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WO2023103543A1 - Procédé de préparation de nucléase p1 - Google Patents

Procédé de préparation de nucléase p1 Download PDF

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Publication number
WO2023103543A1
WO2023103543A1 PCT/CN2022/121569 CN2022121569W WO2023103543A1 WO 2023103543 A1 WO2023103543 A1 WO 2023103543A1 CN 2022121569 W CN2022121569 W CN 2022121569W WO 2023103543 A1 WO2023103543 A1 WO 2023103543A1
Authority
WO
WIPO (PCT)
Prior art keywords
fermentation
preparation
nuclease
retentate
accelerator
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2022/121569
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English (en)
Chinese (zh)
Inventor
陈晓春
张磊
汤亦文
滕红兵
蔡家栋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Biotogether Co Ltd
Nanjing Tech University
Original Assignee
Nanjing Biotogether Co Ltd
Nanjing Tech University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Biotogether Co Ltd, Nanjing Tech University filed Critical Nanjing Biotogether Co Ltd
Publication of WO2023103543A1 publication Critical patent/WO2023103543A1/fr
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Ceased legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/30—Endoribonucleases active with either ribo- or deoxyribonucleic acids and producing 5'-phosphomonoesters (3.1.30)
    • C12Y301/30001—Aspergillus nuclease S1 (3.1.30.1)
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to the field of microbial fermentation, in particular to a preparation method of nuclease P1.
  • Nucleotides are a class of biochemical substances with high added value. They have a wide range of uses and support the development of some large industries, such as high-end dairy and pharmaceutical industries.
  • Nucleotide production mainly adopts three methods: chemical synthesis, microbial fermentation and enzymatic hydrolysis.
  • the enzymatic hydrolysis method can obtain a mixture of four nucleotides at one time by degrading RNA, and the yield of the enzymatic reaction is relatively high.
  • This method is used in the industrial production of nucleotides at home and abroad.
  • the enzyme required in the preparation of nucleotides by enzymatic hydrolysis is nuclease P1, which is a phosphodiesterase that can hydrolyze RNA to obtain four kinds of 5'-nucleotides.
  • the enzymatic hydrolysis product has few impurities and the subsequent separation process is simple. This method is generally used abroad for the production of nucleotides.
  • Penicillium citrinum belongs to the asymmetric Penicillium group, the Penicillium velvetiform subgroup, and the Penicillium citrinum line, and is an important production strain of nuclease P1.
  • 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. How to efficiently produce nuclease P1 and reduce production costs has become the primary task of nucleotide production.
  • the technical problem to be solved by the present invention is to provide a preparation method of nuclease P1 for the deficiencies of the prior art.
  • the present invention discloses a method for preparing nuclease P1.
  • Penicillium citrinum is used for fermentation to prepare a fermentation liquid containing nuclease P1.
  • the preparation method of described fermentation accelerator comprises the steps:
  • step S2 The enzymolysis solution obtained in step S1 is ultrafiltered through an ultrafiltration membrane, and the obtained retentate is a fermentation accelerator.
  • step S1 pH 4-6, 2-6wt% ribonucleic acid aqueous solution and nuclease P1 perform enzymolysis at 50-90°C; preferably, pH 5, 4wt% ribonucleic acid aqueous solution and nuclease P1 perform enzymolysis at 70°C .
  • the pH is adjusted by NaOH.
  • nuclease P1 is added.
  • the dosage of the nuclease P1 is 80-120 U/g ribonucleic acid, preferably 100 U/g ribonucleic acid.
  • the enzymatic activity of the nuclease P1 is 100,000 U/g; the definition of the nuclease P1 enzymatic activity: 1.9 mL of substrate solution (containing RNA with a concentration of about 1%; 0.2M acetic acid buffer at pH5.2 After 0.0005M ZnSO 4 ) in a constant temperature water bath at 70°C for 10min, add 0.1mL of appropriately diluted enzyme solution, keep warm at 70°C for 15min, then add 2.0mL of nucleic acid precipitant (0.25% ammonium molybdate-2.5 perchloric acid), ice Centrifuge after 20 minutes in water bath, take the supernatant and dilute it with distilled water to a certain number of times, and measure the absorbance value at 260nm as A260.
  • substrate solution containing RNA with a concentration of about 1%; 0.2M acetic acid buffer at pH5.2
  • nucleic acid precipitant 0.25% ammonium molybdate-2.5 perchloric acid
  • step S2 the enzymolysis solution obtained in step S1 is cooled to 20°C and ultrafiltered by an ultrafiltration membrane to 1/8 to 1/12 of the original volume to obtain the first retentate; add water to the obtained first retentate to the original volume , and then ultrafiltered to 1/8 to 1/12 of the original volume to obtain the second retentate; add water to the obtained second retentate to the original volume, and then ultrafilter to 1/8 to 1/12 of the original volume to obtain
  • the third retentate, the obtained third retentate is the fermentation accelerator; preferably, each ultrafiltration is performed to 1/10 of the original volume.
  • the ultrafiltration is a roll-type polysulfone ultrafiltration membrane.
  • the molecular weight of the ultrafiltration membrane is 3000-10000 Daltons, preferably 4000-8000 Daltons, more preferably 5000-7000 Daltons, even more preferably 6000 Daltons.
  • the fermentation accelerator prepared by the above method contains oligonucleotide, short-chain RNA, zymosan, protein and yeast content.
  • the dosage of the fermentation accelerator is 1-20g/L fermentation medium, preferably 2-18g/L fermentation medium, more preferably 5-15g/L fermentation medium, even more preferably 8-13g/L fermentation medium.
  • L fermentation medium most preferably 10g/L fermentation medium.
  • the fermentation medium in addition to the fermentation accelerator, also includes 1-100 g/L of glucose, 0.1-10 g/L of peptone, 0.02-10 g/L of potassium dihydrogen phosphate, 0.02-10 g/L of dipotassium hydrogen phosphate, sulfuric acid Magnesium 0.02-10g/L, calcium chloride 0.02-10g/L.
  • the solvent of the fermentation medium is water, and the pH is 5-9.
  • the preservation number of the Penicillium citrinum is CGMCC No.2014, and the strain is recorded in the Chinese patent CN101067116A.
  • the fermentation temperature is 25-35°C, preferably 30°C.
  • the fermentation is a semi-continuous fermentation
  • the semi-continuous fermentation is that Penicillium citrinum is fermented in a fermenter for a period of time, and a part of the fermentation liquid containing the product is periodically released, and then the same volume of fresh medium is added.
  • Fermentation method specific method: a, the enzyme activity of citrus enzyme is stabilized at about 7300-7800U/mL after fermenting for 55 hours at a temperature of 30°C in a fermenter; b, release 85% of the volume of the fermentation broth, and keep As the next batch of fermented seeds, add the same volume of sterile fermentation medium to the fermenter to continue the cultivation, and the enzyme activity is stable at about 7300-8200U/mL for about 18 hours of fermentation; c, repeat the above step b, and so on Feed 5 times, stop the semi-continuous fermentation.
  • the time of single batch fermentation is 18h.
  • the enzyme activity of the fermentation broth containing nuclease P1 is 6500-8500 U/mL, preferably 7300-8200 U/mL, more preferably 8100-8300 U/mL, even more preferably 8200 U/mL.
  • the present invention has the following technical advantages:
  • fermentation is carried out by adding a fermentation accelerator, and the single-batch fermentation time is shortened from 55h to 18h, and the enzyme activity is increased from 2500U/mL to 7300-8200U/mL.
  • the method can significantly shorten the fermentation time, increase the activity of fermentation enzymes, and use a small amount of fermentation accelerators.
  • the fermentation accelerators used in the present invention are wastes from the process of producing nucleotides, with no cost, simple operation, and remarkable effects. .
  • Penicillium citrinum The preservation number of Penicillium citrinum described in the following examples is CGMCC No.2014.
  • the ratio of nuclease P1 and ribonucleic acid described in the following examples is the mass ratio.
  • the obtained ultrafiltration membrane retentate is the fermentation accelerator 1.
  • the obtained ultrafiltration membrane retentate is the fermentation accelerator 2.
  • the obtained ultrafiltration membrane retentate is the fermentation accelerator 3.
  • fermentation medium glucose 25g/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, fermentation Accelerator 110g/L, pH 6.5, sterilized at 121°C for 15 minutes, cooled to about 30°C for later use.
  • the enzyme activity stabilized to 7320U/mL after 18 hours, released 5.1L of fermentation broth again, and added 5.1L of fresh medium in the same way.
  • the feed was fed 5 times in this way, the enzyme activity was all around 7310U/mL, and the semi-continuous fermentation was stopped.
  • fermentation medium glucose 25g/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, fermentation Accelerator 210g/L, pH 6.5, sterilized at 121°C for 15 minutes, cooled to about 30°C for later use.
  • the enzyme activity stabilized to 8180U/mL after 18 hours, and 5.1L of fermentation broth was released again, and 5.1L of fresh medium was added in the same way.
  • the feed was fed 5 times in such a cycle, the enzyme activity was all around 8200U/mL, and the semi-continuous fermentation was stopped.
  • Embodiment 6 is a diagrammatic representation of Embodiment 6
  • fermentation medium glucose 25g/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, fermentation Accelerator 310g/L, pH 6.5, sterilized at 121°C for 15 minutes, cooled to about 30°C for later use.
  • the enzyme activity stabilized to 7620U/mL after 18 hours, released 5.1L of fermentation broth again, and added 5.1L of fresh medium in the same way.
  • the feeding cycle was fed 5 times in this way, and the enzyme activity was all around 7600U/mL, and the semi-continuous fermentation was stopped.
  • Preparation of fermentation medium glucose 25g/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, pH6 .5. Sterilize at 121°C for 15 minutes, then cool down to about 30°C for later use.
  • the enzyme activity stabilized to 2420U/mL after 18 hours, released 5.1L of fermentation broth again, and added 5.1L of fresh medium in the same way. In this way, feeding was circulated 5 times, and the enzyme activity was all around 2500U/mL, and the semi-continuous fermentation was stopped.
  • Embodiment 7 is a diagrammatic representation of Embodiment 7:
  • fermentation medium glucose 25g/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, fermentation Accelerator 22g/L, pH 6.5, sterilized at 121°C for 15 minutes, cooled to about 30°C for later use.
  • the enzyme activity stabilized to 6730U/mL after 18 hours, released 5.1L of fermentation broth again, and added 5.1L of fresh medium in the same way.
  • the feed was fed 5 times in such a cycle, the enzyme activity was all around 6700U/mL, and the semi-continuous fermentation was stopped.
  • Embodiment 8 is a diagrammatic representation of Embodiment 8
  • fermentation medium glucose 25g/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, fermentation Accelerator 218g/L, pH 6.5, sterilized at 121°C for 15 minutes, cooled to about 30°C for later use.
  • the enzyme activity stabilized to 7660U/mL after 18 hours, released 5.1L of fermentation broth again, and added 5.1L of fresh medium in the same way.
  • the feed was fed 5 times in such a cycle, and the enzyme activity was all around 7700U/mL, and the semi-continuous fermentation was stopped.
  • the present invention provides an idea and method for a preparation method of nuclease P1.
  • the above description is only a preferred embodiment of the present invention.
  • some improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention. All components that are not specified in this embodiment can be realized by existing technologies.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
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Abstract

La présente invention concerne un procédé de préparation de nucléase P1. Dans un milieu de fermentation contenant un accélérateur de fermentation, la fermentation est effectuée en utilisant Penicillium citrinum pour préparer un liquide de fermentation contenant la nucléase P1, le procédé de préparation de l'accélérateur de fermentation comprenant les étapes suivantes : (S1) soumission de l'acide ribonucléique à l'enzymolyse pour obtenir un liquide d'enzymolyse ; et (S2) ultrafiltration du liquide d'enzymolyse obtenu en S1 à l'aide d'une membrane d'ultrafiltration, le rétentat obtenu étant l'accélérateur de fermentation.
PCT/CN2022/121569 2021-12-07 2022-09-27 Procédé de préparation de nucléase p1 Ceased WO2023103543A1 (fr)

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CN202111483356.1 2021-12-07
CN202111483356.1A CN113980930B (zh) 2021-12-07 2021-12-07 一种核酸酶p1的制备方法

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Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113980930B (zh) * 2021-12-07 2023-03-03 南京工业大学 一种核酸酶p1的制备方法
CN118109448B (zh) * 2024-03-18 2024-12-06 武汉新华扬生物股份有限公司 一种提高核酸酶稳定性的助剂及其制备方法和应用

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101037681A (zh) * 2007-02-14 2007-09-19 北京燕京中科生物技术有限公司 一种发酵法制备核酸酶p1的方法
CN101418327A (zh) * 2008-11-21 2009-04-29 大连珍奥生物技术股份有限公司 高纯度5’核苷酸的生产新工艺
CN101654677A (zh) * 2009-09-17 2010-02-24 南京工业大学 一种高效表达核酸酶p1的基因及其应用
CN104560913A (zh) * 2014-12-24 2015-04-29 南京工业大学 一种桔青霉半连续发酵制备核酸酶p1的方法
CN107365831A (zh) * 2017-07-18 2017-11-21 三峡大学 一种桔青霉核酸酶p1菌种选育筛选方法
CN109852596A (zh) * 2019-04-04 2019-06-07 江苏省生产力促进中心 一种利用固定化桔青霉发酵制备核酸酶p1的方法
CN110218713A (zh) * 2019-06-24 2019-09-10 南京工业大学 一种提高桔青霉产核酸酶p1酶活的方法
CN113980930A (zh) * 2021-12-07 2022-01-28 南京工业大学 一种核酸酶p1的制备方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100532536C (zh) * 2007-05-25 2009-08-26 南京工业大学 一株高产核酸酶p1的桔青霉菌及其选育方法

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101037681A (zh) * 2007-02-14 2007-09-19 北京燕京中科生物技术有限公司 一种发酵法制备核酸酶p1的方法
CN101418327A (zh) * 2008-11-21 2009-04-29 大连珍奥生物技术股份有限公司 高纯度5’核苷酸的生产新工艺
CN101654677A (zh) * 2009-09-17 2010-02-24 南京工业大学 一种高效表达核酸酶p1的基因及其应用
CN104560913A (zh) * 2014-12-24 2015-04-29 南京工业大学 一种桔青霉半连续发酵制备核酸酶p1的方法
CN107365831A (zh) * 2017-07-18 2017-11-21 三峡大学 一种桔青霉核酸酶p1菌种选育筛选方法
CN109852596A (zh) * 2019-04-04 2019-06-07 江苏省生产力促进中心 一种利用固定化桔青霉发酵制备核酸酶p1的方法
CN110218713A (zh) * 2019-06-24 2019-09-10 南京工业大学 一种提高桔青霉产核酸酶p1酶活的方法
CN113980930A (zh) * 2021-12-07 2022-01-28 南京工业大学 一种核酸酶p1的制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LIAO HONG-DONG, MO XIAO-YAN, SONG WEI: "Purification and Partial Enzymological Properties of Nuclease P1", CHINESE JOURNAL OF PHARMACEUTICALS., vol. 36, no. 9, 10 September 2005 (2005-09-10), pages 536 - 538, 544, XP093071769 *

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