The use of yeast cells or cell cultures of Yarrowia lipolytica, or its extracts containing L-hydroxyproline, and methods for the production of L-hydroxyproline.

Abstract

The present invention relates to a yeast cell of Yarrowia lipolytica, a cell culture product thereof or an extract of the same. The aforesaid yeast cell, cell culture product and extract of the same are characterized by containing L-hydroxyproline, and the ratio [100 × Hyp / (Pro + Hyp)] of the content (μg / mL) of L-hydroxyproline (Hyp) to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline being 35-100.

Description

Cell bodies, cell cultures, or extracts thereof of the yeast Yarrowia lipolytica containing L-hydroxyproline, their uses, and a method for producing L-hydroxyproline The present invention relates to cell bodies, cell cultures, or extracts thereof of yeast containing L-hydroxyproline, their uses, and a method for producing L-hydroxyproline. The present invention also relates to the use of yeast for producing L-hydroxyproline. The present invention also relates to foods and drinks, cosmetics, cosmetic raw materials, and compositions for reinforcing L-hydroxyproline, etc., containing cell bodies, cell cultures, or extracts thereof of yeast. L-Hydroxyproline (hydroxy-L-proline) is an amino acid having a structure in which a hydroxyl group is bonded to the 4-position carbon atom of L-proline. As the effects of L-hydroxyproline, promotion of collagen production in fibroblasts, promotion of epidermal cell proliferation, moisturizing effect equal to or higher than that of collagen, prevention of skin aging, higher transdermal absorbability than tripeptides, improvement effect of wrinkles, improvement effect of atopic dermatitis, etc. can be mentioned. Since L-hydroxyproline is safe for the human body, it can be used by being contained in foods and drinks, cosmetics, pharmaceuticals, etc., and its beneficiality is very high. L-Hydroxyproline can be produced by an organic synthesis method, but a production method using microorganisms is also being studied. For example, Patent Document 1 describes culturing a transformant obtained by introducing a polynucleotide encoding L-proline cis-4-hydroxylase derived from Mesorhizobium loti into a host cell in a medium, generating and accumulating cis-4-hydroxy-L-proline in the culture, and collecting cis-4-hydroxy-L-proline from the culture. A method for producing cis-4-hydroxy-L-proline is described. Japanese Patent No. 5506668 By the way, yeast is a microorganism that has been tried for various industrial uses in the food and drink field and the like for a long time, and cell bodies, cell cultures, or extracts thereof of yeast containing L-hydroxyproline are useful as raw materials for cosmetics, foods and drinks, etc. where the effects of L-hydroxyproline are expected. However, yeast that accumulates L-hydroxyproline in cell bodies or cell cultures has not been reported yet. The main object of the present invention is to provide a yeast cell or cell culture containing L-hydroxyproline, or an extract thereof, and its use, as well as a method for producing L-hydroxyproline. As a result of intensive research to solve the above problems, the present inventors have found that when Yarrowia lipolytica, a yeast, is aerobically cultured, L-hydroxyproline accumulates in its cells or cell culture. Further, it has been found that the cells or cell culture of Yarrowia lipolytica containing the obtained L-hydroxyproline, or an extract thereof, can have a ratio (100×Hyp / (Pro + Hyp)) of the weight content of L-hydroxyproline to the total weight content of L-proline (Pro) and L-hydroxyproline (Hyp) within a specific range. Based on these findings, the present inventors have conducted further research and completed the present invention. The yeast cell or cell culture of the present invention, or an extract thereof, is a cell or cell culture of Yarrowia lipolytica, or an extract thereof, and the yeast cell or cell culture, or an extract thereof, contains L-hydroxyproline, and the ratio (100×Hyp / (Pro + Hyp)) of the content (μg / mL) of L-hydroxyproline to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) is 35 to 100. Preferably, the yeast cell or cell culture of the present invention, or an extract thereof, has an L-hydroxyproline content of 10 μg / mL or more. Another aspect of the yeast cell or cell culture of the present invention, or an extract thereof, is a cell or cell culture of Yarrowia lipolytica, or an extract thereof, and is characterized by having an L-hydroxyproline content of 10 μg / mL or more. The method for producing L-hydroxyproline of the present invention includes a step of accumulating L-hydroxyproline in the cells or cell culture of Yarrowia lipolytica by aerobically culturing Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source, and is characterized in that the nitrogen source is a nitrogen source containing an L-hydroxyproline-containing peptide. In the production method of the present invention, it is preferable that the L-hydroxyproline-containing peptide is a collagen peptide. The average molecular weight of the collagen peptide is preferably from 1,000 to 10,000. In the production method of the present invention, it is preferable that the aerobic culture is carried out for 10 to 100 hours. The present invention also includes the use of Yarrowia lipolytica for producing L-hydroxyproline. The use of the present invention includes accumulating L-hydroxyproline in the cells or cell culture of the above yeast by aerobically culturing the above Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source, and it is preferable that the nitrogen source is a nitrogen source containing an L-hydroxyproline-containing peptide. In the use of the present invention, it is preferable that the L-hydroxyproline-containing peptide is a collagen peptide. The average molecular weight of the collagen peptide is preferably from 1,000 to 10,000. In the use of the present invention, it is preferable that the aerobic culture is carried out for 10 to 100 hours. The composition of the present invention is characterized by containing the cells or cell culture of the yeast of the present invention or an extract thereof. The food or drink of the present invention is characterized by containing the cells or cell culture of the yeast of the present invention or an extract thereof. The cosmetic or cosmetic raw material of the present invention is characterized by containing the cells or cell culture of the yeast of the present invention or an extract thereof. The cosmetic or cosmetic raw material of the present invention is preferably used for applications selected from promoting collagen production, promoting epidermal cell proliferation, moisturizing the skin, preventing skin aging, preventing or improving skin sagging, improving skin firmness, preventing or improving wrinkles, and improving atopic dermatitis. In one aspect, the cosmetic or cosmetic raw material of the present invention is a cosmetic raw material, and preferably has an L-hydroxyproline content of 5 to 300 ppm. The cosmetic or cosmetic raw material of the present invention is also preferably a cosmetic and has an L-hydroxyproline content of 0.01 to 20 ppm. In this specification, ppm means weight ppm. The composition for reinforcing L-hydroxyproline of the present invention is characterized by containing cells or cell cultures of Yarrowia lipolytica containing L-hydroxyproline or extracts thereof. In the composition for reinforcing L-hydroxyproline of the present invention, the ratio (100×Hyp / (Pro + Hyp)) of the content (μg / mL) of L-hydroxyproline to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) in the cells or cell cultures of the above yeast or extracts thereof is preferably 35 to 100. Also, the cells or cell cultures of the above yeast or extracts thereof preferably have an L-hydroxyproline content of 10 μg / mL or more. According to the present invention, it is possible to provide cells or cell cultures of yeast containing L-hydroxyproline or extracts thereof and their uses, as well as a method for producing L-hydroxyproline, etc. The cells or cell cultures of the yeast of the present invention or extracts thereof are suitably used as raw materials for foods and drinks, cosmetics, etc. FIG. 1 is a graph showing the L-hydroxyproline (Hyp) accumulation amount in yeast. FIG. 2 is a graph showing the Hyp accumulation amount in Yarrowia lipolytica cultured in each medium ((a): condition 1, (b): condition 2). FIG. 3 is a graph showing the Hyp accumulation amount in Yarrowia lipolytica cultured for 5 days in PD medium or PD medium supplemented with 0.125% gelatin (white: PD medium containing 0.125% gelatin, black: PD medium). FIG. 4 is a graph showing the Hyp accumulation amount in Yarrowia lipolytica cultured in YPD media with different compositions ((a): Hyp accumulation amount (μg / mL), (b): Hyp amount per cell (μg / mL / OD600)). FIG. 5 is a graph showing the Hyp accumulation amount, Hyp amount per cell, and OD600 of Yarrowia lipolytica cultured in a medium containing peptone or collagen peptide ((a): Hyp accumulation amount (μg / mL), (b): Hyp amount per cell (μg / mL / OD600), (c): OD600)). FIG. 6 is a diagram showing the Hyp accumulation amount in Yarrowia lipolytica cultured with shaking or static culture ((a): Hyp amount per medium in the culture sample (intracellular), (b): Hyp amount in the culture supernatant)). It is a graph showing the ethanol concentration and glucose concentration in the culture supernatant of Yarrowia lipolytica cultured with shaking or static culture ((a): ethanol concentration (v / v%), (b): glucose concentration (wt%)). FIG. 8 is an HPLC chart analyzing a 0.1 N hydrochloric acid solution containing an amino acid mixed standard solution H type and L-hydroxyproline (each amino acid concentration 20 μmol / L) ((a): detected at an excitation wavelength of 350 nm and a fluorescence wavelength of 450 nm in Ch1, (b): detected at an excitation wavelength of 266 nm and a fluorescence wavelength of 305 nm in Ch2). Hereinafter, embodiments of the present invention will be specifically described. However, the present invention is not limited to the following embodiments, and can be appropriately modified and applied without changing the gist of the present invention. In this specification, the genus species of yeast are described by the genus species names described in The Yeasts, a Taxonomic Study Fifth Edition (published by Elsevier, 2011). The cell bodies or cell cultures of yeast or extracts thereof according to the first aspect of the present invention are cell bodies or cell cultures of yeast Yarrowia lipolytica or extracts thereof, and the cell bodies or cell cultures of yeast or extracts thereof contain L-hydroxyproline, and the ratio (100×Hyp / (Pro + Hyp)) of the content (μg / mL) of L-hydroxyproline to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) is 35 to 100. The cell bodies or cell cultures of yeast or extracts thereof according to the second aspect of the present invention are cell bodies or cell cultures of yeast Yarrowia lipolytica or extracts thereof, and the content of L-hydroxyproline is 10 μg / mL or more. Hereinafter, the cell bodies or cell cultures of yeast or extracts thereof according to the first and second aspects of the present invention are also collectively referred to as the cell bodies or cell cultures of yeast or extracts thereof of the present invention. The cell bodies or cell cultures of yeast or extracts thereof of the present invention are cell bodies or cell cultures of yeast Yarrowia lipolytica or extracts thereof. The yeast in the present invention may be any yeast belonging to Yarrowia lipolytica, and only one kind may be used, or two or more kinds may be used. Yarrowia lipolytica can be obtained from various depository institutions and the like. Examples of depository institutions include the National Institute of Technology and Evaluation (2-5-8 Kazusa Kamashima, Kisarazu City, Chiba Prefecture, Japan). It can also be isolated from nature. The cells or cell cultures of yeast of the present invention, or extracts thereof, contain L-hydroxyproline (Hyp). The L-hydroxyproline in the present invention is 4-hydroxy-L-proline. In this specification, the L-hydroxyproline contained in the cells or cell cultures of yeast, or extracts thereof, refers to free L-hydroxyproline. The content or accumulation amount of L-hydroxyproline in the cells or cell cultures of yeast, or extracts thereof, refers to the amount of free L-hydroxyproline. No yeast has been reported to accumulate free L-hydroxyproline in its cells or cell cultures. In the cells or cell cultures of yeast of the first aspect of the present invention, or extracts thereof, the ratio (100×Hyp / (Pro + Hyp)) of the content (μg / mL) of L-hydroxyproline to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) is 35 to 100. Such cells or cell cultures of yeast, or extracts thereof, are suitably used as raw materials for foods, beverages, cosmetics, etc., for which the efficacy of L-hydroxyproline is expected. The "ratio of the content (μg / mL) of L-hydroxyproline to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline (Hyp)" (100×Hyp / (Pro + Hyp)) is hereinafter also referred to as the "(Hyp / (Pro + Hyp)) ratio". In this specification, the L-proline content in the above (Hyp / (Pro + Hyp)) ratio refers to the content of free L-proline contained in the cells or cell cultures of yeast, or extracts thereof. In a preferred embodiment, in the cells or cell cultures of yeast of the second aspect of the present invention, or extracts thereof, the (Hyp / (Pro + Hyp)) ratio is 35 to 100. The above (Hyp / (Pro + Hyp)) ratio is preferably from 40 to 100, more preferably from 50 to 100, still more preferably from 60 to 100, even more preferably from 70 to 100, even more preferably from 80 to 100, and particularly preferably from 90 to 100. The yeast cell bodies or cell body cultures or extracts thereof with such an (Hyp / (Pro + Hyp)) ratio have a high content ratio of Hyp and are particularly suitable as raw materials for foods, beverages, cosmetics, etc. where the efficacy of L-hydroxyproline is expected. The yeast cell bodies or cell body cultures or extracts thereof according to the second aspect of the present invention have an L-hydroxyproline content of 10 μg / mL or more. The yeast cell bodies or cell body cultures or extracts thereof according to the first aspect of the present invention preferably have an L-hydroxyproline content of 10 μg / mL or more. The yeast cell bodies or cell body cultures or extracts thereof with an L-hydroxyproline content within the above range are suitable as raw materials for foods, beverages, cosmetics, etc. where the efficacy of L-hydroxyproline is expected. The L-hydroxyproline content of the yeast cell bodies or cell body cultures or extracts thereof of the present invention is more preferably 15 μg / mL or more, still more preferably 17 μg / mL or more, still more preferably 20 μg / mL or more, even more preferably 30 μg / mL or more, even more preferably 40 μg / mL or more, even more preferably 50 μg / mL or more, even more preferably 70 μg / mL or more, even more preferably 100 μg / mL or more, even more preferably 150 μg / mL or more, particularly preferably 200 μg / mL or more, particularly preferably 250 μg / mL or more, particularly preferably 300 μg / mL or more, particularly preferably 400 μg / mL or more, particularly preferably 450 μg / mL or more, particularly preferably 475 μg / mL or more, and most preferably 500 μg / mL or more. The upper limit of the L-hydroxyproline content of the yeast cell bodies or cell body cultures or extracts thereof is not particularly limited, and a higher content is preferred, but it is usually 6000 μg / mL or less, and may be 3000 μg / mL or less or 2000 μg / mL or less. According to the present invention, it is possible to provide a yeast cell or cell culture or an extract thereof, in which the content of L-hydroxyproline derived from the yeast cell or cell culture is within the above range. The contents (μg / mL) of L-proline and L-hydroxyproline in the yeast cell or cell culture or an extract thereof can be measured by high performance liquid chromatography (HPLC). When the yeast cell or cell culture or an extract thereof contains cells, the cells are disrupted by autodigestion by heating or the like, enzymatic decomposition treatment or the like, and the content of L-proline and L-hydroxyproline is measured using the eluate of the cell contents. As the method for measuring the contents of L-proline and L-hydroxyproline and the measurement conditions of HPLC and the like, the methods and conditions described in the examples may be adopted. Preferably, (1) a method in which amino acids are sequentially derivatized with o-phthalaldehyde (OPA) and 4-chloro-7-nitrobenzofurazan (NBD-Cl) and analyzed by HPLC (detected at an excitation wavelength of 503 nm / fluorescence wavelength of 541 nm), or (2) mercaptopropionic acid, after derivatizing the primary amino group with OPA, derivatizing the secondary amino acid with 9-fluorenylmethyl chloroformate (FMOC) and analyzing by HPLC (detected at an excitation wavelength of 266 nm and a fluorescence wavelength of 305 nm), more preferably by the method (2) above, L-proline and L-hydroxyproline are quantified. The yeast cell or cell culture or an extract thereof of the present invention is obtained by aerobically culturing Yarrowia lipolytica in a liquid medium and, if necessary, performing cell disruption or the like. The yeast cell culture preferably contains yeast cells and / or culture supernatant, and may contain the intracellular contents of the yeast cells. The yeast cells may be viable cells or dead cells. Examples of the cell culture containing yeast cells and / or culture supernatant include a cell culture solution containing yeast cells (cultured cells) and culture supernatant obtained by aerobic culture of the above yeast, a collection of yeast cells from the cell culture solution (cells), or a culture supernatant obtained by removing cells from the cell culture solution. The culture supernatant of the cell culture solution is simply referred to as the culture supernatant. The cell culture is preferably a cell culture solution or a culture supernatant containing yeast cells and culture supernatant. In addition, the extract of the cells or cell culture usually contains intracellular contents of the cells, and preferably contains intracellular contents and culture supernatant. Examples of the extract of the cells or cell culture include an extract obtained by subjecting the cells or cell culture containing the cells (preferably a cell culture solution) to cell disruption treatment such as autolysis treatment or enzymatic decomposition treatment to elute the intracellular contents of the yeast cells into the culture solution, etc. (cell disruption product), and an extract obtained by removing cell residues from the cells or cell culture (cell disruption product) subjected to cell disruption treatment. Preferably, it is an extract containing intracellular contents and culture supernatant obtained by removing cell residues from a cell culture solution (cell disruption product) subjected to cell disruption treatment or the cell disruption product. As described above, the yeast cells, cell culture, or extract thereof of the present invention is usually prepared by subjecting a cell culture solution containing yeast cells and culture supernatant obtained by aerobic culture of yeast Yarrowia lipolytica in a liquid medium to treatments such as cell collection and cell disruption as necessary. The L-hydroxyproline contained in the yeast cells, cell culture, or extract thereof of the present invention preferably originates from the yeast cells or cell culture obtained by the above aerobic culture. It is preferable that the L-hydroxyproline contained in the yeast cells, cell culture, or extract thereof of the present invention is substantially absent before the above aerobic culture. The yeast cells, cell culture, or extract thereof of the present invention can be suitably used, for example, as raw materials for cosmetics, food and beverages including alcoholic beverages. The cell body, cell culture, or extract thereof of the yeast of the present invention preferably has a value (μg / mL / OD600) obtained by dividing the content (μg / mL) of L-hydroxyproline by OD600 of 0.5 or more. The value (μg / mL / OD600) obtained by dividing the content (μg / mL) of L-hydroxyproline by OD600 is hereinafter also referred to as the Hyp / OD600 value. The higher the Hyp / OD600 value, the more preferable it is because the content of L-hydroxyproline per cell is higher. The upper limit of the Hyp / OD600 value of the cell body, cell culture, or extract thereof of yeast is not particularly limited, and the higher the value, the more preferable it is, but it is usually 300 or less. The Hyp / OD600 value is more preferably 1 or more, and for example, 1 to 150 is preferable. The Hyp / OD600 value is even more preferably 25 or more, and even more preferably 50 or more. OD is an abbreviation for optical density and refers to the optical density. OD represents the concentration of cells, etc. Generally, the absorbance OD600 or OD660 at a wavelength of 600 nm or 660 nm of visible light is measured (Bio Experiment Illustrated ▲7▼ Let's Use Yeast Two Hybrid, published by Shujunsha in 2003). OD600 used in the calculation of the Hyp / OD600 value is the absorbance at 600 nm of the cell culture solution (cell culture containing cells and culture supernatant) containing cells and culture supernatant used in the preparation of the cell body, cell culture, or extract thereof. More specifically, when the cell culture solution of yeast obtained by aerobically culturing the above yeast in a liquid medium is used as the cell culture as it is, OD600 is the absorbance OD600 of the cell culture solution (cell culture). When the cell body, cell culture, or extract thereof of yeast is an extract of the cell body or cell culture obtained by disrupting the cell body of yeast and eluting the cell contents, OD600 is the absorbance OD600 of the cell culture solution used for its preparation (cell culture solution containing yeast cell bodies before disruption and culture supernatant). OD600 can be measured, for example, by a spectrophotometer. The cells or cell cultures of the yeast of the present invention, or extracts thereof, preferably have an ethanol content of 1 v / v% or less. When the ethanol content is 1 v / v% or less, it can be particularly preferably used as a raw material for various foods and beverages, cosmetics, and the like. If the ethanol exceeds 1 v / v%, it may have an adverse effect on yeast growth and the like. The ethanol content of the cells or cell cultures of the yeast of the present invention, or extracts thereof, is more preferably 0.8 v / v% or less, and even more preferably 0.5 v / v% or less. The ethanol content can be measured by known methods. The form of the cells or cell cultures of the yeast of the present invention, or extracts thereof, is not particularly limited, and examples thereof include paste form, suspension form, extract form, liquid form, and the like. The cells or cell cultures of the yeast of the present invention, or extracts thereof, can be preferably used as raw materials for cosmetics, foods and beverages, etc., as described below. The cells or cell cultures of the yeast of the present invention, or extracts thereof, can also be used after being powdered by drying or the like. The cells or cell cultures of the yeast of the present invention, or extracts thereof, can be obtained by aerobically culturing Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source, and, if necessary, disrupting the cells. More specifically, the nitrogen source includes a peptide containing L-hydroxyproline. By aerobically culturing Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source containing a peptide containing L-hydroxyproline, L-hydroxyproline accumulates in the cells or cell cultures of the yeast, and cells or cell cultures of yeast containing L-hydroxyproline are obtained. A method including the step of accumulating L-hydroxyproline in the cells or cell cultures of the yeast by aerobically culturing Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source containing a peptide containing L-hydroxyproline is preferable as the method for producing the cells or cell cultures of the yeast of the present invention, or extracts thereof, or the method for producing L-hydroxyproline described above. The method for producing L-hydroxyproline of the present invention includes a step of accumulating L-hydroxyproline in the cells or cell culture of the yeast Yarrowia lipolytica by aerobic culturing the yeast in a liquid medium containing a carbon source and a nitrogen source (hereinafter, also referred to as the Hyp accumulation step). The nitrogen source is a nitrogen source containing an L-hydroxyproline-containing peptide. The production method of the present invention may optionally have steps other than the Hyp accumulation step. For example, it may have one or more steps such as the pre-culture step, cell collection step, cell disruption step, etc. described later. In the method for producing L-hydroxyproline of the present invention, the cells or cell culture of the yeast of the present invention described above or an extract thereof can be obtained. By performing the Hyp accumulation step, cells or a cell culture of yeast containing L-hydroxyproline can be obtained. The obtained cells or cell culture of yeast usually has a (Hyp / (Pro + Hyp)) ratio of 35 to 100. Also, the cells or cell culture of yeast obtained by the Hyp accumulation step usually has an L-hydroxyproline content of 10 μg / mL or more. Such cells or cell culture of yeast can be used as the cells or cell culture of the yeast of the present invention described above. Further, the obtained cells or cell culture can be optionally further treated, such as cell disruption treatment, to prepare an extract of the cells or cell culture of yeast containing L-hydroxyproline. The method for producing L-hydroxyproline including the Hyp accumulation step is also preferable as a method for producing the cells or cell culture of the yeast of the present invention described above or an extract thereof. When obtaining the cells or cell culture of the yeast of the present invention or an extract thereof, the Hyp accumulation step and its preferred embodiments are the same as the Hyp accumulation step and its preferred embodiments in the method for producing L-hydroxyproline. In the Hyp accumulation process, the method of adding the yeast Yarrowia lipolytica to the liquid medium can be achieved by inoculating a small amount of cells directly into the liquid medium containing a carbon source and a nitrogen source for growth. However, in order to increase the cell concentration in a short period, it is preferable to inoculate the pre-cultured bacterial solution. The medium used for pre-culture is not particularly limited, and it may be the same medium as the liquid medium used in the Hyp accumulation process (usually main culture), or a known medium that can be used for yeast may also be used. The time for pre-culture is usually 10 to 72 hours, preferably 12 to 48 hours. The pre-culture temperature is preferably 15 to 40 °C. The inoculation amount of the pre-cultured bacterial solution is usually 1 / 100,000 to 1 / 2 of the amount of the medium used in the Hyp accumulation process, preferably 1 / 1000 to 1 / 10, more preferably 1 / 200 to 1 / 10, and even more preferably 1 / 200 to 1 / 20. If the inoculation amount is within the above range, the growth of Yarrowia lipolytica in the Hyp accumulation process is fast, and L-hydroxyproline can be efficiently accumulated. The nitrogen source of the liquid medium used in the Hyp accumulation process is a nitrogen source containing an L-hydroxyproline-containing peptide. When Yarrowia lipolytica is aerobically cultured using such a nitrogen source, L-hydroxyproline accumulates in the cells or cell cultures. Such a nitrogen source may be used alone or in combination of two or more. The L-hydroxyproline-containing peptide may be one kind or two or more kinds. The L-hydroxyproline-containing peptide may be a peptide containing L-hydroxyproline as a constituent amino acid, but preferably a peptide in which 10% by weight or more of the constituent amino acids is L-hydroxyproline. In one aspect, it is also preferable that the nitrogen source containing the L-hydroxyproline-containing peptide is the L-hydroxyproline-containing peptide. A nitrogen source containing an L-hydroxyproline-containing peptide can be obtained, for example, by hydrolyzing an L-hydroxyproline-containing protein. The L-hydroxyproline-containing protein may be any protein containing L-hydroxyproline as a constituent amino acid, but is preferably a protein in which 10% by weight or more of the constituent amino acids are L-hydroxyproline. As the above L-hydroxyproline-containing protein, a collagenous protein or the like is preferable. Examples of the collagenous protein include proteins prepared from tissues containing collagen such as internal organs, skin, fish scales, and bones; collagen and gelatin. The raw material origin of the collagenous protein is not particularly limited. For example, collagenous proteins derived from animals such as bovine, porcine, and fish can be preferably used. As the collagenous protein, commercially available products can be used. The hydrolysis of the L-hydroxyproline-containing protein can be carried out by a known method using an enzyme or the like. As a nitrogen source containing an L-hydroxyproline-containing peptide, for example, peptone derived from animals can be preferably used. Preferably, it is peptone derived from bovine, porcine, or fish, and more preferably peptone derived from bovine or fish. In one aspect of the present invention, as the peptone, meat peptone, myocardial peptone, and gelatin peptone are also preferable. As an example of a commercially available product of a nitrogen source containing an L-hydroxyproline-containing peptide that can be used in the present invention, for example, product name Pepton (#211677) (Bacto) and the like can be mentioned. In one aspect, the L-hydroxyproline-containing peptide is preferably a collagen peptide. The collagen peptide means hydrolyzed collagen, which may be gelatin obtained by heat-treating natural collagen to denature it, collagen peptide obtained by hydrolyzing natural collagen, or those chemically or enzymatically modified thereof. Hydrolysis can be carried out by an enzyme, an acid, an alkali, etc., and is preferably carried out by an enzyme. Preferably, gelatin obtained by heat-treating natural collagen to denature it or collagen peptide obtained by hydrolyzing natural collagen is used. The raw material origin of the collagen peptide is not particularly limited. For example, collagen peptides derived from animals such as bovine, porcine, and fish can be preferably used. Preferably, it is a collagen peptide derived from fish. As the collagen peptide, commercially available products can be used. As an example of a commercially available product of the collagen peptide that can be used in the present invention, for example, "Collagen Peptide Icos HDL-50SP" (product name) (average molecular weight 5000), "Collagen Peptide Type S" (product name) (average molecular weight 1200), "Super Collagen Peptide SCP-2000" (product name) (average molecular weight 2000) manufactured by Nitta Gelatin Inc., "Collagen Peptide P-5000" (product name) (average molecular weight 5000), "Collagen Peptide F-5000" (product name) (average molecular weight 5000) manufactured by Nozue Chemical Industry Co., Ltd., "Marine Collagen Oligo CF" (product name) (average molecular weight 900-1100), "Marine Collagen Oligo MF" (product name) (average molecular weight 900-1500) manufactured by Nichiyo Co., Ltd., etc. Among them, "Collagen Peptide Type S" (average molecular weight 1200), "Collagen Peptide Icos HDL-50SP" (average molecular weight 5000), etc. are preferable. Among these, for example, "Collagen Peptide Icos HDL-50SP", "Collagen Peptide Type S", "Collagen Peptide F-5000", "Marine Collagen CF", "Marine Collagen Oligo MF" are derived from fish, and "Collagen Peptide P-5000", "Super Collagen Peptide SCP-2000" are derived from pigs. In a preferred embodiment of the present invention, the nitrogen source containing the L-hydroxyproline-containing peptide is preferably a collagen peptide (more preferably a collagen peptide derived from fish) and / or peptone (more preferably derived from cattle, pigs or fish, even more preferably peptone derived from cattle or fish), and particularly preferably a collagen peptide. When such a nitrogen source containing the L-hydroxyproline-containing peptide is used, the accumulation amount of L-hydroxyproline in the yeast cells or cell culture increases. The peptone may be meat peptone, cardiac muscle peptone, or gelatin peptone. An example of a preferred embodiment of the method for producing L-hydroxyproline of the present invention includes a step of aerobically culturing the yeast Yarrowia lipolytica in a liquid medium containing a carbon source and a collagen peptide and / or peptone to accumulate L-hydroxyproline in the yeast cells or cell culture. In one aspect, the nitrogen source containing the L-hydroxyproline-containing peptide preferably has an average molecular weight of 10,000 or less. For example, an average molecular weight of 100 to 10,000 is preferred. The L-hydroxyproline-containing peptide preferably has an average molecular weight of 10,000 or less. For example, an average molecular weight of 100 to 10,000 is preferred. Also, the L-hydroxyproline-containing peptide preferably has a molecular weight of 10,000 or less. The collagen peptide preferably has an average molecular weight of 1,000 to 10,000. When a collagen peptide having an average molecular weight within the above range is used as the nitrogen source, the accumulation amount of L-hydroxyproline in the cells or cell culture increases. The average molecular weight of the L-hydroxyproline-containing peptide is calculated by gel filtration or the like. The average molecular weight of the collagen peptide is usually a value calculated by the method described in the 10th edition of the photographic gelatin test method (PAGI method) "20-2 Average Molecular Weight". The average molecular weight of the peptide refers to the weight average molecular weight. The average molecular weight of the collagen peptide is more preferably from 1,000 to 6,000, still more preferably from 1,000 to 5,500, and particularly preferably from 1,000 to 5,000. When such a collagen peptide is used as a nitrogen source, the accumulation amount of L-hydroxyproline in the cells or cell culture becomes larger. Further, in one aspect, the average molecular weight of the collagen peptide is more preferably from 1,000 to 3,000, and still more preferably from 1,000 to 1,500. In another preferred aspect of the present invention, the average molecular weight of the collagen peptide is more preferably from 2,000 to 5,500, and still more preferably from 3,000 to 5,000. The concentration of the nitrogen source containing the L-hydroxyproline-containing peptide in the liquid medium is preferably from 0.1 to 10% by weight, more preferably from 0.25 to 5% by weight, and still more preferably 1 to 5% by weight. When the concentration of the nitrogen source is within the above range, L-hydroxyproline accumulates in the cells or cell culture. Therefore, for example, cells or cell cultures of yeast having an L-hydroxyproline content of 10 μg / mL or more or extracts thereof can be obtained. Further, cells or cell cultures having a (Hyp / (Pro + Hyp)) ratio of 35 to 100 or extracts thereof can be obtained. The concentration of the nitrogen source in the liquid medium is still more preferably from 1.5 to 4.5% by weight, and particularly preferably from 2 to 4% by weight. The concentration of the nitrogen source may be the above concentration at the start of the culture. In one aspect, it is preferable that the concentration of the collagen peptide or peptone in the liquid medium is within the above range. The carbon source is not particularly limited, and examples thereof include saccharides or sugar alcohols such as glucose, fructose, sucrose, raffinose, mannose, maltose, galactose, mannitol, trehalose, melezitose, cellobiose, starch, molasses, sorbitol, L-sorbose, glycerol, ethanol, glucitol; and organic acids such as acetic acid, citric acid, or gluconic acid. The carbon source may be used alone or in combination of two or more. Among them, saccharides such as glucose, fructose, and sucrose are preferable as the carbon source, and glucose is particularly preferable. The concentration of the carbon source in the liquid medium is preferably 0.1 to 20% by weight, preferably 0.5 to 15% by weight, more preferably 1 to 10% by weight, more preferably 1 to 5% by weight, and even more preferably 2 to 5% by weight. When the concentration of the carbon source in the liquid medium is 1% by weight or more, it is preferable because the growth rate of the cells is high. Note that the concentration of the carbon source may be the above concentration at the start of the culture. In the production method of the present invention, the weight ratio (C / N) of the carbon source (C) to the nitrogen source (N) containing an L-hydroxyproline-containing peptide is preferably 0.25 to 20. When the weight ratio of C / N is within the above range, it is preferable because the accumulation amount of L-hydroxyproline in the cells or cell culture is increased. In one aspect, the weight ratio of C / N is more preferably 0.25 to 5, more preferably 0.3 to 3, and even more preferably 0.4 to 1.5. When the weight ratio of C / N is within the above range, the accumulation amount of L-hydroxyproline in the cells or cell culture becomes even larger. In one aspect, for example, when using a collagen peptide (preferably having an average molecular weight of 1000 to 5000) or peptone as the nitrogen source (N) containing an L-hydroxyproline-containing peptide, the weight ratio of C / N is more preferably 0.25 to 5, more preferably 0.3 to 3, even more preferably 0.4 to 1.5, and particularly preferably 0.5 to 1.3. In another preferred aspect, the weight ratio of C / N is also preferably 0.5 to 20. The weight ratio of C / N may be within the above range at the start of the culture. The liquid medium may contain components other than the above-described carbon source and the nitrogen source containing an L-hydroxyproline-containing peptide. For example, it is preferable to contain a yeast extract. The yeast extract usually does not contain an L-hydroxyproline-containing peptide and is not included in the nitrogen source containing an L-hydroxyproline-containing peptide. The yeast extract may be any one that can be used for culturing yeast, is not particularly limited, and commercially available products can be used. For example, product name Bacto yeast Extract(#212750) (Bacto) etc. can be preferably used. When using yeast extract, the concentration of the yeast extract is preferably 0.1 to 3% by weight, more preferably 0.5 to 3% by weight, based on the liquid medium. The concentration of the yeast extract may be the above concentration at the start of the culture. In one aspect, the pH of the liquid medium is preferably 3 to 9, more preferably 4 to 9, still more preferably more than 4 and 9 or less, even more preferably 4.5 to 8.8, and particularly preferably 5 to 8.7. The pH of the liquid medium may be adjusted as appropriate. For pH adjustment, known acids or alkali agents can be used, such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, glutamic acid, acetic acid, butyric acid, lactic acid, formic acid, succinic acid, maleic acid, malic acid, oxalic acid, citric acid, sodium hydroxide, potassium hydroxide, calcium hydroxide, aqueous ammonia, sodium glutamate, and the like. The culture temperature is preferably 15 to 45°C, more preferably 20 to 40°C, and still more preferably 25 to 35°C. When the culture temperature is within this temperature range, the growth of Yarrowia lipolytica is fast, and the accumulation amount of L-hydroxyproline in the cells or cell culture is increased. The method for performing aerobic culture is not particularly limited. For example, a liquid medium inoculated with Yarrowia lipolytica may be subjected to shaking culture or stirring culture. The speed of shaking or stirring is not particularly limited, but is preferably 30 to 600 rpm. In one aspect, it is more preferably 50 to 600 rpm, and still more preferably 100 to 600 rpm. In another preferred aspect, the speed of shaking or stirring is more preferably 30 to 500 rpm, and still more preferably 50 to 300 rpm. Shaking culture or stirring culture at such a speed is preferable because the accumulation amount of L-hydroxyproline is increased. More preferably, shaking culture is performed at the above speed. Also, bubbling may be performed with sterilized air or oxygen as desired. Also, the culture mode may be any of batch culture, fed-batch culture, and continuous culture, but batch culture is preferable. In the production method of the present invention, static culture may be performed. The culture time is not particularly limited and may be set as appropriate. For example, it is preferable to perform aerobic culture for 10 to 100 hours. When the culture time is within the above range, L-hydroxyproline accumulates in the cells or cell cultures of the above yeast. Also, usually, a yeast cell or cell culture or an extract thereof with a (Hyp / (Pro + Hyp)) ratio of 35 to 100, a yeast cell or cell culture or an extract thereof with an L-hydroxyproline content of 10 μg / mL or more can be obtained. Also, a yeast cell or cell culture or an extract thereof with a low ethanol content (for example, 1 v / v% or less) can be obtained. When the culture time is less than 10 hours, the amount of L-hydroxyproline accumulated may be small, or the (Hyp / (Pro + Hyp)) ratio of the obtained yeast cells or cell cultures or extracts thereof may be less than 35. When the culture time exceeds 100 hours, the ethanol concentration of the obtained yeast cells or cell cultures or extracts thereof may exceed 1 v / v%. Also, contamination may easily occur, or coloring due to autolysis may occur after the death of the yeast. The culture time is more preferably 10 to 80 hours, still more preferably 12 to 72 hours, even more preferably 20 to 60 hours, even more preferably 24 to 55 hours, and particularly preferably 24 to 50 hours. Also, in the production method of the present invention, it is preferable to perform aerobic culture until the L-hydroxyproline content in the cells or cell cultures reaches 10 μg / mL or more. In the production method of the present invention, aerobic culture is usually carried out as main culture, but it may also be pre-culture, or aerobic culture may be carried out in both pre-culture and main culture. By performing the above aerobic culture, L-hydroxyproline accumulates in the cells or cell cultures of Yarrowia lipolytica. The cell culture may be a cell culture solution containing yeast cells and a culture supernatant, may be yeast cells, or may be the culture supernatant of the cell culture solution. By performing the Hyp accumulation step, cells or cell cultures of Yarrowia lipolytica containing L-hydroxyproline and having a (Hyp / (Pro + Hyp)) ratio of 35 to 100 can be obtained. Also, cells or cell cultures of Yarrowia lipolytica having an L-hydroxyproline content of 10 μg / mL or more can be obtained. An extract of yeast cells or cell cultures can be obtained by, for example, subjecting the yeast cells or cell cultures to cell disruption treatment. When preparing a yeast cell or cell culture or an extract thereof containing L-hydroxyproline in the present invention, for example, the cell culture solution containing yeast cells and the culture supernatant obtained in the Hyp accumulation step can be directly used as a yeast cell culture containing L-hydroxyproline. Also, yeast cells can be collected from the cell culture solution, and the obtained cells can be used as yeast cells or cell cultures, or the culture supernatant from which the cells have been removed from the cell culture solution can be used as the cell culture. Furthermore, the above cells or cell culture solution can be subjected to a treatment for disrupting the cells as necessary to elute the cell contents into the culture solution or the like to prepare an extract of the cells or cell cultures. In the preparation of an extract of cells or cell cultures, a step of removing cell residues may be performed after disrupting the cells. Also, the cells or cell cultures or extracts thereof may be subjected to treatments such as sterilization and heating as necessary. The production method of the present invention may include one or more steps such as such a cell collection step, cell disruption step, cell residue removal step, and sterilization step. The method for collecting cells from the cell culture solution is not particularly limited, and a commonly used method can be adopted, for example, centrifugation and the like. The method for disrupting the above-mentioned cells is not particularly limited, and a commonly used method can be adopted. For example, autolysis method, enzymatic degradation method, alkaline extraction method, etc. can be mentioned. Among them, the autolysis method is preferred. In the autolysis method, for example, the cells or cell culture can be heated at 40 - 60 °C for 60 - 180 minutes, or at 95 - 100 °C for 5 - 15 minutes. The method for removing cell residues is not particularly limited. For example, known methods such as filtration and centrifugation can be used to remove cell residues. When sterilization is carried out, it is preferable to heat the yeast cells or cell culture or their extracts at 75 - 90 °C (more preferably 80 °C) for 45 - 90 minutes (more preferably 60 minutes). When removing cell residues and sterilization are carried out, either can be done first. The yeast cells or cell culture or their extracts obtained in the present invention, and their preferred embodiments are the same as the yeast cells or cell culture or their extracts of the present invention described above, and their preferred embodiments. According to the production method of the present invention, for example, the ratio of (Hyp / (Pro + Hyp)) is 35 - 100, and the L-hydroxyproline content is 10 μg / mL. The cells or cell culture or their extracts of Yarrowia lipolytica can be produced. The L-hydroxyproline in the cells or cell culture or their extracts of Yarrowia lipolytica usually comes from the cells or cell culture of the above yeast. L-hydroxyproline derived from natural products or synthesized can be further added to the yeast cells or cell culture or their extracts obtained by the above method. However, in a preferred embodiment, the L-hydroxyproline contained in the yeast cells or cell culture or their extracts consists of L-hydroxyproline derived from the cells or cell culture of Yarrowia lipolytica obtained by the above Hyp accumulation step. The cells or cell cultures of yeast containing L-hydroxyproline obtained in the present invention, or extracts thereof, can be used as raw materials for foods and drinks, cosmetics, etc. described later. Further, in the method for producing L-hydroxyproline of the present invention, a step of purifying L-hydroxyproline from the cells or cell cultures of yeast obtained, or extracts thereof, may be performed. The purification of L-hydroxyproline may be carried out by a known method such as column chromatography, for example. The present invention also encompasses the use of the yeast Yarrowia lipolytica for producing L-hydroxyproline. The above Yarrowia lipolytica is also preferably used for producing cells or cell cultures of yeast containing L-hydroxyproline, or extracts thereof. The cells or cell cultures of yeast containing L-hydroxyproline, or extracts thereof, preferably have a (Hyp / (Pro + Hyp)) ratio of 35 to 100. The cells or cell cultures of yeast containing L-hydroxyproline, or extracts thereof, also preferably have an L-hydroxyproline content of 10 μg / mL or more. Preferred embodiments of the cells or cell cultures of yeast containing L-hydroxyproline, or extracts thereof, are the same as the preferred embodiments of the cells or cell cultures of yeast, or extracts thereof, of the present invention described above. The use of the present invention preferably includes accumulating L-hydroxyproline in the cells or cell cultures of the above yeast by aerobically culturing the yeast Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source. The above nitrogen source is preferably a nitrogen source containing an L-hydroxyproline-containing peptide. In the use of the present invention, the above L-hydroxyproline-containing peptide is preferably a collagen peptide. Further, the average molecular weight of the above collagen peptide is preferably 1000 to 10000. In the use of the present invention, the concentration of the nitrogen source containing the L-hydroxyproline-containing peptide in the above liquid medium is preferably 0.25 to 5% by weight. Further, the weight ratio (C / N) of the carbon source (C) to the nitrogen source (N) containing the L-hydroxyproline-containing peptide is preferably 0.25 to 20. In one aspect, the C / N ratio is also preferably 0.5 to 20. In the use of the present invention, it is preferable to carry out the above aerobic culture for 10 to 100 hours. The liquid medium, carbon source, nitrogen source containing the L-hydroxyproline-containing peptide, and their preferred embodiments in the use of the present invention are the same as those in the above-described method for producing L-hydroxyproline. Further, the conditions for aerobic culture and their preferred embodiments are also the same as those in the above-described method for producing L-hydroxyproline. The use of the present invention may include one or more steps such as the above-described cell collection step, cell disruption step, cell removal step, sterilization step, etc. The cells or cell cultures or extracts thereof of the yeast of the present invention described above can be incorporated into various compositions such as cosmetics, foods and drinks, pharmaceuticals, etc. Compositions containing the cells or cell cultures or extracts thereof of the yeast of the present invention are also included in the present invention. The composition of the present invention may include either the cells or cell cultures or extracts thereof of the yeast of the first aspect and the second aspect of the present invention described above, or may include both. The composition containing the cells or cell cultures or extracts thereof of the yeast of the present invention contains L-hydroxyproline derived from the cells or cell cultures or extracts thereof of the yeast. Examples of the composition of the present invention include cosmetics (cosmetic compositions), foods and drinks (food and drink compositions), pharmaceuticals (pharmaceutical compositions), quasi-drugs (quasi-drug compositions), etc. The composition may be these raw materials. In one aspect, the composition is preferably a cosmetic or a food and drink, or these raw materials. The content of the cells or cell cultures of the yeast or their extracts in the composition of the present invention is not particularly limited and can be appropriately set according to the type and use of the composition. For example, with respect to the composition, it is preferable that the content of the cells or cell cultures of the yeast or their extracts in terms of solid content is 0.00001 to 50% by weight, more preferably 0.00005 to 20% by weight, and even more preferably 0.0001 to 10% by weight. When the composition of the present invention is a cosmetic or a pharmaceutical, its dosage form is not particularly limited, and it can take any dosage form such as a solution, paste, gel, solid, powder, etc. The cosmetics are not particularly limited. For example, they can be cleansing agents, facial washes, lotions, emulsions, creams, beauty serums, hair growth agents, oils, gels, shampoos, hair rinses, hair conditioners, enamels, foundations, lipsticks, face powders, packs, perfumes, powders, colognes, body soaps, soaps, bath salts, sunscreens, etc. The cosmetic or cosmetic raw material containing the cells or cell cultures of the yeast or their extracts of the present invention described above is one of the preferred embodiments in the present invention. The cosmetic and cosmetic raw material may contain components other than the cells or cell cultures of the yeast or their extracts. Various components usually formulated in cosmetics can be formulated in the cosmetic and cosmetic raw material. For example, oil components, fragrances, surfactants, moisturizers, antioxidants, ultraviolet absorbers, preservatives, pigments, dyes, etc. can be appropriately formulated. These blending ratios can be appropriately selected. The cosmetic raw material of the present invention is preferably used for manufacturing the cosmetic of the present invention. The usage and dosage of the cosmetic can be appropriately determined according to the type of the cosmetic, etc. Since the cosmetic or cosmetic raw material of the present invention contains L-hydroxyproline, it is preferably used for applications selected from, for example, promoting collagen production, promoting epidermal cell proliferation, moisturizing the skin, preventing skin aging, preventing or improving skin sagging, improving skin firmness, preventing or improving wrinkles, and improving atopic dermatitis, and is more preferably used for applications selected from improving skin firmness and preventing or improving wrinkles. The content of the cells or cell cultures of the above yeast or extracts thereof in the cosmetic is preferably 0.00001 to 10% by weight, more preferably 0.0001 to 10% by weight, still more preferably 0.0001 to 5% by weight, still more preferably 0.001 to 5% by weight, further preferably 0.01 to 3% by weight, and particularly preferably 0.05 to 2% by weight, in terms of solid content based on the cosmetic. In another preferred embodiment, the content of the cells or cell cultures of the above yeast or extracts thereof in the cosmetic is more preferably 0.00005 to 1% by weight, and still more preferably 0.0001 to 0.5% by weight, in terms of solid content based on the cosmetic. The content of the cells or cell cultures of the above yeast or extracts thereof in the cosmetic raw material is preferably 0.001 to 20% by weight, more preferably 0.01 to 10% by weight, still more preferably 0.05 to 5% by weight, and particularly preferably 0.1 to 2% by weight, in terms of solid content based on the cosmetic raw material. The L-hydroxyproline content in the cosmetic raw material is preferably 5 to 300 ppm, more preferably 10 to 200 ppm, and still more preferably 50 to 100 ppm. In one embodiment, the L-hydroxyproline content in the cosmetic can be, for example, 0.01 to 20 ppm, preferably 0.03 to 15 ppm, and more preferably 0.05 to 10 ppm. It is preferable to blend the cells or cell cultures of the above yeast or extracts thereof so that the L-hydroxyproline content is within the above range. When the composition of the present invention is a food or drink (food or drink composition), the food or drink is not particularly limited. The form of the food or drink may be liquid, semi-liquid, solid, or paste, and may be, for example, any of general foods or drinks, health foods, functional foods, etc. General foods or drinks are not particularly limited and include alcoholic beverages. Health foods refer to foods that are considered healthy or beneficial to health, and include dietary supplements, natural foods, etc. Dietary supplements refer to foods fortified with specific nutritional components. Functional foods refer to foods for supplying nutritional components that fulfill the body's regulatory functions, and include foods for specified health uses, nutritional functional foods. Examples of dietary supplements include beauty drinks, supplements, etc. The food or drink of the present invention may be in the form of pharmaceutical preparations such as capsules, or in the form of drink agents. Food and drink products can contain various ingredients that are recognized as being suitable for incorporation into food and drink products. Such ingredients include, for example, binders, thickeners, colorants, stabilizers, emulsifiers, dispersants, disintegrants, suspending agents, surfactants, preservatives, sweeteners, acidulants, and the like. Food and drink products containing the cells or cell cultures of the yeast of the present invention or extracts thereof are one of the preferred embodiments in the present invention. The content of the cells or cell cultures of the above yeast or extracts thereof in food and drink products is preferably, for example, 0.0001 to 10% by weight in terms of solid content based on the food and drink products, more preferably 0.001 to 5% by weight, and even more preferably 0.01 to 1% by weight. Also, the L-hydroxyproline content in food and drink products is preferably 0.0001 to 0.01% by weight, and it is preferable to incorporate the cells or cell cultures of the above yeast or extracts thereof so that the L-hydroxyproline content falls within the above range. The L-hydroxyproline content in compositions such as the above cosmetics, cosmetic raw materials, food and drink products, etc. is the free L-hydroxyproline content. L-Hydroxyproline preferably originates from the cells or cell cultures of the above yeast or extracts thereof. Compositions such as cosmetics, food and drink products, and raw materials thereof containing the cells or cell cultures of the yeast of the present invention or extracts thereof can be produced by selecting and blending the raw materials, additives, etc. normally used therein according to their types, incorporating the cells or cell cultures of the yeast of the present invention or extracts thereof therein, and using known methods. The present invention also includes a method for producing a cosmetic raw material containing L-hydroxyproline, which comprises a step of accumulating L-hydroxyproline in the cells or cell culture of yeast Yarrowia lipolytica by aerobic culturing the yeast in a liquid medium containing a carbon source and a nitrogen source (Hyp accumulation step). The nitrogen source is a nitrogen source containing an L-hydroxyproline-containing peptide. A preferred embodiment of the method for producing a cosmetic raw material containing L-hydroxyproline of the present invention is the same as the preferred embodiment of the method for producing L-hydroxyproline described above. The method for producing a cosmetic raw material containing L-hydroxyproline is preferred as the method for producing a cosmetic raw material of the present invention. By performing the Hyp accumulation step, cells or cell cultures of yeast containing L-hydroxyproline can be obtained. Further, by performing the above-described cell disruption treatment on the cells or cell cultures of yeast, an extract of cells or cell cultures of Yarrowia lipolytica containing L-hydroxyproline can be obtained. The cells or cell cultures of yeast or extracts thereof thus obtained, and preferred embodiments thereof, are the same as the cells or cell cultures of yeast or extracts thereof of the present invention described above, and preferred embodiments thereof. The obtained cells or cell cultures of yeast containing L-hydroxyproline or extracts thereof can be used as a cosmetic raw material containing L-hydroxyproline by blending additives and the like usually used in cosmetics, if desired. Further, an additive or the like usually used in cosmetics can be blended, if desired, with L-hydroxyproline purified from cells or cell cultures of yeast containing L-hydroxyproline or extracts thereof to produce a cosmetic raw material containing L-hydroxyproline. The cosmetic raw material containing L-hydroxyproline obtained by the present invention is suitably used in cosmetics for uses selected from promotion of collagen production, promotion of epidermal cell proliferation, skin moisturization, prevention of skin aging, prevention or improvement of skin sagging, improvement of skin firmness, prevention or improvement of wrinkles, and improvement of atopic dermatitis. The present invention also encompasses a composition for reinforcing L-hydroxyproline, which comprises cells or cell cultures of Yarrowia lipolytica containing L-hydroxyproline, or extracts thereof. The cells or cell cultures of the above yeast, or extracts thereof, contain L-hydroxyproline, and the ratio (100×Hyp / (Pro + Hyp)) of the content (μg / mL) of L-hydroxyproline to the total content (μg / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) is preferably 35 to 100. The cells or cell cultures of the above yeast, or extracts thereof, preferably have an L-hydroxyproline content of 10 μg / mL or more. The composition for reinforcing L-hydroxyproline containing such cells or cell cultures of yeast, or extracts thereof, can be particularly preferably used as an additive for reinforcing, supplementing or enhancing L-hydroxyproline in cosmetics, food and drink, etc. Preferred embodiments of the cells or cell cultures of yeast, or extracts thereof, are the same as those of the cells or cell cultures of yeast of the present invention described above, or extracts thereof, and their preferred embodiments. The composition for reinforcing L-hydroxyproline can be preferably used in food and drink, cosmetics, etc. as an additive composition for reinforcing L-hydroxyproline. The composition for reinforcing L-hydroxyproline can also be referred to as a composition for supplementing L-hydroxyproline or a composition for enhancing L-hydroxyproline. The composition for reinforcing L-hydroxyproline of the present invention only needs to contain cells or cell cultures of Yarrowia lipolytica containing L-hydroxyproline, or extracts thereof, and the content of the cells or cell cultures, or extracts thereof, may be 100% by weight, but if desired, it may also contain other components. For example, when using the composition for reinforcing L-hydroxyproline as a food additive, it may contain one or more known additives used in food. The L-hydroxyproline-reinforcing composition of the present invention is suitably used, for example, as a cosmetic additive. When the L-hydroxyproline-reinforcing composition is used as a cosmetic additive, it may contain the cells or cell cultures of Yarrowia lipolytica or extracts thereof in the composition, and the content of the cells or cell cultures or extracts thereof may be 100% by weight. Optionally, it may contain one or more known additives used in cosmetics. The method for producing the L-hydroxyproline-reinforcing composition of the present invention preferably includes a step of accumulating L-hydroxyproline in the cells or cell cultures of the above yeast by aerobically culturing Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source. The method for producing the L-hydroxyproline-reinforcing composition including such a step is also included in the present invention. The nitrogen source is a nitrogen source containing an L-hydroxyproline-containing peptide. The method for producing the L-hydroxyproline-reinforcing composition of the present invention and its preferred embodiments are the same as the method for producing L-hydroxyproline and its preferred embodiments described above. The method for producing the L-hydroxyproline-reinforcing composition of the present invention may optionally include a step of adding known food additives, cosmetic additives, etc. to the cells or cell cultures of the yeast or extracts thereof. Another aspect of the method for producing L-hydroxyproline of the present invention includes a step of accumulating L-hydroxyproline in the cells or cell cultures of the above yeast by aerobically culturing Yarrowia lipolytica in a liquid medium containing a carbon source and a nitrogen source. In another aspect of the production method of the present invention, it is preferable that the nitrogen source contains an L-hydroxyproline-containing protein or an L-hydroxyproline-containing peptide. In the production method according to another aspect of the present invention, it is preferable that the L-hydroxyproline-containing protein is a collagenous protein and the L-hydroxyproline-containing peptide is a collagen peptide. In the production method according to another aspect of the present invention, it is preferable that the average molecular weight of the collagenous protein and the collagen peptide is from 1,000 to 100,000. In the production method according to another aspect of the present invention, it is preferable that the concentration of the nitrogen source in the liquid medium is from 0.25 to 5% by weight and the weight ratio (C / N) of the carbon source (C) to the nitrogen source (N) is from 0.5 to 20. Further, it is preferable that the aerobic culture is carried out for 10 to 100 hours. Another use of the present invention is the use of Yarrowia lipolytica for producing L-hydroxyproline, which comprises accumulating L-hydroxyproline in the cells or cell culture of the yeast by aerobically culturing the yeast in a liquid medium containing a carbon source and a nitrogen source, and it is preferable that the nitrogen source contains an L-hydroxyproline-containing protein or an L-hydroxyproline-containing peptide. In another use of the present invention, it is preferable that the L-hydroxyproline-containing protein is a collagenous protein and the L-hydroxyproline-containing peptide is a collagen peptide. In another use of the present invention, it is preferable that the average molecular weight of the collagenous protein and the collagen peptide is from 1,000 to 100,000. In another use of the present invention, it is preferable that the concentration of the nitrogen source in the liquid medium is from 0.25 to 5% by weight and the weight ratio (C / N) of the carbon source (C) to the nitrogen source (N) is from 0.5 to 20. Further, it is preferable that the aerobic culture is carried out for 10 to 100 hours. Examples of the L-hydroxyproline-containing protein include the collagenous proteins described above. The average molecular weight of the L-hydroxyproline-containing protein such as collagenous protein is preferably more than 10,000 and 100,000 or less. The average molecular weight of the protein refers to the weight-average molecular weight. The average molecular weight of the L-hydroxyproline-containing protein such as collagenous protein is calculated by gel filtration or the like. Hereinafter, test examples and the like for more specifically explaining the present invention are shown. Note that the present invention is not limited only to these test examples and the like. In the test examples, unless otherwise specified, “%” means “% by weight”. In the test examples, L-4-hydroxyproline manufactured by Nacalai Tesque, Inc. was used for the preparation of L-hydroxyproline (Hyp) standard solution and the like used for the preparation of the calibration curve. L-proline manufactured by Nacalai Tesque, Inc. was used for the preparation of L-proline (Pro) standard solution and the like. Both the L-hydroxyproline and L-proline measured in the test examples are free L-hydroxyproline and L-proline. All the media used in the test examples are liquid media. The YPD medium used in the test examples was Y:P:D = 1:2:2 (weight ratio) (Y: 1.0% yeast extract, P: 2.0% peptone, D: 2.0% glucose) unless otherwise specified. The yeast extract used for medium preparation is the product named Yeast Extract (#212750) manufactured by Bacto. The peptone used was the product named Pepton (#211677) manufactured by Bacto, which is obtained by decomposing bovine cells with an enzyme derived from porcine pancreas. In the test examples, the pH of the medium before the start of culture was about 6.5 to 8.5. <Test Example 1> Screening for yeast that accumulates L-hydroxyproline (hereinafter, Hyp) Using 60 strains of yeast shown in Table 1, screening for yeast that accumulates Hyp in the culture was performed. Table 1 shows the genus names of each strain. Under the following conditions, yeast was cultured in YPD medium, and then Hyp was quantified by HPLC. (Culture) 1 mL of YPD medium supplemented with 0.5% L-proline (hereinafter referred to as Pro) was inoculated with one platinum loop of each strain and cultured with shaking at 28 °C for 24 hours (300 rpm), and then statically cultured at room temperature for 4 to 7 days to obtain a yeast cell culture solution (cell culture). (Sample Preparation) The following treatments were performed on the yeast cell culture solution to prepare a culture sample and measure the Hyp content. After the above culture, the cells were collected and washed with 1 mL of physiological saline. The cells were suspended in 0.2 mL of 50 mM potassium phosphate buffer (hereinafter referred to as KPB) (pH 6.0) and boiled for 10 minutes to elute the cell contents by autodigestion (boiling method). The cell residue was removed from the obtained cell extract by centrifugation (14,000 rpm, 5 min, 4 °C). Thus, a culture sample for measuring the Hyp accumulation amount was prepared. The culture sample was derivatized by the AccQTag method using the (AccQ-Fluor Reagent Kit) of Waters, and Hyp was quantified by HPLC under the following conditions. The apparatus and conditions used for HPLC analysis are shown below. (Apparatus) High-performance liquid chromatograph: Prominence (manufactured by Shimadzu Corporation) Column: XBridge C18 5μm (2.1 x 150 mm, manufactured by Waters) (Measurement Conditions) Eluent A: Ammonium acetate (10 mM, pH 5) Eluent B: Methanol (0 - 0.5 min. (0% → 1%), 0.5 - 18 min. (1% → 5%), 18 - 19 min. (5% → 9%), 19 - 29.5 min. (9% → 17%), 29.5 - 40 min. (17% → 60%), 40 - 43 min. (60%)) Flow rate: 0.3 mL / min Temperature: 40 °C Detection: Fluorescence detector (excitation wavelength: 250 nm, detection wavelength: 395 nm) The concentration (%) of eluent B is v / v%. The results are shown in Figure 1. Figure 1 is a graph showing the Hyp accumulation amount of yeast. The vertical axis in Figure 1 (Hyp (μg / mL)) indicates the amount of Hyp (μg) per 1 mL of the culture sample. From the above tests, yeast that accumulates Hyp was found. In particular, Yarrowia lipolytica had a large Hyp accumulation amount. <Test Example 2> Regarding the yeast Yarrowia lipolytica, it was examined whether the Hyp accumulation amount changes depending on the medium. Yarrowia lipolytica NBRC0717 was used, and the following liquid media were used. Yarrowia lipolytica NBRC0717 was obtained from the National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan). YPD (1% yeast extract, 2% peptone, 2% glucose) YTD (1% yeast extract, 2% tryptone, 2% glucose) YM (0.3% yeast extract, 0.3% malt extract, 0.5% peptone, 2% glucose) PD (0.4% potato extract, 2% glucose) SD (synthetic medium, 1% glucose) (Pre-culture and main culture) (1) Condition 1 One platinum loop of Yarrowia lipolytica NBRC0717 was inoculated into 1 mL of YPD medium supplemented with 0.5% Pro, and shake-cultured at 28°C for 24 hours (300 rpm) to obtain a pre-culture solution. 0.2 mL of the pre-culture solution was inoculated into 2 mL of each medium supplemented with 0.5% Pro (the above YPD, YTD, YM, PD, or SD), and shake-cultured at 28°C for 36 hours (300 rpm) to obtain a cell culture solution of Yarrowia lipolytica. (2) Condition 2 Pre-culture was carried out under the same conditions as in Condition 1, except that SD medium supplemented with 0.5% Pro was used instead of YPD medium supplemented with 0.5% Pro. 0.2 mL of the pre-culture solution was inoculated into 2 mL of each medium (the above-mentioned YPD, YTD, PD or SD) supplemented with 0.5% Pro, and cultured with shaking at 28 °C for 45 hours (300 rpm) to obtain a cell culture solution of Yarrowia lipolytica. (Sample preparation) After culturing under Condition 1 or 2, the following treatment was performed on the yeast cell culture solution to prepare a culture sample, and the Hyp content was measured. After the above culturing, the cells were collected and the cells were washed with 1 mL of physiological saline. The cells were suspended in 0.2 mL of 50 mM KP buffer (pH 6.0) and boiled for 10 minutes to elute the cell contents by autodigestion (boiling method). The cell residue was removed from the obtained cell extract by centrifugation (14,000 rpm, 5 min, 4 °C). Thus, a culture sample for measuring the Hyp accumulation amount was prepared. (Hyp quantification) The culture sample was derivatized with o-phthalaldehyde (OPA) and 4-chloro-7-nitrobenzofurazan (NBD-Cl), and Hyp was quantified by HPLC. (Derivatization method) Derivatization with 4-Chloro-7-nitrobenzofurazan (NBD-Cl) 50 μL of 0.4 M potassium borate buffer (pH 9.5) was dispensed, and 2 μL of the culture sample was added. 2 μL of 300 mM OPA (Wako 167-09263) (dissolved in methanol) was added and mixed. After reacting at 37 °C for 20 minutes, 50 μL of 2 mM NBD-Cl (Sigma 25455) (dissolved in methanol) was added and mixed. To the solution reacted at 37 °C for 20 minutes, 25 μL of 1 N HCl and 75 μL of 50% methanol were added, and the supernatant centrifuged at 14,000 rpm for 5 minutes was used as a sample for HPLC measurement and transferred to an HPLC vial. The apparatus and conditions used for HPLC analysis are shown below. Column: XBridge C18 column (5 μm; 2.1 mm × 150 mm; Waters) Solution A: 10 mM ammonium acetate (pH 5.0) Solution B: Methanol Gradient: 0 - 0.5 min: B.Conc 0 v / v% → 1 v / v% 0.5 - 18 min: B.Conc 1 v / v% → 5 v / v% 18 - 19 min: B.Conc 5 v / v% → 9 v / v% 19 - 29.5 min: B.Conc 9 v / v% → 17 v / v% 29.5 - 40 min: B.Conc 17 v / v% → 60 v / v% 40 - 43 min: B.Conc 60 v / v% Flow rate: 0.3 mL / min Column temperature: 40 °C Detector: Fluorescence detector, excitation wavelength 503 nm / fluorescence wavelength 541 nm Injection volume: 10 μL The calibration curve was prepared by preparing 50 mM KPB (pH 6.0) solutions of Hyp at 5 μmol / L, 10 μmol / L, 20 μmol / L, 50 μmol / L, 100 μmol / L, and 250 μmol / L. The results are shown in Fig. 2. Fig. 2 is a graph showing the Hyp accumulation amounts of Yarrowia lipolytica cultured in each medium ((a): condition 1, (b): condition 2). Hyp (μg / mL) on the vertical axis of Fig. 2 is the amount of Hyp (μg) per 1 mL of the culture sample. In the culture of condition 1, a large amount of hydroxyproline accumulated in the cells cultured in YPD and YM media (media containing peptone). In the culture of condition 2, Hyp accumulated in the cells cultured in the YPD medium. <Test Example 3> The variation in the accumulation amount depending on the number of culture days was examined. The NBRC1551 strain, NBRC0717 strain, NBRC0746 strain, and NBRC1195 strain of Yarrowia lipolytica were each cultured in YPD medium for 1 day, 2 days, or 3 days, and the Hyp eluted from the yeast cells (cells) was quantified using HPLC. These yeasts identified by the NBRC number were obtained from the National Institute of Technology and Evaluation (2 - 5 - 8 Kazusa Kamashima, Kisarazu City, Chiba Prefecture, Japan). (Pre - culture) 1 mL of YPD medium was inoculated with one loopful of each strain and cultured with shaking (300 rpm) at 28°C for 24 hours to obtain a preculture solution. (Main culture) 0.2 mL of the preculture solution was inoculated into 2 mL of YPD medium and cultured with shaking (300 rpm) at 28°C for 1 to 3 days to obtain a cell culture solution. (Sample preparation and amino acid quantification) Culture samples were prepared in the same manner as in Test Example 2 and derivatized with OPA and NBD-Cl. Analysis by HPLC was performed under the same conditions as in Test Example 2 to quantify Hyp and Pro. The calibration curve for Pro was prepared by preparing 50 mM KPB (pH 6.0) solutions of 5 μmol / L, 10 μmol / L, 20 μmol / L, 50 μmol / L, 100 μmol / L, and 250 μmol / L of Pro. The amounts of Pro and Hyp in each culture sample are shown in Table 2. Also, from the measured values of Pro and Hyp, the ratio (100×Hyp / (Pro + Hyp)) of the content of Hyp (μg / mL) to the total content of Pro and Hyp (μg / mL) was calculated. The results are also shown in Table 2. Yarrowia lipolytica accumulated L-hydroxyproline by culturing. The obtained cell culture contained L-hydroxyproline. Also, the (Hyp / (Pro + Hyp)) ratio was 35 or more. <Test Example 4> The gelatin-assimilating ability of Yarrowia lipolytica was examined. Using the NBRC1551 strain, NBRC0717 strain, and NBRC0746 strain of Yarrowia lipolytica, they were cultured in PD medium or PD medium supplemented with 0.125% gelatin for 5 days, and the cell contents were eluted from the cells and Hyp was quantified by HPLC. The PD medium used was the same medium as that used in Test Example 2. (Preculture) 1 mL of PD medium was inoculated with one loopful of each strain and cultured with shaking (300 rpm) at 28°C for 24 hours to obtain a preculture solution. (Main culture) 2 mL of PD medium or PD medium supplemented with 0.125% gelatin (manufactured by Wako Pure Chemical Industries, Ltd., product name: Gelatin) was inoculated with 0.2 mL of the preculture solution and cultured with shaking at 28°C for 5 days (300 rpm) to obtain a cell culture solution. (Sample Preparation and Hyp Quantification) The preparation of the culture sample and the quantification of Hyp were carried out in the same manner as in Test Example 2. The results are shown in Figure 3. Figure 3 is a graph showing the Hyp accumulation amount of Yarrowia lipolytica cultured in PD medium or PD medium supplemented with 0.125% gelatin for 5 days (white: PD medium containing 0.125% gelatin (PD + gel), black: PD medium). Hyp (μg / mL) on the vertical axis is the amount of Hyp (μg) per 1 mL of the culture sample. From Figure 3, when adding high molecular weight gelatin to the medium as it is, the accumulation of Hyp was less compared to Test Example 3. <Test Example 5> Using Yarrowia lipolytica NBRC0717, it was examined whether the Hyp accumulation amount changes depending on the YPD medium composition. Table 3 shows the YPD compositions of each medium from #1 to #12. In Table 3, Y is yeast extract, P is peptone, and D is glucose. The yeast extract (Y) was kept constant, and peptone (P) and glucose (D) were changed (the numerical values in the table are the final concentrations (wt%)). (Preculture) 1 platinum loop of Yarrowia lipolytica NBRC0717 was inoculated into 1 mL of each YPD medium and cultured with shaking at 28°C for 24 hours (300 rpm) to obtain a preculture solution. (Main Culture) 0.2 mL of the preculture solution was inoculated into 2 mL of each YPD medium and cultured with shaking at 28°C for 4 days (300 rpm) to obtain a cell culture solution. (Sample Preparation and Hyp Quantification) The preparation of the culture sample and the quantification of Hyp were carried out in the same manner as in Test Example 2. The yeast Yarrowia lipolytica cultured in the above YPD medium accumulated Hyp. The Hyp accumulation amount was high when cultured in the media of #1 and #7 - 11. The results when cultured in the media of #1 and #7 - 11 are shown in Figure 4. Figure 4 is a graph showing the Hyp accumulation levels of Yarrowia lipolytica cultured in YPD media with different compositions ((a): Hyp accumulation level (μg / mL), (b): Hyp amount per cell (μg / mL / OD600)). The vertical axis in (a) of Figure 4 represents the amount of Hyp (μg) per 1 mL of the culture sample. The (Hyp / OD) on the vertical axis in (b) of Figure 4 represents the amount of Hyp per cell, which is the value obtained by dividing the amount of Hyp (μg) in 1 mL of the culture sample by the measured value of OD600 (μg / mL / OD600). OD600 was measured using a nucleic acid and protein spectrophotometer (device name: Bio spec mini, manufactured by Shimadzu Corporation). 60 μL of the cell culture solution used for preparing the culture sample was diluted with 1150 μL of ultrapure water, and the absorbance (600 nm) was measured. <Test Example 6> In the YPD media #1 and #10 used in Test Example 5, the peptone used as P was replaced with collagen peptide (product name: Super Collagen Peptide SCP-2000, derived from pig, average molecular weight 2000, manufactured by Nitta Gelatin Inc.), and the Hyp accumulation level was examined. Yarrowia lipolytica NBRC0717 was used as the yeast. In the YPD media, the media in which part or all of the peptone was replaced with collagen peptide can be said to be YPD modified media. Composition of P in YPD media Normal: 100% peptone CP50: 50% peptone, 50% collagen peptide CP100: 100% collagen peptide Medium composition (1) YPD#1 - Normal (YPD#1) (2) YPD#1 - CP50 (in YPD#1, CP50 is used instead of peptone) (3) YPD#1 - CP100 (in YPD#1, CP100 is used instead of peptone) (4) YPD#10 - Normal (YPD#10) (5) YPD#10 - CP50 (in YPD#10, CP50 is used instead of peptone) (6) YPD#10 - CP100 (using CP100 instead of peptone in YPD#10) (Pre - culture) 1 platinum loop of Yarrowia lipolytica NBRC0717 was inoculated into 1 mL of YPD medium, and cultured with shaking at 28 °C for 24 hours (300 rpm) to obtain a pre - culture solution. (Main - culture) 0.1 mL of the pre - culture solution was inoculated into 2 mL of each of the media (1) - (6) above, and cultured with shaking at 28 °C for 4 days (300 rpm) to obtain a cell culture solution. (Sample preparation and Hyp quantification) The preparation of culture samples and Hyp quantification were carried out in the same manner as in Test Example 2. The results are shown in Figure 5. Figure 5 is a graph showing the Hyp accumulation amount, Hyp amount per cell, and OD600 of Yarrowia lipolytica cultured in a medium containing peptone or collagen peptide. ((a): Hyp accumulation amount (μg / mL), (b): Hyp amount per cell (μg / mL / OD600), (c): OD600). In (a) - (c) of Figure 5, N means a medium where P is normal, 50 means a medium where P is CP50, and 100 means a medium where P is CP100. Yarrowia lipolytica can grow and accumulate Hyp even in a medium where peptone is completely replaced by collagen peptide. (Test Example 7) Using Yarrowia lipolytica NBRC0717, the influence of aeration during culture was examined. The media used were YPD media #1 and #10 of Test Example 5. (Pre - culture) 1 platinum loop of Yarrowia lipolytica NBRC0717 was inoculated into 1 mL of YPD medium, and cultured with shaking at 28 °C for 24 hours (300 rpm) to obtain a pre - culture solution. (Main - culture) 0.1 mL of the pre - culture solution was inoculated into 2 mL of each YPD medium (#1 or #10), and cultured with shaking at 28 °C for 3 days (300 rpm) or statically cultured to obtain a cell culture solution. (Sample preparation and Hyp quantification) The preparation of the culture sample and the Hyp quantification were performed in the same manner as in Test Example 2. Also, the culture supernatant was derivatized in the same manner as the culture sample, and Hyp was quantified by HPLC. The results are shown in Fig. 6. Fig. 6 is a diagram showing the Hyp accumulation amount of Yarrowia lipolytica cultured with shaking or static culture ((a): Hyp amount per medium of the culture sample (intracellular), (b): Hyp amount in the culture supernatant). Yarrowia lipolytica had a large Hyp accumulation amount when aerobically cultured. <Test Example 8> The ethanol concentration and glucose concentration in the culture supernatant obtained in Test Example 7 were quantified by HPLC (using a fermentation column (manufactured by Bio-Rad, product name Aminex fermentation monitor column)). The results are shown in Fig. 7. Fig. 7 is a graph showing the ethanol concentration and glucose concentration of the culture supernatant of Yarrowia lipolytica cultured with shaking or static culture ((a): ethanol concentration (v / v%), (b): glucose concentration (wt%)). <Test Example 9> For Yarrowia lipolytica NBRC0717, main culture was performed by changing the composition of the YPD medium (the ratio of Y:P:D and the type of P) and the culture time, and the Hyp content and Pro content in the cell culture were measured. (Pre-culture) One platinum loop of Yarrowia lipolytica NBRC0717 was inoculated into 3 mL of YPD medium and statically cultured at 30 °C for 1 day to obtain a pre-culture solution. (Main culture) 100 μL of the pre-culture solution obtained above was inoculated into 5 mL of the following YPD medium and cultured with shaking (60 rpm) at 30 °C. The culture time was 1 day or 2 days to obtain a cell culture solution (cell culture). In this culture, peptone or collagen peptide (CP) was used as P in the YPD medium. As the collagen peptide, either collagen peptide Ikos HDL-50SP (product name, manufactured by Nitta Gelatin Inc., average molecular weight 5000) (hereinafter referred to as collagen peptide (CP1)) or collagen peptide Type S (product name, manufactured by Nitta Gelatin Inc., average molecular weight 1200) (hereinafter referred to as collagen peptide (CP2)) was used. Also, the ratio of Y:P:D in the YPD medium in this culture was (1) Y:P:D = 1:2:2 (weight ratio) (Y: 1.0% yeast extract, P: 2.0% peptone or collagen peptide, D: 2.0% glucose), or (2) Y:P:D = 1:4:5 (weight ratio) (Y: 1.0% yeast extract, P: 4.0% peptone or collagen peptide, D: 5.0% glucose). Table 4 shows culture conditions 1 to 12 used in this culture. The following autodigestion process and sterilization process were performed on the cell culture broth of Yarrowia lipolytica obtained in this culture to prepare a culture sample, and the Hyp content was measured. (Autodigestion process) The cell culture broth of Yarrowia lipolytica was incubated at 50°C for 2 hours to elute the cell contents into the culture broth by autodigestion. (Sterilization process) The culture broth autodigested above was incubated at 80°C for 1 hour. Cell residues were removed from the obtained extract by centrifugation (3000 rpm, 5 min, 1°C). Thereby, a culture sample for measuring the Hyp accumulation amount was prepared. (Sample preparation) The obtained culture sample was diluted with a 0.1 N hydrochloric acid solution to prepare an HPLC sample. The culture sample prepared from the cell culture broth with a culture time of 1 day was diluted 10-fold, and the culture sample prepared from the cell culture broth with a culture time of 2 days was diluted 40-fold except for condition 12, and condition 12 was diluted 20-fold. (HPLC analysis) In HPLC, a system was used in which primary amino acids (primary amino groups) and secondary amino acids (secondary amino groups) were fluorescently labeled with an autosampler and analyzed by reverse-phase HPLC. o-Phthalaldehyde (OPA) was used for the fluorescent labeling of primary amino groups, and 9-fluorenylmethyl chloroformate (FMOC) was used for the fluorescent labeling of secondary amino groups, respectively. The HPLC sample prepared above was filtered through a 0.45-μm filter into a sample vial and set in the autosampler. 30 μL of MPA (mercapto propionic acid) reagent, 15 μL of OPA reagent, and 5 μL of the above sample were placed in an empty vial, allowed to stand for 1 minute, 5 μL of FMOC reagent was added, and 1 μL of the reacted solution was injected into the HPLC after reacting for 2 minutes. OPA reacts with primary amino acids, and Hyp and Pro with remaining secondary amino groups react with FMOC. By detecting the respective fluorescence wavelengths in two channels, it is possible to simultaneously detect all amino acids. The apparatus and conditions used for HPLC analysis are shown below. (Apparatus) High Performance Liquid Chromatograph Nexera X2 System (product name) manufactured by Shimadzu Corporation System Controller: CBM-20A, Liquid Delivery Unit: LC-30AD (2 units), Degassing Unit: DGU-20A5R, Mixer: MR180μL II, Autosampler: SIL-30AC, Column Oven: CTO-20AC, Fluorescence Detector: RF-20AXS, Workstation: LabSolutions LC / GC (Measurement Conditions) Column: Inertsil ODS-4 100 mm × 3.0 mmφ S-2μm (GL Sciences Inc.) Guard Column: UHPLC Fitting (product name, GL Sciences Inc.) (Max. Pressure: 130 MPa) Mobile Phase Solution A: 15 mmol / L KH ２ PO ４ and 5 mmol / L K ２ HPO ４ (pH 6.5) Solution B: 15 / 45 / 40 (v / v / v) = water / acetonitrile / methanol R0 (rinse solution): water / methanol = 20 / 80 (v / v) R3 (rinse solution): water / acetonitrile = 80 / 20 (v / v) Initial concentration of Solution B: 10 v / v% Flow rate: 0.8 mL / min Column oven temperature: 35 °C Injection volume: 1 μL Detection: Ch1: Excitation wavelength 350 nm, fluorescence wavelength 450 nm Ch2: Excitation wavelength 266 nm, fluorescence wavelength 305 nm Cell temperature: 25 °C, Gain: ×4, Sensitivity: Medium Gradient program 0 - 1.5 min: B.Conc 10 v / v% 1.5 - 6 min: Gradient of B.Conc 10 v / v% → 30 v / v% 6 - 11 min: Gradient of B.Conc 30 v / v% → 40 v / v% 11 - 15 min: B.Conc 100 v / v% 20 - 21.5 min: B.Conc 100 v / v% → 10 v / v% 25 min: Controller stop Calibration curve: For Hyp and Pro respectively, 0.1 N HCl solutions of 6.25 μmol / L, 25 μmol / L, 50 μmol / L, and 100 μmol / L were prepared. Each of these solutions was reacted with the respective reagents of MPA, OPA, and FMOC by the above method, analyzed by HPLC, and a calibration curve was drawn. For the amino acid mixed standard solution H-type solution and the 0.1 N hydrochloric acid solution containing L-hydroxyproline, they were also reacted with the respective reagents and analyzed by HPLC in the same manner. The amino acid mixed standard solution H-type used was manufactured by Wako Pure Chemical Industries, Ltd. Figure 8 is an HPLC chart analyzing an amino acid mixed standard solution H type and a 0.1 N hydrochloric acid solution containing L-hydroxyproline (each amino acid concentration 20 μmol / L) ((a): detected at an excitation wavelength of 350 nm and a fluorescence wavelength of 450 nm for Ch1, (b): detected at an excitation wavelength of 266 nm and a fluorescence wavelength of 305 nm for Ch2). Table 5 shows the quantification results of Hyp and Pro in the culture samples. In the table, CP1 is the above-mentioned collagen peptide (CP1) (product name: Collagen Peptide Icos HDL-50SP), and CP2 is the collagen peptide (CP2) (product name: Collagen Peptide Type S). From the quantification results of Hyp and Pro, the ratio of the content of Hyp to the total content of Pro and Hyp in each culture sample (100×Hyp / (Pro + Hyp)) was calculated. The results are also shown in Table 5. The culture conditions in the table are the conditions of this culture. Note that each medium before the start of the culture contained almost no free Hyp. <Test Example 10> (Pre-culture) 1 platinum loop of Yarrowia lipolytica was inoculated into 3 mL of YPD medium and cultured with shaking at 28°C for 1 day (100 rpm) to obtain a pre-culture solution. (Main culture) The main culture was carried out in the same manner as in Test Example 9, except that a YPD modified medium (1.0% yeast extract, 2.0% collagen peptide (CP1), 1.0% glucose) was used as the medium and cultured with shaking at 28°C for 2 days (200 - 500 rpm). The collagen peptide (CP1) is the same as that in Test Example 9. The cell culture solution of Yarrowia lipolytica obtained in the main culture was subjected to an autolysis process and a sterilization process in the same manner as in Test Example 9 to prepare a culture sample. The Hyp content and Pro content were measured in the same manner as in Test Example 9. The test was performed with n = 4. The results are shown in Table 6 <Test Example 11> Using Yarrowia lipolytica employed in Test Examples 1 to 10, culturing was performed in the same manner as in Test Examples 9 to 10 to obtain a cell culture solution. By incubating this at 50 °C for 2 hours, the cell contents were eluted into the culture solution by autodigestion, and then it was incubated at 80 °C for 1 hour. Thereafter, centrifugation was carried out at 1 °C (3000 rpm, 5 min) to remove the cell residue, and an extract of the L-hydroxyproline-containing cell culture (extract of Hyp-containing cell culture) was obtained. The extract of Hyp-containing cell culture can also be used after appropriate dilution. Hereinafter, an example of a production example of a skin external preparation composition formulated with the extract of each Hyp-containing cell culture obtained in Test Example 11 is shown. <Production Example 1> Soap The blending amounts of the raw materials are shown in Table 7. The soap base was mixed and stirred, and then each extract of Hyp-containing cell culture was added and uniformly mixed, followed by molding. <Production Example 2> Shampoo The blending amounts of the raw materials are shown in Table 8. A preservative dissolved in 1,3-butylene glycol was added to purified water. After uniform stirring, sodium lauryl sulfate and coconut oil fatty acid monoethanolamide were added, and then a dye, a fragrance, and the remaining 1,3-butylene glycol were added. After adding each extract of Hyp-containing cell culture, it was uniformly mixed and stirred. <Production Example 3> Conditioner The blending amounts of the raw materials are shown in Table 9. (1) Stearyl dimethyl benzyl ammonium chloride and sodium chloride were added to purified water and heated to 80 °C to dissolve. (2) Cetostearyl alcohol, hydrogenated polyisobutene, and glycerin monostearate were heated to 80 °C to dissolve. (3) While stirring (1) with a homomixer, (2) was added, and pre-stirring was carried out for 5 minutes after the addition. (4) After completion of the pre-stirring, it was cooled while stirring to 50 °C, the extract of each Hyp-containing cell culture was added, and further stirred and cooled to 35 °C for preparation. <Production Example 4> Hair Tonic The blending amounts of the raw materials are shown in Table 10. Salicylic acid, glycerin, vitamin E dissolved in ethanol, and L-menthol were added to purified water, and dipotassium glycyrrhizinate dissolved in a part of the purified water was further added. Thereafter, the extract of each Hyp-containing bacterial cell culture was added and uniformly mixed to prepare. <Production Example 5> Mist The blending amounts of the raw materials are shown in Table 11. Citric acid and sodium citrate were added to purified water and dissolved. Thereafter, a preservative dissolved in ethanol and polysorbate 80 were added. Thereafter, the extract of each Hyp-containing bacterial cell culture was added and uniformly stirred to prepare. <Production Example 6> Lotion The blending amounts of the raw materials are shown in Table 12. Citric acid and sodium citrate were added to purified water and dissolved. Next, glycerin, 1,3-butylene glycol, and trisodium ethylenediaminetetraacetate were sequentially added, and polyoxyethylene (18) oleyl alcohol ether, vitamin E, and methylparaben dissolved in ethanol were further added and stirred until uniform. Thereafter, the extract of each Hyp-containing bacterial cell culture was added and uniformly stirred to prepare. <Production Example 7> Emulsion The blending amounts of the raw materials are shown in Table 13. (1) Stearic acid, cetyl alcohol, octyldodecyl myristate, and liquid paraffin were heated to 80 ° C and dissolved. (2) Triethanolamine, sodium hyaluronate, glycerin, 1,3-butylene glycol, polyoxyethylene (10) monooleate, and sodium ethylenediaminehydroxy triacetate were added to purified water and heated to 80 ° C. (3) While stirring (1) with a homomixer, (2) was added, and preliminary stirring was performed for 5 minutes after the addition. After completion of the preliminary stirring, it was cooled to 50°C, the extract of each Hyp-containing cell culture was added, and it was further cooled to 35°C for preparation. <Manufacturing Example 8> Cream The blending amounts of the raw materials are shown in Table 14. (1) Stearic acid, glyceryl monostearate, sorbitan sesquistearate, polyoxyethylene sorbitan monostearate, cetostearyl alcohol, squalane, hexakis(hydroxystearic acid / stearic acid / rosin acid) dipentaerythritol, olive oil, octyldodecyl myristate and methylpolysiloxane were heated to 80°C and dissolved. (2) Glycerin, 1,3-butylene glycol, sodium hydroxide and methylparaben were added to purified water and heated to 80°C for dissolution. (3) While stirring (1) with a homomixer, (2) was added, and preliminary stirring was carried out for 5 minutes after the addition. (4) After completion of the preliminary stirring, it was cooled to 50°C, the extract of each Hyp-containing cell culture was added, and it was further cooled to 35°C for preparation. The cells, cell cultures and extracts thereof of Yarrowia lipolytica containing L-hydroxyproline of the present invention are useful as raw materials for cosmetics, foods and drinks, etc.

Claims

------04 / 02 / 2019------(OCR) Page 1 of 3 pages. Claims 1. Yeast cells or cell cultures of Yarrowialipolytica or its extracts containing: L-hydroxyproline in which the ratio of the amount ((formula) g / mL) of L-hydroxyproline to the total amount ((formula) g / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) as determined by the formula (100 x Hyp / (Pro + Hyp)) is 35 to 100.

2. Yeast cells or cell cultures or its extracts as per Claims 1 in which the amount of L-hydroxyproline is 10 JLlg / mL or more.

3. Yeast cells or cell cultures or its extracts as per Claims 1 in which the amount of L-hydroxyproline is 10 JLlg / mL or more.

4. A method for the production of L-hydroxyproline consisting of the steps of: accumulation of L-hydroxyproline in yeast cells or Yarrowialipolytica cell cultures by aerobic cultivation of yeast in a liquid medium containing a carbon source and a nitrogen source, where the nitrogen source contains peptides containing L-hydroxyproline.

5. A production method according to claim 4, where the peptides containing L-hydroxyproline are collagen peptides. 6.

7. A production method under claim 5 in which collagen peptides have an average molecular weight of 1000 to 10000.

8. A production method under any of claims 4 through 6 in which aerobic cultivation is performed for 10 to 100 hours.

9. The use of Yarrowialipolytica for the production of L-hydroxyproline.

10. The use under claim 8 which includes: accumulation of L-hydroxyproline in yeast cells or cell cultures of Yarrowialipolytica by aerobic cultivation of yeast in a liquid medium containing a carbon source and a nitrogen source in which the nitrogen source contains peptides containing L-hydroxyproline.

11. The use under claim 9 in which peptides containing L-hydroxyproline are collagen peptides. (Page 2 of 3) 11. The use under claim 10 in which collagen peptides have an average molecular weight of 1000 to 10000.

12.

13. The composition includes: yeast cells or cell cultures or their extracts as per any of the claims 1 through 3. 14.Food or beverages containing: yeast cells or cell cultures or their extracts as per any one of the claims of Rights 1 through 315. Cosmetics or cosmetic raw materials containing: yeast cells or cell cultures or their extracts as per any one of the claims of Rights 1 through 316. Cosmetics or cosmetic raw materials as per Rights 15 which have been used in selected applications for the promotion of collagen production, promotion of epidermal cell growth, skin hydration, skin aging, skin firmness or reduction, improvement of skin tightening, skin firmness or reduction, and relief of atopic dermatitis.

17. Cosmetics or cosmetic raw materials as per Rights 15 or 16 where the cosmetic or cosmetic raw material is a cosmetic raw material; and the amount of L-hydroxyproline in the cosmetic raw material is 5 to 300 ppm.18.Cosmetics or cosmetic raw materials under claim 15 or 16, where cosmetics or cosmetic raw materials are cosmetics; and the amount of L-hydroxyproline in cosmetics is 0.01 to 20 ppm19. L-hydroxyproline enhancing compounds comprising: yeast cells or Yarrowial ipolytica cell cultures or their extracts containing L-hydroxyproline.

20. L-hydroxyproline enhancing compounds according to Patent 19, page 3 of page 3 of page 3, in which, in yeast cells or cell cultures or their extracts, the ratio of the amount (|g / mL) of L-hydroxyproline to the total amount (|g / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) as determined by the formula (100XHyp / (Pro+Hyp)) is 35 to 100.

21. L-hydroxyproline enhancing compounds according to Patent 19 or 20 in which the amount of L-hydroxyproline in yeast cells or cell cultures or their extracts is 10|g / mL or more. ------------ Page 1 of page 3 of Patent 1.Yeast cells or cell cultures of Yarrowialipolytica or its extracts containing: L-hydroxyproline where the ratio of the amount (chemical formula g / mL) of L-hydroxyproline to the total amount (chemical formula g / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) as determined by the formula (100xHyp / (Pro+Hyp)) is 35 to 100.

2. Yeast cells or cell cultures or its extracts according to claim 1 where the amount of L-hydroxyproline is 10 chemical formula g / mL or more.

3. Yeast cells or cell cultures of Yarrowialipolytica or its extracts containing: L-hydroxyproline.

4. A method for the production of L-hydroxyproline consisting of the following steps: accumulation of L-hydroxyproline in yeast cells or Yarrowial ipolytica cell cultures by aerobic cultivation of yeast in a liquid medium containing a carbon source and a nitrogen source, where the nitrogen source contains peptides containing L-hydroxyproline.

5. A production method according to claim 4 where the peptides containing L-hydroxyproline are collagen peptides.

6. A production method according to claim 5 where the collagen peptides have an average molecular weight of 1000 to 10000. 7.

7. The production method under any one of the claims 4 through 6 where aerobic cultivation has been performed for 10 to 100 hours.

8. The use of Yarrowialipolytica for the production of L-hydroxyproline.

9. The use under claim 8 which includes: accumulation of L-hydroxyproline in yeast cells or cell cultures of Yarrowialipolytica by aerobic cultivation of yeast in a liquid medium containing a carbon source and a nitrogen source, where the nitrogen source contains peptides containing L-hydroxyproline.

10. The use under claim 9 where the peptides containing L-hydroxyproline are collagen peptides. Page 2 of 3.

11. The use under claim 10 where the collagen peptides have an average molecular weight of 1000 to 10000.

12.

13. Use under any of the claims 9 through 11 where aerobic cultivation has been performed for 10 to 100 hours.

14. Composition consisting of: yeast cells or cell culture media or their extracts under any of the claims 1 through 3.

15. Food or beverage consisting of: yeast cells or cell culture media or their extracts under any of the claims 1 through 3.Cosmetics or cosmetic raw materials incorporating: yeast cells or cell cultures or their extracts as per any one of the claims of Claim 1 through 316. Cosmetics or cosmetic raw materials as per Claim 15 which have been used in selected beneficial applications for promoting collagen production, promoting epidermal cell growth, moisturizing the skin, delaying skin aging, preventing or reducing skin sagging, improving skin firmness, preventing or reducing wrinkles and relieving atopic dermatitis.

17. Cosmetics or cosmetic raw materials as per Claim 15 or 16 where the cosmetic or cosmetic raw material is a cosmetic raw material; and the amount of L-hydroxyproline in the cosmetic raw material is 5 to 300 ppm.

18. Cosmetics or cosmetic raw materials as per Claim 15 or 16 where the cosmetic or cosmetic raw material is a cosmetic; and the amount of L-hydroxyproline in the cosmetic raw material is 0.01 to 20 ppm. 19.L-hydroxyproline enhancing compounds comprising: yeast cells or cell cultures of Yarrowial ipolytica or its extracts containing L-hydroxyproline (page 3 of 3, page 20). L-hydroxyproline enhancing compounds according to claim 19 where in yeast cells or cell cultures or their extracts the ratio of the amount (chemical formula g / mL) of L-hydroxyproline to the total amount (chemical formula g / mL) of L-proline (Pro) and L-hydroxyproline (Hyp) as determined by the formula (100XHyp / (Pro+Hyp)) is 35 to 100.

21. L-hydroxyproline enhancing compounds according to claim 19 or 20 where the amount of L-hydroxyproline in yeast cells or cell cultures or their extracts is 10 (chemical formula g / mL or more).

Images