JP3138071B2 - Algae growth promoter - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an algae growth promoter, and more particularly, to an algae growth promoter, a pigment content improver and a carbon dioxide fixer containing 5-aminolevulinic acid or a salt thereof as an active ingredient. And a chemical improving agent.

[0002]

BACKGROUND OF THE INVENTION Algae are important not only as foods but also as artificial foods such as animals, fish, shrimp, crab and sericulture as well as important foods. I have. In addition, phycocyanin, carotenoids and other pigment raw materials, organic acid raw materials such as eicosapentaenoic acid, polysaccharide raw materials such as carrageenan, cosmetic cosmetics and hair restorer, are widely used as industrial raw materials, and play an important role for humanity. I have.

[0003] These algae are collected as natural products,
It is common to culture artificially in a pond or the like. Also, research on industrial algae cultivation methods has been partially advanced, for example, JP-A-47-43388, JP-A-48-87077, JP-A-52-34295.
No. 4,972,088, etc., have proposed a medium component, a method of irradiating light, etc., but at present they are not at an industrially sufficient level, and there are only examples of practical use of microalgae such as chlorella.

There are several problems in attempts to artificially cultivate and culture algae. Among them, one of the major problems is that algae have a slower growth rate than microorganisms used for fermentation and the like. . In addition, phycocyanins, pigments derived from algae typified by carotenoids, as well as their use as natural pigments, are also attracting attention for their physiological activities, and are also expected to improve the pigment content in algae. I have. Furthermore, in recent years, an increase in the concentration of carbon dioxide in the atmosphere has promoted global warming, and has become a major social problem.The use of algae has attracted attention as a means of fixing carbon dioxide. The ability to fix carbon dioxide gas was not sufficient for practical use.

[0005] Accordingly, there has been a demand for the development of a substance that more effectively promotes the growth of algae, a substance that improves the pigment content in the algae, and a substance that improves the algae's ability to fix carbon dioxide.

[0006]

Under such circumstances, the present inventors have conducted intensive studies and as a result, have found that 5-aminolevulinic acid or a salt thereof has a strong growth promoting action on algae,
In addition, the inventors have found that the content of the pigment in algae is particularly improved, and the ability to fix carbon dioxide is also improved. Thus, the present invention has been completed.

That is, the present invention provides an algal growth promoting agent containing 5-aminolevulinic acid or a salt thereof at a concentration of 0.03-700 ppm in terms of 5-aminolevulinic acid, and 5-aminolevulinic acid or a salt thereof as an active ingredient. It is intended to provide an algae phycocyanin or carotenoid content improver and an algae carbon dioxide immobilization ability improver.

[0008] It is known that 5-aminolevulinic acid or a salt thereof used as an active ingredient of the algae growth promoter of the present invention is useful as a herbicide and an insecticide. , JP-A-2-138201), it is not known at all that it has a growth promoting effect on algae.

[0009] 5-Aminolevulinic acid or a salt thereof is a known compound and can be produced by any method of chemical synthesis, production by microorganisms, and production by enzymes. When a production method using a microorganism or an enzyme is used, the product can be used as it is without separation and purification as long as it does not contain a substance harmful to algae.

Examples of the salt of 5-aminolevulinic acid include hydrochloride, phosphate, nitrate, sulfate, acetate, propionate, butyrate, valerate, citrate, fumarate and maleate. And metal salts such as sodium, potassium and calcium salts. These salts are used as aqueous solutions at the time of use, and their actions are the same as those of 5-aminolevulinic acid. 5-Aminolevulinic acid and a salt thereof can be used alone or in combination of two or more.

The algae growth promoter of the present invention may contain, as necessary, other drugs, sugars, amino acids, organic acids, alcohols, vitamins, minerals, etc., in addition to 5-aminolevulinic acid or a salt thereof. .

The dosage forms of the algae growth promoter of the present invention include powders, granules, liquids and the like. In order to make these dosage forms, a solvent, a dispersion medium, a bulking agent and the like are used. It can be manufactured according to the method.

[0013] 5-Aminolevulinic acid and its salts are substances that are widely present in nature, are also present in the human body, and have extremely low toxicity. In addition, while being rapidly absorbed and used by algae, such as when culturing algae, it is degraded by microorganisms and the like in the natural world, and therefore has very little persistence and high safety.

The use concentration of the algal growth promoter of the present invention is as follows:
It is 0.03 to 700 ppm in terms of 5-aminolevulinic acid. If the concentration is too low, a sufficient effect cannot be obtained, while if it is too high, the effect is not so much improved and the growth may be suppressed, which is not preferable.

The algae growth promoter of the present invention exhibits a growth promotion effect of about one month under typical conditions after being incorporated into the algae. However, when culturing continuously over a long period of time, when a greater effect is desired, or when culturing in a pond or the like, multiple or continuous additions are effective.

[0016] As the algae become subject to the growth promoter of the algae present invention is not particularly limited, for example Zenokokkasu (Xenococcus), Afanotesu (Aphanothece)
, Rivularia , Cylindrosperum ( Cyl
indrospermum ), blue- green algae such as Spirulina ;
Euglena and other Euglena ; Botrydium
Yellow flagellates such as ( Botrydium ) and Chromulina ; Red algae such as Porphyridium ; Diatoms such as Bacillaria ; Chlamydomonas ( Ch)
lamydomonas), Dunaliella (Dunaliella), Kurorozaruchina (Chlorosarcina), and the like green algae such as Chlorella (Chlorella).

5-Aminolevulinic acid or a salt thereof is also effective as an algae pigment content improver. The use form, addition method, addition concentration, and the like of the algae as a pigment content improver are all the same as those of the algae growth promoter.

Further, 5-aminolevulinic acid or a salt thereof is also effective as an agent for improving the algae's ability to fix carbon dioxide. The use form, addition method, addition concentration, etc. of the algae as a carbon dioxide immobilizing ability improving agent are all the same as those of the algae growth promoter.

[0019]

The algae growth promoter of the present invention can be used simply by adding a very small amount to the culture solution during the cultivation of the algae, exhibits an excellent growth promoting action of the algae, and exhibits photosynthetic activity. To improve the pigment content of algae and the ability to fix carbon dioxide gas.

[0020]

EXAMPLES Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples. Example 1 1 l of SOT medium having the composition shown in Table 1
Sterilized at 1 ° C for 15 minutes. After cooling, Spirulina and cultured in the usual way platensis (spirulina platensis) IAM
A 10% culture of M-135 was aseptically inoculated. In addition, 5
-Aminolevulinic acid (5-ALA) at 0, 100 ppm,
Add 500ppm, 700ppm, stand in a water tank, 30
C., and the cells were cultured by irradiating light to 3000 lux with an incandescent bulb. Table 2 shows OD ₅₆₀ values indicating the culturing time and the amount of cells.

[0021]

TABLE 1 (Composition of SOT _{medium) NaHCO 3 1680mg K 2 HPO 4} 50mg NaNO 3 250mg K 2 SO 4 100mg NaCl 100mg MgSO 4 · 7H 2 O 20mg CaCl 2 · 2H 2 O 4mg FeSO 4 · 7H 2 O 1mg Na _{2 EDTA · 2H 2 O 8mg a} 5 solution * 0.1ml distilled water 99.9ml *) the composition of _{a 5 solution H 3 BO 3 286mg} MnSO 4 · 7H 2 O 250mg ZnSO 4 · 7H 2 O 22.2mg CuSO 4 · 5H 2 O 7.9mg Na ₂ MoO ₄ 2.1mg distilled water 100ml

[0022]

[Table 2]

As is clear from Table 2, it was recognized that the addition of the algae growth promoter of the present invention promoted the growth of algae and increased the production amount.

Example 2 A cultivation was carried out in the same manner as in Example 1 except that the SOT medium used in Example 1 was further supplemented with 1 g of glucose, and the 5-ALA concentration was changed to 0 or 100 ppm. Table 3 shows the results.

[0025]

[Table 3]

As is clear from Table 3, it was found that the use of the algal growth promoter of the present invention promoted the growth of algae.

[0027] Spirulina Example 3 Algae platensis (spirulina plate
nsis ) Using NIES-39, culture scale to 50ml large tube
Using a medium containing 30 ml of the medium, the mixture was stirred with a magnetic stirrer at 200 rpm, and cultured in the same manner as in Example 1 except that the 5-ALA concentration was set to 0 or 100 ppm. Table 4 shows the results.

[0028]

[Table 4]

As is clear from Table 4, it was found that the use of the algal growth promoter of the present invention promoted the growth of algae.

[0030] Spirulina Example 4 algae platensis (spirulina plate
nsis ) Using NIES-39, culture scale to 50ml large tube
Using a medium containing 30 ml, the mixture was stirred with a magnetic stirrer at 200 rpm, and the 5-ALA concentration was reduced to 0, 100, 500
Culture was carried out in the same manner as in Example 2 except that 700 ppm was used. Table 5 shows the results.

[0031]

[Table 5]

As is clear from Table 5, it was found that the use of the algae growth promoter of the present invention promoted the growth of algae.

Example 5 Spirulina cultured in the same manner as in Example 3 was cultured.
After the collection, the dye was quantified according to a conventional method (Algae Research Method, edited by Nishizawa and Chihara, Kyoritsu Shuppan, pp. 474-507). Table 6 shows the results.

[0034]

[Table 6]

As is clear from Table 6, it was found that the addition of 5-ALA increased the pigment content in algae.

Example 6 Spirulina cultured in the same manner as in Example 4 was
After the collection, the dye was quantified in the same manner as in Example 5. Table 7 shows the results.

[0037]

[Table 7]

As is clear from Table 7, it was confirmed that the addition of 5-ALA increased the pigment content in algae.

Example 7 100 ml of MBM medium having the composition shown in Table 8 was placed in a 200 ml Erlenmeyer flask, and sterilized at 121 ° C. for 15 minutes. After cooling, Chlorella vulgaris ( chlorella vul
garis ) 5% of the culture solution of IAM C-2 was aseptically inoculated. To this, 5-aminolevulinic acid (5-ALA) was added to 0,
The solution was added to 100 ppm, placed in a water bath temperature-controlled to 30 ° C., and irradiated with light from an incandescent bulb so that the surface of the flask became 5000 lux. As the ventilation, a mixture of carbon dioxide: air = 1: 1, which was filtered with a sterilizing filter, was bubbled twice a morning and evening at a rate of 1 liter per day. Table 9 shows OD ₅₆₀ values indicating the number of days of culture and the amount of cells.

[0040]

TABLE 8 (Composition of MBM _{medium) KNO 3 5.0g MgSO 4 · 7H} 2 O 2.5g KH 2 PO 4 1.25g K 2 HPO 4 0.1g FeSO 4 · 7H 2 O 0.0028g trace components Arron's A ₅ soln. * 1ml distilled water 999ml *) trace component Arron's a ₅ soln. of the composition _{H 3 BO 3 2.86g MnCl 2 ·} 4H 2 O 1.81g ZnSO 4 · 7H 2 O 0.22g CuSO 4 · 5H 2 O 0.08g Na 2 MoO 4 0.021 g distilled water 1000 ml 1 drop of concentrated sulfuric acid

[0041]

[Table 9]

As is clear from Table 9, it was recognized that the addition of 5-ALA promoted the growth of algae.

Example 8 Culture was carried out in the same manner as in Example 7 except that air was continuously ventilated at a rate of 100 ml / min. Table 10 shows the results.

[0044]

[Table 10]

As is clear from Table 10, it was confirmed that the addition of 5-ALA promoted the growth of algae.

Example 9 In Example 8, after 8 days, the exhaust gas was subjected to an infrared absorption type differential carbon dioxide concentration meter (Shimadzu carbon dioxide concentration difference measuring device, IRA-
102), and as a control, the relative amount of carbon dioxide absorbed and the relative amount of bacterial cells were measured when the concentration of 5-ALA was 0 ppm (5-ALA was not added) as 100%. Table 11 shows the results.

[0047]

[Table 11]

As is evident from Table 11, it was found that the addition of 5-ALA improved the carbon dioxide fixation rate of algae.

Example 10 250 ml of the SOT medium having the composition shown in Example 1 was placed in a 500 ml bottle and sterilized at 121 ° C. for 15 minutes. After cooling, Spirulina platensis ( Spirulina pla
tensis ) 10% culture of IAM M-135, NIES-39 and NIES-46
Was aseptically inoculated. Then, add 5-ALA to 0,5
0, 100, and 500 ppm, and stand in a water tank.
C., and the cells were cultured by irradiating light to 5,000 lux with an incandescent bulb. Tables 12, 13 and 14 show OD ₅₆₀ values indicating the number of days of culture and the amount of cells.

[0050]

[Table 12]

[0051]

[Table 13]

[0052]

[Table 14]

As is clear from Tables 12, 13 and 14, it was confirmed that the addition of the algal growth promoter of the present invention promoted the growth of algae and increased the production amount. Although it was found that there was an effect on any of the strains, there was a difference in the sensitivity, and it showed an effect on IAM M-135 at a lower concentration than the other two strains.

Example 11 Into a medium-containing bottle prepared in the same manner as in Example 10,
The composition shown in 15 B ₆ solution was added in an amount of 0.1% by volume, and sets the magnetic stirrer rotor and dissolved oxygen meter, and sterilized 15 minutes at 121 ° C.. After cooling, Spirulina platensis ( Spirulina platen)
sis) OD ₅₆₀ value after inoculation of NIES-39 was inoculated to a range of from 0.08 to 0.09. Add 5-ALA to 0 and 50 respectively.
Add it to a concentration of 0 ppm, stand in a water tank, control the temperature to 30 ° C, and stir with a magnetic stirrer. Both sides of the bottle were irradiated. Table 16 shows the value of OD ₅₆₀ indicating the amount of bacterial cells with respect to the number of culture days, the amount of phycocyanin measured by the same method as in Example 5, and the photosynthetic activity (oxygen generation rate) measured by a dissolved oxygen meter.

[0055]

[Table 15] B ₆ Composition of the solution _{NH 4 VO 3 229.6mg Cr 2 K} 2 (SO 4) 4 · 24H 2 O 960.2mg NiSO 4 · 6H 2 O 447.8mg Co (NO 3) 2 · 6H 2 O 493.8mg Na ₂ WO ₄・ 2H ₂ O 179.4mg Ti ₂ (SO ₄ ) ₃ 785.7mg 1 / 10N H ₂ SO ₄ 1000ml

[0056]

[Table 16]

As is evident from Table 16, it was found that the addition of 5-ALA promoted proliferation, improved photosynthetic activity, and improved pigment content.

[0058] Except for using the SOT medium without added Example 12 B ₆ solution was prepared as in Example 11. Table 17 shows the results.

[0059]

[Table 17]

As is clear from Table 17, it was found that the addition of 5-ALA promoted proliferation, improved photosynthetic activity, and improved pigment content. Further, effect compared to Table 16 is large, if they do not contain a B ₆ solution, i.e. it has been found that the effect of adding 5-ALA as when growth conditions are poor is large.

Example 13 Cultivation was carried out for 3 days under the same conditions as in Examples 11 and 12, and the dye content was measured in the same manner as in Example 5. The results are shown in Table 18.

[0062]

[Table 18]

[0063] As apparent from Table 18, and the dye content is increased by the addition of 5-ALA, this effect, if not containing B ₆ solution, i.e. be effective as when bad growth conditions is large all right.

──────────────────────────────────────────────────続 き Continuation of front page (56) References JP-A-4-338305 (JP, A) Plant & Cell Physiol. , 11, 663-673 (1979) Plant Physiol. , 55, 791-5 (1975) Plant Science Letters, 37, 213-6 (1985) (58) Fields investigated (Int. Cl. ⁷ , DB name) A01N 37/44 CA (STN)

Claims (3)

(57) [Claims] (1) At a working concentration in terms of 5-aminolevulinic acid
An algal growth promoter containing 0.03-700 ppm of 5-aminolevulinic acid or a salt thereof. 2. An algae phycocyanin or carotenoid content improver comprising 5-aminolevulinic acid or a salt thereof as an active ingredient. 3. An agent for improving algae's ability to fix carbon dioxide, comprising 5-aminolevulinic acid or a salt thereof as an active ingredient.