CN103173389A - Method for culturing hair weed cells industrially - Google Patents
Method for culturing hair weed cells industrially Download PDFInfo
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- CN103173389A CN103173389A CN201310083166XA CN201310083166A CN103173389A CN 103173389 A CN103173389 A CN 103173389A CN 201310083166X A CN201310083166X A CN 201310083166XA CN 201310083166 A CN201310083166 A CN 201310083166A CN 103173389 A CN103173389 A CN 103173389A
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Abstract
The invention relates to the field of industrial cultivation, in particular to a method for culturing hair weed cells industrially; the method comprises the steps of a. manufacturing of a hair weed cultivation device: the cultivation device comprises a sampling device, a real-time monitoring device, an air inlet and outlet device and a liquid discharge device; b. nutrient solution preparation: preparing hair weed nutrient solution, and conveying the prepared nutrient solution into the cultivation device by a liquid preparation pipe; c. cultivation conditions: adding original hair weed cell seeds into the cultivation device, filling oxygen into the nutrient solution, and adjusting the environment of the nutrient solution to enable the nutrient solution to reach certain index; carrying out aerobic cultivation for 10-15 days, sampling by a sampling pipe at fixed period within the 10-15 days and detecting the light absorption value of the samples; and d. harvest: when the light absorption value OD 750nm is 0.6, harvesting. The method can be used for continuously culturing and harvesting the hair weeds; and under the condition that the hair weeds in the nutrient solution can be maintained to grow at the highest growth speed, the growth of the hair weeds can not be inhibited due to higher cell density, or can not be blocked due to lower cell density.
Description
Technical field
The present invention relates to batch production and cultivate the field, relate in particular to a kind of method that hair weeds cells is cultivated in batch production.
Background technology
Deliver vegetables, for sending out being commonly called as of shape beads cyanobacteria, have another name called and send out the shape nostoc, it is the edible blue-green algae of a kind of famous and precious land natural disposition, because of nutritious, also have certain pharmaceutical use and pronunciation and an approaching favor that is subject to consumers in general, especially Fujian, Guangdong, Hong Kong, Macao and Taiwan and south east asia human consumer of " making a good deal of money " two words.
On the world market, the supply of delivering vegetables is mainly from Mongolia, but the Japan that only has of product is sold in processing, and its raw material is affected by raw material supply for to deliver vegetables from Mongolian import is wild, and output is little.Delivering vegetables for a long time is subject to liking of human consumer as a kind of famous and precious food, but the growth of delivering vegetables is very slow, and natural standing stock limited causes the hair-like seaweed resource subject to severe risks of damage to immoderate the gathering of delivering vegetables, so that is on the verge of exhaustion.
Play State Council in July, 2000 and gather, process and sell in what forbid delivering vegetables, delivering vegetables is listed in country-level focused protection plant, therefore can't carry out as the production of raw material to deliver vegetables, and therefore is badly in need of carrying out the artificial mass-producing of delivering vegetables and cultivates.Realize at present hair weeds cells laboratory artificial culture on a small scale, but there is not yet the report of the device and method of artificial large scale culturing hair weeds cells.
On January 22nd, 2003, China national Department of Intellectual Property discloses the patent of invention of " a kind of method of cultivating hair weeds cells " by name, the disclosed technical scheme of this patent is soaked wild delivering vegetables and homogenized through nutrient solution, then get hair weeds cells through illumination cultivation, this method is successfully delivered vegetables by changing under terrestial enviornment in liquid nutrient medium wild, makes the artificial culture hair weeds cells become possibility.
On June 28th, 2006, China national Department of Intellectual Property discloses the patent of invention of " hair weeds cells process for solid culture " by name, the disclosed technical scheme of this patent is with the hair weeds cells suspension of activation, be inoculated in solid medium matter top layer, spray a certain amount of nutrient solution for preparing in advance, cultivate hair weeds cells under Artificial Control or natural condition, and do the wet rhythm and pace of moving things and cultivate, control suitable culture condition, realize complete artificial culture or open-air enlarged culturing.This technological breakthrough traditional training method, accelerated value-added speed.
On October 31st, 2007, Chinese State Intellectual Property Office disclosed the patent of invention of " stablizing the method for the hair weeds cells high-density culture of high yield Nostoc flagelliforme Polysaccharides " by name, the disclosed technical scheme of this patent is taked mixotrophism or heterotrophism training mode, the interpolation organic carbon source has improved the speed of growth of hair weeds cells, has shortened the culture cycle of hair weeds cells.
On September 29th, 2010, China national Department of Intellectual Property discloses the patent of invention of " a kind of method of two stage High-efficient Production hair weeds cells and exocellular polysaccharide thereof " by name, the disclosed technical scheme steps of this patent is inoculated in culture vessel for (1) gets the hair weeds cells seed liquor, substratum is the BG11 substratum of improvement, cultivated 8 ~ 10 days, (2) be the glucose of 1 ~ 20g/L as adding middle concentration in above-mentioned nutrient solution, cultivated 2 ~ 8 days, collect hair weeds cells, get the exocellular polysaccharide in centrifuged supernatant extraction nutrient solution.The culture cycle that the method that this invention relates to has effectively shortened hair weeds cells has improved the output of exocellular polysaccharide simultaneously.
But above these technical schemes only limit to the laboratory and cultivate on a small scale, and culture vessel used also only limits to the glass pot of triangular flask and several decaliters, do not have so far batch production to cultivate the method for hair weeds cells.
Summary of the invention
The object of the invention is to overcome the problems referred to above, provide a kind of and can adapt to various culture environment, can produce the method that hair weeds cells is cultivated in high quality hair weeds cells and a kind of batch production that can large scale continuous prod.
To achieve these goals, the technical solution adopted in the present invention is: the method for hair weeds cells is cultivated in a kind of batch production, it is characterized in that, comprises the steps:
A makes the culture apparatus of delivering vegetables: culture apparatus is made of sample adding device, real-time monitoring device, air inlet and gas barrier and pumping equipment;
B prepares nutrient solution: prepare the nutrient solution of delivering vegetables, by liquid distribution pipe, the nutrient solution for preparing is transported in culture apparatus;
C culture condition: add original hair weeds cells kind in culture apparatus, make nutrient solution reach following index to the logical oxygen of nutrient solution, adjustment nutrient solution environment, i.e. 24 ~ 28 ℃ of temperature, illumination 3200 ~ 4000lux, light dark period 12h:12h, pH value 8.0 ~ 9.5; Aerated culture 10 ~ 15 days, during in regularly take a sample by stopple coupon and detect its light absorption value;
D gathers: when light absorption value reaches OD
750nmBe 0.6 o'clock, gather.
The method of hair weeds cells is cultivated in aforesaid a kind of batch production, and in step b, the composition of nutrient solution and part by weight are: anhydrous CaCl
226g, MgSO
47H
2O 75g, NaSiO
39H
2O 58g, K
2HPO
43H
2O 67g, NaNO
31.5kg, 1 ton of micro-10g, membrane filtration pure water.
The method of hair weeds cells is cultivated in aforesaid a kind of batch production, in step c, leads to the oxygen pump by Scroll-tupe and realizes logical oxygen.
The method of hair weeds cells is cultivated in aforesaid a kind of batch production, in step c, when gathering, 350 order filtering nets is tiled on the plastic wire bed plastic wire bed below placement liquid collector; Open draining valve, the hair weeds cells nutrient solution is slowly flowed into filtering net filter, valve-off after the hair weeds cells nutrient solution of 1/3 volume in the culture apparatus of at every turn gathering injects the nutrient solution of equal volume simultaneously by liquid distribution pipe.
The method of hair weeds cells is cultivated in aforesaid a kind of batch production, after the moisture of hair weeds cells nutrient solution is filtered dry, thoroughly clean hair weeds cells with deionized water, hair weeds cells after flushing acidity instrumentation pH value, show that neutrality is qualified, hair weeds cells after rinsing is well changed in pallet and puts into-20 ℃ of refrigerator freezings, treat that the freezing vacuum freeze drier that adopts fully carries out drying.
The present invention takes cultured continuously and gathers in the crops continuously to delivering vegetables, delivering vegetables in nutrient solution remained under the fastest density conditions of the speed of growth, can not suppress because hair weeds cells density is too high its speed of growth, or hinder its growth because hair weeds cells density is too low, be applicable to the large-scale industrialized production of hair weeds cells.
Description of drawings
Fig. 1 is the structure iron of single cylindrical tube;
Fig. 2 is the structure iron of the cultivation unit that consists of of some cylindrical tubes;
Wherein, 1-T type folder; The 2-vapor pipe; The 3-PH meter; The 4-thermometer; The 5-oxygen electrode; The 6-sample feeding pipe; The 7-valve; The 8-dosing is in charge of; 9-dosing house steward; The 10-threeway; The 11-upper cover; The 12-cylindrical tube; The 13-Bubbled stone; The 14-bottom; The 15-screw thread; The 16-valve; The 17-thief hole; The 18-threeway; The 19-liquid header; The 20-discharge opeing is in charge of; The 21-non-return valve; The 22-air inlet is in charge of; 23-inlet manifold; The 24-stainless steel frame; The 25-wheel.
Embodiment
The invention will be further described below in conjunction with the drawings and specific embodiments.
The method of hair weeds cells is cultivated in a kind of batch production, described cultural method comprises the culture apparatus of delivering vegetables customized, preparation nutrient solution, controls culture condition, gathers and dry hair weeds cells, and it is characterized in that: (1) culture apparatus of delivering vegetables customized: described device comprises a cylindrical tube 12, a circular upper cover and a circular bottom.Described cylindrical tube 12 reaches upward, bottom is transparent PC plastics and makes.The internal diameter of described cylindrical tube 12 is 30cm, high 150cm.The inner wall thickness of described cylindrical tube 12 is 1cm, and TBE (threaded both ends) place, up and down thickness is 2cm, and the thickness of circular upper cover is 7cm, and the thickness of circular bottom is 10cm.Mainly formed by sample adding device, real-time monitoring device, air inlet and gas barrier and pumping equipment four parts, also be provided with the wheel 25 that is positioned at cylindrical tube 12 bottoms, the stainless steel shelf 24 of support cylinder shape cylindrical shell 12, the PH meter 3 of detection nutrient solution pH value, the thermometer 4 of detection culture-liquid temp.
(2) preparation nutrient solution: at first prepare nutrient solution in the dosing pond, by liquid distribution pipe (comprise dosing house steward 9 and dosing are in charge of 8), the nutrient solution for preparing is transported in culture apparatus, the composition of described nutrient solution and part by weight are: anhydrous CaCl
226g, MgSO
47H
2O 75g, NaSiO
39H
2O 58g, K
2HPO
43H
2O 67g, NaNO
31.5kg, micro-10g, 1 ton of membrane filtration pure water.
(3) culture condition: at first add nutrient solution and original hair weeds cells kind in culture apparatus, then open the logical oxygen of the logical oxygen pump of Scroll-tupe, diffuser comprises that air inlet is in charge of 22 and inlet manifold 23, detect the oxygen electrode 5 of oxygen level, gas is transported to Bubbled stone by inlet pipe, gas forms numerous tiny bubbles through Bubbled stone, nutrient solution and the seed of delivering vegetables are stirred, the seed of delivering vegetables can be circulated be suspended in nutrient solution, be beneficial to photosynthetic efficiency.By real-time monitoring device, the culture environment in cylindrical tube 12 are carried out real-time monitoring and regulation and control, guarantee to deliver vegetables can be in a constant environment healthy growth.Adjusting at last the interior environment of nutrient solution makes it reach following index: 24 ~ 28 ℃ of temperature (being preferably 24 ~ 26 ℃), illumination 3200 ~ 4000lux or 40 ~ 50 μ molm
-2S
-1, light dark period 12h:12h(is illumination 12h, the dark 12h of light), pH value 8.0 ~ 9.5 (being preferably 9.0 ~ 9.5).
(4) gather and dry: after kind of liquid and nutrient solution are delivered vegetables in inoculation, control temperature and the pH value of nutrient solution, aerated culture 10 ~ 15 days, at this moment between in regularly take a sample by stopple coupon and detect its light absorption value, when light absorption value reaches OD
750nmBe 0.6 o'clock, can gather, when gathering, 350 purpose filtering nets are tiled on the plastic wire bed, plastic wire bed below placement liquid collector recycles nutrient solution; Open draining valve, pumping equipment is provided with liquid header 19 and discharge opeing is in charge of 20, hair weeds cells liquid is slowly flowed into filtering net filter, the nutrient solution of delivering vegetables of 1/3 volume in the culture apparatus of at every turn gathering, valve-off injects the nutrient solution of equal volume simultaneously by liquid-inlet pipe; After moisture is filtered dry, thoroughly clean hair weeds cells with deionized water, hair weeds cells after flushing records its pH value with acidometer, show that neutrality is for qualified, hair weeds cells after rinsing is well changed in pallet put into-20 ℃ of refrigerator freezings, treat that the freezing vacuum freeze drier that fully it adopted carries out drying.
Cultural method of the present invention culture apparatus (cylindrical tube 12) used is totally enclosed, can cultivate in factory building, can cultivate in plastic greenhouse or glasshouse, also cultivates under condition in the open.
The device of hair weeds cells is cultivated in a kind of mass-producing, apparatus main body comprises cylindrical tube 12, circular upper cover 11, circular bottom 14, it is characterized in that: described upper cover 11 is provided with some holes, circular upper cover 11 has five holes, be respectively PH meter, thermometer, oxygen electrode open holes, exhaust duct aperture, nutrient solution and plant liquid sample introduction pore, being connected with on-line real time monitoring device, sample feeding pipe 6, vapor pipe 2 respectively; Described bottom 14 is provided with some holes, is respectively air inlet pipe hole and discharge opeing pore, is connected with liquid discharge pipe with inlet pipe (comprising that inlet manifold 23 is connected 22 with air inlet) respectively and comprises that liquid header 19 is connected 20 with discharge opeing) be connected.
The on-line real time monitoring device comprises PH meter 3, thermometer 4 and oxygen electrode 5, is respectively used to monitor pH value, temperature and the oxygen level of hair weeds cells nutrient solution (being that hair weeds cells kind liquid and nutrient solution form).The inlet pipe end is equipped with Bubbled stone 13, and Bubbled stone 13 is positioned at the bottom of cylindrical tube 12 and above circular bottom 14.Cylindrical tube 12 is installed on stainless steel frame 24, and each stainless steel frame 24 is installed some (more than 4) cylindrical tube 12, and some cylindrical tubes 12 horizontally are combined as a unit successively.
T-shaped folder 1 is housed on vapor pipe 2, and T-shaped folder 1 opens and closes with the need.Sample feeding pipe 6 and cylindrical tube 12 junctions install threeway 10, one ends additional and connect nutrient solutions and be in charge of, and an end connects hair weeds cells kind liquid injection port, and the other end connecting cylinder shape cylindrical shell 12 is equipped with valve 7 in threeway 10.
Inlet pipe (comprise inlet manifold 23 and air inlet are in charge of 22) is communicated with an end of cylindrical tube 12 non-return valve 21 is installed, and prevents that the nutrient solutions in cylindrical tube 12 from blowing back into inlet pipe.Air in inlet pipe is provided by the logical oxygen machine of Scroll-tupe, installs air filter additional between the logical oxygen machine of Scroll-tupe and inlet pipe.Liquid discharge pipe and cylindrical tube 12 junctions install threeway 18, one end connected drainage house steward 19, one ends additional and connect thief holes 17, and the other end connecting cylinder shape cylindrical shell 12 is equipped with valve 16 in threeway 19.
The method of the invention also is applicable to the cultivation of little algaes such as spirulina, chlorella, salt algae, also is applicable to the New Energy Industries such as biofuel simultaneously.
Above-described embodiment does not limit the present invention in any form, and all employings are equal to replaces or technical scheme that the mode of equivalent transformation obtains, all drops on protection scope of the present invention.
Claims (5)
1. the method that hair weeds cells is cultivated in batch production, is characterized in that, comprises the steps:
A makes the culture apparatus of delivering vegetables: culture apparatus is made of sample adding device, real-time monitoring device, air inlet and gas barrier and pumping equipment;
B prepares nutrient solution: prepare the nutrient solution of delivering vegetables, by liquid distribution pipe, the nutrient solution for preparing is transported in culture apparatus;
C culture condition: add original hair weeds cells kind in culture apparatus, make nutrient solution reach following index to the logical oxygen of nutrient solution, adjustment nutrient solution environment, i.e. 24 ~ 28 ℃ of temperature, illumination 3200 ~ 4000lux, light dark period 12h:12h, pH value 8.0 ~ 9.5; Aerated culture 10 ~ 15 days, during in regularly take a sample by stopple coupon and detect its light absorption value;
D gathers: when light absorption value reaches OD
750nmBe 0.6 o'clock, gather.
2. the method for hair weeds cells is cultivated in a kind of batch production according to claim 1, it is characterized in that:
In step b, the composition of nutrient solution and part by weight are: anhydrous CaCl
226g, MgSO
47H
2O 75g, NaSiO
39H
2O 58g, K
2HPO
43H
2O 67g, NaNO
31.5kg, 1 ton of micro-10g, membrane filtration pure water.
3. the method for a kind of batch production cultivation hair weeds cells according to claim 2, is characterized in that: in step c, lead to the oxygen pump by Scroll-tupe and realize logical oxygen.
4. the method for according to claim 2 or 3 described a kind of batch production cultivation hair weeds cells, is characterized in that: in step c, when gathering, 350 order filtering nets are tiled on the plastic wire bed plastic wire bed below placement liquid collector; Open draining valve, the hair weeds cells nutrient solution is slowly flowed into filtering net filter, valve-off after the hair weeds cells nutrient solution of 1/3 volume in the culture apparatus of at every turn gathering injects the nutrient solution of equal volume simultaneously by liquid distribution pipe.
5. the method for hair weeds cells is cultivated in a kind of batch production according to claim 4, it is characterized in that: after the moisture of hair weeds cells nutrient solution is filtered dry, thoroughly clean hair weeds cells with deionized water, hair weeds cells after flushing acidity instrumentation pH value, show that neutrality is qualified, hair weeds cells after rinsing is well changed in pallet and puts into-20 ℃ of refrigerator freezings, treat that the freezing vacuum freeze drier that adopts fully carries out drying.
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| CN201310083166XA CN103173389A (en) | 2013-03-15 | 2013-03-15 | Method for culturing hair weed cells industrially |
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| CN201310083166XA CN103173389A (en) | 2013-03-15 | 2013-03-15 | Method for culturing hair weed cells industrially |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI802999B (en) * | 2021-09-23 | 2023-05-21 | 鴻海精密工業股份有限公司 | Method and system for cell harvesting suggestion |
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| CN86102421A (en) * | 1986-04-10 | 1987-10-21 | 余兆丰 | A kind of liquid culture method of edible fungi |
| CN1392245A (en) * | 2002-07-26 | 2003-01-22 | 中国科学院水生生物研究所 | Method for cultivating hair weeds cell |
| CN1530439A (en) * | 2003-03-14 | 2004-09-22 | 天津科技大学 | Cell culture of hair vegetable and its culture product |
| CN101845395A (en) * | 2010-05-11 | 2010-09-29 | 天津科技大学 | Method for efficiently producing long thread moss cells and extracellular polysaccharide thereof in two stages |
-
2013
- 2013-03-15 CN CN201310083166XA patent/CN103173389A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN86102421A (en) * | 1986-04-10 | 1987-10-21 | 余兆丰 | A kind of liquid culture method of edible fungi |
| CN1392245A (en) * | 2002-07-26 | 2003-01-22 | 中国科学院水生生物研究所 | Method for cultivating hair weeds cell |
| CN1530439A (en) * | 2003-03-14 | 2004-09-22 | 天津科技大学 | Cell culture of hair vegetable and its culture product |
| CN101845395A (en) * | 2010-05-11 | 2010-09-29 | 天津科技大学 | Method for efficiently producing long thread moss cells and extracellular polysaccharide thereof in two stages |
Non-Patent Citations (2)
| Title |
|---|
| 刘明志: "发菜人工培养条件探索", 《湖南农业科学》, no. 1, 31 January 2005 (2005-01-31), pages 36 - 39 * |
| 李运广,等: "发菜(N os toc f lagellif orm e) 培养条件的研究", 《武汉植物学研究》, vol. 21, no. 5, 31 December 2003 (2003-12-31), pages 411 - 414 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI802999B (en) * | 2021-09-23 | 2023-05-21 | 鴻海精密工業股份有限公司 | Method and system for cell harvesting suggestion |
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Application publication date: 20130626 |