JP2010168284A - Method for producing fucoxanthin and/or fucosterol - Google Patents
Method for producing fucoxanthin and/or fucosterol Download PDFInfo
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本発明は、フコキサンチンおよび／またはフコステロールの製造方法に関し、更に詳細には、ハバノリまたはモズクの盤状体または糸状体を原料とし、高収量のフコキサンチンおよび／またはフコステロールを得ることが可能なこれらの化合物の製造方法に関する。 The present invention relates to a process for producing fucoxanthin and / or fucosterol, and more specifically, it is possible to obtain a high-yield fucoxanthin and / or fucostosterol using a havanori or mozuku disc or filament as a raw material. The present invention relates to a method for producing these compounds.
褐藻類はカロテノイドの１種であるフコキサンチンやステロールの１種であるフコステロール等の機能性成分を有している。フコキサンチンは下記構造を有する既知の物質であり、その効果としては体内摂取による抗腫瘍効果（特許文献１）や神経細胞保護効果（特許文献２）、血糖値上昇抑制効果（特許文献３）等が報告されている。 Brown algae have functional components such as fucoxanthin, which is a kind of carotenoid, and fucosterol, which is a kind of sterol. Fucoxanthin is a known substance having the following structure, and its effects include an antitumor effect (Patent Document 1), a nerve cell protective effect (Patent Document 2), a blood glucose level increase suppressing effect (Patent Document 3), etc. Has been reported.
一方、フコステロールは下記構造を有する既知の物質であり、その効果としては体内摂取による抗動脈硬化作用（特許文献４）やコレステロール低減作用、血栓予防作用等が一般的に知られている。 On the other hand, fucosterol is a known substance having the following structure, and its effects are generally known as an anti-arteriosclerosis action (patent document 4), a cholesterol reduction action, a thrombus prevention action, etc. by ingestion.
従来、これらフコキサンチンやフコステロールを製造するにあたっては、ナガマツモ科（Chordaceae）のオキナワモズク（Cladosiphon okamuranus）の成熟体や盤状体・糸状体が原料として使用されていた。しかしながら、成熟体におけるこれらの機能性成分の含有量は少なく、また、盤状体や糸状体でも、含有量は成熟体に比べて高いものの実用的に十分な量とはいえず、また培養も難しいという問題があった。 Conventionally, in the production of these fucoxanthins and fucostosterols, mature forms, discoids and filaments of Chordaceae's Chordaceae were used as raw materials. However, the content of these functional components in the mature body is small, and even in the plate-like body and the filamentous body, although the content is higher than that of the mature body, it cannot be said to be a practically sufficient amount, and the culture is also not possible. There was a problem that it was difficult.
従って、フコキサンチンおよびフコステロールをより高収量で取得することが可能な製造方法の開発が求められており、本発明は、そのような製造方法を提供することを課題とするものである。 Therefore, development of a production method capable of obtaining fucoxanthin and fucostosterol at higher yields has been demanded, and an object of the present invention is to provide such a production method.
本発明者らは上記課題を解決するために鋭意研究をした結果、原料の褐藻類としてカヤノモリ科（Scytosiphonaceae）のハバノリ（Petalonia binghamiae）またはモズク科（Spermatochnaceae）のモズク（Nemacystus decipiens）の盤状体または糸状体を使用すると、培養による増体量が大きく、またフコキサンチンおよびフコステロールの含有量を高め得ることを見出し、本発明を完成させた。 As a result of diligent research to solve the above-mentioned problems, the present inventors have found that as a raw brown algae, Scytosiphonaceae's Petalonia binghamiae or Spermatochnaceae's Nemacystus decipiens discoid Alternatively, the present inventors have found that the use of filaments increases the amount of increase in culture and can increase the contents of fucoxanthin and fucosterol, thereby completing the present invention.
すなわち、本発明はハバノリ（Petalonia binghamiae）およびモズク（Nemacystus decipiens）よりなる群から選ばれた褐藻類の盤状体または糸状体の抽出物からフコキサンチンおよび／またはフコステロールを分離することを特徴とするフコキサンチンおよび／またはフコステロールの製造方法である。 That is, the present invention is characterized by separating fucoxanthin and / or fucosterol from an extract of a brown algal plate or filament selected from the group consisting of Petalonia binghamiae and Nemacystus decipiens. This is a method for producing fucoxanthin and / or fucosterol.
本発明の製造方法によれば、培養により、褐藻類を著しく増体させるとともに、褐藻類中のフコキサンチンおよび／またはフコステロールの含有量を高めることができるため、より経済的に高収量のフコキサンチンおよび／またはフコステロールを製造することが可能である。 According to the production method of the present invention, brown algae can be significantly increased by culturing, and the content of fucoxanthin and / or fucosterol in the brown algae can be increased. It is possible to produce xanthine and / or fucosterol.
本発明においては、原料の褐藻類として、カヤノモリ科（Scytosiphonaceae）のハバノリ（Petalonia binghamiae）またはモズク科（Spermatochnaceae）のモズク（Nemacystus decipiens）が用いられる。また、これらの褐藻類は、成熟体から遊走子の放出、遊走子から盤状体または糸状体の発生、盤状体または糸状体の着床による直立体の形成、直立体から成熟体への成長という生育サイクルのうちの、盤状体または糸状体を用いる。なお、盤状体または糸状体は、上記生育サイクルのうちの遊走子と直立体の中間に位置するものであり、別名で呼ばれることもがあるが、本発明においてはこれらの何れも含む。 In the present invention, as the raw brown algae, Scytosiphonaceae's Petalonia binghamiae or Spermatochnaceae's Mozuku (Nemacystus decipiens) is used. In addition, these brown algae can release zoospores from matured bodies, generate discoids or filaments from the zoospores, form solid bodies by implantation of the boarders or filamentous bodies, and convert from solid bodies to mature bodies. Of the growth cycle of growth, a disk or filament is used. In addition, although a board | plate-like body or a filamentous body is located in the middle of the zoospore and a direct solid in the said growth cycle, and may be called with another name, in the present invention, all of these are included.
上記盤状体または糸状体は、天然由来のものであってもよいが、人工的に培養されたものを使用することが好ましく、特に盤状体または糸状体の種苗を直立体を形成させない条件で培養、増加されたものを利用することにより、多量に盤状体または糸状体を得ることができるので好ましい。 The above plate-like body or filamentous body may be of natural origin, but it is preferable to use an artificially cultured one, and particularly the condition that the seedlings of the plate-like body or the filamentous body do not form a straight solid. It is preferable to use the ones cultured and increased in (1) because a large amount of discs or filaments can be obtained.
上記の直立体を形成させない条件での培養は、褐藻類の盤状体または糸状体の種苗を、培養容器へ着床させない条件で培養することにより行われ、例えば、連続攪拌培養したり、着床しにくい材料で形成した培養容器中で培養すれば良い。 Cultivation under conditions that do not form a straight solid is performed by culturing brown seedlings or seedlings of brown algae under conditions that do not allow implantation in a culture vessel. What is necessary is just to culture | cultivate in the culture container formed with the material which is hard to floor.
より具体的には、連続攪拌により直立体を形成させずに培養する場合は、攪拌子や攪拌機による機械攪拌や、通気等による攪拌を行い、培養液全体を攪拌しながら培養すれば良い。攪拌条件は容器の大きさ等により適宜変化するが、例えば、培養容器として１Ｌの容量の平底フラスコを用い、通気による攪拌を行う場合には、培養容器内に空気を１．０Ｌ／分〜２．５Ｌ／分、好ましくは１．５Ｌ／分〜２．３Ｌ／分で導入すれば良い。 More specifically, when culturing without forming a solid body by continuous stirring, the culture may be performed while stirring the whole culture solution by performing mechanical stirring with a stirrer or a stirrer, stirring by aeration, or the like. The stirring conditions vary as appropriate depending on the size of the container. For example, when a 1 L flat bottom flask is used as a culture container and stirring is performed by aeration, air is supplied in the culture container from 1.0 L / min to 2. It may be introduced at a rate of 0.5 L / min, preferably 1.5 L / min to 2.3 L / min.
上記培養の際に用いる培養液は、褐藻類の盤状体または糸状体を培養することができるものであれば特に制限されないが、滅菌された人工海水または海水に、窒素源およびリン源等の栄養成分を含有したものであることが好ましい。窒素源としては、硝酸塩、硫酸アンモニウム等のアンモニウム塩、尿酸等が例示できる。このうち、褐藻類中のフコキサンチンおよびフコステロールの含有量を向上できるため、硫酸アンモニウム等のアンモニウム塩を添加することが好ましい。リン源としては、グリセロリン酸２ナトリウム、β−グリセロリン酸ナトリウム等の有機リン酸塩、リン酸３ナトリウム、リン酸３カリウム等の無機リン酸塩が例示できる。これらのうち、無機リン酸塩がフコキサンチンおよびフコステロールの含有量を向上できるため好ましい。窒素源の添加量は、窒素が好ましくは４００〜１６００μＭ、より好ましくは７００〜９００μＭとなる量である。一方、リン源の添加量は、リンが好ましくは１５〜６０μＭ、より好ましくは２０〜４０μＭとなる量である。培地中に添加可能なその他の栄養成分としては、ビタミンＢ１２、ビオチン、チアミンが例示できる。これらの栄養成分を含有する培養液として、例えば公知のＰＥＳＩ培地（Ｍａｓａｋａｚｕ Ｔａｔｅｗａｋｉ、Ｆｏｒｍａｔｉｏｎ ｏｆ ａ ｃｒｕｓｔａｃｅｏｕｓ ｓｐｏｒｏｐｈｙｔｅ ｗｉｔｈ ｕｎｉｌｏｃｕｌａｒ ｓｐｏｒａｎｇｉａ ｉｎ Ｓｃｙｔｏｓｉｐｈｏｎ ｌｏｍｅｎｔａｒｉａ、Ｐｈｙｃｏｌｏｇｉａ
６（１）１９６６．）等を使用することができ、また、ＫＷ−２１（第一製網製）の商品名で市販されている藻類培養液を用いることもできる。これらの培養液を窒素源およびリン源が上記濃度となるよう添加すれば良い。
The culture solution used for the culture is not particularly limited as long as it can cultivate brown algal discoids or filamentous bodies. However, sterilized artificial seawater or seawater may contain nitrogen sources and phosphorus sources. It is preferable that it contains a nutrient component. Examples of the nitrogen source include nitrates, ammonium salts such as ammonium sulfate, uric acid and the like. Among these, it is preferable to add an ammonium salt such as ammonium sulfate because the content of fucoxanthin and fucostosterol in brown algae can be improved. Examples of the phosphorus source include organic phosphates such as disodium glycerophosphate and sodium β-glycerophosphate, and inorganic phosphates such as trisodium phosphate and tripotassium phosphate. Of these, inorganic phosphates are preferable because they can improve the contents of fucoxanthin and fucosterol. The amount of nitrogen source added is such that nitrogen is preferably 400-1600 μM, more preferably 700-900 μM. On the other hand, the amount of the phosphorus source added is such that phosphorus is preferably 15 to 60 μM, more preferably 20 to 40 μM. Examples of other nutrient components that can be added to the medium include vitamin B12, biotin, and thiamine. As a culture solution containing these nutrient components, for example, known PESI media (Masakazu Tatewaki, Formation of a crusaceous sporophys with unipolar spirala
6 (1) 1966. ) And the like, and an algae culture solution marketed under the trade name of KW-21 (manufactured by Daiichi Seimitsu) can also be used. What is necessary is just to add these culture solutions so that a nitrogen source and a phosphorus source may become the said density | concentration.
また、この培養は恒温条件下で行うことが好ましい。ハバノリは、通常１２〜２９℃、好ましくは１５〜２５℃、より好ましくは２０〜２５℃の温度である。一方、モズクは、通常１２〜３０℃、好ましくは１５〜３０℃、より好ましくは１５〜２５℃である。この範囲であると、増体量が大きく、またフコキサンチンおよびフコステロールの含有量が高くなる。 In addition, this culture is preferably performed under a constant temperature condition. Habanori is usually at a temperature of 12 to 29 ° C, preferably 15 to 25 ° C, more preferably 20 to 25 ° C. On the other hand, a mozuku is 12-30 degreeC normally, Preferably it is 15-30 degreeC, More preferably, it is 15-25 degreeC. Within this range, the weight gain is large, and the fucoxanthin and fucosterol content is high.
更に、この培養は光照射条件下で行うことが好ましい。ハバノリは、好ましくは１０〜１６０μｍｏｌ・ｍ−２・ｓ−１であり、より好ましくは２０〜８０μｍｏｌ・ｍ−２・ｓ−１である。一方、モズクは、好ましくは１０〜１６０μｍｏｌ・ｍ−２・ｓ−１であり、より好ましくは１０〜８０μｍｏｌ・ｍ−２・ｓ−１である。この範囲であると、増体量が大きく、またフキキサンチンおよびフコステロールの含有量が高くなる。このような光量により、８Ｌ：１６Ｄ〜２４Ｌ：０Ｄ、好ましくは１２Ｌ：１２Ｄ〜１６Ｌ：８Ｄの光周期で照射を行えば良い。 Furthermore, this culture is preferably performed under light irradiation conditions. Habanori is preferably 10 to 160 μmol · m −2 · s −1 , more preferably 20 to 80 μmol · m −2 · s −1 . On the other hand, the mozuku is preferably 10 to 160 μmol · m −2 · s −1 , more preferably 10 to 80 μmol · m −2 · s −1 . When the amount is within this range, the weight gain is large, and the contents of fuxanthine and fucosterol are high. Irradiation with such an amount of light may be performed with an optical period of 8L: 16D to 24L: 0D, preferably 12L: 12D to 16L: 8D.
上記した培養方法により、褐藻類の盤状体または糸状体のみを繰り返し、増殖させて培養することができる。なお、盤状体または糸状体は、培養容器中に着床すると、すぐに直立体を形成し、成熟体にまで成長してしまうので、盤状体または糸状体を着床させることは盤状体または糸状体を多く得るためには適さない。 By the culture method described above, it is possible to repeat and grow only the plate or filamentous body of brown algae. In addition, since a plate-like body or a filamentous body immediately forms a solid body and grows to a mature body when it is placed in a culture vessel, it is discotic to place a plate-like body or a filamentous body on a plate. It is not suitable for obtaining many bodies or filaments.
上記の培養方法に使用する褐藻類の盤状体または糸状体の種苗は、天然から採取してもよいが、例えば、次のようにして得ることもできる。すなわち、まず、単子嚢を形成していない母藻を数本程度と、盤状体または糸状体が着床することのできる着床担体とを共に滅菌海水に入れ、恒温条件で培養し、遊走子から変化した盤状体または糸状体を着床担体に着床させる。 The seedlings or seedlings of brown algae used in the above culture method may be collected from nature, but can also be obtained, for example, as follows. That is, first, put together about several mother algae that do not form a monocyst and an implantation carrier on which a discoid or filament can be implanted, in sterile seawater, and culture under constant temperature conditions, The disc-like body or the filamentous body changed from the zoospore is placed on the landing carrier.
ここで着床担体としては、ガラス、アクリル、プラスチック、ポリカーボネート、繊維、岩石、砂等が好ましく、特にガラス板（スライドガラス）、アクリル板が好ましい。また、滅菌海水は海水または人工海水をオートクレーブあるいはろ過することにより得られる。この滅菌海水には、上記盤状体または糸状体の培養と同様に栄養成分を適宜添加することができる。 Here, as the flooring carrier, glass, acrylic, plastic, polycarbonate, fiber, rock, sand and the like are preferable, and a glass plate (slide glass) and an acrylic plate are particularly preferable. Sterile seawater can be obtained by autoclaving or filtering seawater or artificial seawater. Nutritional components can be appropriately added to the sterilized seawater in the same manner as in the culture of the plate or filament.
盤状体または糸状体が着床担体に着床するまでの培養は、上記盤状体または糸状体の培養と同様の条件で行えば良い。また、前記培養と同時に、盤状体または糸状体が着床担体に着床できる程度に通気等の攪拌を行っても良い。 The culture until the plate-like body or filamentous body is deposited on the implantation carrier may be performed under the same conditions as the culture of the above-mentioned plate-like body or filamentous body. At the same time as the culture, stirring such as aeration may be performed to such an extent that the plate-like body or the filamentous body can be deposited on the implantation carrier.
次いで、盤状体または糸状体が着床した着床担体を取りだし、滅菌海水で洗浄した後、上記盤状体または糸状体の培養と同様の条件で再度培養する。 Next, after the landing carrier on which the plate-like body or the filamentous body is deposited is taken out, washed with sterilized seawater, it is cultured again under the same conditions as the culture of the above-mentioned disk-like body or the filamentous body.
上記盤状体または糸状体が着床した着床担体の培養においては、培養２、３日後から１日おきに平筆よる洗浄を行う。この洗浄に用いる水としては、滅菌海水および水道水が使用されるが、水道水での洗浄は１０秒以内で行うことが好ましい。 In culturing the implantation carrier on which the plate-like body or the filamentous body is deposited, washing with a flat brush is performed every other day after 2 or 3 days of culturing. As water used for this washing, sterilized seawater and tap water are used, but washing with tap water is preferably performed within 10 seconds.
上記盤状体または糸状体が着床した着床担体の培養開始後８日〜１０日の十分に成長した盤状体または糸状体を、検鏡（１００倍）により雑藻の少ない部分を選んで掻き取って盤状体または糸状体の種苗とすることができる。また、必要により上記で掻き取った盤状体または糸状体を試験管に入れて滅菌海水を使用して３回程度ピペッティングで洗浄し、その後、寒天平板（海水に０．５〜１０％の寒天と０．０１〜１０％の栄養成分（藻類培養液）を添加した寒天培地）に塗りつけて、温度１５〜３５℃、好ましくは２０〜３０℃、１０〜１６０μｍｏｌ・ｍ−２・ｓ−１、好ましくは１０〜８０μｍｏｌ・ｍ−２・ｓ−１、光周期８Ｌ：１６Ｄ〜２４Ｌ：０Ｄ、好ましくは１２Ｌ：１２Ｄ〜２４Ｌ：０Ｄの条件下で培養しても良い。この培養から２０〜３０日後、寒天平板に増殖してきた盤状体または糸状体のうち、雑藻が混入していないコロニーを選択し、これを掻き取って盤状体または糸状体の種苗とすることができる。 Select a portion with less miscellaneous algae by speculum (100 times) from a fully grown plate-like body or filamentous body 8 to 10 days after the start of culturing of the above-mentioned plate-like body or filamentous body. Can be used as seedlings of a disk-like body or a filamentous body. Also, if necessary, the disc-like body or filamentous body scraped above is put in a test tube and washed by pipetting about three times using sterilized seawater, and then agar plates (0.5 to 10% in seawater). Agar and an agar medium supplemented with 0.01 to 10% nutrient components (algae culture solution), and a temperature of 15 to 35 ° C., preferably 20 to 30 ° C., 10 to 160 μmol · m −2 · s −1. The culture may be performed under the conditions of 10 to 80 μmol · m −2 · s −1 , photoperiod 8L: 16D to 24L: 0D, preferably 12L: 12D to 24L: 0D. After 20 to 30 days from the culture, a colony not contaminated with miscellaneous algae is selected from the plate-like body or filamentous body that has grown on the agar plate, and this is scraped to form a seedling of the plate-like body or filamentous body. be able to.
一方、盤状体または糸状体から、フコキサンチンおよび／またはフコステロールを抽出するために使用される溶媒は、フコキサンチンおよび／またはフコステロールを抽出することのできるものであれば特に制限されないが、例えば、水、メタノール、エタノール、プロパノール等のアルコール類、アセトン等のケトン類、酢酸エチル等のエステル類、ヘキサン等の脂肪族炭化水素類、ベンゼン等の芳香属炭化水素類、クロロホルム等のハロゲン系有機溶媒、もしくはこれらの混合溶媒が好ましく、フコキサンチンおよび／またはフコステロールが食品等にも使用されることからエタノールもしくは含水エタノールがより好ましい。含水エタノールは、エタノール：水の混合比が５０：１〜１：１、好ましくは１０：１〜２：１（質量比）の範囲である。含水エタノールを用いると、クロロフィル等の不純物の含有量が少なく、フコキサンチン等の収率を向上できるために好ましい。これらの溶媒は、盤状体または糸状体に対して４：１〜１０００：１、好ましくは４：１〜２００：１の質量比で添加すれば良い。 On the other hand, the solvent used for extracting fucoxanthin and / or fucosterol from the plate or filament is not particularly limited as long as it can extract fucoxanthin and / or fucostol. For example, water, alcohols such as methanol, ethanol and propanol, ketones such as acetone, esters such as ethyl acetate, aliphatic hydrocarbons such as hexane, aromatic hydrocarbons such as benzene, halogens such as chloroform An organic solvent or a mixed solvent thereof is preferable. Since fucoxanthin and / or fucosterol is also used for foods and the like, ethanol or hydrous ethanol is more preferable. Hydrous ethanol has a mixing ratio of ethanol: water of 50: 1 to 1: 1, preferably 10: 1 to 2: 1 (mass ratio). Use of hydrous ethanol is preferable because the content of impurities such as chlorophyll is small and the yield of fucoxanthin and the like can be improved. These solvents may be added at a mass ratio of 4: 1 to 1000: 1, preferably 4: 1 to 200: 1, with respect to the disk or filament.
上記溶媒を用いる抽出は常法により行うことができ、例えば、溶媒としてエタノールを用いる場合であれば、１５℃〜４０℃、好ましくは３０℃〜４０℃の温度で、０．５時間〜１．５時間、好ましくは１時間〜１．５時間抽出を行えば良い。また、抽出に当たっては、必要により、超音波、攪拌機等により攪拌を行っても良い。 Extraction using the above solvent can be performed by a conventional method. For example, when ethanol is used as the solvent, the temperature is 15 ° C. to 40 ° C., preferably 30 ° C. to 40 ° C., for 0.5 hour to 1. Extraction may be performed for 5 hours, preferably 1 to 1.5 hours. In extraction, if necessary, stirring may be performed with an ultrasonic wave, a stirrer, or the like.
以上のようにして得られる有機溶媒抽出物から、フコキサンチンおよび／またはフコステロールを分離、取得するには、有機溶媒抽出物をそのまま、あるいは残渣を取り除いてからＨＰＬＣ等に付し、これにより分離・精製を行えばよい。 In order to separate and obtain fucoxanthin and / or fucosterol from the organic solvent extract obtained as described above, the organic solvent extract can be subjected to HPLC or the like after removing the residue as it is or after removing the residue.・ Purification may be performed.
斯くして得られるフコキサンチンおよび／またはフコステロールは上記したコレステロール低減作用や血栓予防作用を期待した各種健康食品や、抗腫瘍効果、神経細胞保護効果、血糖値上昇抑制効果等を目的とした医薬品等の用途に使用することができる。 Fucoxanthin and / or fucosterol thus obtained are various health foods that are expected to have the above-described cholesterol-reducing action and thrombus-preventing action, and pharmaceuticals aimed at antitumor effects, nerve cell protection effects, blood glucose level increase effects, etc. It can be used for such applications.
本発明で用いるハバノリおよびモズクの糸状体等は、従来使用されているオキナワモズクと比べ、容器への附着性が弱く成熟しにくいため、容器を用いた攪拌培養を容易に行えるという利点も有する。附着性が高いと、容器壁面にコケ状に附着し、光の照射が阻害されたり、通気による攪拌培養が滞るため生産量が低下する。また、ハバノリおよびモズクの糸状体等はオキナワモズクと比べて真水に強く、夾雑物を真水処理して洗い流すことができるため培養がより容易である。 The havanori and mozuku filaments used in the present invention have an advantage that they can be easily agitated and cultured in a container because they are less sticky to the container and difficult to mature than the Okinawa mozuku used conventionally. When the attachment property is high, it adheres to the wall surface of the container in a moss-like manner, and light irradiation is hindered, and stirring culture by aeration is delayed, resulting in a decrease in production. In addition, habanori and mozuku filaments are more resistant to fresh water than Okinawa mozuku, and impurities can be washed away by treating with fresh water, so that culture is easier.
以下に、実施例を挙げて本発明をより詳細に説明するが、本発明はこれら実施例に何ら限定されるものではない。 Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples.
実 施 例 １
ハバノリ糸状体の培養（１）：水温の影響
下記表１の組成のＰＥＳＩ培養液を２％添加したろ過海水を１２１℃４０分間オートクレーブしたものを培養液とした。この培養液９００ｍｌを１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．２ｇ ｄｒｙ ｗｅｉｇｈｔ （以下、ｄ．ｗ．））加え、空気を２．３Ｌ／分で導入し、温度１５、２０、２５、３０℃の４条件により、光量８０μｍｏｌ・ｍ−２・ｓ−１、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後乾燥重量を測定した。
Example 1
Cultivation of Habanori filamentous body (1): Influence of water temperature A solution obtained by autoclaving filtered seawater to which 2% of PESI culture solution having the composition shown in Table 1 below was added at 121 ° C. for 40 minutes was used as a culture solution. 1 g (dry weight: 0.2 g dry weight (hereinafter referred to as dw)) of 900 ml of this culture broth placed in a 1 L flat bottom flask and dehydrated for 6 minutes with a commercially available dehydrator after filtering through a 30 μm mesh. In addition, air was introduced at 2.3 L / min, and the cells were cultured for 1 week under a light amount of 80 μmol · m −2 · s −1 and a light cycle of 12 L: 12D under four conditions of temperatures of 15, 20, 25, and 30 ° C. The dry weight was measured after culture.
実 施 例 ２
ハバノリ糸状体の培養（２）：光量の影響
実施例１と同じ組成のＰＥＳＩ培養液を２％添加したろ過海水を１２１℃４０分間オートクレーブしたものを培養液とした。この培養液９００ｍｌを１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．２ｇ ｄ．ｗ）加え、空気を２．３Ｌ／分で導入し、光量１０、２０、４０、８０、１６０μｍｏｌ・ｍ−２・ｓ−１、水温２５℃、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後乾燥重量を測定した。
Example 2
Cultivation of Habanori filamentous body (2): Effect of light intensity A culture solution was obtained by autoclaving filtered seawater to which 2% of PESI culture solution having the same composition as in Example 1 was added at 121 ° C. for 40 minutes. 900 g of this culture solution was put into a 1 L flat bottom flask, 1 g (dry weight: 0.2 g d.w) of the filamentous material filtered through a 30 μm mesh for 6 minutes with a commercially available dehydrator was added, and 2.3 L of air was added. Per minute, and cultured for one week under light amounts of 10, 20, 40, 80, 160 μmol · m −2 · s −1 , a water temperature of 25 ° C., and a photoperiod of 12 L: 12D. The dry weight was measured after culture.
実 施 例 ３
ハバノリ糸状体の培養（３）：培養液組成の影響
実施例１と同じ組成および濃度のＰＥＳＩ培養液（試験区１）と、この組成において窒素源を硝酸ナトリウムから硫酸アンモニウムに代えたもの（試験区２）、リン源をグリセロリン酸２ナトリウムからリン酸３ナトリウムに代えたもの（試験区３）、窒素、リン源ともに硫酸アンモニウム、リン酸３ナトリウムに代えたもの（試験区４）、の４種類の培養液を作成した。培養液９００ｍｌをそれぞれ１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．２ｇ ｄ．ｗ）加え、空気を２．３Ｌ／分で導入し、水温２５℃、光量８０μｍｏｌ・ｍ−２・ｓ−１、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後乾燥重量を測定した。
Example 3
Cultivation of Habanori filamentous body (3): Influence of culture solution composition PESI culture solution (test group 1) having the same composition and concentration as in Example 1, and in this composition, the nitrogen source was changed from sodium nitrate to ammonium sulfate (test group) 2) Four types of phosphorous source: glycerophosphate disodium to trisodium phosphate (test group 3), nitrogen and phosphorus source both ammonium sulfate and trisodium phosphate (test group 4) A culture solution was prepared. Place 900 ml of each culture solution in a 1 L flat bottom flask, add 1 g (dry weight: 0.2 g d.w) of the filamentous material filtered through a 30 μm mesh for 6 minutes with a commercially available dehydrator, and add 2.3 L of air. Per minute, and cultured for 1 week under a water temperature of 25 ° C., a light intensity of 80 μmol · m −2 · s −1 , and a photoperiod of 12 L: 12D. The dry weight was measured after culture.
実 施 例 ４
ハバノリ糸状体からのフコキサンチンおよびフコステロールの製造：
（フコキサンチンおよびフコステロールの抽出）
実施例１ないし３で得られたハバノリ糸状体を凍結乾燥し、２００倍容のメタノールにて４０℃で１時間抽出を行なった。抽出液は残渣を除去した後、減圧下にて溶媒を留去し、残留物を一定量のメタノールに再溶解し、分析用サンプルとした。下記方法により、フコキサンチンおよびフコステロールを測定した。結果を乾燥重量と併せて表２に示す。
Example 4
Production of fucoxanthin and fucosterol from habanori filaments:
(Extraction of fucoxanthin and fucosterol)
The habanori filaments obtained in Examples 1 to 3 were lyophilized and extracted with 200 volumes of methanol at 40 ° C. for 1 hour. After the residue was removed from the extract, the solvent was distilled off under reduced pressure, and the residue was redissolved in a fixed amount of methanol to obtain a sample for analysis. Fucoxanthin and fucosterol were measured by the following method. The results are shown in Table 2 together with the dry weight.
（フコキサンチンの定量方法）
フコキサンチンはＨＰＬＣにより定量を行った。カラムはＣＯＳＭＯＳＩＬ ５Ｃ２２−ＡＲ−ＩＩ（ナカライテスク製、内径４．６ｘ２５０ｍｍ）を用い、移動相はアセトニトリル／水の混合液（８０：２０）を用いた。フコキサンチンの検出は波長４５０ｎｍの吸収で行い、流速は１．５ｍｌ／ｍｉｎで分析を行った。
(Quantification method of fucoxanthin)
Fucoxanthin was quantified by HPLC. The column was COSMOSIL 5C 22 -AR-II (manufactured by Nacalai Tesque, inner diameter 4.6 × 250 mm), and the mobile phase was a mixture of acetonitrile / water (80:20). Fucoxanthin was detected by absorption at a wavelength of 450 nm, and analysis was performed at a flow rate of 1.5 ml / min.
（フコステロールの定量方法）
フコステロールはＧＣ−ＭＳ（ガスクロマトグラフ質量分析装置）により定量を行った。カラムはＪ＆Ｗ社製ＤＢ−５ＭＳ（内径０．２５ｍｍｘ３０ｍ、膜厚０．２５ｍ）を用い、キャリアガスとしてヘリウムを用いた。フコステロールの検出はＳＩＭモードでフコステロール由来フラグメントイオンｍ／ｚ＝３１４で行った。
(Fucosterol quantification method)
Fucosterol was quantified by GC-MS (gas chromatograph mass spectrometer). The column was DB-5MS (inner diameter 0.25 mm × 30 m, film thickness 0.25 m) manufactured by J & W, and helium was used as a carrier gas. The fucosterol was detected in SIM mode with a fragment ion derived from fucosterol m / z = 314.
２０、２５℃条件下では、培養開始時の２倍の重量となった。また検鏡の結果、糸状体が伸長していたことから、この重量の増加は糸状体の生長によるものであった。培養液中に直立体の形成は認められなかった。さらにフコキサンチン含量およびフコステロール含量は水温が高くなるほど含量が多くなった。 Under 20 and 25 ° C. conditions, the weight was twice that at the start of the culture. Further, as a result of microscopic examination, the filamentous body was elongated, and this increase in weight was due to the growth of the filamentous body. The formation of a straight solid was not observed in the culture solution. Further, the fucoxanthin content and fucosterol content increased as the water temperature increased.
また、光量が強くなるほど増体量が大きく、２０μｍｏｌ・ｍ−２・ｓ−１では培養開始時の２倍、４０μｍｏｌ・ｍ−２・ｓ−１では２．５倍、８０μｍｏｌ・ｍ−２・ｓ−１では３．５倍、１６０μｍｏｌ・ｍ−２・ｓ−１では４倍の重量となった。また検鏡の結果、糸状体が伸長していたことから、この重量の増加は糸状体の生長によるものであった。培養液中に直立体の形成は認められなかった。フコキサンチン含量については、極端に光量の強い１６０μｍｏｌ・ｍ−２・ｓ−１で含量が少なくなった。 In addition, as the amount of light increases, the amount of gain increases, and 20 μmol · m −2 · s −1 is double that at the start of culture, 40 μmol · m −2 · s −1 is 2.5 times, and 80 μmol · m −2. The weight was 3.5 times as high as s −1 and 4 times as high as 160 μmol · m −2 · s −1 . Further, as a result of microscopic examination, the filamentous body was elongated, and this increase in weight was due to the growth of the filamentous body. The formation of a straight solid was not observed in the culture solution. Regarding the fucoxanthin content, the content decreased at 160 μmol · m −2 · s −1 with extremely strong light.
さらに培養液の組成については、試験区１で０．５６ｇ ｄ．ｗ．、試験区２で０．６４ｇ ｄ．ｗ．、試験区３で０．５７ｇ ｄ．ｗ．、試験区４で０．５６ｇ ｄ．ｗ．となり、全ての条件で約３倍の重量となった。また検鏡の結果、糸状体が伸長していたことから、この重量の増加は糸状体の生長によるものであった。培養液中に直立体の形成は認められなかった。さらにフコキサンチン含量について、試験区１では２．３１ｍｇ／ｇ ｄ．ｗ．であるのに対し、試験区２で３．２０ｍｇ／ｇ ｄ．ｗ、試験区３で３．８５ｍｇ／ｇ ｄ．ｗ．、試験区４で４．５３ｍｇ／ｇ ｄ．ｗ．となり、窒素源、リン源として硫酸アンモニウム、リン酸３ナトリウムを用いると、相加的に含量が多くなった。フコステロール含量は試験区１で２．６８ｍｇ／ｇ ｄ．ｗ．、試験区２で３．０７ｍｇ／ｇ ｄ．ｗ．、試験区３で３．２２ｍｇ／ｇ ｄ．ｗ．、試験区４で３．８０ｍｇ／ｇ ｄ．ｗ．となり、窒素源、リン源として硫酸アンモニウム、リン酸３ナトリウムを用いると、相加的に含量が多くなった。 Furthermore, regarding the composition of the culture solution, 0.56 g in test group 1 d. w. , 0.64 g in test section 2 d. w. In test zone 3, 0.57 g d. w. In test section 4, 0.56 g d. w. It became about 3 times the weight under all conditions. Further, as a result of microscopic examination, the filamentous body was elongated, and this increase in weight was due to the growth of the filamentous body. The formation of a straight solid was not observed in the culture solution. Furthermore, about the fucoxanthin content, it was 2.31 mg / g d. w. On the other hand, 3.20 mg / g in test group 2 d. w, 3.85 mg / g in test group 3 d. w. 4.53 mg / g in test group 4 d. w. When ammonium sulfate and trisodium phosphate were used as the nitrogen source and phosphorus source, the contents increased in addition. The fucosterol content was 2.68 mg / g d. w. In test area 2, 3.07 mg / g d. w. In the test group 3, 3.22 mg / g d. w. In test group 4, 3.80 mg / g d. w. When ammonium sulfate and trisodium phosphate were used as the nitrogen source and phosphorus source, the contents increased in addition.
実 施 例 ５
モズク糸状体の培養（１）：水温の影響
実施例１の組成のＰＥＳＩ培養液を２％添加したろ過海水を１２１℃４０分間オートクレーブしたものを培養液とした。この培養液９００ｍｌを１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．２ ｇ ｄ．ｗ．）加え、空気を２．３Ｌ／分で導入し、温度１５、２０、２５、３０℃の４条件、それ以外の条件として光量８０μｍｏｌ・ｍ−２・ｓ−１、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後、乾燥重量を測定した。
Example 5
Cultivation of Mozuku filamentous body (1): Influence of water temperature A culture solution was obtained by autoclaving filtered seawater to which 2% of PESI culture solution having the composition of Example 1 was added at 121 ° C. for 40 minutes. 900 g of this culture broth was placed in a 1 L flat bottom flask, 1 g (dry weight: 0.2 g dw) of a filamentous material filtered through a 30 μm mesh and dehydrated for 6 minutes with a commercially available dehydrator was added, and 2 air was added. The sample was introduced at 3 L / min, and cultured under 4 conditions of temperatures of 15, 20, 25, and 30 ° C., and under other conditions, with a light amount of 80 μmol · m −2 · s −1 and a photoperiod of 12 L: 12D for 1 week. After incubation, the dry weight was measured.
実 施 例 ６
モズク糸状体の培養（２）：光量の影響
実施例１の組成のＰＥＳＩ培養液を２％添加したろ過海水を１２１℃４０分間オートクレーブしたものを培養液とした。この培養液９００ｍｌを１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．２ ｇ ｄ．ｗ）加え、空気を２．３Ｌ／分で導入し、光量０、１０、２０、４０、８０、１６０μｍｏｌ・ｍ−２・ｓ−１、それ以外の条件として水温２５℃、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後、乾燥重量を測定した。
Example 6
Cultivation of mozuku filamentous body (2): Effect of light quantity A culture solution was obtained by autoclaving filtered seawater to which 2% of PESI culture solution having the composition of Example 1 was added at 121 ° C for 40 minutes. 900 g of this culture broth was placed in a 1 L flat bottom flask, 1 g (dry weight: 0.2 g d.w) of the filamentous material filtered through a 30 μm mesh for 6 minutes with a commercially available dehydrator was added, and air was added in an amount of 2. The mixture was introduced at 3 L / min, and cultured for 1 week under a light temperature of 0, 10, 20, 40, 80, 160 μmol · m −2 · s −1 , and a water temperature of 25 ° C. and a photoperiod of 12 L: 12 D as other conditions. After incubation, the dry weight was measured.
実 施 例 ７
モズク糸状体の培養（３）：培養液組成の影響
実施例１と同じ組成・濃度のＰＥＳＩ培養液（試験区１）と、組成中の窒素源を硝酸ナトリウムから硫酸アンモニウムに代えたもの（試験区２）、リン源をグリセロリン酸２ナトリウムからリン酸３ナトリウムに代えたもの（試験区３）、窒素、リン源ともに硫酸アンモニウム、リン酸３ナトリウムに代えたもの（試験区４）、の４種類の培養液を作成した。作成した培養液９００ｍｌをそれぞれ１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．２ｇ ｄ．ｗ）加え、空気を２．３Ｌ／分で導入し、水温２５℃、光量８０μｍｏｌ・ｍ−２・ｓ−１、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後乾燥重量を測定した。
Example 7
Cultivation of mozuku filamentous body (3): Influence of culture solution composition PESI culture solution (test group 1) having the same composition and concentration as in Example 1 and a nitrogen source in the composition changed from sodium nitrate to ammonium sulfate (test group) 2) Four types of phosphorous source: glycerophosphate disodium to trisodium phosphate (test group 3), nitrogen and phosphorus source both ammonium sulfate and trisodium phosphate (test group 4) A culture solution was prepared. Place 900 g of the prepared culture solution in a 1 L flat bottom flask, add 1 g (dry weight: 0.2 g d.w) of the filamentous material filtered through a 30 μm mesh for 6 minutes with a commercial dehydrator, and add 2 air. The solution was introduced at 3 L / min, and cultured for 1 week under a water temperature of 25 ° C., a light intensity of 80 μmol · m −2 · s −1 , and a photoperiod of 12 L: 12D. The dry weight was measured after culture.
実 施 例 ８
モズク糸状体からのフコキサンチンおよびフコステロールの製造：
（フコキサンチンおよびフコステロールの抽出）
実施例５ないし７で得られたモズク糸状体を凍結乾燥し、２００倍容のメタノールにて４０℃で１時間抽出を行なった。抽出液は残渣を除去した後、減圧下にて溶媒を留去し、残留物を一定量のメタノールに再溶解し、分析用サンプルとした。実施例４と同様の方法により、フコキサンチンおよびフコステロールを測定した。結果を乾燥重量と併せて表３に示す。
Example 8
Production of fucoxanthin and fucosterol from mozuku filaments:
(Extraction of fucoxanthin and fucosterol)
Mozuku filaments obtained in Examples 5 to 7 were lyophilized and extracted with 200 volumes of methanol at 40 ° C. for 1 hour. After the residue was removed from the extract, the solvent was distilled off under reduced pressure, and the residue was redissolved in a fixed amount of methanol to obtain a sample for analysis. Fucoxanthin and fucosterol were measured by the same method as in Example 4. The results are shown in Table 3 together with the dry weight.
水温について、１週間培養後、１５℃で０．３９ｇ ｄ．ｗ．、２０℃で０．５３ｇ ｄ．ｗ．、２５℃で０．５８ｇ ｄ．ｗ．、３０℃で０．５５ｇ ｄ．ｗ．となり、１５℃以外の水温で約２．８倍の増重が認められた。また検鏡の結果、糸状体が伸長していたことから、この重量の増加は糸状体の生長によるものであった。培養液中に直立体の形成は認められなかった。またフコキサンチン含量およびフコステロール含量は水温が低くなるほど含量が多くなった。 Regarding water temperature, after culturing for 1 week, 0.39 g at 15 ° C. d. w. 0.53 g at 20 ° C. d. w. 0.58 g at 25 ° C. d. w. 0.55 g at 30 ° C. d. w. Thus, a weight increase of about 2.8 times was observed at water temperatures other than 15 ° C. Further, as a result of microscopic examination, the filamentous body was elongated, and this increase in weight was due to the growth of the filamentous body. The formation of a straight solid was not observed in the culture solution. The fucoxanthin content and fucosterol content increased as the water temperature decreased.
また光量が強くなるほど増重した。２０μｍｏｌ・ｍ−２・ｓ−１では培養開始時の２．３倍、４０μｍｏｌ・ｍ−２・ｓ−１では２．７倍、８０μｍｏｌ・ｍ−２・ｓ−１では３倍、１６０μｍｏｌ・ｍ−２・ｓ−１では３．１倍の重量となった。検鏡の結果、糸状体が伸長していたことから、この重量の増加は糸状体の生長によるものであった。培養液中に直立体の形成は認められなかった。さらにフコキサンチン含量およびフコステロール含量については、低光量の１０μｍｏｌ・ｍ−２・ｓ−１で含量が最も多くなった。 The weight increased as the light intensity increased. 20 μmol · m −2 · s −1 is 2.3 times the start of culture, 40 μmol · m −2 · s −1 is 2.7 times, 80 μmol · m −2 · s −1 is 3 times, 160 μmol · m -2 · s -1 was 3.1 times the weight. As a result of microscopic examination, the filament was elongated, and this increase in weight was due to the growth of the filament. The formation of a straight solid was not observed in the culture solution. Further, the fucoxanthin content and fucosterol content were highest at a low light intensity of 10 μmol · m −2 · s −1 .
さらに培養液については、試験区１で０．４２ｇ ｄ．ｗ．、試験区２で０．４５ｇ ｄ．ｗ．、試験区３で０．４６ｇ ｄ．ｗ．、試験区４で０．５０ｇ ｄ．ｗ．となり、窒素源、リン源として硫酸アンモニウム、リン酸３ナトリウムを用いることにより、増体量が大きくなった。また検鏡の結果、糸状体が伸長していたことから、この重量の増加は糸状体の生長によるものであった。培養液中に直立体の形成は認められなかった。さらにフコキサンチン含量について、試験区１では０．３４ｍｇ／ｇ ｄ．ｗ．であるのに対し、試験区２で０．９４ｍｇ／ｇ ｄ．ｗ．、試験区３で１．３１ｍｇ／ｇ ｄ．ｗ．、試験区４で０．８６ｍｇ／ｇ ｄ．ｗ．となり、リン源をリン酸３ナトリウムに代えたもので含量が最も多くなった。フコステロール含量は試験区１では０．４４ｍｇ／ｇ ｄ．ｗ．であるのに対し、試験区２で１．２４ｍｇ／ｇ ｄ．ｗ．、試験区３で１．０１ｍｇ／ｇ ｄ．ｗ．、試験区４で１．０３ｍｇ／ｇ ｄ．ｗ．となり、窒素源、リン源として硫酸アンモニウム、リン酸３ナトリウムを用いると含量が多くなった。 Furthermore, regarding the culture solution, 0.42 g d. w. , 0.45 g in test section 2 d. w. 0.46 g in test section 3 d. w. In test section 4, 0.50 g d. w. Thus, the amount of gain increased by using ammonium sulfate and trisodium phosphate as the nitrogen source and phosphorus source. Further, as a result of microscopic examination, the filamentous body was elongated, and this increase in weight was due to the growth of the filamentous body. The formation of a straight solid was not observed in the culture solution. Furthermore, regarding the fucoxanthin content, 0.34 mg / g d. w. In contrast, in test group 2, 0.94 mg / g d. w. 1.31 mg / g in test group 3 d. w. In test group 4, 0.86 mg / g d. w. Thus, the content was highest when the phosphorus source was replaced with trisodium phosphate. Fucosterol content was 0.44 mg / g in test group 1 d. w. On the other hand, 1.24 mg / g d. w. 1.01 mg / g in test group 3 d. w. 1.03 mg / g in test group 4 d. w. Thus, when ammonium sulfate or trisodium phosphate was used as the nitrogen source and phosphorus source, the content increased.
比 較 例 １
オキナワモズク糸状体の培養：光量の影響
実施例１の組成のＰＥＳＩ培養液を２％添加したろ過海水を１２１℃４０分間オートクレーブしたものを培養液とした。この培養液９００ｍｌを１Ｌの平底フラスコに入れ、３０μｍメッシュで濾過した糸状体を市販の脱水機で６分間脱水したものを１ｇ（乾燥重量：０．１３ｇ ｄ．ｗ）加え、空気を２．３Ｌ／分で導入し、光量０、１０、２０、４０、８０、１６０μｍｏｌ・ｍ−２・ｓ−１、それ以外の条件として水温２５℃、光周期１２Ｌ：１２Ｄ下で１週間培養した。培養後、乾燥重量を測定した。
Comparative Example 1
Culture of Okinawa Mozuku filamentous body: Influence of light quantity The culture solution was obtained by autoclaving filtered seawater to which 2% of PESI culture solution having the composition of Example 1 was added at 121 ° C for 40 minutes. 900 g of this culture broth was placed in a 1 L flat bottom flask, 1 g (dry weight: 0.13 g d.w) of the filamentous material filtered through a 30 μm mesh for 6 minutes with a commercial dehydrator was added, and 2.3 L of air was added. / introduced in quantity of the light 0,10,20,40,80,160μmol · m -2 · s -1, water temperature 25 ° C. as other conditions, photoperiod 12L: were cultured for one week under 12D. After incubation, the dry weight was measured.
増体量は最大でも２．５倍にとどまり、ハバノリやモズクと比べて小さいものであった。 The amount of gain was only 2.5 times at the maximum, which was small compared to Habanori and Mozuku.
本発明によれば、褐藻類からフコキサンチンおよびフコステロールを高収量で得ることができるため、健康食品や医薬品等に用いる原料の製造方法として極めて有用である。
以 上
According to the present invention, fucoxanthin and fucosterol can be obtained from brown algae in a high yield, which is extremely useful as a method for producing raw materials used for health foods, pharmaceuticals, and the like.
more than
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| CN105012647A (en) * | 2015-08-27 | 2015-11-04 | 济南邦文医药科技有限公司 | Traditional Chinese medicine composition for treating cold-dampness blocking collaterals type diabetic limb early-stage arteriosclerosis obliterans |
| JP2018068245A (en) * | 2016-11-01 | 2018-05-10 | 日本粉末薬品株式会社 | Method for extracting fat-soluble component from alga |
| JP2020074733A (en) * | 2018-11-09 | 2020-05-21 | 株式会社サウスプロダクト | Method for producing algae high in fucoxanthin |
| CN111235207A (en) * | 2020-04-16 | 2020-06-05 | 杭州巴洛特生物科技有限公司 | Process for improving efficiency of synthesizing fucosterol from sargassum thunbergii |
| CN111410676A (en) * | 2020-04-22 | 2020-07-14 | 杭州巴洛特生物科技有限公司 | Method for producing fucosterol pharmaceutical intermediate and algal polysaccharide |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105012647A (en) * | 2015-08-27 | 2015-11-04 | 济南邦文医药科技有限公司 | Traditional Chinese medicine composition for treating cold-dampness blocking collaterals type diabetic limb early-stage arteriosclerosis obliterans |
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| JP2020074733A (en) * | 2018-11-09 | 2020-05-21 | 株式会社サウスプロダクト | Method for producing algae high in fucoxanthin |
| JP2021045120A (en) * | 2019-09-12 | 2021-03-25 | 理研食品株式会社 | Germination method of upright nemacystus decipiens from discal nemacystus decipiens |
| CN111235207A (en) * | 2020-04-16 | 2020-06-05 | 杭州巴洛特生物科技有限公司 | Process for improving efficiency of synthesizing fucosterol from sargassum thunbergii |
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