JP7495066B2 - 栄養価が改善した藻類オイル - Google Patents
栄養価が改善した藻類オイル Download PDFInfo
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- JP7495066B2 JP7495066B2 JP2021519825A JP2021519825A JP7495066B2 JP 7495066 B2 JP7495066 B2 JP 7495066B2 JP 2021519825 A JP2021519825 A JP 2021519825A JP 2021519825 A JP2021519825 A JP 2021519825A JP 7495066 B2 JP7495066 B2 JP 7495066B2
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- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 1
- KKCBUQHMOMHUOY-UHFFFAOYSA-N sodium oxide Chemical compound [O-2].[Na+].[Na+] KKCBUQHMOMHUOY-UHFFFAOYSA-N 0.000 description 1
- 229910001948 sodium oxide Inorganic materials 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 235000020712 soy bean extract Nutrition 0.000 description 1
- 235000011496 sports drink Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- JBQYATWDVHIOAR-UHFFFAOYSA-N tellanylidenegermanium Chemical compound [Te]=[Ge] JBQYATWDVHIOAR-UHFFFAOYSA-N 0.000 description 1
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- IHIXIJGXTJIKRB-UHFFFAOYSA-N trisodium vanadate Chemical compound [Na+].[Na+].[Na+].[O-][V]([O-])([O-])=O IHIXIJGXTJIKRB-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 239000004034 viscosity adjusting agent Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 description 1
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- A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
- A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
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- A61K8/00—Cosmetics or similar toiletry preparations
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Description
本出願は、2018年11月2日出願の米国仮出願第62/754,896号の優先権を主張し、当該文献は、その全体が参照によって本明細書に組み込まれる。
本明細書では、微生物オイルの栄養価及び物理的特性を改善する発酵方法が提供される。具体的には、オメガ7脂肪酸の含量が増加したオイルの生産方法が提供される。この方法は、亜鉛含量が0.3mg/L未満の発酵培地においてオイル生産微生物を培養することを含み、この培養によって、脂肪酸を含むオイルが生産され、オイルのオメガ7脂肪酸の含量は、対照オイルと比較して増加する。任意選択で、オイルは、培養物の微生物から単離される。任意選択で、オイルの脂肪酸に占める飽和脂肪酸の割合は35%以下である。
この実施例では、炭素消費速度の制御を伴う微生物発酵によって達成可能な典型的な脂肪酸プロファイルが示される。30Lの発酵槽において仕込体積を20L~30Lとして発酵を実施し、発酵の間にグルコースシロップを供給したことにより内容物体積は増加した。対照発酵培地には通常量の亜鉛を含めた(リットル当たり):グルコース60g、大豆ペプトン2g、塩化ナトリウム1.65g、硫酸マグネシウム七水和物4g、リン酸二水素カリウム2.2g、リン酸水素二カリウム2.4g、硫酸アンモニウム20g、塩化カルシウム二水和物0.1g、塩化鉄3mg、硫酸銅五水和物3mg、モリブデン酸ナトリウム二水和物1.5mg、硫酸亜鉛七水和物3mg、塩化コバルト六水和物1.5mg、塩化マンガン四水和物1.5mg、硫酸ニッケル六水和物1.5mg、ビタミンB12 0.03mg、ビオチン0.03mg、及びチアミン塩酸塩6mg。シリコーンベースの消泡剤を必要に応じて控え目に使用して発泡を抑制し、発酵全体を通じてこの消泡剤の使用量を0.3g/L未満とした。培養における非制限的な炭素消費速度が最大で3g/L/時間となるように発酵槽の撹拌及びエアレーションを制御した。培養物が消費する利用可能なグルコースが培地中に常に存在するように、培養または発酵を通じてグルコースシロップの形態の炭素を発酵槽に追加供給した。
この実施例では、亜鉛が、MUFA合成の抑制を担う微量元素であることが示される。7Lの発酵槽において発酵を実施した。対照発酵培地には、実施例1と同じ栄養材料を含めた。硫酸銅五水和物、モリブデン酸ナトリウム二水和物、及び硫酸亜鉛七水和物を含めない発酵培地を調製した。この発酵培地は、図4及び図5ならびに表2において「TEなし」と称される。発酵培地を追加で3つ調製した。1つ目は銅を含めないもの(「Cuなし」)とし、2つ目はモリブデン酸を含めないもの「Mbなし」とし、3つ目は亜鉛を含めないもの(「Znなし」)とした。しかしながら、「Znなし」培地は、水由来の残留亜鉛を約0.1mg/Lでそのまま含むものとした。こうした培地は、これら3つのうちのどれが、非添加時にMUFA生産を引き起こすのかを特定するために調製した。実験は2連で実施し、平均値を示す。亜鉛に曝露されなかった培養物(「Znなし」)のオイルプロファイルは、微量元素を添加しなかった培養物(「TEなし」)のものと非常に類似していた(表2)。微量元素を含めなかった培養物中のMUFA含量、及び亜鉛を含めなかった培養物中のMUFA含量は、それぞれ25.31%及び22.85%であった一方で、銅を含めなかった培養物中のMUFA含量(5.76%)、またはモリブデンを含めなかった培養物中のMUFA含量(3.36%)は、微量元素を完全に補充して含めた培養物のもの(6.73%)と非常に類似していた。脂質プロファイルにおける主要な飽和脂肪酸は、ミリスチン酸(C14:0)及びパルミチン酸(C16:0)の2つであった。脂質プロファイルに存在した他の飽和脂肪酸(C12:0、C13:0、C15:0、C17:0、C18:0、及びC20:0)は、濃度がはるかに低く、「他の脂肪酸」カテゴリー下のデータを構成する。ミリスチン酸濃度は変化せずそのままであり(11.21~12.22%)、どの発酵培地を使用したかとは無関係であった。亜鉛を含めなかった培養物(「Znなし」及び「TEなし」)では、亜鉛を添加して含めた培養物のもの((28.21%(「Cuなし」)、27.62%(「Mbなし」)、及び26.69%(「完全TE」)、表2)と比較してパルミチン酸の大幅な減少が見られた(14.9%及び16.37%、表2)。その結果、亜鉛を含めなかった培養物中の飽和脂肪酸含量は、亜鉛を含めた培養物のものと比較してはるかに少なかった(31.21%(「TEなし」)及び31.72%(「Znなし」)、表2)。そのようなオイルプロファイルでは、低温流動特性が改善している。銅及びモリブデン酸を添加せずに増殖を行った培養物からは、飽和脂肪酸含量がそれぞれ43.72%及び44.05%である脂質プロファイルが得られ、これらの脂質プロファイルは、飽和脂肪酸含量が42.97%である対照培養脂質と非常に類似している。これら3つの培養物のいずれからも、低温流動特性(例えば、室温でのオイル流動性)が改善したオイルは得られなかった。銅及びモリブデン酸を添加しないようにしても、微量元素を完全に補充して得られた培養物と比較して脂質プロファイルの変化は生じない(図4)。亜鉛を添加せずに増殖を行った培養物中の一価不飽和脂肪酸の主要メンバーは、C16:1(n-7)パルミトレイン酸及び18:1(n-7)バクセン酸であり(図5、「Znなし」及び「TEなし」)、オメガ7脂肪酸の総量は7.27g/Lから20.03g/Lとなり、175%増加した(図5、「Znなし」及び「完全TE」)。
亜鉛制限培養物でのMUFAの生産は、時間依存的様式で生じる。流加発酵においてスラウストキトリッドを培養し、発酵の過程にわたって約440g/Lのグルコースが消費された。消費速度を2~3g/L/時間に維持した。発酵制御の差異に起因して、この実施例及び前の実施例に記載のプロセスに要する時間は100~300時間となり得る。そのため、この実施例で論じる発酵を、時間ではなく総消費炭素のパーセントに標準化した。亜鉛元素を0.104mg/L、0.195mg/L、0.286mg/L、及び0.651mg/Lとして培養物を得た。0.104mg/Lの亜鉛は水中に存在する亜鉛の量であり、この培養培地に亜鉛を追加で添加することは行わなかった。利用可能な炭素が80%消費された後、亜鉛の濃度が0.65mg/L未満の培養物ではMUFAの生産が始まる一方で、亜鉛を過剰に含む培養物では全MUFA含量の増加は見られない(表4)。発酵プロセスの最後の数時間の間に、すべての発酵においておよそ23g/Lのバイオマスが生産され、新たなバイオマスの80~100%が脂質であった。表4は、炭素供給の最後の20%が消費される間の脂肪酸プロファイルの変化を示す。亜鉛の濃度を0.65mg/L未満とした培養物では、新たに生産された脂質の大部分がパルミトレイン酸(C16:1(n-7))、バクセン酸(C18:1(n-7))、及びドコサヘキサエン酸(C22:6(n-3))の形態であった(表4及び図8)。対照培養物は、亜鉛を添加して含めて0.65mg/Lとし、完全微量元素を含めたものであるが、新たに生産されたものは、ほどんどがパルミチン酸(C16:0)及びドコサヘキサエン酸であった。添加した亜鉛が0.65mg/L未満の培養物では、新たに生産されたすべての脂質に占める一価不飽和脂肪酸の割合は32%、60%、及び56%であった一方で、対照培養物によって新たに生産されたすべての脂質に占めるMUFAの割合は11%にすぎなかった。
Claims (24)
- オメガ7脂肪酸の含量が増加したオイルの生産方法であって、前記方法が、亜鉛含量が0.104~0.3mg/Lの発酵培地においてオイル生産微生物を培養することを含み、前記培養によって、脂肪酸を含むオイルが生産され、前記オイルがDHAを含み、前記オイル生産微生物がスラウストキトリッド株である、方法。
- 前記脂肪酸が飽和脂肪酸を含み、前記飽和脂肪酸が、C16:0(パルミチン酸)及びC14:0(ミリスチン酸)を含む、請求項1に記載の方法。
- 前記オイルが、10~30%のオメガ7脂肪酸を含む、請求項1又は2に記載の方法。
- 前記オメガ7脂肪酸が、パルミトレイン酸(C16:1(n-7))、バクセン酸(C18:1(n-7))、またはそれらの組み合わせを含む、請求項1~3のいずれか1項に記載の方法。
- 前記オイルが、5~15%のバクセン酸(C18:1(n-7))を含む、請求項1~4のいずれか1項に記載の方法。
- 前記オイルが、5~15%のパルミトレイン酸(C16:1(n-7))を含む、請求項1~5のいずれか1項に記載の方法。
- 前記微生物から前記オイルを抽出することをさらに含む、請求項1~6のいずれか1項に記載の方法。
- 前記培地が、0.104~0.15mg/Lの亜鉛を含む、請求項1~7のいずれか1項に記載の方法。
- 前記培地が、0.104~0.2mg/Lの亜鉛を含む、請求項1~7のいずれか1項に記載の方法。
- 前記培地が、0.104~0.270mg/Lの亜鉛を含む、請求項1~7のいずれか1項に記載の方法。
- 前記オイルが、20%未満のパルミチン酸(C16:0)を含む、請求項1~10のいずれか1項に記載の方法。
- 炭素消費速度が、1~4g/L/時間に制御される、請求項1~11のいずれか1項に記載の方法。
- オメガ7脂肪酸の含量が増加したオイルの生産方法であって、前記方法が、亜鉛含量が0.104~0.3mg/Lの発酵培地においてオイル生産微生物を培養することを含み、前記培養によって、脂肪酸を含むオイルが生産され、前記脂肪酸に占める飽和脂肪酸の割合が35%以下であり、前記オイルが、5~15%のバクセン酸(C18:1(n-7))を含み、前記オイル生産微生物がスラウストキトリッド株である、方法。
- 前記オイル中の前記脂肪酸に占める飽和脂肪酸の割合が30%未満である、請求項13に記載の方法。
- 前記オイル中の前記脂肪酸に占める飽和脂肪酸の割合が25%未満である、請求項13に記載の方法。
- 前記飽和脂肪酸が、C16:0(パルミチン酸)及びC14:0(ミリスチン酸)を含む、請求項13~15のいずれか1項に記載の方法。
- 前記オイルが、10~30%のオメガ7脂肪酸を含む、請求項13~16のいずれか1項に記載の方法。
- 前記オメガ7脂肪酸が、パルミトレイン酸(C16:1(n-7))を含む、請求項13~17のいずれか1項に記載の方法。
- 前記オイルが、5~15%のパルミトレイン酸(C16:1(n-7))を含む、請求項18に記載の方法。
- 前記微生物から前記オイルを抽出することをさらに含む、請求項13~19のいずれか1項に記載の方法。
- 前記培地が、0.104~0.15mg/Lの亜鉛を含む、請求項13~20のいずれか1項に記載の方法。
- 前記培地が、0.104~0.2mg/Lの亜鉛を含む、請求項13~20のいずれか1項に記載の方法。
- 前記培地が、0.104~0.270mg/Lの亜鉛を含む、請求項13~20のいずれか1項に記載の方法。
- 前記オイルが、20%未満のパルミチン酸(C16:0)を含む、請求項13~23のいずれか1項に記載の方法。
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US20170356018A1 (en) | 2016-06-10 | 2017-12-14 | MARA Renewables Corporation | Method of making lipids with improved cold flow properties |
JP2018520636A (ja) | 2016-04-01 | 2018-08-02 | 中国科学院青島生物能源与過程研究所 | ω−7脂肪酸合成物、及び黄緑色藻を培養して該合成物を生産する方法と応用 |
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