CN109234268B - Method for treating yeast ribonucleic acid mother liquor - Google Patents
Method for treating yeast ribonucleic acid mother liquor Download PDFInfo
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- CN109234268B CN109234268B CN201811092810.9A CN201811092810A CN109234268B CN 109234268 B CN109234268 B CN 109234268B CN 201811092810 A CN201811092810 A CN 201811092810A CN 109234268 B CN109234268 B CN 109234268B
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- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
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Abstract
The invention relates to a method for treating yeast ribonucleic acid mother liquor, which comprises the steps of adding quicklime into the yeast ribonucleic acid mother liquor after ribonucleic acid sedimentation separation to adjust the pH value to be near neutral, concentrating clear liquid obtained after solid-liquid separation by using a nanofiltration membrane to ensure that ribonucleic acid is enriched to reach sedimentation concentration for secondary sedimentation, and improving the total yield of ribonucleic acid products. The method not only recovers PO4 3‑The calcium phosphate salt is prepared, the RNA loss is reduced, the product yield is improved, the saline water is recycled, and the production cost is reduced. The method is simple to operate, saves cost, protects environment and is suitable for large-scale production.
Description
Technical Field
The invention belongs to the technical field of bioseparation, and relates to a method for treating yeast ribonucleic acid mother liquor.
Background
RNA is an important material basis for the existence of life. RNA and its nuclease decomposition products adenylic acid, guanylic acid, uridylic acid, cytidylic acid, etc., have important uses in the industries of medicine, food, cosmetics, agriculture, etc. RNA also has multiple physiological functions of maintaining the immune function of the body (the research progress [ J ] of ribonucleic acid prepared by a yeast fermentation method in the Wen army, Rozhou sphere, 2008, 19 (4): 638-. Therefore, research and development of RNA are increasingly receiving attention. But its use is limited due to the quality and price of current RNA products. The existing process for extracting RNA by yeast fermentation is an effective way for producing RNA and corresponding nucleotide.
Extraction of RNA from microorganisms is the most practical and efficient method in industry. The RNA in the yeast is not only high in content, but also suitable for the research of extracting ribonucleic acid from beer yeast (Zhujundong, Huang Guo Rong)]Food science, 2006, (27) 2: 160-162). Yeast RNA extraction was mainly performed in 3 steps: 1) breaking cell wall, and dissolving RNA from cells by high-temperature cooking with high-concentration saline water; 2) centrifuging to separate RNA from yeast body to obtain RNA clear solution; 3) settling by H3PO4Adjusting pH to isoelectric point of RNA to precipitate RNAPrecipitating, purifying and drying to obtain a finished product.
However, RNA can not be settled completely in the process of sedimentation, and the sedimentation mother liquor also contains a small amount of RNA. If discarded directly, it is wasted. The secondary sedimentation is direct, and the secondary sedimentation cannot be realized because of low concentration. In addition, the settled mother liquor also contains a large amount of PO4 3-And NaC1, which would cause environmental pollution and cost waste if not disposed of for direct discharge.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method which is simple to operate, reduces waste, saves cost, protects environment and is suitable for large-scale production of ribonucleic acid.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a method for treating yeast ribonucleic acid mother liquor comprises adding calx to adjust pH to near neutral, and precipitating PO4 3-And concentrating clear liquid obtained after solid-liquid separation by using a nanofiltration membrane to ensure that the ribonucleic acid is enriched to reach the sedimentation concentration for secondary sedimentation, thereby improving the total yield of the ribonucleic acid product.
Wherein, quicklime is added to adjust the pH value to 7.0 +/-0.1.
Wherein, the solid-liquid separation adopts any one or more of a horizontal screw centrifuge, a disc separator, a three-leg centrifuge and a plate-and-frame filter, and the three-leg centrifuge is preferably used.
Wherein, the solid-liquid separation is carried out, and the phosphate is recovered from the obtained solid matter.
Wherein the nanofiltration membrane is concentrated, and the concentration multiple is 3-10 times, preferably 6 times.
Wherein the concentration of the enriched ribonucleic acid is 9-40 g/L, preferably 14-17 g/L.
Wherein the nanofiltration membrane is a polysulfone nanofiltration membrane, and the molecular weight cut-off of the membrane is 300-1000 Dalton, preferably 300-600 Dalton.
The nanofiltration membrane is a hollow fiber membrane, a plate-frame membrane, a roll-up membrane or a tubular membrane, and preferably a hollow fiber membrane.
Wherein NaCl contained in the nanofiltration permeating liquid is recycled to the yeast wall breaking step.
Has the advantages that: the method of the invention recovers PO through simple operation4 3-The calcium phosphate byproduct is prepared, the loss of RNA is reduced, the product yield is improved, the saline water is recycled to reduce the production cost, and the purpose of protecting the environment is achieved. The method is simple to operate, saves cost and is suitable for large-scale production.
Detailed Description
The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.
Example 1
Adding H into 10L solution with RNA content of 15 g/L3PO4Adjusting pH to 2.01, washing the precipitated RNA with ethanol, filtering, oven drying, and settling to obtain a mother liquor containing 3.2 g/L RNA and 21.3 g/L PO4 3-Adding quicklime to adjust the pH value of the mother liquor to 7.01, centrifuging at 6000rpm for 5min by using a laboratory centrifuge, drying a centrifugal solid phase to prepare calcium phosphate, performing nanofiltration concentration on a centrifugal clear phase by using a polysulfone hollow fiber nanofiltration membrane with the molecular weight cutoff of 300 channels for 4 times, increasing the RNA concentration to 11.3 g/L, performing secondary sedimentation to obtain RNA, wherein the primary sedimentation yield of the RNA is 78.7%, and the total yield of the RNA is 93.3%, and recycling nanofiltration permeate for the yeast cell disruption process.
Example 2
Adding H into 100L solution with RNA content of 13.5 g/L3PO4Adjusting pH to 2.03, washing the precipitated RNA with ethanol, filtering, oven drying, and settling to obtain a mother liquor containing 2.8 g/L RNA and 19.6 g/L PO4 3-Adding quicklime to adjust the pH value of the mother liquor to 6.98, centrifuging by a three-leg centrifuge, drying a centrifugal solid phase to prepare calcium phosphate, performing nanofiltration concentration on a centrifugal clear phase by 6 times through a polysulfone hollow fiber nanofiltration membrane with the molecular weight cutoff of 600, increasing the RNA concentration to 14.8 g/L, and performing secondary sedimentation to obtain RNA, wherein the primary sedimentation yield of the RNA is 79.3%, and the total RNA yield is 93.7 percent. And recycling nanofiltration permeate for the yeast wall breaking process.
Example 3
5m RNA content 14.6 g/L3Solution, adding H3PO4Adjusting pH to 2.05, washing the precipitated RNA with ethanol, filtering, oven drying, and settling to obtain a mother liquor containing 3.1 g/L RNA and 19.3 g/L PO4 3-Adding quicklime to adjust the pH value of the mother liquor to 7.03, centrifuging by a horizontal spiral centrifuge, drying a centrifugal solid phase to prepare calcium phosphate, performing nanofiltration concentration on a centrifugal clear phase by a polysulfone hollow fiber nanofiltration membrane with the molecular weight cutoff of 1000 by 10 times, increasing the RNA concentration to 27.1 g/L, and performing secondary sedimentation to obtain RNA, wherein the primary sedimentation yield of the RNA is 78.8%, and the total yield of the RNA is 92.6%.
Claims (8)
1. A method for processing yeast ribonucleic acid mother liquor is characterized in that the pH value of the yeast ribonucleic acid mother liquor after ribonucleic acid sedimentation separation is adjusted to be near neutral by adding quick lime, clear liquid obtained after solid-liquid separation is concentrated by a nanofiltration membrane, ribonucleic acid is enriched to reach sedimentation concentration for secondary sedimentation, and the total yield of ribonucleic acid products is improved;
the nanofiltration membrane is a polysulfone nanofiltration membrane, and the molecular weight cut-off of the membrane is 300-1000 daltons.
2. The method for treating yeast ribonucleic acid mother liquor according to claim 1, characterised in that quicklime is added to adjust the pH to 7.0. + -. 0.1.
3. The method for treating yeast ribonucleic acid mother liquor according to claim 1, wherein the solid-liquid separation is performed by any one or more of horizontal screw centrifuge, disc separator, three-leg centrifuge and plate-and-frame filter.
4. The method for treating yeast ribonucleic acid mother liquor according to claim 1, characterised in that phosphate is recovered from the solid-liquid separation obtained solid matter.
5. The method for treating yeast ribonucleic acid mother liquor according to claim 1, wherein the nanofiltration membrane is concentrated by a factor of 3-10.
6. The method for processing yeast ribonucleic acid mother liquor according to claim 1, wherein the concentration of the enriched ribonucleic acid is 9-40 g/L.
7. The method for treating yeast ribonucleic acid mother liquor according to claim 1, wherein the nanofiltration membrane is a hollow fiber membrane, a plate-and-frame membrane, a roll-type membrane or a tubular membrane.
8. The method for treating yeast ribonucleic acid mother liquor according to claim 1, wherein NaCl contained in the nanofiltration permeate is recycled to the yeast cell wall breaking step.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811092810.9A CN109234268B (en) | 2018-09-19 | 2018-09-19 | Method for treating yeast ribonucleic acid mother liquor |
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| CN201811092810.9A CN109234268B (en) | 2018-09-19 | 2018-09-19 | Method for treating yeast ribonucleic acid mother liquor |
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| CN109234268A CN109234268A (en) | 2019-01-18 |
| CN109234268B true CN109234268B (en) | 2020-07-10 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104371000A (en) * | 2013-08-14 | 2015-02-25 | 梅乐和 | Method for extracting glutathione and ribonucleic acid in beer waste yeast with high efficiency |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104371000A (en) * | 2013-08-14 | 2015-02-25 | 梅乐和 | Method for extracting glutathione and ribonucleic acid in beer waste yeast with high efficiency |
Non-Patent Citations (2)
| Title |
|---|
| Improvement in RNA Extraction from S. cerevisie by Optimization in the Autolysis and NH3 Hydrolysis;Antonio Martins Oliveira等;《Braz. Arch. Biol. Technol.》;20111031;第54卷(第5期);全文 * |
| 酵母RNA超声辅助提取工艺研究;任国艳等;《食品工业科技》;20091231;第30卷(第8期);155-157 * |
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