WO2010058558A1 - アラニン高含有酵母の製造方法 - Google Patents
アラニン高含有酵母の製造方法 Download PDFInfo
- Publication number
- WO2010058558A1 WO2010058558A1 PCT/JP2009/006165 JP2009006165W WO2010058558A1 WO 2010058558 A1 WO2010058558 A1 WO 2010058558A1 JP 2009006165 W JP2009006165 W JP 2009006165W WO 2010058558 A1 WO2010058558 A1 WO 2010058558A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- yeast
- alanine
- rich
- extract
- rich yeast
- Prior art date
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/20—Synthetic spices, flavouring agents or condiments
- A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/14—Yeasts or derivatives thereof
- A23L33/145—Extracts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
Definitions
- the present invention relates to a method for producing alanine-rich yeast, alanine-rich yeast, alanine-rich yeast extract, seasoning composition, and food and drink containing alanine-rich yeast extract.
- Patent Document 1 describes a yeast extract characterized in that the content of free amino acids is 25% by weight or more and the total content of nucleic acid-based taste-imparting components is 2% by weight or more.
- Patent Document 2 discloses that the total free amino acid content in the yeast extract is 3.0% or more, the alanine content in the total free amino acid content is 10% or more, the glutamic acid content is 25% or more, and A yeast extract composition derived from a yeast belonging to Candida tropicalis, Candida lipolytica or Candida utilis having a histidine content of 10% or more is described.
- JP 2007-49989 A Japanese Patent No. 3519572
- Patent Document 1 Many amino acids exhibit astringency, but it can be expected that the astringency of the yeast extract can be suppressed by adjusting the alanine content in the yeast extract.
- Patent Document 2 the operation is complicated such as using an enzyme.
- Patent Document 2 is inferior in safety, palatability, and the like as a food because yeast that has been subjected to gene mutation treatment is used in addition to complicated operations such as the use of enzymes.
- the present invention has been made in view of the above circumstances, a method for producing alanine-rich yeast containing alanine at a higher concentration than before, alanine-rich yeast, alanine-rich yeast extract, seasoning composition, and It aims at providing the alanine containing food / beverage products.
- alanine is contained in yeast by raising the culture solution to a specific pH (shifting to an alkaline region) during the cultivation of yeast in the stationary phase. We found that the amount increased. And it discovered that a yeast extract with a high alanine content could be manufactured by manufacturing a yeast extract using this yeast, and completed this invention. That is, the present invention adopts the following configuration.
- a method for producing yeast having a high alanine content comprising subjecting a yeast in a stationary phase to liquid culture under conditions where the pH of the liquid medium is 7.5 or more and less than 11.
- the liquid culturing step includes a step of adjusting the pH of a yeast liquid medium in a stationary growth phase to 7.5 or more and less than 11, and a step of further culturing the yeast within the pH range.
- an alanine-rich yeast with a markedly increased alanine content can be produced simply by shifting the pH of the liquid medium of the stationary phase yeast to alkali. .
- An alanine-rich yeast extract containing alanine at a high concentration can be obtained by performing extraction from the alanine-rich yeast of the present invention.
- FIG. 1 shows an increase curve of the number of bacteria with respect to culture time in Example 1.
- FIG. 2 shows an increase curve of dry yeast cell weight with respect to culture time in Example 1.
- FIG. 3 shows the change in pH of the liquid medium with respect to the culture time in Example 1.
- the method for producing a high alanine yeast according to the present invention is characterized in that liquid culture is performed under a condition that the pH of a liquid medium of yeast in a stationary phase of growth is 7.5 or more and less than 11.
- the yeast may be a unicellular fungus, and specifically, Saccharomyces spp., Shizosaccharomyces spp., Pichia spp., Candida spp., Kluyveromyces spp., Williopsis spp., Debaryomyces spp., Galactomyces spp., Torulaspora spp. And bacteria belonging to the genus Zygosaccharomyces.
- Candida tropicalis, Candida lipolytica, Candida utilis, Candida sake, and Saccharomyces cerevisiae are edible.
- Saccharomyces cerevisiae and the like are preferable, and Saccharomyces cerevisiae and Candida utilis that are widely used are more preferable.
- the pH of the yeast liquid medium in the stationary stationary phase is 7.5 or more. What is necessary is just to carry out liquid culture on the conditions which are less than 11.
- the culture medium composition of these strains is not particularly limited, and those used in conventional methods can be used.
- one or more selected from the group consisting of glucose, sucrose, acetic acid, ethanol, molasses, sulfite pulp waste liquid and the like used for normal microorganism culture as a carbon source is used, and as a nitrogen source, urea
- one or more selected from the group consisting of inorganic salts such as ammonia, ammonium sulfate, ammonium chloride or ammonium phosphate, and nitrogen-containing organic substances such as corn steep liquor (CSL), casein, yeast extract or peptone used.
- CSL corn steep liquor
- casein casein
- yeast extract or peptone used.
- phosphoric acid component, potassium component, and magnesium component may be added to the medium.
- the raw material can be used.
- inorganic salts such as zinc, copper, manganese, and iron ions may be used.
- vitamins and nucleic acid-related substances may be added.
- the culture format may be batch culture, fed-batch culture or continuous culture, but industrially fed-batch culture or continuous culture is employed.
- Culture conditions in the logarithmic growth phase or culture conditions before pH adjustment may be in accordance with general yeast culture conditions.
- the temperature is 20 to 40 ° C., preferably 25 to 35 ° C., and the pH is 3.5 to 7.5, especially 4.0 to 6.0 is desirable.
- it is preferable that it is aerobic conditions.
- it is preferable to culture while aeration and stirring.
- the amount of aeration and the conditions for stirring can be appropriately determined in consideration of the culture volume and time, and the initial concentration of bacteria.
- the ventilation is 0.2-2V. V. M.M. (Volume per volume per minute) and stirring can be performed at about 50 to 800 rpm.
- the method of liquid culture under the condition that the pH of the liquid medium of yeast in the stationary phase of growth is 7.5 or more and less than 11 is not particularly limited.
- the pH of the liquid medium may be adjusted to 7.5 or more and less than 11, and urea or the like is added to the medium in advance, so that the pH naturally falls to 7.5 or more and less than 11 as the culture time passes.
- the liquid medium may be alkali-shifted.
- the amount of urea or the like added to the medium is not particularly limited, but is preferably about 0.5 to 5% with respect to the medium, although it depends on the cell concentration of the yeast to be cultured.
- the method of adjusting the pH of the liquid medium to 7.5 or more and less than 11 when the cultured yeast enters the stationary phase is not particularly limited, and for example, an alkaline component is appropriately added to adjust the pH of the liquid medium. May be adjusted to 7.5 or more and less than 11, preferably 7.5 or more and 10 or less.
- the pH adjustment may be performed at any time during the stationary phase, but is preferably performed immediately after entering the stationary phase. This is because it is possible to sufficiently increase the alanine concentration in the yeast and to reduce the time required until the end of the entire process.
- the pH of the yeast liquid medium in the logarithmic growth phase is 7.5 or more and less than 11 because the yeast growth is suppressed and the free alanine content does not increase.
- the alkaline component is not particularly limited, and examples thereof include the following components; NH 4 OH (ammonia water), ammonia gas, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, and the like.
- NH 4 OH ammonia water
- ammonia gas sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, and the like.
- Inorganic alkalis, alkaline bases such as sodium carbonate and potassium carbonate, organic alkalis such as urea, and the like. Of these, ammonia water, ammonia gas, and urea are preferred.
- the temperature and other conditions for culturing yeast in a stationary phase in a liquid medium having a pH of 7.5 or more and less than 11 may be in accordance with general yeast culture conditions.
- the temperature is preferably 25 to 35 ° C.
- the alanine content in the yeast after the pH is shifted to 7.5 or more and less than 11 tends to decrease with the passage of the culture time after reaching the peak. This also depends on conditions such as the cell concentration, pH and temperature of the yeast to be cultured. This is presumably because the influence of alkali on the yeast becomes too great if the culture is carried out for an excessively long time under alkaline conditions. Therefore, in the present invention, an optimal culture time can be appropriately selected for each culture condition, particularly for each pH after alkali shift.
- the alanine content is 2.0% by weight or more per dry weight, preferably 2.0 to 12.0% by weight. Is obtained.
- “alanine content per dry weight of yeast extract” means the proportion (% by weight) of alanine contained in the solid content obtained by drying the yeast extract.
- alanine content for example, it can be measured by the Accutag Ultra (AccQ-Tag Ultra) labeling method using an Acquity UPLC analyzer manufactured by Waters (USA).
- the calibration curve may be created using, for example, an amino acid mixed standard solution H type (manufactured by Wako Pure Chemical Industries, Ltd.).
- an amino acid automatic analyzer JLC-500 / V type manufactured by JEOL Ltd. it is not particularly limited.
- a yeast extract rich in free alanine which is a good taste component, can be easily obtained by extracting and producing a yeast extract from the yeast. Since the yeast extract thus obtained has a high free alanine content, the astringency peculiar to amino acids is suppressed, and the yeast extract is a better yeast extract.
- the present invention can produce alanine-rich yeast by a simple process using only an alkali shift of a liquid medium. Further, as described above, it is not necessary to use a special medium as the medium, and it can be manufactured from inexpensive raw materials such as ammonia.
- a high alanine-containing yeast containing a high concentration of alanine in the yeast is obtained, but a fraction containing alanine may be obtained from the alanine-rich yeast.
- a method for fractionating a fraction containing alanine from alanine-rich yeast any method may be used as long as it is a commonly used method.
- an alanine-rich yeast extract can be produced from the alanine-rich yeast cultured by the above method.
- a method for producing alanine-rich yeast extract any method can be used as long as it is a conventional method. Autolysis method, enzymatic decomposition method, acid decomposition method, alkaline extraction method, hot water extraction method Etc. are adopted.
- the alanine in the yeast extract obtained only by the hot water extraction method is considered to be almost entirely free alanine, unlike the yeast extract obtained by an enzyme reaction method such as an autolysis method.
- the alanine-rich yeast of the present invention has a large amount of free amino acids. Therefore, even if the yeast extract is extracted only by hot water treatment, a yeast extract with good taste can be obtained. Conventionally, in order to increase the content of tasty amino acids such as free alanine, it has been common to perform hydrolysis treatment using acids, alkalis and the like using plant and animal proteins. However, there is a problem that the hydrolyzed product of protein contains MCP (chloropropanols) suspected to be carcinogenic.
- MCP chloropropanols
- the high alanine-containing yeast produced by the method of the present invention has a high free amino acid content in the first place, after the yeast is extracted by a hot water method or the like, it is decomposed or treated with an acid or alkali. Even if not performed, a yeast extract having a sufficiently high free alanine content can be prepared. That is, by using the high alanine-containing yeast of the present invention, a yeast extract excellent in both taste and safety can be easily produced.
- the yeast extract obtained according to the present invention has a very high taste and can be used in foods and drinks, etc., and can produce foods and drinks with a rich taste.
- alanine-rich yeast extract powder can be obtained by powdering the alanine-rich yeast extract of the present invention, and yeast containing 5.0% by weight or more of alanine per dry weight can be obtained by appropriately selecting yeast. Extract powder is obtained.
- dry yeast cells may be prepared from the alanine-rich yeast cultured by the above method.
- any method for preparing dry yeast cells any method can be used as long as it is a usual method, but industrially, freeze-drying method, spray-drying method, drum-drying method and the like are adopted.
- the alanine-rich yeast of the present invention may be used as a seasoning composition.
- the seasoning composition may consist of only the yeast extract of the present invention, and contains other components such as a stabilizer and a preservative in addition to the yeast extract of the present invention. May be.
- the seasoning composition can be appropriately used for various foods and drinks as in the case of other seasoning compositions.
- the present invention relates to a high alanine-containing yeast obtained by the above method, and a food or drink containing the alanine-rich yeast extract extracted from the alanine-rich yeast.
- a food or drink containing alanine at a high concentration can be efficiently produced.
- These foods and drinks may be any foods and drinks that can normally be added with dry yeast, yeast extract, and seasoning compositions containing these, for example, alcoholic beverages, soft drinks, fermented foods, seasonings, soups. , Breads and confectionery.
- a preparation obtained from the above alanine-rich yeast or a fraction containing alanine-rich yeast may be added in the production process of the food or drink.
- alanine-rich yeast may be used as it is as a raw material.
- Yeast Sacharomyces cerevisiae AB9813 strain was cultured by the method described in ⁇ 1> to ⁇ 8> below, and extract extraction and alanine analysis were performed from the yeast culture solution.
- pH shift an alkaline region
- pH shift NH 4 OH water (10%)
- FIG. 1 shows an increasing curve of the number of bacteria with respect to culture time.
- FIG. 2 shows an increase curve of dry yeast cell weight with respect to culture time.
- FIG. 3 shows the change of the pH of the culture solution with respect to the culture time.
- the increase in the number of bacteria ⁇ 10 6 cells / ml
- the dry yeast cell weight g / L
- the pH of the culture solution was measured, as shown in FIG. 3, after entering the stationary phase of growth, the pH shifted to alkali (7.5 or more and less than 11).
- ⁇ 8> Measuring method of alanine content The alanine content per each dry yeast cell weight of the yeast before pH shift and the yeast after pH shift was measured. Specifically, the alanine content was measured by the Accutag Ultra (AccQ-Tag Ultra) labeling method using an Acquity UPLC analyzer manufactured by Waters (USA). In this measurement method, free alanine in a sample can be selectively quantified. A calibration curve was prepared using an amino acid mixed standard solution H type (manufactured by Wako Pure Chemical Industries, Ltd.). The measurement results are shown in Table 1.
- the alanine content (wt%) in the dry cell weight was increased from 1.4 wt% to 2.2 wt% due to pH shift.
- the alanine content (% by weight) per dry weight was increased from 5.7% to 8.9% by weight. That is, it was confirmed that a yeast extract having a high alanine content can be obtained by preparing a yeast extract from alanine-rich yeast produced using the production method of the present invention.
- Example 3 shows the alanine content (% by weight) per dry weight of each yeast extract obtained as a result of the measurement.
- the yeast extract of the present invention has significantly more alanine, which is an amino acid having a taste, and this yeast extract is suitable as a seasoning.
- miso soup and consomme soup were prepared using yeast extract powder (derived from Saccharomyces cerevisiae) obtained by pulverizing yeast extract produced from yeast (pH 9.0) prepared in the same manner as in Example 1.
- the compounding quantity of the yeast extract with respect to miso soup and consomme soup is 0.2%.
- miso soup and consomme soup were prepared in the same manner using Mist Powder N (manufactured by Asahi Food and Health Co., Ltd.), and sensory evaluation was performed by the following method.
- miso soup had a difference in the average value of salty taste and umami, and was significantly different in richness.
- results of Table 5 in the consomme soup, there was a difference in the average value of saltiness and richness, and there was a significant difference in umami. This is considered because the alanine content of the yeast extract of the present invention is significantly higher than before.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was Saccharomyces cerevisiae ABS1 strain, and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 5 shows the measured values of alanine content before and after the pH shift.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was Saccharomyces cerevisiae ABS2 strain, and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 6 shows measured values of alanine content before and after the pH shift.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was Saccharomyces cerevisiae ABS3 strain, and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 6 shows measured values of alanine content before and after the pH shift.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was camellia (bakery yeast), and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 6 shows measured values of alanine content before and after the pH shift.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was Candida utilis ABC1 strain, and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 6 shows measured values of alanine content before and after the pH shift.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was Candida utilis ABC2 strain, and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 6 shows the measured values of alanine content before and after the pH shift.
- the yeast was cultured in the same manner as described in Example 1 except that the yeast to be cultured was Candida utilis ABC3 strain, and extract extraction and alanine analysis were performed from the yeast culture solution.
- Table 6 shows measured values of alanine content before and after the pH shift.
- the yeast is cultivated in the same manner as in Example 1 except that the yeast to be cultured is Saccharomyces cerevisiae ABS4 and the pH shift is set at a pH of 7.0 to 9.5.
- the extract was extracted and analyzed for alanine.
- Table 6 shows measured values of alanine content before and after the pH shift.
- the yeast having a high concentration of alanine in the microbial cells can be obtained by the method for producing alanine-rich yeast of the present invention, it can be used in the food field such as the production of yeast extract.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Biotechnology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Botany (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Seasonings (AREA)
- Seeds, Soups, And Other Foods (AREA)
Abstract
Description
グルタミン酸は、従来からグルタミン酸ナトリウムが化学調味料などとして普及しているが、近年は、グルタミン酸だけでなくその他のアミノ酸をも豊富に含む酵母エキスが、呈味性のみならず、健康上の点からも好まれている。
また、特許文献2には、酵母エキス中の全遊離アミノ酸含有量が3.0%以上であり、全遊離アミノ酸含有量中のアラニン含有量が10%以上、グルタミン酸含有量が25%以上、かつヒスチジン含有量が10%以上であるキャンディダ トロピカリス、キャンディダ リポリティカ又はキャンディダ ユーティリスに属する酵母由来の酵母エキス組成物が記載されている。
特許文献1については、酵素を使用するなど操作が煩雑である。
また、特許文献2については、酵素を使用するなど操作が煩雑であるのに加え、遺伝子変異処理を行なった酵母を用いているため、食品としての安全性、嗜好性等に劣る。
[2] 前記の液体培養する工程が、増殖の定常期にある酵母の液体培地のpHを7.5以上11未満に調整する工程、前記酵母を前記pHの範囲内でさらに培養する工程を含む、前記[1]に記載のアラニン高含有酵母の製造方法。
[3] 前記酵母がサッカロマイセス・セレビシエ(Saccharomyces cerevisiae)、キャンディダ・ユティリス(Candida utilis)である前記[1]又は[2]に記載のアラニン高含有酵母の製造方法。
[4] 前記[1]~[3]の何れか1項に記載のアラニン高含有酵母の製造方法によって得られたアラニン高含有酵母。
[5] 前記[4]に記載のアラニン高含有酵母から抽出されたアラニン高含有酵母エキス。
[6] 前記[5]に記載のアラニン高含有酵母エキスを含有することを特徴とする調味料組成物。
[7] 前記[4]に記載のアラニン高含有酵母、前記[5]に記載のアラニン高含有酵母エキス、又は前記[6]に記載の調味料組成物を含有することを特徴とするアラニン含有飲食品。
以下に、本発明の実施形態について詳細に説明する。
酵母としては、単細胞性の真菌類であればよく、具体的には、サッカロマイセス(Saccharomyces)属菌、シゾサッカロマイセス(Shizosaccharomyces)属菌、ピキア(Pichia)属菌、キャンディダ(Candida)属菌、クリベロマイセス(Kluyveromyces)属菌、ウィリオプシス(Williopsis)属菌、デバリオマイセス(Debaryomyces)属菌、ガラクトマイセス(Galactomyces)属菌、トルラスポラ(Torulaspora)属菌、ロドトルラ(Rhodotorula)属菌、ヤロウィア(Yarrowia)属菌、ジゴサッカロマイセス(Zygosaccharomyces)属菌などが挙げられる。
これらの中でも、可食性であることから、キャンディダ・トロピカリス(Candidatropicalis)、キャンディダ・リポリティカ(Candida lypolitica)、キャンディダ・ユティリス(Candida utilis)、キャンディダ・サケ(Candida sake)、サッカロマイセス・セレビシエ(Saccharomyces cerevisiae)などが好ましく、より好ましくは汎用されているサッカロマイセス・セレビシエ、キャンディダ・ユティリスである。
また、通気・攪拌を行ないながら培養することが好ましい。通気の量と攪拌の条件は、培養の容量と時間、菌の初発濃度を考慮して、適宜決定することができる。例えば、通気は0.2~2V.V.M.(Volume per volume per minuts)程度、攪拌は50~800rpm程度で行なうことができる。
培地に添加する尿素などの量は、特に限定されるものではなく、培養する酵母の菌体濃度にもよるが、培地に対して0.5~5%程度が好ましい。
pH調整は、定常期であればいつ行なってもよいが、定常期に入った直後に行なうことが好ましい。酵母内のアラニン濃度を十分に高めることが可能である上に、全工程終了時までに要する時間を短縮することができるためである。なお、対数増殖期にある酵母の液体培地のpHを7.5以上11未満にすると、酵母の増殖が抑制され遊離アラニン含有量が増加しないため好ましくない。
上記のうち、アンモニア水、アンモニアガス、尿素が好ましい。
なお、本発明において、「酵母エキスの乾燥重量当たりのアラニン含有量」酵母エキスを乾燥させて得られる固形分中に含まれるアラニンの割合(重量%)を意味する。
また、日本電子社製アミノ酸自動分析装置JLC―500/V型などを用いて測定することも可能であるが、特に限定されるものではない。
アラニン高含有酵母からアラニンを含有する分画物を分画する方法としては、通常行われている方法であればいずれの方法でもよい。
従来、遊離アラニン等の呈味性アミノ酸の含有量を高めるために、植物性、動物性タンパク質を用いて、酸やアルカリ等を用いた加水分解処理が行われることが一般的であった。しかしながら、タンパク質の加水分解処理物は、発ガン性の疑いのあるMCP(クロロプロパノール類)を含む、という問題がある。
これに対して、本発明の方法により製造された高アラニン含有酵母は、そもそも遊離アミノ酸含有量が高いため、該酵母を熱水方法等により抽出した後、酸やアルカリ等による分解処理や酵素処理を行わずとも、遊離アラニン含有量が十分に高い酵母エキスを調製することができる。すなわち、本発明の高アラニン含有酵母を用いることにより、呈味性と安全性の両方に優れた酵母エキスを、簡便に製造することができる。
以下の組成からなる培地を、容量350mL(2Lバッフル付き三角フラスコ)で2本作製した。
(培地組成)
糖蜜 8%
尿素 0.6%
(NH4)2SO4 0.16%(硫酸アンモニウム)(NH4)2HPO4 0.08%(リン酸水素2アンモニウム)
(1)糖蜜(糖度36%)167mlをミリQ水にて750mlにメスアップ後、2Lバッフル付き三角フラスコに350mlずつ分注した。
(2)オートクレーブ処理(121℃、15min)を行なった。
(3)使用時に糖蜜のみの培地に無菌的に窒素成分混液(×100)を1/50量添加(各7mL)した。
培養温度 30℃
振とう 160rpm(ロータリー)
培養時間 24h
(植菌量 300mL)
以下の組成からなる培地を、容量2000mL(流加終了時3Lの設定)作製した。
(培地組成)
塩化アンモニウム 0.18%(流加終了時3L換算)5.3g
(NH4)2HPO4 0.04%(リン酸水素2アンモニウム、流加終了換算)1.2g
(培養条件)
培養温度 30℃
通気 3L/min
撹拌 600rpm
pH制御 下限制御pH5.0(10%アンモニア水にて)、上限制御なし
消泡剤 アデカネート原液
流加培地 糖蜜(糖度36%)、容量800mL(1Lメジウム瓶にて、最終8%)
次に、培養した酵母が定常期に入った直後に、NH4OH水(10%)にて培養液のpHをアルカリ性域にシフト(以下、pHシフトという。)させて(設定pH9.0)、更に酵母を培養した。本培養開始後48時間で終了した。
図1に示すように、菌数(×106cells/ml)の増加は、培養18時間後には定常状態に達し、増殖の定常期に入ったことが確認された。また、乾燥酵母菌体重量(g/L)も培養後24時間後にはほぼ定常状態になっており、増殖の定常期であることが確認された。培養液のpHを測定したところ、図3に示すように、増殖の定常期に入った後に、pHがアルカリ(7.5以上11未満)にシフトした。
(1)酵母を本培養した培養液を50mlプラスチック遠心チューブ(ファルコン2070)へ移し、遠心分離(3,000g、20℃、5min、HP―26)を行なった。
(2)上清を捨て、ペレットをミリQ水20mlに懸濁し、遠心分離(3,000g、20℃、5min、HP―26)を行なった。これを2回繰り返した。
(3)上清を捨て、ペレットをミリQ水20mlに懸濁した。
あらかじめ秤量しておいたアルミ皿(直径5cm)に、酵母懸濁液2mlとり、105℃にて4時間乾燥させた。
乾燥後の重量(酵母乾燥後重量)を測定し、以下の式(1)により固形分の重量(乾燥酵母菌体重量、単位g/L)を算出した。
酵母乾燥後重量 - アルミ皿重量 = 乾燥酵母菌体重量 ・・・(1)
(1)残りの酵母懸濁液(約18ml)を遠心分離(3,000g、20℃、5min、HP―26)した。
(2)残りの懸濁液1.5mlをエッペンドルフチューブに移して、チューブをブロックヒーターに移し、80℃にて30分加熱した(エキス化)。または、温浴中100℃にて10分間過熱してもよい(エキス化)。
(3)その後、遠心分離(6,000g、4℃、5min)にて上清液(エキス溶液)を分離した。
エキス溶液500μlをアルミ皿にとり、105℃、4時間乾燥させた。その後、エキス化前の乾燥重量から、エキス重量比(w/w)を算出した。
pHシフト前の酵母及びpHシフト後の酵母の、それぞれの乾燥酵母菌体重量あたりのアラニン含有量を測定した。具体的には、アラニン含有量は、(米国)ウォーターズ社製Acquity UPLC分析装置を用いて、アキュタグウルトラ(AccQ-Tag Ultra)ラベル化法により測定した。該測定法では、試料中の遊離アラニンを選択的に定量することができる。また、検量線は、アミノ酸混合標準液H型(和光純薬社製)を用いて作成した。測定結果を表1に示す。
以上の結果から、定常期後に7.5以上11未満にpH調整してさらに培養を行なうことによって、酵母中のアラニンが増加することが示された。
なお、前述したように、本実施例では、試料中の遊離アラニンを測定しており、よって、表1に示す含有量は、遊離アラニンの含有量である。
比較例として、ミーストパウダーN(アサヒフードアンドヘルス株式会社製)を用い、同様にみそ汁とコンソメスープを作製し、以下の方法で官能評価を行なった。
専門パネラー10名によるブラインド2点比較により、比較官能検査を実施した。2対比較テストとして、t-検定を行なった。
(評価基準)
塩味(減塩効果)、旨味、コクの3項目について、基準のみそ汁または基準となるコンソメスープを0とし、以下のように5段階で評価した。
「強い」=+2、
「やや強い」=+1、
「どちらでもない」=0、
「やや弱い」=-1、
「弱い」=-2。
みそ汁の結果を表4に示し、コンソメスープの結果を表5に示す。
(実施例番号:シフト前->シフト後)
(実施例 5:4.05->10.03)、
(実施例 6:1.17-> 5.99)、
(実施例 7:1.55-> 6.03)、
(実施例 8:0.96-> 4.20)、
(実施例 9:1.05->10.10)、
(実施例10:0.54-> 3.98)、
(実施例11:0.61-> 8.46)。
Claims (7)
- 増殖の定常期にある酵母を、液体培地のpHが7.5以上11未満である条件下で液体培養する工程を含む、アラニン高含有酵母の製造方法。
- 前記の液体培養する工程が、
増殖の定常期にある酵母の液体培地のpHを7.5以上11未満に調整する工程;及び
当該酵母を当該pHの範囲内でさらに培養する工程;
を含む、請求項1に記載のアラニン高含有酵母の製造方法。 - 前記酵母がサッカロマイセス・セレビシエ(Saccharomyces cerevisiae)、キャンディダ・ユティリス(Candida utilis)である請求項1又は2に記載のアラニン高含有酵母の製造方法。
- 請求項1~3の何れか1項に記載のアラニン高含有酵母の製造方法によって得られたアラニン高含有酵母。
- 請求項4に記載のアラニン高含有酵母から抽出されたアラニン高含有酵母エキス。
- 請求項5に記載のアラニン高含有酵母エキスを含有することを特徴とする調味料組成物。
- 請求項4に記載のアラニン高含有酵母、請求項5に記載のアラニン高含有酵母エキス、又は請求項6に記載の調味料組成物を含有することを特徴とするアラニン含有飲食品。
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2010539141A JP5693231B2 (ja) | 2008-11-18 | 2009-11-17 | アラニン高含有酵母の製造方法 |
BRPI0921010A BRPI0921010B1 (pt) | 2008-11-18 | 2009-11-17 | método para produzir uma levedura tendo uma quantidade de alanina de 2,0 a 12,0% em peso em uma base de peso seco de um extrato de levedura |
AU2009318675A AU2009318675B2 (en) | 2008-11-18 | 2009-11-17 | Method for producing alanine-rich yeast |
CN2009801454850A CN102216443A (zh) | 2008-11-18 | 2009-11-17 | 产生富含丙氨酸的酵母的方法 |
US13/128,683 US20110223288A1 (en) | 2008-11-18 | 2009-11-17 | Method for producing alanine-rich yeast |
EP09827339A EP2351829A4 (en) | 2008-11-18 | 2009-11-17 | METHOD FOR THE PRODUCTION OF ALAN-rich YEAST |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008-294643 | 2008-11-18 | ||
JP2008294643 | 2008-11-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2010058558A1 true WO2010058558A1 (ja) | 2010-05-27 |
Family
ID=42198001
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2009/006165 WO2010058558A1 (ja) | 2008-11-18 | 2009-11-17 | アラニン高含有酵母の製造方法 |
Country Status (7)
Country | Link |
---|---|
US (1) | US20110223288A1 (ja) |
EP (1) | EP2351829A4 (ja) |
JP (1) | JP5693231B2 (ja) |
CN (1) | CN102216443A (ja) |
AU (1) | AU2009318675B2 (ja) |
BR (1) | BRPI0921010B1 (ja) |
WO (1) | WO2010058558A1 (ja) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011145525A1 (ja) * | 2010-05-17 | 2011-11-24 | アサヒビール株式会社 | アラニン高含有調味料組成物 |
JP2012140348A (ja) * | 2010-12-28 | 2012-07-26 | Maruzen Pharmaceut Co Ltd | 抗炎症剤、抗老化剤、及び育毛剤、並びに化粧料 |
JP2015216888A (ja) * | 2014-05-19 | 2015-12-07 | キリン株式会社 | 醸造酵母の培養方法および培地 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5431076B1 (ja) * | 1973-09-07 | 1979-10-04 | ||
JPS60133892A (ja) * | 1983-10-31 | 1985-07-17 | ジエネツクス・コ−ポレイシヨン | L−フエニルアラニンの製造方法 |
JPS63123390A (ja) * | 1986-11-10 | 1988-05-27 | Idemitsu Kosan Co Ltd | L−フエニルアラニンの製造方法 |
JPH09313169A (ja) * | 1996-05-31 | 1997-12-09 | Ajinomoto Co Inc | 酵母エキスの製造法 |
JP3519572B2 (ja) | 1997-05-27 | 2004-04-19 | 日本たばこ産業株式会社 | 酵母エキス組成物およびそれを得るための酵母変異株 |
JP2007049989A (ja) | 2005-07-20 | 2007-03-01 | Nippon Paper Chemicals Co Ltd | 酵母エキス及びその製造方法 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS4725B1 (ja) * | 1968-01-24 | 1972-01-05 | ||
US3616234A (en) * | 1969-08-11 | 1971-10-26 | Ajinomoto Kk | Method of preparing protease from candida lipolytica |
US3888839A (en) * | 1972-11-29 | 1975-06-10 | Anheuser Busch | Isolated yeast protein product with intact rna and a process for making same |
US3914450A (en) * | 1973-04-09 | 1975-10-21 | Anheuser Busch | Concentrated extract of yeast and processes of making same |
JPS63112965A (ja) * | 1986-06-09 | 1988-05-18 | Takeda Chem Ind Ltd | 酵母エキスの製造法 |
ATE346909T1 (de) * | 1997-09-29 | 2006-12-15 | Japan Tobacco Inc | Hefeextraktzusammensetzung, hefe zur herstellung derselben und verfahren zur herstellung einer hefeextraktzusammensetzung |
JP2006246791A (ja) * | 2005-03-10 | 2006-09-21 | Toray Ind Inc | D−アラニンの製造法 |
-
2009
- 2009-11-17 AU AU2009318675A patent/AU2009318675B2/en active Active
- 2009-11-17 JP JP2010539141A patent/JP5693231B2/ja active Active
- 2009-11-17 US US13/128,683 patent/US20110223288A1/en not_active Abandoned
- 2009-11-17 WO PCT/JP2009/006165 patent/WO2010058558A1/ja active Application Filing
- 2009-11-17 CN CN2009801454850A patent/CN102216443A/zh active Pending
- 2009-11-17 BR BRPI0921010A patent/BRPI0921010B1/pt active IP Right Grant
- 2009-11-17 EP EP09827339A patent/EP2351829A4/en not_active Withdrawn
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5431076B1 (ja) * | 1973-09-07 | 1979-10-04 | ||
JPS60133892A (ja) * | 1983-10-31 | 1985-07-17 | ジエネツクス・コ−ポレイシヨン | L−フエニルアラニンの製造方法 |
JPS63123390A (ja) * | 1986-11-10 | 1988-05-27 | Idemitsu Kosan Co Ltd | L−フエニルアラニンの製造方法 |
JPH09313169A (ja) * | 1996-05-31 | 1997-12-09 | Ajinomoto Co Inc | 酵母エキスの製造法 |
JP3519572B2 (ja) | 1997-05-27 | 2004-04-19 | 日本たばこ産業株式会社 | 酵母エキス組成物およびそれを得るための酵母変異株 |
JP2007049989A (ja) | 2005-07-20 | 2007-03-01 | Nippon Paper Chemicals Co Ltd | 酵母エキス及びその製造方法 |
Non-Patent Citations (1)
Title |
---|
See also references of EP2351829A4 |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011145525A1 (ja) * | 2010-05-17 | 2011-11-24 | アサヒビール株式会社 | アラニン高含有調味料組成物 |
JP2012140348A (ja) * | 2010-12-28 | 2012-07-26 | Maruzen Pharmaceut Co Ltd | 抗炎症剤、抗老化剤、及び育毛剤、並びに化粧料 |
JP2015216888A (ja) * | 2014-05-19 | 2015-12-07 | キリン株式会社 | 醸造酵母の培養方法および培地 |
Also Published As
Publication number | Publication date |
---|---|
US20110223288A1 (en) | 2011-09-15 |
AU2009318675A1 (en) | 2010-05-27 |
JP5693231B2 (ja) | 2015-04-01 |
EP2351829A1 (en) | 2011-08-03 |
CN102216443A (zh) | 2011-10-12 |
EP2351829A4 (en) | 2012-09-05 |
BRPI0921010A2 (pt) | 2015-08-18 |
AU2009318675B2 (en) | 2013-01-10 |
JPWO2010058558A1 (ja) | 2012-04-19 |
BRPI0921010B1 (pt) | 2020-01-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5717478B2 (ja) | 酵母 | |
JP5730579B2 (ja) | アミノ酸高含有酵母の製造方法 | |
JP5693231B2 (ja) | アラニン高含有酵母の製造方法 | |
JP6008505B2 (ja) | Gaba高含有酵母の製造方法 | |
JP2015154788A (ja) | コク味を増強する調味料の製造方法 | |
WO2015141531A1 (ja) | 酵母エキスの製造方法 | |
WO2012067106A1 (ja) | 酵母エキスの製造方法 | |
WO2010058527A1 (ja) | グルタミン酸高含有酵母の製造方法 | |
EP2572591B1 (en) | Alanine-rich seasoning composition |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 200980145485.0 Country of ref document: CN |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 09827339 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2010539141 Country of ref document: JP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2009318675 Country of ref document: AU |
|
REEP | Request for entry into the european phase |
Ref document number: 2009827339 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2009827339 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 13128683 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2009318675 Country of ref document: AU Date of ref document: 20091117 Kind code of ref document: A |
|
ENP | Entry into the national phase |
Ref document number: PI0921010 Country of ref document: BR Kind code of ref document: A2 Effective date: 20110513 |