JP4777702B2 - Method for producing taurine-rich yeast and method for adjusting taurine content - Google Patents
Method for producing taurine-rich yeast and method for adjusting taurine content Download PDFInfo
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- 244000144972 livestock Species 0.000 claims description 4
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Description
本発明は、タウリンを高濃度に含有する酵母及びその製造法に関する。より詳細には、培養条件を適宜設定することにより、任意のタウリン含量を有する酵母を製造する方法、及びそれにより得られる酵母に関する。 The present invention relates to a yeast containing a high concentration of taurine and a method for producing the same. More specifically, the present invention relates to a method for producing a yeast having an arbitrary taurine content by appropriately setting culture conditions, and a yeast obtained thereby.
近年、人間も含む陸上哺乳類から魚等の水棲動物まで、タウリンの重要性、特に健康面における重要性が明らかにされつつある。タウリンとはアミノ酸の一種で、体内でメチオニンやシステインなどの含硫アミノ酸から作られ、生命維持に必須な心臓、脳、網膜に多く存在する。タウリンには細胞内浸透圧調節因子という、細胞に栄養素や情報を取り入れ、細胞膜を安定させ、細胞の機能を維持する役割がある。 In recent years, the importance of taurine, particularly in terms of health, has been clarified from land mammals including humans to aquatic animals such as fish. Taurine is a type of amino acid that is made in the body from sulfur-containing amino acids such as methionine and cysteine, and is found in many in the heart, brain, and retina essential for life support. Taurine has a role called an intracellular osmotic pressure regulator that takes nutrients and information into cells, stabilizes cell membranes, and maintains cell functions.
タウリンは、疲労やストレス下に置かれた状態では欠乏しがちになるため、食餌により補給する必要がある。現在最も安価なタウリン源として合成タウリンが候補として考えられるが、合成タウリンは一部の医薬品に使用が認められているのみで、一般の食品や飼料安全法に規定されている配合飼料への使用は認められていない。 Taurine tends to be deficient when placed under fatigue or stress and must be supplemented by diet. Synthetic taurine is currently considered as a candidate for the cheapest taurine source, but synthetic taurine is only approved for use in some pharmaceuticals, but it is used for mixed foods stipulated in general food and feed safety laws. Is not allowed.
上記背景技術に鑑み、本発明では、一般の食品や配合飼料への使用が可能な程に安全なタウリン源、及びその製造方法を提供することである。 In view of the above background art, the present invention is to provide a taurine source that is safe enough to be used for general foods and blended feeds, and a method for producing the same.
発明者による鋭意研究の結果、酵母の処理液中にタウリンを含有させ、その処理液中で一定の条件下で酵母を処理すると、酵母がタウリンを菌体内に効率的に取込むこと、特に、酵母を栄養培地ではなくむしろ任意のタウリン濃度の処理液中で非増殖的に酵母を嫌気的並びに好気的に攪拌することにより、酵母菌体中に効率的にタウリンが取込まれることを見出し、本発明を完成させるに至った。 As a result of intensive research by the inventor, when taurine is contained in the yeast treatment solution and the yeast is treated under certain conditions in the treatment solution, the yeast efficiently incorporates taurine into the cells, We found that taurine is efficiently incorporated into yeast cells by aerobically and aerobically stirring the yeast non-proliferatively in a treatment solution with an arbitrary taurine concentration rather than a nutrient medium. The present invention has been completed.
上記課題を解決するための手段は、以下の通りである。
＜１＞タウリン含量が０.１〜１２％（乾物重量）の範囲であることを特徴とする、タウリン高含有酵母。
＜２＞タウリンを含有する溶液中に酵母を懸濁し、処理することを特徴とする、タウリン高含有酵母の製造法。
＜３＞タウリンが、合成タウリン、畜産物由来タウリン、水産物由来タウリン、畜産物エキス又は海産物エキスのいずれか一つ若しくは二つ以上に由来することを特徴とする、上記＜２＞に記載の製造法。
＜４＞溶液中のタウリン含量が０．１〜９％（w/v）であることを特徴とする、上記＜２＞又は＜３＞のいずれかに記載の製造法。
＜５＞処理時間が３〜６０時間であることを特徴とする、上記＜２＞〜＜４＞のいずれかに記載の製造法。
＜６＞処理温度が２５〜４０℃であることを特徴とする、上記＜２＞〜＜５＞のいずれかに記載の製造法。
＜７＞溶液のpHが３〜７であることを特徴とする、上記＜２＞〜＜６＞のいずれかに記載の製造法。
＜８＞タウリンを含有する溶液が、非栄養的であることを特徴とする、上記＜２＞〜＜７＞に記載の製造法。
＜９＞処理が、非増殖的な攪拌又は振とうであることを特徴とする、上記＜２＞〜＜８＞に記載の製造法。
＜１０＞酵母を懸濁する溶液に含まれるのタウリン濃度を調節することにより、得られるタウリン高含有酵母中のタウリン含量を調節することを特徴とする、上記＜２＞〜＜９＞のいずれかに記載の製造法。
＜１１＞タウリン高含有酵母中のタウリン含量が０.１〜１２％（乾物重量）の範囲であることを特徴とする、上記＜２＞〜＜１０＞のいずれかに記載の製造法。
Means for solving the above problems are as follows.
<1> Taurine-rich yeast, wherein the taurine content is in the range of 0.1 to 12% (dry matter weight).
<2> A method for producing a high taurine-containing yeast, wherein the yeast is suspended and treated in a solution containing taurine.
<3> The production according to <2>, wherein the taurine is derived from any one or more of synthetic taurine, livestock-derived taurine, marine product-derived taurine, livestock product extract, or marine product extract. Law.
<4> The method according to any one of <2> or <3> above, wherein the taurine content in the solution is 0.1 to 9% (w / v).
<5> The process according to any one of <2> to <4>, wherein the treatment time is 3 to 60 hours.
<6> The process according to any one of <2> to <5>, wherein the treatment temperature is 25 to 40 ° C.
<7> The production method according to any one of <2> to <6>, wherein the solution has a pH of 3 to 7.
<8> The method according to <2> to <7>, wherein the solution containing taurine is non-nutritive.
<9> The process according to <2> to <8> above, wherein the treatment is non-proliferative stirring or shaking.
<10> Any one of <2> to <9> above, wherein the taurine content in the high taurine-containing yeast is adjusted by adjusting the taurine concentration contained in the solution in which the yeast is suspended. The manufacturing method of crab.
<11> The method according to any one of <2> to <10> above, wherein the taurine content in the yeast having a high taurine content is in the range of 0.1 to 12% (dry matter weight).
本発明のタウリン高含有酵母の製造法においては、処理条件を適宜調節することにより任意のタウリン含量を有する酵母を製造することが可能となる。 In the method for producing a high taurine-containing yeast of the present invention, a yeast having an arbitrary taurine content can be produced by appropriately adjusting the treatment conditions.
本発明において、タウリン高含有酵母とは、０．１〜１２％（乾物重量）の範囲でタウリンを含有する酵母のことを指す。本発明におけるタウリン高含有酵母は、タウリンを含有する溶液に酵母を懸濁、攪拌及び／又は振とうし、酵母にタウリンを取り込ませることにより調製される。本発明タウリン含有酵母の基となる酵母としては、特に限定はされないが食用酵母であるのが好ましい。食用酵母としては、特に制限はなく公知のものの中から選択することができ、パン酵母、ビール酵母、ワイン酵母、清酒酵母及び味噌醤油酵母から選択されるのが好ましく、パン酵母であるのが特に好ましい。食用酵母の菌株としては、例えばサッカロミセス（Ｓａｃｃｈａｒｏｍｙｃｅｓ）属、トルロプシス（Ｔｏｒｕｌｏｐｓｉｓ）属、ミコトルラ（Ｍｙｃｏｔｏｒｕｌａ）属、トルラスポラ（Ｔｏｒｕｌｏｐｓｉｓ）属、キャンディダ（Ｃａｎｄｉｄａ）属、ロードトルラ（Ｒｈｏｄｏｔｏｒｕｌａ）属、ピキア（Ｐｉｃｈｉａ）属などが挙げられる。 In the present invention, a high taurine-containing yeast refers to a yeast containing taurine within a range of 0.1 to 12% (dry matter weight). The yeast with a high taurine content in the present invention is prepared by suspending, stirring and / or shaking yeast in a solution containing taurine, and allowing the yeast to incorporate taurine. Although it does not specifically limit as yeast used as the base of this invention taurine containing yeast, It is preferable that it is an edible yeast. The edible yeast is not particularly limited and can be selected from known ones, preferably selected from baker's yeast, brewer's yeast, wine yeast, sake yeast and miso soy sauce yeast, particularly baker's yeast. preferable. The strain of edible yeast, for example Saccharomyces (Saccharomyces) genus Torulopsis (Torulopsis) genus Mikotorura (Mycotorula) genus Torulaspora (Torulopsis) genus Candida (Candida) genus Rhodotorula (Rhodotorula) genus Pichia (Pichia) sp Etc.
また食用酵母の菌株の具体例としては、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｃｅｒｅｖｉｓｉａｅ、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｃａｒｌｓｂｅｒｇｅｎｓｉｓ、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｕｖａｒｕｍ、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｒｏｕｘｉｉ、Ｔｏｒｕｌｏｐｓｉｓ ｕｔｉｌｉｓ、Ｔｏｒｕｌｏｐｓｉｓ ｃａｎｄｉｄａ、Ｍｙｃｏｔｏｒｕｌａ ｊａｐｏｎｉｃａ、Ｍｙｃｏｔｏｒｕｌａ ｌｉｐｏｌｙｔｉｃａ、Ｔｏｒｕｌａｓｐｏｒａ ｄｅｌｂｒｕｅｃｋｉｉ、Ｔｏｒｕｌｏｐｓｉｓ ｆｅｒｍｅｎｔａｔｉ、Ｃａｎｄｉｄａ ｓａｋｅ、Ｃａｎｄｉｄａ ｔｒｏｐｉｃａｌｉｓ、Ｃａｎｄｉｄａ ｕｔｉｌｉｓ、Ｈａｎｓｅｎｕｌａ ａｎｏｍａｌａ、Ｈａｎｓｅｎｕｌａ ｓｕａｖｅｏｌｅｎｓ、Ｓａｃｃｈａｒｏｍｙｃｏｐｓｉｓ ｆｉｂｌｉｇｅｒａ、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｌｉｐｏｌｙｔｉｃａ、Ｒｈｏｄｏｔｏｒｕｌａ ｒｕｂｒａ、Ｐｉｃｈｉａ ｆａｒｉｎｏｓａ、などが挙げられる。これらの中でも、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｃｅｒｅｖｉｓｉａｅ、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｃａｒｌｓｂｅｒｇｅｎｓｉｓが好ましく、Ｓａｃｃｈａｒｏｍｙｃｅｓ ｃｅｒｅｖｉｓｉａｅ が特に好ましい。 Specific examples of strains of edible yeast also, Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Saccharomyces uvarum, Saccharomyces rouxii, Torulopsis utilis, Torulopsis candida, Mycotorula japonica, Mycotorula lipolytica, Torulaspora delbrueckii, Torulopsis fermentati, Candida sake, Candida tropicalis, Candida utilis, Hansenula anomala , Hansenula suaveolens , Saccharo mycopsis fibrigera , Saccharomyces lipolytica , Rhodotorula rubra , Pichia farinosa , and the like. Among these, Saccharomyces cerevisiae and Saccharomyces carlsbergensis are preferable, and Saccharomyces cerevisiae is particularly preferable.
本発明において使用するタウリンとしては、医薬品グレードの合成タウリン、畜産物由来タウリン、水産物由来タウリンをはじめ、タウリンを高含有する畜産物エキス、海産物エキスでも使用可能である。本発明の用途に使用しうるタウリン原料としては、例えば豚内臓エキス、オキアミエキス、あるいはイカエキス等が挙げられる。 As taurine used in the present invention, pharmaceutical grade synthetic taurine, animal product-derived taurine, marine product-derived taurine, animal product extracts and marine product extracts containing a high amount of taurine can be used. Examples of the taurine raw material that can be used in the application of the present invention include pork viscera extract, krill extract, squid extract and the like.
本発明のタウリン高含有酵母の製造を実施するための処理としては、タウリンを添加した栄養培地で酵母を増殖的に培養しても差し支えないが、非栄養的なタウリン溶液中で酵母を懸濁せしめ、非増殖的に攪拌及び／又は振とう処理するのが望ましい。なお、本発明において「非栄養的」とは、培地中に酵母の増殖に必要な栄養成分のいずれかが欠けている状態であることを意味する。また、本発明において「非増殖的」とは、培地中に酵母の増殖に必須な栄養成分のいずれかが欠けている為、酵母が増殖出来ない状態であることを意味する。 As a treatment for carrying out the production of the taurine-rich yeast of the present invention, the yeast may be cultured in a nutrient medium supplemented with taurine, but the yeast is suspended in a non-nutritive taurine solution. It is desirable to squeeze and agitate and / or shake non-proliferatively. In the present invention, “non-nutritive” means that any nutrient component necessary for the growth of yeast is lacking in the medium. Further, in the present invention, “non-proliferative” means that the yeast cannot grow because any nutrient component essential for the growth of the yeast is lacking in the medium.
また、酵母をタウリン溶液により処理する場合、最初に一定量のタウリン溶液を調製してその中で酵母を処理するバッチ式の処理方法、あるいはタウリン溶液を少量ずつ流下させて処理する流加式の処理方法等、いずれの処理法によっても本発明を実施することが可能であるが、望ましくはバッチ式の処理方法である。 In addition, when treating yeast with a taurine solution, a batch type treatment method in which a fixed amount of taurine solution is first prepared and the yeast is treated therein, or a fed-batch type method in which the taurine solution is flowed down little by little. Although the present invention can be carried out by any processing method such as a processing method, it is preferably a batch processing method.
タウリン溶液により酵母を処理する時間としては、３〜６０時間、好ましくは３５〜５５時間、更に望ましくは４５〜５０時間である。なお、３時間未満の余りに短時間ではタウリンが酵母菌体内に十分に取り込まれず、また５０時間以上ではその時間の増加に見合うだけの効果が見られなくなる。 The time for treating the yeast with the taurine solution is 3 to 60 hours, preferably 35 to 55 hours, and more preferably 45 to 50 hours. It should be noted that taurine is not sufficiently taken into the yeast cells in a too short time of less than 3 hours, and that the effect sufficient for the increase in the time is not seen in 50 hours or more.
タウリン溶液により酵母を処理する温度としては、２５〜４０℃の範囲が適用可能であるが、望ましくは３０〜３７℃、より望ましくは３５℃である。なお、２５℃未満ではタウリンが菌体内に効率的に取り込まれず、一方４０℃以上では酵母菌体の自己消化が起こり、望ましくない。 The temperature at which the yeast is treated with the taurine solution can be in the range of 25-40 ° C, preferably 30-37 ° C, more preferably 35 ° C. In addition, if it is less than 25 degreeC, a taurine will not be efficiently taken in into a microbial cell, On the other hand, self-digestion of a yeast microbial cell will occur at 40 degreeC or more, and is undesirable.
タウリン溶液により酵母を処理する際のpHは、３〜８の範囲が適用可能であるが、望ましくは５〜７、より望ましくは５である。なお、pHが３以下あるいは８以上の条件になると酵母菌体にとってストレスとなり、効率的にタウリンを菌体内に取り込まなくなるため、望ましくない。 The pH at the time of treating yeast with a taurine solution can be in the range of 3 to 8, preferably 5 to 7, and more preferably 5. It should be noted that when the pH is 3 or less or 8 or more, it is not desirable because it causes stress for yeast cells and does not efficiently incorporate taurine into the cells.
タウリン溶液により処理する際の酵母濃度としては、開始時菌体量が１０〜３０％の範囲が適用可能である。なお、酵母の細胞当たりのタウリン含量を高くするためには酵母濃度が少ない方が望ましく、開始時菌体量を１０％程度に低くするのが望ましい。一方、溶液中タウリンの利用率を高くするためには、酵母濃度が高い方が望ましく、開始時菌体量を３０％程度に高くするのが望ましい。 As a yeast concentration at the time of processing with a taurine solution, the range of 10-30% of the amount of cells at the start is applicable. In order to increase the taurine content per yeast cell, it is desirable that the yeast concentration is low, and it is desirable to reduce the starting cell mass to about 10%. On the other hand, in order to increase the utilization rate of taurine in the solution, it is desirable that the yeast concentration is high, and it is desirable to increase the amount of starting cells to about 30%.
酵母を処理する際の、処理液中のタウリン濃度としては、０．１〜９．０％（w/v）の範囲が適用可能であり、また、左記タウリン濃度の範囲内において、酵母菌体中のタウリン含量を任意に調節することが可能である。なお、処理液中のタウリン濃度が９％以上になると、溶解したタウリンが処理中に結晶を形成して析出するため、望ましくない。 As the taurine concentration in the treatment liquid when treating yeast, a range of 0.1-9.0% (w / v) is applicable, and the yeast cells are within the taurine concentration range shown on the left. It is possible to arbitrarily adjust the taurine content therein. If the taurine concentration in the treatment liquid is 9% or more, the dissolved taurine is not desirable because it forms crystals and precipitates during the treatment.
以下に本発明の実施例を示すが、本発明は以下の実施例に限定されるものではない。
まず、溶液中のタウリンを効率良く酵母菌体に取込むための処理条件の検討を、３Lの小型ジャーファーメンタを用いて行った。
＜試験例１：処理時間の検討＞
酵母菌体に効率的にタウリンを取込ませるのに必要な処理時間の検討を行った。
タウリンが0.5、1.0及び2.0％（ｗ／ｖ）となるように必要量の合成タウリン（潜江永葯業製）を５０〜６０℃の温水に溶解した溶液を調製し、このタウリン溶液にパン酵母菌体（オリエンタル酵母工業（株）製レギュラーイースト）２００ｇ（１ｘ１０９個／ｍｌ）を懸濁し、更に水を添加して総量を１Lに調整した。この懸濁液を３Lジャーファーメンタにて３０℃で５１時間、１００ｒｐｍで攪拌処理した。途中一定時間経過毎に少量の菌体を回収して遠心分離（３０００ｒｐｍ、１５分）によって集菌し、イオン交換水で２回洗浄した。洗浄後の菌体は超音波処理あるいはガラスビーズによって菌体破砕し、１０％（ｗ／ｖ）になるようにTCA（トリクロロ酢酸）を添加して１０分間熱湯抽出した。左記抽出液の濾過液を検体とし、HPLCによってタウリン含量を測定した。なお、今回の試験ではタウリンの溶解、酵母の懸濁、洗浄には脱イオン水を用いたが、水道水でも代替可能である。測定結果を表１に示す。
First, treatment conditions for efficiently incorporating taurine in a solution into yeast cells were examined using a 3 L small jar fermenter.
<Test Example 1: Examination of processing time>
The treatment time required for efficiently incorporating taurine into yeast cells was examined.
Prepare a solution prepared by dissolving the required amount of synthetic taurine (manufactured by Yukie Nagai Kogyo Co., Ltd.) in hot water at 50-60 ° C. so that the taurine is 0.5, 1.0 and 2.0% (w / v). 200 g (1 × 10 9 cells / ml) of yeast cells (Oriental Yeast Co., Ltd. regular yeast) were suspended, and water was added to adjust the total amount to 1 L. This suspension was stirred with a 3 L jar fermenter at 30 ° C. for 51 hours at 100 rpm. On the way, a small amount of cells were collected every certain time, collected by centrifugation (3000 rpm, 15 minutes), and washed twice with ion-exchanged water. The washed cells were crushed by sonication or glass beads, and TCA (trichloroacetic acid) was added to 10% (w / v), followed by extraction with hot water for 10 minutes. The taurine content was measured by HPLC using the filtrate of the left extract as a sample. In this test, deionized water was used for dissolving taurine, suspending yeast, and washing, but it can be replaced with tap water. The measurement results are shown in Table 1.
いずれの時間においても処理液のタウリン濃度が高い区ほど酵母菌体のタウリン含量も高い値を示した。また、酵母のタウリン含量はいずれの区も経時的に増加しているが、４５時間でほぼ平衡に達している。よって、いずれのタウリン濃度の処理液においても最大のタウリン含量の酵母を得るには４５時間以上処理を行えば良いことが明らかとなった。 At any time, the higher the taurine concentration of the treatment solution, the higher the taurine content of the yeast cells. In addition, the taurine content of yeast increased with time in all sections, but almost reached equilibrium in 45 hours. Therefore, it was revealed that in any treatment solution having any taurine concentration, it is sufficient to carry out the treatment for 45 hours or longer in order to obtain a yeast having the maximum taurine content.
＜試験例２：処理温度の検討＞
タウリン溶液の温度が酵母のタウリン取り込みに大きく影響を及ぼすことが考えられるため、酵母にタウリンを効率的に取込ませるための処理温度の検討を行った。
処理液中のタウリン濃度は2.0％（w/v）とし、処理温度は２５、３０および３５℃の３区を設定した。その他の処理条件及びタウリンの分析法は試験例１と同じであるが、24時間の処理で明確な差が認められたので、24時間で処理を終了した。結果を表２に示す。
Since it is considered that the temperature of the taurine solution has a great influence on the taurine uptake of the yeast, the treatment temperature for efficiently incorporating the taurine into the yeast was examined.
The taurine concentration in the treatment liquid was 2.0% (w / v), and the treatment temperature was set to 3 zones of 25, 30 and 35 ° C. The other treatment conditions and the method for analyzing taurine were the same as in Test Example 1, but a clear difference was observed after the treatment for 24 hours, so the treatment was completed in 24 hours. The results are shown in Table 2.
本試験の結果、２５℃の低温でも酵母によるタウリンの取り込みは起こるが、３０℃と３５℃に比べて少ないことが明らかとなった。また培養温度が高いほど酵母のタウリン取り込みは高く、特に３５℃で高い値を示していることが見られた。なお、パン酵母は４０℃に近くなると自己消化が激しくなることが知られている。ため、今回の試験結果と合わせ考え、培養温度は３５℃付近が望ましいことが明らかとなった。また、実施例１のタウリン２.０％区の２４時間目の値が２３７０ｍｇ／Ｋｇ湿酵母、今回試験の３０℃区２４時間目の値が２６７７ｍｇ／Ｋｇ湿酵母であるため、使用する酵母の活性に大きな違いが無い場合、処理条件を厳密に管理すれば酵母によるタウリンの取り込み量に関し、再現性が高いと考えられる。 As a result of this test, it was revealed that taurine uptake by yeast occurs even at a low temperature of 25 ° C., but it is less than that at 30 ° C. and 35 ° C. It was also found that the higher the culture temperature, the higher the taurine uptake of the yeast, and in particular, a high value at 35 ° C. In addition, it is known that baker's yeast will become self-digesting intensely when it approaches 40 degreeC. For this reason, it has been clarified that the culture temperature is preferably around 35 ° C. in consideration of the test results. In addition, since the value at 24 hours for the 2.0% taurine group in Example 1 is 2370 mg / Kg wet yeast and the value at 24 hours at 30 ° C. in this test is 2677 mg / Kg wet yeast, If there is no significant difference in activity, it is considered that reproducibility is high with respect to the amount of taurine taken up by yeast if the treatment conditions are strictly controlled.
＜試験例３：タウリン溶液のpHの検討＞
酵母に効率良くタウリンを取込ませるための、タウリン溶液のpHを検討した。
酵母の処理開始時のpHを硫酸と水酸化ナトリウムを用いて３、５および７に調整し、処理液中のタウリン濃度を2.0％（w/v）、処理温度を３５℃とし、その他の処理条件とタウリンの分析法は試験例１と同様とした。また、２４時間の処理で明確な結果が得られたので、２４時間で処理を終了した。結果を表３に示す。pH5以下で酵母のタウリンの取り込みが若干少なかったが、pH５及び７においては比較的多いことが明らかとなった。
The pH of the taurine solution was investigated to allow yeast to efficiently incorporate taurine.
The pH at the start of yeast treatment is adjusted to 3, 5 and 7 using sulfuric acid and sodium hydroxide, the taurine concentration in the treatment solution is 2.0% (w / v), the treatment temperature is 35 ° C., and other treatments The conditions and the method for analyzing taurine were the same as in Test Example 1. Moreover, since the clear result was obtained by the process for 24 hours, the process was completed in 24 hours. The results are shown in Table 3. It was revealed that the taurine uptake of yeast was slightly less at
＜試験例４：酵母濃度の検討＞
酵母に効率良くタウリンを取込ませるための、処理開始時の菌体濃度を検討した。処理開始時の酵母湿菌体量は、処理液総量の１０、２０および３０％の３区とし、処理液のタウリン濃度は2.0％（w/v）、pHは５とし、その他の処理条件とタウリンの分析法は試験例１と同じである。結果を表４に示す。
In order to allow yeast to efficiently incorporate taurine, the cell concentration at the start of treatment was examined. The amount of wet yeast cells at the start of the treatment is 3 sections of 10, 20 and 30% of the total amount of the treatment solution, the taurine concentration of the treatment solution is 2.0% (w / v), the pH is 5, and other treatment conditions and The method for analyzing taurine is the same as in Test Example 1. The results are shown in Table 4.
本試験により、開始時菌体量が少ない区ほど高い傾向を示すことが明らかとなった。よって、酵母のタウリン含量を高くするためには開始時菌体量を１０％程度に低くするのが望ましい。
但し、溶液中のタウリンの酵母への取り込み絶対量で比較すると、菌体中に含まれるタウリンの各区間比は、１０％区では１０８８３×１＝１０８８３、２０％区では８２２６×２＝１６４５２、３０％区では７８９５×３＝２３６８５となり、３０％、２０％、１０％の順となる。よって、溶液中タウリンの利用率を高くすることを目的とする場合には、開始時菌体量を３０％程度に高くするのが望ましいということになる。すなわちその時々の目的によって培養開始時の菌体濃度を調整すれば良い。なお、以下の試験例及び実施例では、酵母のタウリン含量を高くすることを第１の目標としたため、開始時菌体量を１０％とした。
From this test, it was found that the smaller the starting bacterial mass, the higher the tendency. Therefore, in order to increase the taurine content of yeast, it is desirable to reduce the amount of cells at the start to about 10%.
However, when compared with the absolute amount of taurine in the yeast in the solution, the ratio of each section of taurine contained in the microbial cells is 1083 × 1 = 10883 in the 10% section, 8226 × 2 = 16452, in the 20% section, In the 30% ward, 7895 × 3 = 23685, which is the order of 30%, 20%, and 10%. Therefore, when the purpose is to increase the utilization rate of taurine in the solution, it is desirable to increase the amount of bacterial cells at the start to about 30%. That is, the bacterial cell concentration at the start of the culture may be adjusted according to the purpose at that time. In the following test examples and examples, since the first goal was to increase the taurine content of yeast, the amount of cells at the start was set to 10%.
＜試験例５：処理液中のタウリン濃度の検討＞
試験例１の結果では処理液のタウリン濃度が０.５〜２.０％（ｗ／ｖ）の範囲ではタウリン濃度が高い区ほど酵母中のタウリン含量が高い傾向が見られたため、酵母のタウリン取り込みに最も効率の良いタウリン濃度の検討を行った。
処理温度は３５℃とし、処理液のタウリン濃度は２、３、６および９％（ｗ／ｖ）の４区とした。その他の処理条件とタウリンの分析法は試験例１と同じであるが、処理時間は４８時間とした。結果を表５及び図１に示す。
In the results of Test Example 1, since the taurine concentration in the yeast tended to be higher in the section where the taurine concentration was higher in the range where the taurine concentration in the treatment solution was 0.5 to 2.0% (w / v), The most effective taurine concentration for uptake was examined.
The treatment temperature was 35 ° C., and the taurine concentration of the treatment solution was 2, 3, 6, and 9% (w / v). Other treatment conditions and the method for analyzing taurine were the same as in Test Example 1, but the treatment time was 48 hours. The results are shown in Table 5 and FIG.
処理液のタウリン濃度が２〜６％（ｗ／ｖ）の範囲では、酵母のタウリン含量も直線的に増加するが、６％（ｗ／ｖ）以上になると傾きが小さくなり、取込み効率が悪くなる傾向が見られた。なお、処理液のタウリン濃度が２〜６％（ｗ／ｖ）の範囲で、処理液のタウリン濃度X（％）及び酵母のタウリン含量Y（ｍｇ／Ｋｇ湿酵母）の相関を解析した結果、Y＝１５８０X＋３４０９の直線式が得られ、R２は０．９９８７であった。なお、試験例１の処理温度が３０℃、処理液中のタウリン濃度が０.５〜２．０％（ｗ／ｖ）の条件においては、Y=1514X＋４２４、R２が０．９９７８という相関式が得られた。この結果も合わせて図１に示す。以上より、一定の処理条件下であれば酵母のタウリン取り込みは温度と処理液中のタウリン濃度によって規定されることが示唆される。 When the taurine concentration of the treatment liquid is in the range of 2 to 6% (w / v), the taurine content of the yeast also increases linearly, but when it exceeds 6% (w / v), the slope becomes small and the uptake efficiency is poor. The tendency to become was seen. As a result of analyzing the correlation between the taurine concentration X (%) of the treatment liquid and the taurine content Y (mg / Kg wet yeast) of the yeast in the range of 2-6% (w / v) of the taurine concentration of the treatment liquid, Y = 1580X + 3409 linear equation of is obtained, R 2 was 0.9987. In addition, in the conditions where the processing temperature of Test Example 1 is 30 ° C. and the taurine concentration in the processing solution is 0.5 to 2.0% (w / v), the correlation equation that Y = 1514X + 424 and R 2 is 0.9978. was gotten. The results are also shown in FIG. From the above, it is suggested that the taurine uptake of yeast is defined by the temperature and the taurine concentration in the treatment solution under certain treatment conditions.
次に、処理開始後４８時間目の酵母の水分含量を測定し、更に定法に従って菌体を加水分解して酵母乾燥物当りのタウリン含量を求めた。結果を表６及び図２に示す。
両者の間にはY（酵母のタウリン含量：％（乾物重量））＝１．２４７７X（処理液のタウリン濃度：％（ｗ／ｖ））＋２．６３８５の直線式が得られ、R２＝０．９９８８であった。また、乾物酵母の総タウリン含量Y（％（乾物重量））と湿物酵母の酸抽出可能タウリン含量X（％（湿物重量））の間にはY=７．８９０６X−０．０４８６の直線式が得られた。処理液のタウリン濃度が６〜９％（ｗ／ｖ）の範囲では、Y（酵母のタウリン含量：ｍｇ／Ｋｇ湿酵母）＝５２７X（処理液のタウリン濃度：％（ｗ／ｖ））＋９６９３の直線式が得られる。この式を乾物のタウリン含量の式に直すと、Y（酵母のタウリン含量：％（乾物重量））＝０．４０X（処理液のタウリン濃度：％（ｗ／ｖ））＋７．７となる。この式を用いて、処理温度３５℃、処理液中タウリン濃度０〜９％（ｗ／ｖ）の範囲において試験例１〜５で検討した処理条件下で４８時間酵母を処理した場合に得られる酵母中のタウリン含量を推定すると表７のようになる。
試験例１〜５より、最適な処理条件として、温度３５℃、pH５、処理時間４５時間以上、処理開始時湿菌体濃度１０％（但し、培養液タウリンの利用率を高くする場合には１０％以上にする）が考えられる。なお、通気の必要性についても検討したが、菌体が沈降しない程度に攪拌（回転羽根を用いて１００ｒｐｍで攪拌）すれば十分であることが明らかとなったため、通気は行わないこととした。この条件下で培養液タウリン濃度０〜９％（ｗ／ｖ）の範囲で酵母を処理すれば、ほぼ任意のタウリン含量の酵母が得られることとなる。
From Test Examples 1 to 5, the optimum treatment conditions were as follows: temperature 35 ° C.,
（３KL大型タンクを用いたタウリン富化条件の検討）
上記実施例１において得られた検討結果を基に、３KLの実生産用大型タンクで処理を行った場合の再現性を検討した。
処理液中タウリン濃度を、３５℃での溶解可能限界量である９％（ｗ／ｖ）とし、以下の手順で処理液の調製を行った。
− １６００Lの熱水に合成タウリン２２５Kgを添加、溶解する。
− ３KLタンクに上記のタウリン溶液の全量を入れ、４０℃前後に保温する。
− ２５０Kg湿菌体相当量の濃縮クリーム状酵母を添加する。
− 水で全体量を２５００Lに調整する（以上の処理で酵母濃度は１０％、処理液のタウリン濃度は９％（ｗ／ｖ）となる）。
(Examination of taurine enrichment condition using 3KL large tank)
Based on the examination results obtained in Example 1 above, the reproducibility when the treatment was performed in a 3 KL large production tank was examined.
The taurine concentration in the treatment liquid was 9% (w / v), which is the limit amount of solubility at 35 ° C., and the treatment liquid was prepared according to the following procedure.
-Add and dissolve 225 kg of synthetic taurine in 1600 L of hot water.
-Put all of the above taurine solution in a 3KL tank and keep it warm at around 40 ° C.
Add 250 kg wet cell equivalent of concentrated creamy yeast.
-Adjust the total volume to 2500 L with water (with the above treatment, the yeast concentration is 10% and the taurine concentration in the treatment solution is 9% (w / v)).
上記処理液を３５±２℃、pHは５.０±０.５、攪拌速度１００ｒｐｍで４４時間処理し、処理後の酵母はノズルセパレータで２回以上洗浄後濃縮し、菌体濃度を湿菌体で５０％以上とした。更に濃縮菌体の水分含量を定法により算出し、総タウリン含量を試験例５と同じ方法により算出した。その結果、得られた酵母中のタウリン含量は１１．５％（乾物重量）であった。この値は３Lジャーファーメンタで得られた結果（１１．３％（乾物重量））とほぼ一致しており、３KLの大型タンクで培養した場合にも高い再現性が認められた。 The above treatment solution was treated at 35 ± 2 ° C., pH of 5.0 ± 0.5, and stirring speed of 100 rpm for 44 hours, and the treated yeast was washed with a nozzle separator at least twice and concentrated. 50% or more by body. Furthermore, the water content of the concentrated cells was calculated by a conventional method, and the total taurine content was calculated by the same method as in Test Example 5. As a result, the taurine content in the obtained yeast was 11.5% (dry matter weight). This value almost coincided with the result (11.3% (dry matter weight)) obtained with the 3 L jar fermenter, and high reproducibility was observed even when cultured in a large tank of 3 KL.
３Lジャーファーメンタで得られた結果を基に、３KLの大型タンクでも目的とするタウリン含量の酵母を得ることが可能か否かにつき検討を行った。なお、本実施例においては、酵母のタウリン含量の目標を５％乾物とした。 Based on the results obtained with the 3L jar fermenter, it was examined whether it was possible to obtain the target taurine-containing yeast even in a 3 KL large tank. In this example, the target for the taurine content of yeast was 5% dry matter.
試験例５で示したように、処理液のタウリン濃度X（％（ｗ／ｖ））と酵母のタウリン含量Y（％（乾物重量））に間にはY=１．２４７７X＋２．６３８５の関係があるので、５．０−２．６３８５＝１．２４７７X、X=１．８９となり、約１．９％（ｗ／ｖ）のタウリン濃度が必要と予想されるため、２５００Lの処理液量であれば４７．５Kgのタウリンが必要となる。なお、処理液の調製にあたり、１６００Lの熱水に合成タウリン４８Kg添加し、その後２５０Kg湿菌体相当量の濃縮クリーム状酵母と水を加えて総量を２５００Lとし、タウリン濃度１．９２％（ｗ／ｖ）の処理液を得た。 As shown in Test Example 5, there is a relationship of Y = 1.2477X + 2.6385 between the taurine concentration X (% (w / v)) of the treatment liquid and the taurine content Y (% (dry weight)) of the yeast. Therefore, 5.0-2.6385 = 1.2477X, X = 1.89, and a taurine concentration of about 1.9% (w / v) is expected. For example, 47.5 kg of taurine is required. In preparation of the treatment liquid, 48 kg of synthetic taurine was added to 1600 L of hot water, and then 250 kg wet cell equivalent of concentrated creamy yeast and water were added to make the total volume 2500 L, with a taurine concentration of 1.92% (w / The processing liquid of v) was obtained.
実施例２と同様の処理条件、菌体の洗浄濃縮、タウリンの分析を行った結果、酵母のタウリン含量は５．４％（乾物重量）で、当初の目的をほぼ達成することができた。以上の結果から、３Lの小型ジャーファーメンタでの試験によって得られた最適処理条件は、３KLの大型タンクでにおいても適用可能であることが明らかとなった。 As a result of performing the same treatment conditions as in Example 2, washing and concentration of cells, and analysis of taurine, the taurine content of the yeast was 5.4% (dry weight), and the initial purpose was almost achieved. From the above results, it has been clarified that the optimum processing conditions obtained by the test using the 3L small jar fermenter can be applied to the 3KL large tank.
Claims (7)
（ａ）群：サッカロミセス（Saccharomyces）属、トルロプシス（Torulopsis）属、ミコトルラ（Mycotorula）属、トルラスポラ（Torulopsis）属、キャンディダ（Candida）属、ロードトルラ（Rhodotorula）属又はピキア（Pichia）属の酵母。
（ｂ）群：Saccharomyces cerevisiae、Saccharomyces carlsbergensis、Saccharomyces uvarum、Saccharomyces rouxii、Torulopsis utilis、Torulopsis candida、Mycotorula japonica、Mycotorula lipolytica、Torulaspora delbrueckii、Torulopsis fermentati、Candida
sake、Candida tropicalis、Candida
utilis、Saccharomycopsis fibligera、Saccharomyces lipolytica、Rhodotorula rubra又はPichia farinosaである酵母。 A method for producing a taurine-rich yeast comprising suspending and treating a yeast corresponding to at least one of the following groups (a) and (b) in an aqueous taurine solution , wherein the taurine-rich yeast has a taurine content of 6: A method for producing a high taurine-containing yeast, characterized by being in the range of 48 to 11.3% (dry matter weight) .
(A) Group: Yeast of the genus Saccharomyces, the genus Torulopsis, the genus Mycotorula, the genus Torulopsis, the genus Candida, the genus Rhodotorula or the genus Pichia.
(B) Group: Saccharomyces cerevisiae, Saccharomyces carlsbergensis, Saccharomyces uvarum, Saccharomyces rouxii, Torulopsis utilis, Torulopsis candida, Mycotorula japonica, Mycotorula lipolytica, Torulaspora delbrueckii, Torulopsiment, Torulopsi
sake, Candida tropicalis, Candida
Yeast which is utilis, Saccharomycopsis fibligera, Saccharomyces lipolytica, Rhodotorula rubra or Pichia farinosa.
（１）処理時間を４５時間以上とする。
（２）処理温度を３０℃〜３５℃とする。
（３）タウリン水溶液のpHをpH５〜７とする。
（４）タウリン水溶液の酵母濃度を、酵母のタウリン含量を高くするためには１０％とし、タウリン利用率を高めるためには３０％とする。
（５）Ｙを酵母の目標タウリン含量（ｍｇ／Ｋｇ湿酵母）、Ｘをタウリン水溶液のタウリン濃度（％）としたとき、Ｘが０．５〜２．０％（ｗ／ｖ）の範囲ではＹ＝１５１４Ｘ＋４２４の直線式に従い、Ｘが２〜６％（ｗ／ｖ）の範囲ではＹ＝１５８０Ｘ＋３４０９の直線式に従ってＸを設定する。 In performing the method for producing a taurine-rich yeast according to claim 1 , the taurine content of the yeast is increased by the treatment according to one or more of the following conditions (1) to (5), or the taurine utilization rate by the yeast A method for adjusting the taurine content, characterized by increasing the content.
(1) The processing time is 45 hours or more.
(2) The treatment temperature is 30 ° C to 35 ° C.
(3) The pH of the aqueous taurine solution is adjusted to pH 5-7.
(4) The yeast concentration of the taurine aqueous solution is 10% for increasing the taurine content of the yeast, and 30% for increasing the taurine utilization rate.
(5) When Y is the target taurine content of yeast (mg / Kg wet yeast) and X is the taurine concentration (%) of the taurine aqueous solution, X is in the range of 0.5 to 2.0% (w / v). According to the linear formula of Y = 1514X + 424, X is set according to the linear formula of Y = 1580X + 3409 when X is in the range of 2 to 6% (w / v).
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