JPS60110285A - Method for culturing yeast with carotenoid biosynthesis ability - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for culturing yeast capable of carotenoid biosynthesis, and more specifically, the amount of carotenoids produced is enhanced by culturing yeast capable of carotenoid biosynthesis under specific conditions. Concerning how to obtain yeast.

Yeast is edited by Yoshitaka Hashitani, “Yeast Science”: pp. 3-9 (196
As described in ``December 15, 1977, 1st edition, published by Iwanami Shoten,'' most of the life is carried out in single cells such as spherical or rabbit-shaped, and the vegetative cells are used as reproductive organs, and they are not sexually active. It is a microorganism that belongs to the fungus chicken, which becomes ascus and asexually multiplies mainly by budding, and some of these microorganisms have the ability to biosynthesize yellow or red thread pigments called carotenoids within their cells. has also been found. Hereinafter, such yeast will be referred to as yeast having carotenoid biosynthesis ability.

Carotenoids, such as α-carotene, β-carotene, and r-carotene, are useful as provitamin A in yellow dyes for prize feed additives, food additives, etc., and are currently mainly produced synthetically. For example, by mixing citral and acetone, it is made into gusoidyonone.
Erycarotinide is synthesized by treating this with sulfuric acid at a low temperature to give ff'+i to obtain ionone, and then dimerizing the ionone thus obtained.

Conventionally, regarding the cultivation of yeast with carotenoid biosynthesis ability,
Examples of research aimed at promoting carotenoid biosynthesis include those related to culture medium composition [Journal of the Japanese Society of Agricultural Chemistry].
, 199 and 810 (1960)], there are also research reports on the relationship with light, and Hikari F! It has been reported that carotenoid biosynthesis is not promoted by σ injection [Journal of the Japanese Society of Agricultural Chemistry, 34.195 (1960)]; on the other hand, b
The biosynthesis of carotenoids is accelerated by irradiation with J-visual light, especially in the 400 nm to 500 nm range [Okayama University Graduate School Bulletin, Journal, 6 B-68, (197
8)] etc. have also been reported.

However, no literature has been found that reports on the relationship between f.

The present inventors have long been interested in the relationship between microorganisms and light, especially ultraviolet rays, and have continued to study the effects that appear when various microorganisms are cultured under ultraviolet irradiation. This time, completely unexpectedly, when yeast with carotenoid biosynthesis ability is cultured under irradiation with a specific ultraviolet light containing gold, carotenoid biosynthesis by the yeast becomes active, and carotenoid production is marked and enhanced. This discovery led to the completion of the present invention.

Therefore, according to the present invention, yeast having carotenoid biosynthesis ability is obtained, which is characterized in that yeast having a carotenoid biosynthesis ability of at least 280 to 400% is obtained.
A method for cultivating yeast is provided.

The carotenoid biosynthetic pathway by microorganisms has been studied in considerable detail, for example, T, F-G.
oodwin (1971), In: l5Ler, O.
, (ad), Carotinoids.

pp 577-686. Birkh; arbses
-Verlag.

In Ba5el Stwttgart, the biosynthesized acetic acid passes through mevalonic acid in the microbial cell to produce the skeleton phytoene, which is then converted into lycopene and xanthophyll through processes such as dehydrogenation, ionone formation, and oxidation. It has been reported that carotenoids form many types of carotenoids, including carotenoids and carotenes. However, the im species of carotenoid that are biosynthesized differ depending on the type of microorganism [Rho, Kozo Hayashi, ed., "Plant Pigments", pp. 57-58. (1st edition published by Yokendo on August 28, 1980)]
From yeast, α-carotene, β-carotene, γ-carotene,
The production of carotenoid pigments such as ζ-carotene, δ-carotene, neurosporene, tolulene, torralodin, β-cliftoxatin, rubixanthin, and spiryloxanthin is known.

In addition, various microorganisms other than yeast, such as photosynthetic bacteria and fungi, use visible light, especially 400 nm, for carotenoid biosynthesis.
Many studies have reported that the above blue light has no effect, and the photoreceptors are thought to be frappin protein, porphyrin pigments, and carotenoid pigments.
The photoreceptors differ depending on the type of microorganism, and a unified opinion has not yet been reached, and the details of the effects of light on carotenoid biosynthesis were unknown.

According to the research conducted by the present inventors using yeast, unlike the previous findings shown above, yeast bacteria having carotenoid biosynthesis ability can be stimulated by ultraviolet rays having a wavelength in the range of at least 280 to 400 nm. When cultured in a nutrient medium under light irradiation, β-
It was found that carotene and γ- were significantly enhanced.

In particular, the amount of tolulene and torralozin produced was significantly increased, and judging from the metabolic pathway of these products, the irradiated light of this wavelength is involved in the dehydrogenation and dispersion reaction process in the pathway from neurosporene to torralozin. However, it is thought that the progress of this metabolic process is controlled, and it is presumed that an unknown photoreceptor exists.

To which the method of the present invention can be applied, the carotenoid is not particularly limited as long as it has the ability to produce carotenoid-C1 from the carotenoid biosynthetic pathway described above, and can be selected from a wide range. Yeasts that have the ability to produce carotenoids have so far been discovered to belong to Rhodotorula maple, Rhodosporideum genus, and Sporopolomyces genus, but the method of the present invention can be applied to yeasts belonging to any of these genera. Among them, yeast having carotenoid biosynthesis ability belonging to the Rhodosporideum genus is particularly suitable. Specific examples of yeast having carotenoid biosynthesis ability to which the method of the present invention can be applied are as follows.

(1) Rhodotorula practice (i (hodotorula) example:
Rhodotorula minuta (Rh, m1tsbta) IF
O1102, Rhodotorula rubra (Rh, rubrα) IPO158
6. Rhodotorla gluti-nis
IFOO895, Rhodotorula graminis (Rh, grami-nis
J IFOO190゜Rhodotorula lactosa (Rh, 1actosα) IPo
142 B, Rhodotorula marina (Rh, marine α) IF014
82, φ Rhodotorula barida (Rh, paled α) IFOO7
15゜(2)a)” Suboriteum (Rhodospor
idium) Example: Rhodosborideum toruuroites (R
hs.

torutoides) IFOo 559, Rhodosporideum derobobum (Rhs.

diobovatum) IFol 829, Rhodosporideum infilmorminium (Rhs, in
firm, o-miniatrbm, ) IFO105'
l, (3) Sporobolo CI Myces sp.
my-ces) Example Nisboroboromyces no450 ceus C5p.

pararosews ) IFOl 104, Sporopolomycetes phorsatechus C5p.

holosaticus) IFo 1082, Sporobolomyces gracilis C5p.

gracilis) IFO122'l, Sporobolomyces odorus <Sp.

odorus) IFO1085 Sporobolomyces roseus (Sp.

roseu, s) IFo 1087, yeast cells described above, and those to which the method of the present invention can be particularly preferably applied include Rhodotorula minuku, Rhodotorula rubra, Rhodotorula glutenis, Rhodosboridium φ-toruloides, and Sporozolomyces Examples include Pararoseus.

The yeast having carotenoid biosynthesis ability as described above can be cultured using known yeast culture techniques, except that it is carried out under specific light irradiation according to the present invention. Therefore, as the nutrient medium, the medium conventionally used for culturing yeast can be similarly used, and as the carbon source, for example, shields such as glucose, maltose, sucrose, sucrose, xylose, etc.
Examples of nitrogen sources include sodium glutamate, glycine, asparagine, leucine, valine, methionine, tryptophan, and amino acids such as histidine hydrochloride; potassium nitrate, calcium nitrate, and ammonium nitrate. nitrates such as; ammonium salts such as ammonium sulfate, ammonium chloride, ammonium phosphate; urea; nucleobases such as adenine, guanine, uracil; yeast extract, pegtone,
Vinegar mother powder, corn stave liquor, meat extract, casein,
Examples include wheat gluten, NZ amine, f5 seed oil, malt extract, and the like.

In addition to the above carbon and nitrogen sources, the nutrient medium may contain inorganic salts, such as potassium dihydrogen phosphate, magnesium sulfate, sodium chloride, calcium chloride, boric acid, copper sulfate, potassium iodide, chloride, if necessary. Ferric iron, manganese sulfate, zinc sulfate, sodium molybdate and/or trace elements such as calcium pantonate, biotin, thiamine hydrochloride, p-ami7benzoic acid, #acid, inositate, niamin, riboflavin, pyridoxine hydrochloride, etc. as appropriate. May be added.

Other β-carotene precursors, mevalonic acid, etc. can also be added.

Suitable culture temperatures are generally about 25 to about 82°C, preferably about 26 to about 28°C, and
It is generally desirable to maintain the pH of the medium within the range of about 4.5 to about 6.5, preferably about 5 to about 6. Cultivation is usually carried out under aerobic conditions with stirring or shaking, and it is appropriate to blow sterilized air in an amount approximately equal to the volume of the culture medium every minute. In addition, the culture period is the period until the proliferation of bacterial cells is almost completed, for example, after inoculation of bacterial cells.
IXXX for about 8 to about 5 days, and it is preferable to continue culturing for 2 to 8 days thereafter.

The present invention is characterized in that yeast having carotenoid biosynthesis ability is cultured at a wavelength of at least 280 to 400 nm.
The light quality, irradiation timing, and irradiation intensity formula will be described in more detail below.

In this specification, the term "light" is used in a broad sense, and includes not only visible light but also ultraviolet and infrared light.

The light irradiated according to the method of the present invention is particularly limited as long as it contains ultraviolet light having a wavelength within the above wavelength range.
It is also possible to use substantially only the ultraviolet rays without l-1. The visible light that can be irradiated in combination with the ultraviolet light may be white light or a spectral light thereof, such as blue light, green light, yellow light, orange light, red light or any mixed color light thereof. However, for visible light, the wavelength is particularly 4.
Blue light in the range of 00 to 500 nm or one containing blue light is suitable.

In addition, the irradiated ultraviolet rays range from 280 tLm to 400 tLm.
It is not necessary to consist of ultraviolet rays in the entire wavelength range up to 280 nm, but it is also possible to use ultraviolet rays in a part of the wavelength range.
Those containing ultraviolet light having a wavelength within the range of ~860 nm are preferred. More preferably at least 290
Light containing ultraviolet radiation having a wavelength in the range ˜820% fi is advantageous.

The source of such light (i.e., the light source) may be either natural light or artificial light, and examples of the artificial light source include fluorescent lamps, mercury lamps, sunlight lamps, general light bulbs, floodlight bulbs, and special light bulbs (such as light bulbs). Classifications include Toshiba Lamp Catalog, Toshiba Electric Materials Co., Ltd.), etc. More specifically, the fluorescent lamp [0 indicates the product name. ], 8 daylight FLR40
5IV-E/M, Toshiba), Plantorx (FL40S
BR/N, bundle 9, Fish Lux (FL40SBR.

F/NL, Toshiba), blue (FL40B/NL, Toshiba),
White color (FL40BfF'/ML, Toshiba), white color (FL
40SD/ML, Toshiba), daylight color (FL40SD/NL
, Toshiba), Deluxe (FL40SIF'-DL-X/
ML, Toshiba), warm white (FL4oSW/ML, FL4o
SW-A/NL, Toshiba), 610,0'K for leaf tobacco
(FL 405RD-5DL6100° to 1 Toshiba),
Fluorescent lamp for health line (PL light lamp (FL20S BL-B, Matsushita Electric), fluorescent chemical lamp (FL20S-BL, Toshiba), fluorescent lamp for photography (FL40SD-5DL-CP)
/NL, i Shiba), high color rendering s fluorescent lamp (FL20SIF-E
DL-50, Toshiba), sunlight lamp (B125, B25
0, Toshiba), etc.

In particular, fluorescent lamps for health lines, fluorescent lamps for catching insects, black light blue, and fluorescent chemical lamps are preferred.

The above irradiation sources are equipped with colored glass filters, if necessary.
A combination of plastic filters, colored *lophanes, etc. may be used. Specifically, colored glass made by Toshiba Glass.
Examples of ultraviolet transmitting and visible absorbing filters include UV-D88S, UV-B85, and UV-DB6A.
%UV-B86BSUV-186C etc., UV-transmitting filter 1-Toshite UV-25, UV-29, UV-8
1. There are UV-88, UV-5s, UV-81, ultraviolet filters such as P-89A-P-89E, and cobalt blue filters such as C-89A and C-89B.

C-40A~C-4ocH actual purple and blue filter! :
F-40, r-42, r-44, B-.

46, B-47, B2S3: IK as a light blue filter
$'11 Trillion T-42A, T-42BST-44 L-505 as Nisian filter: LB-B2, LB-B4, LB-B8, LB-B1 as color temperature concealing variant filter
1, LB-B18; F as a fluorescent light color correction filter
There are L-41', CF-D, CF-B, etc.

The intensity of the ultraviolet rays within the wavelength range to be irradiated is not strictly limited, and can be used depending on the type of yeast to be cultured, the placement of the bacterial cells in the medium, etc., but in general, approximately 1 to about too, oooμW/cd1 good 1
or about 10 to about 80,000 ttW/cd, more preferably about 10 to about 20,000 ttW/cd, and most preferably about 80 to about 2,000 μF/cd. It is. It should be noted that the optimal irradiation intensity can be easily determined by those skilled in the art by repeating laboratory-scale flask shaking culture and the like for the yeast to be used and its culture conditions.

In addition, among the various irradiation wavelength range lights, the irradiation light intensity in the ultraviolet region of 280 to 400 nm is measured using, for example, a microvoltmeter (model PM-16, 4) manufactured by Toa Denpa Kogyo ■ and Kipp.
and Zonen Co., Ltd. (D, elf.

Nettrlands) I) Can be measured by combining a radiant heat sensor (model A knee 1),
In addition, the irradiation light intensity in the visible range of 400 nm or more is, for example, p
It can be measured using a spectra photometer/radiometer (model 80.1) manufactured by Hoto 5osearch (U, S, A, .).

The light irradiation method as described above is not particularly limited and may be any method as long as the cultured yeast cells are sufficiently irradiated with ultraviolet rays within the wavelength range mentioned above. Methods such as those listed can be used.

Artificial light irradiation methods include a method in which a conventional fermentation tank is used as is, and a light source is inserted into the space between the rotary blade and the inner wall of the fermentation tank for irradiation; a method in which the light source is also used as a barrier plate for the fermentation tank, etc. is possible. In addition, when installing a light source outside the fermentation tank, it is possible to irradiate a part of the fermentation tank wall by making it into a glass tank that transmits ultraviolet rays, and also to remove a part of the culture lees from the fermentation tank and place it outside the tank. Another possible method is to circulate the irradiation to the fermentation tank through a glass tube or tank equipped with a light source.

On the other hand, methods of using sunlight as a light source include cultivating in a glass fermentation tank installed outdoors using sunlight f, or combining this method with the above artificial irradiation method to irradiate artificial light at night. Another possible method is to install a solar concentrator outdoors and introduce sunlight into the fermentation tank using glass fiber or the like to irradiate it.

When irradiating the light containing ultraviolet radiation as described above according to the invention, the irradiation can be started at any stage of the logarithmic growth phase of the yeast to be cultured, advantageously starting from the middle of the logarithmic growth phase or even after the logarithmic growth phase. It is preferable to start irradiation late in the growth phase.

Until the irradiation with light is started, it is desirable that the yeast be cultured in a substantially dark state where it is not irradiated with light as much as possible.
Here, "logarithmic growth phase" is "Yeast Science" 192, edited by Yoshitaka Otoya.
198 (first edition published by Iwanami Shoten, December 15, 1967), it refers to a specific growth phase in a series of growth processes of yeast cells grown by the budding method in a culture system. That is, cells grown in a new culture system usually have a period of stagnant proliferation, followed by a lag phase during which they prepare for division, then gradually accelerate, and then undergo exponential division or budding and doubling. This is called the total logarithmic growth phase. Thereafter, proliferation stops and enters a stationary phase in which the number of proliferating cells becomes constant. Eventually, the number of bacterial cells decreases due to the depletion of nutrients in the system, leading to a death stage.

Note that the "middle phase" of the logarithmic growth phase refers to the period from immediately after the growth reaches the logarithmic growth phase until it enters the stationary phase. It means the period immediately before entering the steady period after the middle period, that is, the deceleration period.

Irradiation may be carried out continuously from the above-mentioned starting point, or may be carried out intermittently depending on the case. When carried out intermittently, the light (irradiation)/dark (non-irradiation) cycle includes, for example, a cycle of 12 hours of irradiation followed by 12 hours of darkness;
Preferably, a cycle of 6 hours of irradiation and 6 hours of darkness, more preferably a cycle of 2 hours of irradiation and 2 hours of darkness, and particularly preferably a cycle of 1 hour of irradiation and 1 hour of darkness.

Further, the irradiation can be carried out for at least 2 to 8 days from the above-mentioned irradiation start time, preferably for at least 2 to 5 days, and more preferably until the end of breeding.

According to the method of the present invention described above, it is possible to obtain and recover yeast cells with a significantly increased production amount of carotenoids compared to the cultivation in the dark that is normally used in the conventional fermentation industry. can.

The carotenoids synthesized within the yeast cells cultured by the method of the present invention include α-carotene, β-2 carotene, γ-carotene, δ-carotene, toluene, torralodin, and carotenoid acid, and the amount of production thereof Although the production ratio differs depending on the type of yeast, one thing that can be said in common is that, among these carotenoids, the amount of tolulene and carotenoid acid produced is significantly greater (usually about 2 to 17 times more) than when cultured in the dark. ).

After the yeast cultured by the method of the present invention is separated and recovered from the culture medium using a conventional method, it can be used as a live cell or dried as a coloring agent, feed additive, medicinal yeast, yeast extract, biocatalyst, etc. be able to.

Alternatively, if necessary, the yeast may be further processed, e.g.
After the yeast cells are crushed with a cell crusher such as a crusher, a ball mill, an ultrasonic crusher, or a compressor, they may be used for the above-mentioned purposes, and carotenoids can also be extracted and purified therefrom. Extraction and purification of carotenoids can be carried out using methods known per se.
229 pages (published by Yokendo on August 28, 1981), T, W
.

Goodwin, Chemistry and Bio
che-mistry of plant pigme
nts, Acad.

press (1g76), Chromatogr,
Rev.

This can be carried out by the method described in literature such as No. 1 Sat., 188-29B (1971). Specifically, for example,
Extraction can be performed by extracting the crushed bacterial cells using an organic solvent, such as acetone, methanol, ethanol, benzene, ether, or a mixture thereof; 1. The extract can be concentrated by distilling off the solvent under reduced pressure, etc. The carotenoid concentrate thus obtained may be saponified using caustic soda or aqueous caustic acid solution (concentration: about 5%), if necessary.

Isolation and purification of carotenoids from the carotenoid concentrate is described, for example, in Kozo Hayashi, ed., "Botanical Color Thread", pages 205-216 (1976 edition, August 28, No. 1', published by Yokendo). Alternatively, it can be carried out by means such as adsorption column chromatography, thin layer chromatography, filter paper chromatography, and countercurrent distribution method.

The carotenoids thus obtained can be used as dyes, colorants,
It can be used for food additives, feed additives, animal drugs, medical products, etc.

Next, the present invention will be explained in more detail with reference to Examples.

Examples 1-8 and Comparative Examples 1-8 Glucose 80JV, L-parin L2iXKH.

P ()41 EXKg S 04 ・4B, 011/
SMn SO+ 4H, 00, 18F! , NaC
L O,1y, Fe C1a・6H,00,01,9
, Thiamine hydrochloride 0.05 J? , p-aminobenzoic acid 0
．． 01 g and distilled water 17! An aqueous solution consisting of (pH &
Prepare 6), dispense 100 g of this aqueous solution into a 500-volume shake flask (Pyrex), and
After autoclaving at 0°C for 18 minutes, the rhodosporidium torroides (Rhodosporidium, tor.
utoides, IF00559) was inoculated with one platinum loop.
Preculture was performed for 80 hours under dark conditions at 27° C. with a shaking frequency of 200 times/min.

Prepare 1,200 ml of water bath solution having exactly the same composition as the above aqueous solution used for pre-culture, and add 50 ml of the water bath solution to 500 mL of water bath solution.
Dispense into 24 shake flasks (made of Pyrex glass) and sterilize at 110°C for 18 minutes.
1 was inoculated and cultured in the dark for about 48 hours at a culture temperature of 27° C. and a shaking frequency of 200 plates/min.

When the number of viable bacterial cells reached 7 x 10'/- and the growth of bacterial cells reached the late logarithmic growth phase, the cells were continuously irradiated with various light beams shown in Table 1 below at an irradiation intensity of approximately 800 μW/cd for 5 days. Culture was performed while

Carotenoid oxygen was extracted and quantified by the method described in "Plant Pigments" edited by Kozo Hayashi, pages 205-229 (first edition published by Yokendo, August 28, 1972).

That is, the vacuum freeze-dried bacterial cells were mixed with quartz sand and ground, and then extracted several times with an acetone-methanol mixture (7:2). Extraction [--combined in a nitrogen stream for 45 minutes
It was dried under reduced pressure at °C, dissolved in about 5 ml of n-hexane, and further divided into two layers by adding 10% NaCL.
It was used as a hexane electrolytic solution for quantitative determination of carotenoids.

The wavelength and 11% value of the lowest absorption used for quantification are c
IrL was 486 nm and 8240, and was calculated by converting the total rotinoid pigment amount per dry bacterial weight as drenin equivalent. As for drenine and carotenoid acids (mainly containing torralodin), a part of the above n-hexane electrolyte solution was isolated and purified by thin layer chromatography (thin layer chromatography: Merck & Co., Ltd., product number 5626).

Drenin uses a wavelength with a maximum absorption of 486 nm and an El cm number of 8240, and carotenoid acids have a maximum absorption of 5101.
It was quantified using a wavelength of m and a 51% value of 1982.

a The culture results are shown in Table 2 below. Note that the numerical values for each test group are the average values obtained from four shake flasks.

Examples 4 to 6 and Comparative Examples 4 to 5 An aqueous solution G having exactly the same composition as the aqueous solution used in Examples 1 to 8 was prepared, and 100 - of this aqueous solution was dispensed into a 500 ml photographic flask. After autoclaving for 13 min at °C, Rhodotorula mista (Rhodo-1 orrbl°a
m1nrbta, IFO1102) was inoculated with a platinum loop,
The cells were precultured for 40 hours at 27° C. under dark conditions with shaking 200 times/min.

Prepare a water bath solution with exactly the same composition as that used for pre-culture, dispense 50-mL of the aqueous solution into 20 500-volume shaker flasks (11).
After sterilization at 0° C. for 18 minutes, the cells were cultured in the dark for about 48 hours under the above-mentioned conditions of 200 times/min.

When the number of viable bacterial cells reached 5 x 10''/- and the growth of bacterial cells reached the late logarithmic growth phase, they were continuously irradiated with various light beams shown in Table 1 above at a light intensity of approximately 300 μW/crit for 5 days. Culture was performed while

Extraction and quantification of carotenoid colored threads were carried out in the above-mentioned Examples 1-
This was done using method 3. The culture results are shown in Table 8 below. Note that the numerical values for each test plot are the average values obtained from four shake flasks.

Table 3 Examples 7 to 9 and Comparative Examples 6 to 7 Rhodotorula rubra (Rh, rubra
, IFD1536) was cultured. The number of viable bacteria is 4.
In the late logarithmic growth phase when the axiO"/g is reached, the various light beams shown in Table 1 above are applied at an intensity of about 300 μm/crA for 5
It was continuously irradiated for days. The culture results are shown in Table 4 below.

Table-4 Examples 10-12 and Comparative Examples 8-9 Glucose 30y1 Asparagine 2.5N, KE.

Po, 1,! v, MgSO4-4H201El, Mn
5O, 4H, OO, 19, Fe C13-GH200
, 019, thiamine hydrochloride o, osg, p-ami7benzoic acid (lit O, 01, y.

Biotin 0. OO5,! i/, sodium pantothenate o, o s, lit, pyridoxine hydrochloride o, osg
, nicotinamide o, osy, riboflavin 0.00
An aqueous solution consisting of 2.9 and distilled water 11 was adjusted to pH 5.6.
This aqueous solution (100°C) was dispensed into a 500° shake flask (made of Pyrex glass) and heated at 100°C.
After autoclaving for 18 minutes, Sporobolomyses pararose
One platinum loop of ro-8eulJ, I7''01104) was inoculated, and preculture was performed for 80 hours under dark culture conditions at 25° C. at a pregnancy rate of 200 times/min.

Prepare 1 aqueous solution having exactly the same composition as the water bath used for preculture, dispense 50 ml of the aqueous solution into 20 500-volume shake flasks (made of Pyrex glass),
After sterilizing at 100°C for 18 minutes, inoculate 5 of the above pre-cultured bacterial powder suspension (number of viable bacteria: 3.9xto'/*),
Dark culture was carried out for about 88 hours at a culture temperature of 27° C. and a shaking frequency of 200 times/min. Number of viable bacteria is &8X10'
/m in the middle of the logarithmic growth phase, the cells were continuously irradiated with the various light beams shown in Cloth-1 for 5 days at a light intensity of about 800 μW/cd.

Extraction and quantification of hand-made dyes were carried out in Examples 1 to 8 above.
It was done using the same method. The culture results are shown in Table 5 below.

Table-5

Claims (1)

[Scope of Claims] 1. An enzyme having carotenoid biosynthesis ability is cultured in a nutrient medium under irradiation with light containing ultraviolet light having a wavelength in the range of at least 280 to 400 nm, and the amount of carotenoid produced from the culture is 1. A method for culturing yeast possessing carotenoid biosynthetic ability, the method comprising obtaining yeast with enhanced carotenoid biosynthesis ability. 2. The method according to claim 1, wherein the irradiation with light is started at any point in the logarithmic growth phase of the yeast. & The method according to claim 1, wherein the light irradiation is started in the latter half of the logarithmic growth phase of the yeast. 4. The method of claim 1, wherein the light contains ultraviolet radiation having a wavelength in the range of at least 280-860 nm. 5. The method according to claim 1, wherein the yeast is a yeast that belongs to the genus Rhodotoral, Rhodosporideum, or Sporopolomyces and has the ability to biosynthesize carotenoids. 6. The intensity of the irradiated ultraviolet rays is approximately 1 on the medium surface.
~100,000μF/c! The method of claim 1 within the range t. +2