CN1699548A - Method for improving algae biomass and yield of biologicalactivity product thereof - Google Patents
Method for improving algae biomass and yield of biologicalactivity product thereof Download PDFInfo
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Abstract
The invention relates to a method for improving algae biomass and yield of biologicalactivity product, wherein homogeneous design chart experiment scheme is employed, and DPS software is used for regression analysis and the establishment of the related models, the current flow, processing time and dosage of impulse time during micro-algae ultrasonic treatment are determined for different time period of the micro-algae growth process, the method comprses culturing the micro-algae culture liqid in 250 ml triangular bottle, whihin 1-6 days during the culture cycle, carrying out ultrasonic treatment, which can substancially increase the content of biomass, polysaccharides, phycobili protein and unsaturated fatty acid.
Description
Technical field
The present invention relates to adopt ultrasonication to improve the method for micro algae biomass and physiological active products output thereof.
Background technology
The interaction mechanism of ultrasonic wave in the material medium can be divided into heat effect, cavatition and mechanical mass transfer effect, and the power of its effect is relevant with frequency of ultrasonic and intensity.Low-intensity ultrasonic relies on mechanical vibration and stable cavitation effect to make mass transfer frictional belt attenuate, and the motion of solute particle is quickened, and improves biological respinse speed; Suitable low-intensity ultrasonic acts on rotation and the vortex motion that animal and plant cells can produce miniflow, intracellular organic matter in the born of the same parents, improves cytolemma and cell walls penetrance, and these effects can improve the metabolic function of cell.Low frequency, lower powered ultrasonication can cause microtrauma at cell surface, make the cell walls partial fracture, thereby change the permeability of cytoplasmic membrane, make intracellular organic matter release or make the outer material of born of the same parents enter (ChistiY etc., Trends Biotechnol, 2003 in the cell; Wang Wu etc., Wuxi Light Industry Univ.'s journal, 2002).Produce the cereuisiae fermentum of ATP with ultrasonication after, the permeability of cytolemma is handled De Genggao (Duan Xuehui etc., East China University of Science's journal, 2000) than additive method.In the waste paper fermentative production alcohol process, can make alcohol output improve nearly 20% with supersound process.Penicillin fermentation process is through ultrasonication, and penicillin yield can improve (Wang Wu etc., Wuxi Light Industry Univ.'s journal, 2002) more than 10%.Ultrasonic wave is used to improve anaerobic sludge digestion treatment effect (Hogan F etc., Water Sci Technol, 2004), with oxygen demand (the Gronroos A etc. of soluble compound in the digestion of the auxiliary method raising of ultrasonic wave sewage sludge, Ultrason Sonochem, 2005).Ultrasonic intensity and reactor scale are to the enzyme glycolysis effect kinetics of various waste paper in the continuous illumination stirred pot exert an influence (Li C etc., Ultrason Sonochem, 2005).It is rare that ultrasonic wave is used for the report of metabolic regulation aspect of little algae meta-bolites, and it is remarkable to the biological effect of Zhanjiang Isochrysis galbana, makes algae growth rate constant, index of unsaturated fatty acid improve (Li Wenquan etc., ocean journal, 2002).Be used to control blue-green alga bloom (Lee TJ etc., Water Sci Technol, 200 of eutrophication lake water; Ahn CY etc., Environ Sci Technol, 2003), be used to divide spirulina cells and carry out extracting fast and optionally on a large scale Phycocyanins, C-(Furuki T etc., J Appl Phycol, 2003), biopurification water quality (Mason TJ etc., Ultrason Sonochem, 2003), gather in the crops little algae (BosmaR etc., J Appl Phycol, 2003).High frequency (1.7MHz), low strength (0.6Wcm
-2) ultrasonic energy effectively suppress the fast breeding of blue-green algae cell, the cell growth rate of crossing through ultrasound wave irradiation is reduced to 38.9% (Tang JW etc., J Appl Phycol, 2003) of control group.Utilize ultrasonic energy to promote the microorganism cells growth, strengthen the biocatalysis ability, developed or conceptual ultrasonic wave bio-reactor emerge (Chisti Y etc., Trends Biotechnol, 2003) arranged.
Porphyridium cruentum (Porphyridium cruentum) belongs to the single-cell algae of rhodophyta, is rich in multiple biologically active substance, and its protein content is 32.1%, and aminoacids content is 29.54%; Carbohydrate 39.4%, total dietary fiber 21.7%; Ash 17.6%, moisture 7.8%, every kilogram of dry algae powder calcium, magnesium, potassium and zinc content are respectively 20000mg, 4100mg, 9500mg and 92mg; Every kilogram of dry algae powder β-Hu Luobusu 240mg, vitamin B6 18mg; Toxic heavy metal lead, arsenic and mercury content comply with relevant regulations (Huang Jian etc., food and fermentation industries, 2005).In recent years studies show that Porphyridium cruentum and exocellular polysaccharide thereof have (1) antivirus action; (2) transfer blood fat (Chen Bilian etc. use and the environmental organism journal 2004); (3) anti-inflammatory action or antitumor cell effect; (5) radiation resistance; (6) hypoglycemic activity (Liu Li equality, Chinese marine drug, 2005).Because the research of Porphyridium cruentum and exocellular polysaccharide pharmacodynamics thereof makes great progress, therefore improve Porphyridium cruentum biomass and physiological active products output, reducing production costs becomes the focus of current research and difficult point.
Rhodella reticulata (Rhodella reticulata) is the single-cell algae that belongs to rhodophyta with Porphyridium cruentum together, and the cell exoperidium one deck cement sheath, is grown into by cell to produce the outer water-soluble mucopolysaccharide of born of the same parents after stationary phase and form.Rhodella reticulata contains natural products such as abundant phycobiliprotein, exocellular polysaccharide and unsaturated fatty acids.Especially the physico-chemical property of exocellular polysaccharide and very similar (the Geresh S etc. of Porphyridium cruentum, J Biochem Biophys Methods, 2002), therefore, the functions such as antiviral, hypoglycemic and reducing blood-fat that may have similar Porphyridium cruentum and exocellular polysaccharide thereof, external many scholars have carried out studying (Mihova SG etc., J Biotechnol around aspects such as the exocellular polysaccharide of Rhodella reticulata, phycobiliprotein, photosynthetical systems, 1996), obtained good progress.
Nannochloropsis oceanica (Nannochloropsis sp) is the unicellular little algae in a kind of ocean of Chrysophyta, have easy cultivation, rapid, the nutritious characteristics of breeding, in aquaculture, use comparatively extensive, (written quiet etc. mud blood clam, crab (Shen and fixed etc., Shanghai Aquatic Products Univ. 9CN)'s journal, 1999), shrimp in recent years, water conservancy related fisheries, 2002) etc. grow seedlings and wheel animalcule (Liu Xinfu etc., ocean science, 2002; Lubzens E etc., Aquaculture, 1995; Chebil L etc., Aquacultural Eng, 1998) use preferable good bait in cultivating.The algae powder contains 37.6% can utilize carbohydrate, 28.8% crude protein and 18.4% total fat, also contain Ca, K, Na, Mg, Zn, Fe, Mn, Cu, Ni, fatty acid content: 5.0%16: 0, ω 7,3.8%18 in 4.7%16: 1: 1 ω 9, ω 6 in 0.4%18: 2,0.7%20: 4 ω 6 and 2.2%20: 5 ω 3 (Rebolloso-Fuentes MM etc., AgricFood Chem, 2001).In the substratum that is rich in nitrogen, phosphorus, this algae EPA content can reach 35% (Wei Dong etc., ocean science, 2000) of total fatty acids.At present, the world market upper density is 12 * 10
976 Euros/L of cells/mL algae liquid price (http://www.bluebiotech.de).Therefore, how to improve Nannochloropsis oceanica culture density and fatty acid content, become one of focus of phycology research in the world.
Summary of the invention
Purpose of the present invention is exactly with reference to " 7 groups of 3 factors, the 7 levels " experimental program in the uniform designs table, by homogeneous design optimization, utilization DPS software carries out regression analysis and sets up corresponding model the homogeneous design result, the dosage of " 3 factors, 7 levels " in electric current, treatment time and burst length during the little algae of ultrasonication determining to adopt in the different time period of micro algae growth process realizes improving the purpose of micro algae biomass and physiological active products amount (exocellular polysaccharide, phycobiliprotein or unsaturated fatty acids).
The value of 3 factors, 7 levels of the present invention is respectively electric current: 21%～39%, and treatment time: 5s～245s, pulse: 0.5s～9.5s.
The time period that ultrasonic wave is handled in the micro algae growth process of the present invention is: handle every day 1 time, every day handle 2 times, handle and handled 1 time or the 6th day in 1 time, the 4th day and handle 1 time every other day.
Common vetch algae of the present invention is meant the Nannochloropsis oceanica of Porphyridium cruentum, Rhodella reticulata and the Chrysophyta of the unicellular monoid of rhodophyta.
As follows for realizing the technical scheme that purpose of the present invention adopts:
Micro algae culturing liquid is loaded in the 250mL triangular flask cultivates, in 1-6 in culture cycle days sometime the section, with electric current is that a certain factor value of 21-39%, pulse 5.0-9.5s and treatment time 45-205s carries out ultrasonication, and the content of its biomass, exocellular polysaccharide, phycobiliprotein, unsaturated fatty acids is greatly improved respectively.
1, when growing biomass to improve with the ultrasonication micro algae culturing liquid, micro algae culturing liquid is loaded in the 250mL triangular flask in first day that cultivates, and adopting dosage is that electric current 27-39%, pulse 6.5-9.5s and treatment time 125-165s handled 1 time micro algae culturing liquid on the 4th day.After culture cycle finished, the biomass during with results was compared with undressed little algae, has improved 13-31.5%.
2, when using the ultrasonication micro algae culturing liquid with raising exocellular polysaccharide content, micro algae culturing liquid is loaded in the 250mL triangular flask cultivates, adopting dosage is that electric current 21-30%, pulse 5.0-9.5s and treatment time 125-205s handle 1 time or the 6th day every other day to micro algae culturing liquid and handle 1 time.After culture cycle finished, the biomass during with results was compared with undressed little algae, and exocellular polysaccharide has improved 38-46.5%.
3, when using the ultrasonication micro algae culturing liquid with raising phycobiliprotein content, micro algae culturing liquid is loaded in the 250mL triangular flask cultivates, to adopt dosage be electric current 21-36%, pulse 5.0-8.0s and treatment time 45-165s to micro algae culturing liquid is cultured to the 4th day and handles 1 time or the 6th day and handle 1 time.After culture cycle finished, the biomass during with results was compared with undressed little algae, and phycobiliprotein has improved 13-18.5%.
4, when using the ultrasonication micro algae culturing liquid with the raising unsaturated fatty acids, micro algae culturing liquid is loaded in the 250mL triangular flask cultivates, adopting dosage is that electric current 21-36%, pulse 5.0-6.5s and treatment time 85-125s handle 1 time the 1st day micro algae culturing liquid.After culture cycle finished, the biomass during with results was compared with undressed little algae, and unsaturated fatty acids has improved 13-18.5%.
Ultrasonication treatment process of the present invention, have that equipment is simple, easy to implement the method, operational safety, can make the physiological active products output such as biomass, exocellular polysaccharide, phycobiliprotein, unsaturated fatty acids of little algae improve 10%-60%, bigger promotional value is arranged.
Specific embodiments
The present invention is described further below in conjunction with concrete embodiment.
Embodiment 1
Porphyridium cruentum (Porphyridium cruentum) nutrient solution is loaded in the 250mL triangular flask and cultivates, in first day that cultivates, according to dosage is electric current 39%, pulse 9.5s and treatment time 125s factor index, with ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid the 4th day is handled 1 time.After culture cycle finished, the biomass during with results was compared with undressed Porphyridium cruentum, has improved 25.5%.Simultaneously, the content of exocellular polysaccharide, phycobiliprotein, unsaturated fatty acids is also corresponding increases.
Embodiment 2
Rhodella reticulata (Rhodella reticulata) nutrient solution is loaded in the 250mL triangular flask and cultivates, in first day that cultivates, according to dosage is electric current 27%, pulse 8.0s and treatment time 165s factor index, with ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid the 4th day is handled 1 time.After culture cycle finished, the biomass during with results was compared with undressed Rhodella reticulata, has improved 28.5%.Simultaneously, the content of exocellular polysaccharide, phycobiliprotein, unsaturated fatty acids is also corresponding increases.
Embodiment 3
Nannochloropsis oceanica (Nannochloropsis sp) nutrient solution is loaded in the 250mL triangular flask and cultivates, in first day that cultivates, according to dosage is electric current 27%, pulse 6.5s and treatment time 165s factor index, ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid is handled 1 time every other day.After culture cycle finished, the biomass during with results was compared with undressed Nannochloropsis oceanica, and biomass has improved 32.5%.Simultaneously, the content of unsaturated fatty acids and phycobiliprotein is also corresponding increases.
Embodiment 4
Porphyridium cruentum (Porphyridium cruentum) nutrient solution is loaded in the 250mL triangular flask and cultivates, according to dosage is electric current 24%, pulse 9.5s and treatment time 205s factor index, with ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid is handled 1 time every other day.After culture cycle finished, the exocellular polysaccharide content during with results was compared with undressed Porphyridium cruentum, and exocellular polysaccharide has improved 45.5%.Simultaneously, the content of biomass, phycobiliprotein, unsaturated fatty acids is also corresponding increases.
Embodiment 5
Rhodella reticulata (Rhodella reticulata) nutrient solution is loaded in the 250mL triangular flask and cultivates, according to dosage is electric current 30%, pulse 5.5s and treatment time 135s factor index, with ultrasonoscope (VC750, Sonicsand materials, Inc, USA) adjust after, the micro algae culturing liquid that is cultured to the 6th day is handled 1 time.After culture cycle finished, the exocellular polysaccharide content during with results was compared with undressed Rhodella reticulata, and exocellular polysaccharide has improved 42.5%.Simultaneously, the content of biomass, phycobiliprotein, unsaturated fatty acids is also corresponding increases.
Embodiment 6
Porphyridium cruentum (Porphyridium cruentum) nutrient solution is loaded in the 250mL triangular flask and cultivates, according to dosage is electric current 33%, pulse 6.5s and treatment time 45s factor index, with ultrasonoscope (VC750, Sonicsand materials, Inc, USA) adjust after, the micro algae culturing liquid that is cultured to the 6th day is handled 1 time.After culture cycle finished, the phycobiliprotein content during with results was compared with undressed Porphyridium cruentum, and phycobiliprotein has improved 18.5%.Simultaneously, the content of biomass, exocellular polysaccharide, unsaturated fatty acids is also corresponding increases.
Embodiment 7
Rhodella reticulata (Rhodella reticulata) nutrient solution is loaded in the 250mL triangular flask and cultivates, according to dosage is electric current 21%, pulse 7.5s and treatment time 165s factor index, with ultrasonoscope (VC750, Sonics andmaterials, Inc, USA) adjust after, handle 1 time being cultured to the 4th day micro algae culturing liquid; After culture cycle finished, the phycobiliprotein content during with results was compared with undressed Rhodella reticulata, and phycobiliprotein has improved 14.5%.Simultaneously, the content of biomass, exocellular polysaccharide, unsaturated fatty acids is also corresponding increases.
Embodiment 8
Porphyridium cruentum (Porphyridium cruentum) nutrient solution is loaded in the 250mL triangular flask and cultivates, in first day that cultivates, according to dosage is electric current 30%, pulse 6.5s and treatment time 85s factor index, with ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid is handled 1 time.After culture cycle finished, the unsaturated fatty acid content during with results was compared with undressed Porphyridium cruentum, and unsaturated fatty acids has improved 13.5%.Simultaneously, the content of biomass, exocellular polysaccharide, phycobiliprotein is also corresponding increases.
Embodiment 9
Rhodella reticulata (Rhodella reticulata) nutrient solution is loaded in the 250mL triangular flask and cultivates, in first day that cultivates, according to dosage is electric current 33%, pulse 5.0s and treatment time 100s factor index, with ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid is handled 1 time.After culture cycle finished, the unsaturated fatty acid content during with results was compared with undressed Rhodella reticulata, and unsaturated fatty acids has improved 18.5%.Simultaneously, the content of biomass, exocellular polysaccharide, phycobiliprotein is also corresponding increases.
Embodiment 10
Nannochloropsis oceanica (Nannochloropsis sp) nutrient solution is loaded in the 250mL triangular flask and cultivates, in first day that cultivates, according to dosage is electric current 21%, pulse 6.5s and treatment time 125s factor index, with ultrasonoscope (VC750, Sonics and materials, Inc, USA) adjust after, micro algae culturing liquid is handled 1 time.After culture cycle finished, unsaturated fatty acid content was compared with undressed Nannochloropsis oceanica in the Nannochloropsis oceanica during with results, and unsaturated fatty acids has improved 18.5%.Simultaneously, biomass and phycobiliprotein are also corresponding increases.
Claims (6)
1, a kind of method that adopts the little algae of ultrasonication, it is characterized in that in 1-6 days in little algae culture cycle, with electric current is that the factor value of 21-39%, pulse 5.0-9.5s and treatment time 45-205s carries out ultrasonication, to improve the content of its biomass, exocellular polysaccharide, phycobiliprotein, unsaturated fatty acids.
2, the method for the little algae of ultrasonication according to claim 1 is characterized in that described common vetch algae is meant the Nannochloropsis oceanica of the Porphyridium cruentum of the unicellular monoid of rhodophyta, Rhodella reticulata and Chrysophyta.
3, the method for the little algae of ultrasonication according to claim 1, when it is characterized in that improving the growth biomass, the 4th day of cultivating of Yu Weizao, adopting dosage is that electric current 27-39%, pulse 6.5-9.5s and treatment time 125-165s handle 1 time micro algae culturing liquid.
4, the method for the little algae of ultrasonication according to claim 1, when it is characterized in that improving exocellular polysaccharide content, adopting dosage is that electric current 21-30%, pulse 5.0-9.5s and treatment time 125-205s handle 1 time or the 6th day every other day to micro algae culturing liquid and handle 1 time.
5, the method for the little algae of ultrasonication according to claim 1, when it is characterized in that improving phycobiliprotein content, to adopt dosage be electric current 21-36%, pulse 5.0-8.0s and treatment time 45-165s to micro algae culturing liquid is cultured to the 4th day and handles 1 time or the 6th day and handle 1 time.
6, the method for the little algae of ultrasonication according to claim 1, when it is characterized in that improving unsaturated fatty acid content, adopting dosage is that electric current 21-36%, pulse 5.0-6.5s and treatment time 85-125s handle 1 time the 1st day micro algae culturing liquid.
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| CN104894102A (en) * | 2015-05-14 | 2015-09-09 | 邹宁 | Ultrasonic reinforcement culture method of microalgae |
| CN106801072A (en) * | 2017-01-20 | 2017-06-06 | 哈尔滨工业大学 | The method that ultrasound promotes microalgae grease synthesis |
| CN112608956A (en) * | 2020-12-10 | 2021-04-06 | 浙江大学 | Method for improving yield of extracellular polysaccharide of bacteria by using ultrasonic waves |
| CN114891777A (en) * | 2022-05-19 | 2022-08-12 | 江苏大学 | Device and method for improving microalgae extracellular polysaccharide secretion by high-voltage electric field discharge stimulation |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102559651A (en) * | 2011-12-14 | 2012-07-11 | 浙江科技学院 | Method for improving propagation rate of microalgae by using acoustic frequency technology |
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2005
- 2005-06-28 CN CNB2005100808599A patent/CN100344746C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104894102A (en) * | 2015-05-14 | 2015-09-09 | 邹宁 | Ultrasonic reinforcement culture method of microalgae |
| CN106801072A (en) * | 2017-01-20 | 2017-06-06 | 哈尔滨工业大学 | The method that ultrasound promotes microalgae grease synthesis |
| CN112608956A (en) * | 2020-12-10 | 2021-04-06 | 浙江大学 | Method for improving yield of extracellular polysaccharide of bacteria by using ultrasonic waves |
| CN112608956B (en) * | 2020-12-10 | 2024-03-15 | 浙江大学 | Method for improving bacterial extracellular polysaccharide yield by utilizing ultrasonic waves |
| CN114891777A (en) * | 2022-05-19 | 2022-08-12 | 江苏大学 | Device and method for improving microalgae extracellular polysaccharide secretion by high-voltage electric field discharge stimulation |
| CN114891777B (en) * | 2022-05-19 | 2024-05-14 | 江苏大学 | Device and method for improving secretion of microalgae extracellular polysaccharide by using high-voltage electric field discharge stimulation |
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