for 3 days, and use Eluted with 50ml sterile water to obtain spore suspension.

[0091] 2. Seed preparation: inoculate the spore suspension into a 2L seed shake flask culture medium according to the inoculum size of 2% and carry out shake flask culture, and cultivate it at a temperature of 29°C for 20 hours to obtain the seed bacterial liquid, wherein the seed shake flask culture medium The formula is the same as that of the seed medium in step 3.

[0092] 3. Seed tank cultivation: according to the inoculum size of 0.25%, the seed bacteria solution was inserted into a seed tank (1 ton storage capacity) equipped with 750L seed culture medium, and aerated and stirred at a temperature of 29°C for 24 hours to obtain seeds Culture solution, wherein the stirring speed is 100r...

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Abstract

The invention relates to the field of a microbe fermenting enzyme preparation, and more specifically provides a method for producing nuclease P1 through fermentation of penicillium citrinum. The method comprises the following steps: 1) bacterial strain activation; 2) seed preparation: inoculating the seeds in a seed shake culture medium for culture according to the inoculation amount of 1-2.5%; 3) seeding tank culture: inoculating the seeds in a seeding tank for culturing for 24-28 hours according to the inoculation amount of 0.1-0.5%, wherein the rotating speed is 50-150 rpm, and a ventilation ratio is 1: 0.5-1:1; and 4) fermentation cylinder production: inoculating the seeds in a fermentation cylinder for culturing for 100-200 hours according to the inoculation amount of 3-8%, wherein the rotating speed is 100-200 rpm, and a ventilation ratio is 1: 0.5-1:1.2, and feeding a culture medium containing a nitrogen source, a trace element and a carbon source after fermentation for 45-48 hours. The enzyme activity level of the nuclease can reach 3000-4100 [mu] / ml, solves the problems of low nuclease production level and high cost in the prior art, and has large-scale industrial value.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation enzyme preparations, in particular to a production method for producing nuclease P1 by fermentation of Penicillium citrinum. Background technique [0002] Nuclease P1 (Nuclease P1), also known as 5'-phosphodiesterase, acts to hydrolyze 3',5'-phosphodiester bonds in nucleic acids to obtain four 5'-nucleotides. Nucleotides are important low-molecular compounds in organisms and have many physiological functions. At present, with the wide application in the field of infant milk powder and health care products, the market for nucleotides is getting bigger and bigger. Among them, nuclease P1 plays a vital role in the industrial production of nucleotides. [0003] According to current domestic research reports, nuclease P1 is mainly obtained through liquid submerged fermentation to obtain crude enzyme liquid, and then refined by downstream processing to obtain finished products. Liquid ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/22C12R1/80
CPCC12N9/22C12Y301/04001
Inventor 喻晨俞学锋李知洪姚鹃吴尧董先有李汉文余华顺
Owner ANGELYEAST CO LTD