CN109355194A - A method of improving chlorella chlorophyll content - Google Patents
A method of improving chlorella chlorophyll content Download PDFInfo
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- CN109355194A CN109355194A CN201811416377.XA CN201811416377A CN109355194A CN 109355194 A CN109355194 A CN 109355194A CN 201811416377 A CN201811416377 A CN 201811416377A CN 109355194 A CN109355194 A CN 109355194A
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Abstract
The invention discloses a kind of methods for improving chlorella chlorophyll content, and the method includes the steps of: (1) chlorella algae being placed in BG11 culture solution and carry out enrichment culture, when its cell density reaches 108When a cell/mL order of magnitude, chlorella seed liquor is obtained；(2) regulation and control substance is added in BG11 culture solution, improvement BG11 culture solution is made；(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement BG11 culture solution of step (2) production by 1% volume ratio, obtains chlorella culture；(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 6~8 days.Method provided by the present invention is remarkably improved the chlorophyll content of chlorella, improves chlorella quality, improves the comprehensive utilization value of chlorella.Moreover, this method has the characteristics that easy to operate, practical, easy realization continuous production.
Description
Technical field
The invention belongs to technical field of microalga biology, in particular to a kind of method for improving chlorella chlorophyll content.
Background technique
In structure, chlorophyll and the intracorporal ferroheme structure of people are extremely similar, therefore are easily absorbed by the body, and have tax to live
The effect of cell has extensive purposes in medicine, food and daily chemical industry.In field of food, chlorophyll is as additive
It has been applied to the processing of the food such as chewing gum, hard candy, fruit juice, agar, cake；In field of medicaments, chlorophyll, which has, to be changed
Kind constipation reduces cholesterol, anti-aging, anti-inflammatory, promotes the functions such as toxin discharge, anti-anemia action and protection liver, to skin group
The regeneration knitted also has certain effect, can be used for treating scald, chronic ulcer etc..The derivative application of chlorophyll is also relatively broad,
If sodium copper chlorophyllin can be used as coloring agent, deodorant in food additives and daily use chemicals process, as medical material,
Sodium copper chlorophyllin can be used as the drug of liver-protecting and stomach-protecting and anti-anemia action, for treating catarrhal jaundice, duodenal ulcer, pancreas
The diseases such as scorching and leukaemia.
Chlorophyll is the primary pigments that chlorella carries out photosynthetic important substance and chlorella.According to pertinent literature
Report confirms: the solar energy that can be utilized in water is few many compared to land, to complete identical energy conversion, plant in water
The content that object participates in photosynthetic chlorophyll necessarily then is greater than terrestrial plant.The chlorophyll content of terrestrial plant is about in nature
It is 0.3~0.5%, and the content of chlorella Determination of Chlorophyll is about 0.5~2%.In addition, chlorella, which has, grows rapid, Yi Pei
It supports, limited by place and season and the features such as production scale is controllable, be not only advantageous to save the cost, it is also ensured that resource
Supply.Therefore, chlorella is considered as the important source material for developing chlorophyll and Related product.
In recent years, with the gradually development of science and technology, the increasingly raising of people's health consciousness, to the leaf from chlorella
The understanding of green element and developmental research are also more deep, and application range is also increasingly wider, therefore produces chlorophyll using chlorella
With biggish market value and application potential.But the type and content of chlorella chlorophyll can and environmental condition different because of algae
Variation and it is different, the content for how significantly improving chlorella chlorophyll has become current chlorella field of biotechnology
Important subject.
Summary of the invention
In order to solve the above technical problems, specifically being adopted the present invention provides a kind of method for improving chlorella chlorophyll content
Technical solution is as follows:
A method of improving chlorella chlorophyll content comprising the steps of:
(1) chlorella algae is placed in BG11 culture solution and carries out enrichment culture, when its cell density reaches 108It is a thin
When born of the same parents/mL order of magnitude, chlorella seed liquor is obtained；
(2) regulation and control substance is added in BG11 culture solution, improvement BG11 culture solution is made；
(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement that step (2) make by 1% volume ratio
In BG11 culture solution, chlorella culture is obtained；
(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 6~8 days.
Preferably, BG11 culture solution described in step (1), ingredient are as follows: NaNO3(1.5g/L), K2HPO4(40mg/
L), MgSO4·7H2O (70mg/L), CaCl2·2H2O (40mg/L), citric acid (6mg/L), ferric citrate (6mg/L),
EDTANa2(1mg/L), Na2CO3(20mg/L), H3BO3(2.86mg/L), MnCl2·4H2O (1.86mg/L), ZnSO4·7H2O
(0.22mg/L), Na2MoO4·2H2O (0.39mg/L), CuSO4·5H2O (0.08mg/L), Co (NO3)2·6H2O(0.05mg/
L), pH=7.10.
Preferably, regulation and control substance described in step (2) is sodium selenite (0.25~0.75g/L), Indomethacin (0.25
~0.75g/L), inositol (0.1~0.6g/L) or FeCl3(0.05~0.25g/L).
Preferably, the cell density of chlorella culture described in step (3) is 106A cell/mL the order of magnitude.
Preferably, chlorella condition of culture described in step (4) are as follows: 25 ± 1 DEG C of temperature, 60 ± 5 μm of ol of illumination
photons/m2/ s, photoperiod are the dark 12h of light 12h/, shaking speed: 140 ± 10 revs/min.
The beneficial effects of the present invention are:
A kind of method improving chlorella chlorophyll content provided by the present invention, not only can induce chlorella chlorophyll
Synthesis, hence it is evident that improve the chlorophyll content of chlorella, can also improve chlorella quality, improve the comprehensive utilization value of chlorella.
In addition, this method has the characteristics that easy to operate, practical, easy realization continuous production, it is very beneficial for promoting on a large scale
And application.
Specific embodiment
According to following embodiments, the present invention may be better understood.However, as it will be easily appreciated by one skilled in the art that real
It applies content described in example and is merely to illustrate the present invention, without sheet described in detail in claims should will not be limited
Invention.
Embodiment 1:
A method of chlorella chlorophyll content being improved, the present embodiment is the present invention further explains according to the following steps
It states:
(1) chlorella algae is placed in BG11 culture solution first and carries out enrichment culture, when the cell density of chlorella reaches
To 108When a cell/mL order of magnitude, chlorella seed liquor is obtained.Wherein, the ingredient of BG11 culture solution are as follows: NaNO3(1.5g/L),
K2HPO4(40mg/L), MgSO4·7H2O (70mg/L), CaCl2·2H2O (40mg/L), citric acid (6mg/L), ferric citrate
(6mg/L), EDTANa2(1mg/L), Na2CO3(20mg/L), H3BO3(2.86mg/L), MnCl2·4H2O (1.86mg/L),
ZnSO4·7H2O (0.22mg/L), Na2MoO4·2H2O (0.39mg/L), CuSO4·5H2O (0.08mg/L), Co (NO3)2·
6H2O (0.05mg/L), pH=7.10.
(2) sodium selenite of 0.25~0.75g/L is added in BG11 culture solution, improvement BG11 culture solution is made.
(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement that step (2) make by 1% volume ratio
In BG11 culture solution, obtaining cell density is 106A cell/mL order of magnitude chlorella culture.
(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 7 days, condition of culture are as follows: temperature 25
± 1 DEG C, 60 ± 5 μm of ol photons/m of illumination2/ s, the photoperiod control be the dark 12h of light 12h/, shaking speed: 140 ± 10 turns/
Minute.
After culture in 7 days, chlorella is in the improvement BG11 culture solution of the sodium selenite Han 0.25,0.5 and 0.75g/L
Chlorophyll content significantly improves, and chlorophyll content is respectively 2.89 μ g/107A cell, 3.10 μ g/107A cell and 2.51 μ
g/107A cell is control group (1.04 μ g/10 respectively7A cell) 2.78 times, 2.98 times and 2.41 times.
Embodiment 2:
A method of chlorella chlorophyll content being improved, the present embodiment is the present invention further explains according to the following steps
It states:
(1) chlorella algae is placed in BG11 culture solution first and carries out enrichment culture, when the cell density of chlorella reaches
To 108When a cell/mL order of magnitude, chlorella seed liquor is obtained.Wherein, the ingredient of BG11 culture solution are as follows: NaNO3(1.5g/L),
K2HPO4(40mg/L), MgSO4·7H2O (70mg/L), CaCl2·2H2O (40mg/L), citric acid (6mg/L), ferric citrate
(6mg/L), EDTANa2(1mg/L), Na2CO3(20mg/L), H3BO3(2.86mg/L), MnCl2·4H2O (1.86mg/L),
ZnSO4·7H2O (0.22mg/L), Na2MoO4·2H2O (0.39mg/L), CuSO4·5H2O (0.08mg/L), Co (NO3)2·
6H2O (0.05mg/L), pH=7.10.
(2) Indomethacin of 0.25~0.75g/L is added in BG11 culture solution, improvement BG11 culture solution is made.
(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement that step (2) make by 1% volume ratio
In BG11 culture solution, obtaining cell density is 106A cell/mL order of magnitude chlorella culture.
(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 7 days, condition of culture are as follows: temperature 25
± 1 DEG C, 60 ± 5 μm of ol photons/m of illumination2/ s, the photoperiod control be the dark 12h of light 12h/, shaking speed: 140 ± 10 turns/
Minute.
After culture in 7 days, chlorella is in the improvement BG11 culture solution of the Indomethacin Han 0.25,0.5 and 0.75g/L
Chlorophyll content significantly improves, and chlorophyll content is respectively 2.15 μ g/107A cell, 2.23 μ g/107A cell and 1.51 μ
g/107A cell is control group (1.04 μ g/10 respectively7A cell) 2.07 times, 2.14 times and 1.45 times.
Embodiment 3:
A method of chlorella chlorophyll content being improved, the present embodiment is the present invention further explains according to the following steps
It states:
(1) chlorella algae is placed in BG11 culture solution first and carries out enrichment culture, when the cell density of chlorella reaches
To 108When a cell/mL order of magnitude, chlorella seed liquor is obtained.Wherein, the ingredient of BG11 culture solution are as follows: NaNO3(1.5g/L),
K2HPO4(40mg/L), MgSO4·7H2O (70mg/L), CaCl2·2H2O (40mg/L), citric acid (6mg/L), ferric citrate
(6mg/L), EDTANa2(1mg/L), Na2CO3(20mg/L), H3BO3(2.86mg/L), MnCl2·4H2O (1.86mg/L),
ZnSO4·7H2O (0.22mg/L), Na2MoO4·2H2O (0.39mg/L), CuSO4·5H2O (0.08mg/L), Co (NO3)2·
6H2O (0.05mg/L), pH=7.10.
(2) inositol of 0.1~0.6g/L is added in BG11 culture solution, improvement BG11 culture solution is made.
(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement that step (2) make by 1% volume ratio
In BG11 culture solution, obtaining cell density is 106A cell/mL order of magnitude chlorella culture.
(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 8 days, condition of culture are as follows: temperature 25
± 1 DEG C, 60 ± 5 μm of ol photons/m of illumination2/ s, the photoperiod control be the dark 12h of light 12h/, shaking speed: 140 ± 10 turns/
Minute.
After culture in 8 days, leaf of the chlorella in the improvement BG11 culture solution of the inositol Han 0.1,0.2,0.4 and 0.6g/L
Chlorophyll contents significantly improve, and chlorophyll content is respectively 2.08 μ g/107A cell, 2.87 μ g/107A cell, 3.42 μ g/
107A cell and 2.73 μ g/107A cell is control group (1.04 μ g/10 respectively7A cell) 2.00 times, 2.76 times, 3.29
Times and 2.63 times.
Embodiment 4:
A method of chlorella chlorophyll content being improved, the present embodiment is the present invention further explains according to the following steps
It states:
(1) chlorella algae is placed in BG11 culture solution first and carries out enrichment culture, when the cell density of chlorella reaches
To 108When a cell/mL order of magnitude, chlorella seed liquor is obtained.Wherein, the ingredient of BG11 culture solution are as follows: NaNO3(1.5g/L),
K2HPO4(40mg/L), MgSO4·7H2O (70mg/L), CaCl2·2H2O (40mg/L), citric acid (6mg/L), ferric citrate
(6mg/L), EDTANa2(1mg/L), Na2CO3(20mg/L), H3BO3(2.86mg/L), MnCl2·4H2O (1.86mg/L),
ZnSO4·7H2O (0.22mg/L), Na2MoO4·2H2O (0.39mg/L), CuSO4·5H2O (0.08mg/L), Co (NO3)2·
6H2O (0.05mg/L), pH=7.10.
(2) FeCl of 0.05~0.25g/L is added in BG11 culture solution3, improvement BG11 culture solution is made.
(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement that step (2) make by 1% volume ratio
In BG11 culture solution, obtaining cell density is 106A cell/mL order of magnitude chlorella culture.
(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 6 days, condition of culture are as follows: temperature 25
± 1 DEG C, 60 ± 5 μm of ol photons/m of illumination2/ s, the photoperiod control be the dark 12h of light 12h/, shaking speed: 140 ± 10 turns/
Minute.
After culture in 6 days, chlorella is in the FeCl containing 0.05,0.10,0.15,0.20 and 0.25g/L3Improvement BG11
Chlorophyll content in culture solution significantly improves, and chlorophyll content is respectively 1.17 μ g/107A cell, 1.54 μ g/107It is a thin
Born of the same parents, 2.07 μ g/107A cell, 2.43 μ g/107A cell and 2.46 μ g/107A cell is control group (1.04 μ g/10 respectively7
A cell) 1.13 times, 1.48 times, 1.99 times, 2.34 times and 2.37 times.
Comparative example 1:
A method of chlorella chlorophyll content being improved, this comparative example is the present invention further explains according to the following steps
It states:
(1) chlorella algae is placed in BG11 culture solution first and carries out enrichment culture, when the cell density of chlorella reaches
To 108When a cell/mL order of magnitude, chlorella seed liquor is obtained.Wherein, the ingredient of BG11 culture solution are as follows: NaNO3(1.5g/L),
K2HPO4(40mg/L), MgSO4·7H2O (70mg/L), CaCl2·2H2O (40mg/L), citric acid (6mg/L), ferric citrate
(6mg/L), EDTANa2(1mg/L), Na2CO3(20mg/L), H3BO3(2.86mg/L), MnCl2·4H2O (1.86mg/L),
ZnSO4·7H2O (0.22mg/L), Na2MoO4·2H2O (0.39mg/L), CuSO4·5H2O (0.08mg/L), Co (NO3)2·
6H2O (0.05mg/L), pH=7.10.
(2) the chlorella seed liquor that step (1) obtains is inoculated into BG11 culture solution by 1% volume ratio, is obtained
Cell density is 106A cell/mL order of magnitude chlorella culture.
(3) chlorella culture made from step (2) is placed on illumination shaking table and is cultivated 7 days, condition of culture are as follows: temperature 25
± 1 DEG C, 60 ± 5 μm of ol photons/m of illumination2/ s, the photoperiod control be the dark 12h of light 12h/, shaking speed: 140 ± 10 turns/
Minute.
After culture in 7 days, the chlorophyll content of chlorella is 1.04 μ g/107A cell.
Claims (5)
1. a kind of method for improving chlorella chlorophyll content, which is characterized in that comprise the steps of:
(1) chlorella algae is placed in BG11 culture solution and carries out enrichment culture, when its cell density reaches 108A cell/mL number
When magnitude, chlorella seed liquor is obtained；
(2) regulation and control substance is added in BG11 culture solution, improvement BG11 culture solution is made；
(3) the chlorella seed liquor that step (1) obtains is inoculated into the improvement BG11 that step (2) make by 1% volume ratio
In culture solution, chlorella culture is obtained；
(4) chlorella culture made from step (3) is placed on illumination shaking table and is cultivated 6~8 days.
2. a kind of method for improving chlorella chlorophyll content according to claim 1, which is characterized in that in step (1)
The pH=7.10 of the BG11 culture solution, composition are as follows: the NaNO of 1.5g/L3, 40mg/L K2HPO4, 70mg/L MgSO4·
7H2O, the CaCl of 40mg/L2·2H2O, the EDTANa of the citric acid of 6mg/L, the ferric citrate of 6mg/L, 1mg/L2、20mg/L
Na2CO3, 2.86mg/L H3BO3, 1.86mg/L MnCl2·4H2O, the ZnSO of 0.22mg/L4·7H2O, 0.39mg/L
Na2MoO4·2H2O, the CuSO of 0.08mg/L4·5H2O, the Co (NO of 0.05mg/L3)2·6H2O。
3. a kind of method for improving chlorella chlorophyll content according to claim 1, which is characterized in that in step (2)
The regulation and control substance is the sodium selenite of 0.25~0.75g/L, the Indomethacin of 0.25~0.75g/L, 0.1~0.6g/L
The FeCl of inositol or 0.05~0.25g/L3。
4. a kind of method for improving chlorella chlorophyll content according to claim 1, which is characterized in that in step (3)
The cell density of the chlorella culture is 106A cell/mL the order of magnitude.
5. a kind of method for improving chlorella chlorophyll content according to claim 1, which is characterized in that in step (4)
The chlorella condition of culture are as follows: 25 ± 1 DEG C of temperature, 60 ± 5 μm of ol photons/m of illumination2/ s, photoperiod are light 12h/
Dark 12h, shaking speed are 140 ± 10 revs/min.
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| CN103045481A (en) * | 2013-01-08 | 2013-04-17 | 南京师范大学 | Method for promoting growth of Nannochlorisoculata and increasing contents of chlorophyll and mycoprotein of Nannochlorisoculata |
| CN104152357A (en) * | 2014-08-06 | 2014-11-19 | 华南理工大学 | High-density culture method for improving chlorophyll and protein content of chlorella at same time |
| CN105779293A (en) * | 2016-05-18 | 2016-07-20 | 中国计量大学 | Chlorella preservation method |
| CN106701588A (en) * | 2016-11-17 | 2017-05-24 | 中盐工程技术研究院有限公司 | Culture method of selenium-enriched chlorella |
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2018
- 2018-11-26 CN CN201811416377.XA patent/CN109355194A/en active Pending
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|---|---|---|---|---|
| CN103045481A (en) * | 2013-01-08 | 2013-04-17 | 南京师范大学 | Method for promoting growth of Nannochlorisoculata and increasing contents of chlorophyll and mycoprotein of Nannochlorisoculata |
| CN104152357A (en) * | 2014-08-06 | 2014-11-19 | 华南理工大学 | High-density culture method for improving chlorophyll and protein content of chlorella at same time |
| CN105779293A (en) * | 2016-05-18 | 2016-07-20 | 中国计量大学 | Chlorella preservation method |
| CN106701588A (en) * | 2016-11-17 | 2017-05-24 | 中盐工程技术研究院有限公司 | Culture method of selenium-enriched chlorella |
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Application publication date: 20190219 |