Method for producing cosmetic and chlorella extract

Abstract

The present invention provides a cosmetic comprising a Chlamydomonas reinhardtii extract, the aforementioned Chlamydomonas reinhardtii extract comprising one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine, and isoleucine-glycine.

Description

Technical Field

[0001] This invention relates to a cosmetic and a method for manufacturing Chlamydomonas reinhardtii extract. Background Technology

[0002] As dipeptides derived from the pig placenta, glycine-leucine, leucine-glycine, glycine-isoleucine, and isoleucine-glycine are known.

[0003] Patent document 1 describes a cosmetic material composition containing porcine placental extract.

[0004] On the other hand, as an example of Chlamydomonas reinhardtii, Patent Document 2 describes the Honda DREAMO strain (accession number FERM BP-22306).

[0005] [Existing technical documents]

[0006] [Patent Literature]

[0007] Patent Document 1: Japanese Patent Application Publication No. 2018-203628

[0008] Patent Document 2: International Publication No. 2017 / 217116 Summary of the Invention

[0009] [The problem the invention aims to solve]

[0010] However, organic cosmetics cannot be made using pig placenta.

[0011] The present invention aims to provide an organic cosmetic product containing at least one of dipeptides derived from porcine placenta.

[0012] [Technical means to solve the problem]

[0013] (1) A cosmetic product comprising Chlamydomonas reinhardtii extract, wherein the Chlamydomonas reinhardtii extract comprises one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine and isoleucine-glycine.

[0014] (2) The cosmetic according to (1), wherein the aforementioned Chlamydomonas reinhardtii is Honda DREAMO strain (accession number FERMBP-22306).

[0015] (3) A method for manufacturing Chlamydomonas reinhardtii extract, the method comprising one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine and isoleucine-glycine, and the method comprising the step of culturing the aforementioned Chlamydomonas reinhardtii under intermittent light irradiation.

[0016] (4) The method for manufacturing Chlamydomonas reinhardtii extract according to (3), wherein the irradiation time of the aforementioned intermittent light every 24 hours is more than 8 hours and less than 10 hours.

[0017] [Invention Effects]

[0018] According to the present invention, it is possible to provide an organic cosmetic containing at least one of dipeptides derived from porcine placenta. Attached Figure Description

[0019] none Detailed Implementation

[0020] The embodiments of the present invention will be described below.

[0021] [cosmetic]

[0022] The cosmetic of this embodiment contains Chlamydomonas reinhardtii extract. Here, the Chlamydomonas reinhardtii extract contains one or more dipeptides selected from the group consisting of glycine-leucine (Gly-Leu), leucine-glycine (Leu-Gly), glycine-isoleucine (Gly-Ile), and isoleucine-glycine (Ile-Gly). Thus, an organic cosmetic containing at least one dipeptide from porcine placenta can be obtained. As a result, if the cosmetic of this embodiment is used, it is expected to achieve the same effects as cosmetics containing porcine placental extract.

[0023] There are no specific limitations on what constitutes *Chlamydomonas reinhardtii*; for example, UTEX 90 strain and Honda DREAMO strain (accession number FERM BP-22306) can be cited. Here, UTEX 90 strain is the original strain of Honda DREAMO strain.

[0024] Here, the extract of the Honda DREAMO strain contains Gly-Leu, Leu-Gly, and Ile-Gly.

[0025] The cosmetic form described in this embodiment is not particularly limited; for example, a beauty serum can be cited. A beauty serum may contain, for example, a base containing butylene glycol and water, and Chlamydomonas reinhardtii extract, wherein the content of the Chlamydomonas reinhardtii extract is 1 ppm or more and 10,000 ppm or less. Here, the ratio of butylene glycol to water is not particularly limited.

[0026] [Manufacturing method of Chlamydomonas reinhardtii extract]

[0027] The method for producing Chlamydomonas reinhardtii extract according to this embodiment includes a step of culturing Chlamydomonas reinhardtii under intermittent light irradiation. As a result, the content of one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine, and isoleucine-glycine in the Chlamydomonas reinhardtii extract increases.

[0028] The photoperiod duration per 24 hours is preferably 8 hours or more but less than 10 hours, and more preferably 8.5 hours or more but less than 9.5 hours. When the photoperiod duration per 24 hours is 8 hours or more but less than 10 hours, the content of one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine, and isoleucine-glycine in the Chlamydomonas reinhardtii extract will increase.

[0029] There are no particular restrictions on the wavelength of the light irradiated during the bright period; for example, it can be above 300 nm and below 800 nm. There are also no particular restrictions on the photosynthetically active photon flux density of the light irradiated during the bright period; for example, it can be 60 μmol / m³. 2 / s or higher and 400 μmol / m 2 / s or less. There are no particular limitations on the light source used for illumination during the bright period; for example, white light-emitting diodes (LEDs) can be listed.

[0030] There are no particular limitations on the cultivation methods for Chlamydomonas reinhardtii, such as static culture, shaking culture, submerged culture, and aeration culture.

[0031] There are no particular limitations on the culture medium used for culturing Chlamydomonas reinhardtii. Examples include Tris-Acetate-Phosphate (TAP) medium, N1 medium, urea medium, High Salt Medium (HSM) medium, and BG11 medium.

[0032] There are no particular restrictions on the temperature for culturing Chlamydomonas reinhardtii; for example, it can be above 25°C and below 28°C.

[0033] When culturing *Chlamydomonas reinhardtii* using the aeration method, there is no particular limit to the gas throughput per unit volume; for example, it can be above 0.001 VVM and below 10 VVM. Furthermore, VVM indicates how many times the volume of gas passes through the culture medium per minute.

[0034] When culturing *Chlamydomonas reinhardtii* using the aeration method, it is preferable to supply carbon dioxide to the culture medium. There are no particular restrictions on the concentration of carbon dioxide in the aeration gas; for example, it can be 0.1% by volume or more and 10% by volume or less. Furthermore, there are no particular restrictions on the source of carbon dioxide; examples include carbon dioxide cylinders, gases emitted from factories, etc., and concentrated carbon dioxide recovered from the atmosphere using direct air capture (DAC).

[0035] There are no particular limitations on the methods for recovering cultured Chlamydomonas reinhardtii; for example, centrifugation and filtration can be used.

[0036] In addition, the recovered Chlamydomonas reinhardtii can be dried as needed.

[0037] There are no particular limitations on the drying methods for Chlamydomonas reinhardtii, such as freeze drying, spray drying, hot air drying, and sun drying.

[0038] There are no particular limitations on the extraction methods used for extracting Chlamydomonas reinhardtii; for example, ultrasonic extraction can be cited.

[0039] There are no particular limitations on the solvents used for extracting Chlamydomonas reinhardtii, and examples include: lower alcohols such as methanol, ethanol, propanol, and isopropanol; lower aliphatic ketones such as acetone and methyl ethyl ketone; polyols such as 1,3-butanediol, propylene glycol, and glycerol; and hydrophilic organic solvents such as dimethyl sulfoxide (DMSO). DMSO is preferred.

[0040] There are no particular limitations on the recovery methods for Chlamydomonas reinhardtii extract; for example, centrifugation and filtration can be used.

[0041] In addition, the recovered Chlamydomonas reinhardtii extract can also be dried as needed.

[0042] There are no particular limitations on the drying method for Chlamydomonas reinhardtii extract; freeze drying is an example.

[0043] [Example]

[0044] The following describes embodiments of the present invention, but the present invention is not limited to these embodiments.

[0045] [Example 1]

[0046] [Cultivation of Chlamydomonas reinhardtii]

[0047] The Honda DREAMO strain of Chlamydomonas reinhardtii was recovered from urea medium and cultured for 3–4 days with aeration (seed culture) using 6 L of urea medium under the following culture conditions.

[0048] Temperature: 25~28℃

[0049] Light-dark cycle: Light period 8-10 hours, Dark period 16-14 hours

[0050] Photosynthetically active photon flux density of light irradiated during the Ming period: 60–400 μmol / m 2 / s

[0051] Light source for illumination during the bright period: white LED

[0052] Gas throughput per unit volume: 0.15 VVM

[0053] Concentration of carbon dioxide in the gas: 5% by volume

[0054] Next, after centrifuging 500 mL of the seed culture at 3000 rpm for 5 minutes, the culture supernatant was removed, and the Honda DREAMO strain obtained from the seed culture was recovered.

[0055] Secondly, the recovered HondaDREAMO strain was cultured under aerated conditions for 7 days (pre-culture) using 6 L of UREA medium under the same conditions as the seed culture.

[0056] Then, using 1 L of N1 medium, the pre-cultured HondaDREAMO strain was cultured under the same conditions as the seed culture for 7 days with aeration (formal culture). The aeration conditions are shown below.

[0057] Gas throughput per unit volume: 0.15 VVM

[0058] Concentration of carbon dioxide in the gas: 5% by volume

[0059] Next, after centrifuging 5 L of the formal culture at 6000 rpm for 10 minutes, the culture supernatant was removed, and the formally cultured Honda DREAMO strain was recovered.

[0060] [extract]

[0061] Using an ultrasonic homogenizer, approximately 20 mL of the recovered Honda DREAMO strain was ultrasonically extracted in 100 mL of DMSO for 5 minutes. The ultrasonic extraction was performed on ice.

[0062] Next, the mixture of extract and residue was centrifuged at 3000 rpm for 5 minutes to remove the residue and recover about 100 mL of extract.

[0063] The extract was then freeze-dried to obtain an extract of the Honda DREAMO strain.

[0064] [Comparative Example 1]

[0065] Except for irradiating with continuous light instead of intermittent light, i.e., setting the light period to 24 hours, the extract of Honda DREAMO strain was obtained in the same manner as in Example 1.

[0066] [Content of dipeptides in Honda DREAMO strain extract]

[0067] The content of dipeptides (Gly-Leu, Leu-Gly, and Ile-Gly) in each 1 g (dry weight) of Honda DREAMO strain extract was measured using high performance liquid chromatography (HPLC) [g].

[0068] Table 1 shows the evaluation results of dipeptide content in Honda DREAMO strain extract.

[0069] [Table 1]

[0070] Example 1 Comparative Example 1 Gly-Leu <![CDATA[8.70×10 -5 ]]> <![CDATA[4.20×10 -5 ]]> Leu-Gly <![CDATA[7.00×10 -6 ]]> <![CDATA[7.00×10 -7 ]]> Ile-Gly <![CDATA[2.90×10 -5 ]]> <![CDATA[2.40×10 -6 ]]> total <![CDATA[1.23×10 -4 ]]> <![CDATA[4.51×10 -5 ]]>

[0071] As shown in Table 1, the dipeptide content in the Honda DREAMO strain extract of Example 1 is higher than that in the Honda DREAMO strain extract of Comparative Example 1.

Claims

1. A cosmetic product comprising Chlamydomonas reinhardtii extract, The aforementioned Chlamydomonas reinhardtii extract contains one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine, and isoleucine-glycine.

2. The cosmetic product according to claim 1, wherein, The aforementioned Chlamydomonas reinhardtii strain is Honda DREAMO (accession number FERMBP-22306).

3. A method for manufacturing Chlamydomonas reinhardtii extract, wherein the method is a method for manufacturing Chlamydomonas reinhardtii extract. The aforementioned Chlamydomonas reinhardtii extract contains one or more dipeptides selected from the group consisting of glycine-leucine, leucine-glycine, glycine-isoleucine, and isoleucine-glycine. The manufacturing method includes the step of culturing the aforementioned Chlamydomonas reinhardtii under intermittent light irradiation.

4. The method for manufacturing Chlamydomonas reinhardtii extract according to claim 3, wherein, The aforementioned intermittent light exposure time is more than 8 hours and less than 10 hours per 24 hours.