JP2009095304A - Monascus purpureus strain excellent in monakolin k productivity - Google Patents
Monascus purpureus strain excellent in monakolin k productivity Download PDFInfo
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- 244000113306 Monascus purpureus Species 0.000 title abstract description 5
- 235000002322 Monascus purpureus Nutrition 0.000 title abstract description 5
- 229930188430 monakolin Natural products 0.000 title 1
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 claims abstract description 32
- 241000030999 Monascus pilosus Species 0.000 claims abstract description 11
- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 claims description 29
- 238000011156 evaluation Methods 0.000 claims description 3
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- 230000001488 breeding effect Effects 0.000 abstract description 2
- 229940026314 red yeast rice Drugs 0.000 description 31
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 16
- 240000007594 Oryza sativa Species 0.000 description 11
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- PLUBXMRUUVWRLT-UHFFFAOYSA-N Ethyl methanesulfonate Chemical compound CCOS(C)(=O)=O PLUBXMRUUVWRLT-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
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- VZUNGTLZRAYYDE-UHFFFAOYSA-N N-methyl-N'-nitro-N-nitrosoguanidine Chemical compound O=NN(C)C(=N)N[N+]([O-])=O VZUNGTLZRAYYDE-UHFFFAOYSA-N 0.000 description 1
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- 229960004844 lovastatin Drugs 0.000 description 1
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 description 1
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Abstract
Description
本発明は、モナコリンＫの生産能力に優れた紅麹菌株及び紅麹に関する。 The present invention relates to a red yeast strain and a red yeast rice strain excellent in the ability to produce monacolin K.
紅麹は穀類にモナスカス属の菌株を繁殖させた麹で、中国、台湾などでは紅酒、老酒、紅乳腐などの醸造原料として利用されており、また古来より生薬として「消食活血」「健脾燥胃」などの効果が知られている。（李時珍「本草綱目」（１５９０年））。また、血圧降下作用、コレステロール改善作用等、様々な機能を有することが知られ、味噌、醤油、食酢等の醸造食品をはじめ、パン、麺類等、色々な食品に使われている。 Red rice cake is a rice cake made by breeding Monascus strains on cereals, and it has been used as a raw material for brewing red wine, old sake, and red milk tofu in China and Taiwan. Effects such as “stomach” are known. (Lee Jinjin "Hongoku Tuname" (1590)). It is also known to have various functions such as blood pressure lowering action and cholesterol improving action, and is used in various foods such as brewed foods such as miso, soy sauce and vinegar, bread and noodles.
紅麹のコレステロール改善作用については、モナスカス属の菌株によって生産されるモナコリンＫ（ロバスタチンとも呼ばれる）が、体内のコレステロール合成の律速酵素であるHMG-CoAレダクターゼを阻害することにより血中コレステロールを低下させることが明らかにされている（非特許文献１、２、特許文献１〜３）。それに伴い、紅麹はコレステロール調節用のサプリメント素材としての利用も増加している。
As for the cholesterol-improving effect of red yeast rice, monacolin K (also called lovastatin) produced by a strain of Monascus lowers blood cholesterol by inhibiting HMG-CoA reductase, the rate-limiting enzyme for cholesterol synthesis in the body. (
紅麹菌が生産するモナコリンKの量は製麹条件により調節することが可能で、原料に米胚芽を添加し、水分率および培養温度を一定条件に調節することにより、モナコリンK含量が多い紅麹を生産できることが明らかにされている（特許文献４）。さらに紅麹からモナコリンＫを抽出し、高濃度のモナコリンＫを含むエキスが得られることが報告されている（特許文献５）。 The amount of monacolin K produced by koji mold can be adjusted according to the koji making conditions. By adding rice germ to the raw material and adjusting the moisture content and culture temperature to a certain condition, the koji koji with high monacholine K content (Patent Document 4). Furthermore, it has been reported that monacolin K is extracted from red yeast rice so that an extract containing a high concentration of monacolin K can be obtained (Patent Document 5).
ところが現在使用している菌株では、最適製麹条件下でもサプリメント用原料として必要なモナコリンＫ含量（２％）を得ることはできず、サプリメント用の原料としては紅麹ではなくエキス加工した製品が用いられている。しかしエキスは、紅麹に本来含まれているアミノ酸、ミネラルなどの栄養成分の一部が消失しており、直接サプリメント用原料として利用できるモナコリンＫ含量が確保された紅麹が望まれていた。
本発明は、モナコリンKの生産性が高い紅麹菌株、及び当該菌株によって得られる紅麹を提供することを主な目的とするものである。 The main object of the present invention is to provide a red yeast rice strain having high productivity of monacolin K and a red yeast rice obtained by the strain.
本発明は、紅麹菌株モナスカス・ピローサス(Monascus pilosus) NBRC4520に変異処理を行って、モナコリンKの生産性が高い菌株のスクリーニングを試み、該紅麹菌株の取得に至った。 In the present invention, mutation treatment was performed on Monascus pilosus NBRC4520 to try to screen for a strain having high productivity of monacolin K, and the strain was obtained.
本発明は、以下の紅麹菌株及び紅麹を提供する。
項１．モナスカス・ピローサス（Monascus pilosus）に属するモナコリンK高産生性紅麹菌株。
項２．受託番号：NITE P-412によって独立行政法人製品評価技術基盤機構特許微生物寄託センターに受託された項１に記載の紅麹菌株。
項３．項１又は２に記載の紅麹菌株を用いて得られる紅麹。
The present invention provides the following red yeast strain and red yeast rice.
本発明により取得した紅麹菌株を利用すれば、従来菌株では製造できなかった高濃度のモナコリンKを含有する、例えば、紅麹重量に対し２％程度の濃度で含まれる紅麹の製造ができる。また、例えば、モナコリンＫ含量が０．２重量％程度の従来紅麹を生産する場合も、従来菌株を使った場合に比べ、約１週間程度培養期間を短縮することが可能になる。 If the red yeast strain obtained according to the present invention is used, it is possible to produce a red yeast rice containing a high concentration of monacolin K that could not be produced by a conventional strain, for example, at a concentration of about 2% with respect to the weight of the red yeast rice. . In addition, for example, when producing conventional red yeast having a monacolin K content of about 0.2% by weight, the culture period can be shortened by about one week compared to the case of using a conventional strain.
従来は、モナコリンＫを含む紅麹をサプリメント等に調製する場合、通常１０倍程度に濃縮（エキス化）する必要があり、濃縮工程は必須であった。しかしながら、本発明によれば、モナコリンＫ含量が高い紅麹の生産が可能になることから、エキスに代わるサプリメント用原料としての提供が可能になり、従来よりも少量の紅麹で必要量のモナコリンＫを摂取することができ（モナコリンＫの１日摂取推奨量は約2mgとされている）、サプリメントとして調製されやすいことから、より簡便にモナコリンＫによる効果（血中コレステロール値の低下等）が得られる。また、濃縮工程を省略できることから、紅麹に本来含まれているアミノ酸、ミネラルなどの栄養成分の消失を回避できるのみならず、生産コストの低減も可能である。 Conventionally, when preparing red yeast rice cake containing monacolin K as a supplement or the like, it is usually necessary to concentrate (extract) about 10 times, and the concentration step is essential. However, according to the present invention, it becomes possible to produce red yeast rice with a high monacholine K content, so that it can be provided as a raw material for supplements in place of the extract. K can be ingested (the recommended daily intake of monacolin K is about 2 mg), and since it is easy to prepare as a supplement, the effects of monacolin K (such as lowering blood cholesterol levels) can be more easily can get. Further, since the concentration step can be omitted, not only the disappearance of nutrient components such as amino acids and minerals originally contained in red yeast rice can be avoided, but also the production cost can be reduced.
さらに、本発明の紅麹菌株は、目的とする量のモナコリンＫを含量する紅麹を作るのに要する製麹日数が、従来の紅麹菌株に比べて少なくて済み、生産性に優れたものである。 Furthermore, the strain of red yeast rice of the present invention requires less days for making red yeast rice containing the target amount of monacolin K than the conventional strain of red yeast rice and has excellent productivity. It is.
本発明のモナコリンＫ高生産性紅麹菌株は、従来紅麹菌株であるモナスカス属に属する菌(Monasucus. pilosus)を親菌株として、これに対して変異処理を行い、目的菌株をスクリーニングすることにより取得することができる。 The Monacolin K highly productive red yeast strain of the present invention is obtained by screening a target strain by subjecting a strain belonging to the genus Monascus (Monasucus. Can be acquired.
モナコリンＫは紅麹菌の培養物から発見されたコレステロール合成阻害剤で、下記式(1)で示される構造をもつ物質であり、肝臓での生合成を抑制することによって、血中のコレステロールを減少させる働きがある。 Monacholine K is a cholesterol synthesis inhibitor discovered from the culture of Monascus and is a substance with the structure represented by the following formula (1). It reduces blood cholesterol by suppressing biosynthesis in the liver. There is a work to make.
変異処理を行うモナスカス属菌種としては、モナスカス・プルプレウス（Monascus purpureus）、モナスカス・アンカ（Monascus anka）、モナスカス・ピローサス（Monascus pilosus）等が挙げられ、好ましくはモナスカス・ピローサスである。 Examples of the species of the genus Monascus to be mutated include Monascus purpureus, Monascus anka, Monascus pilosus, and the like, preferably Monascus pyrosus.
変異処理法は紫外線(UV)照射、X線照射、γ線照射などの物理的方法、NTG（N-メチル-Ｎ’-ニトロ-N-ニトロソグアニジン）、EMS（エチルメタンスルホネート）などの突然変異誘起処理による化学的方法のいずれでもよい。 Mutation treatment methods include physical methods such as ultraviolet (UV) irradiation, X-ray irradiation, and γ-ray irradiation, mutations such as NTG (N-methyl-N'-nitro-N-nitrosoguanidine) and EMS (ethyl methanesulfonate). Any chemical method by induction treatment may be used.
高モナコリンＫ生産菌株のスクリーニング方法としては、例えば以下のような方法が挙げられる。
変異処理の終了した紅麹菌胞子を基本平板培地上に塗抹し、出現してきたコロニーをランダムに分離する。次に分離した菌株を約５日間、斜面培地で培養し増殖させた後、フラスコスケールの紅麹製麹を無菌的に行う。製麹条件としては、原料の精白米に3〜10％量の米胚芽を添加し、水分率を３５〜５０％に維持、培養温度は２０〜３５℃、ただし製麹後期3日以上の期間は２７℃以下として製麹日数１４〜１７日で行えばよい（特許文献２）。こうして得られた紅麹を１１０℃、２０分間の熱処理により菌および酵素を失活させた後、通風乾燥機で水分率が１０％以下になるまで乾燥し、粉砕機にて粉末化、調製した後、以下の方法でモナコリンＫの生産性を評価する。
Examples of screening methods for high-monacolin K-producing strains include the following methods.
The gonococcal spores that have undergone the mutation treatment are smeared on a basic plate medium, and the colonies that have appeared are randomly separated. The isolated strain is then cultured for about 5 days on a slant medium and allowed to grow, and then flask-scale soy sauce making is performed aseptically. As the koji-making conditions, 3 to 10% rice germ is added to the polished rice as a raw material, the moisture content is maintained at 35 to 50%, and the culture temperature is 20 to 35 ° C. May be carried out at a temperature of 27 ° C. or less for 14 to 17 days of iron making (Patent Document 2). The red yeast rice thus obtained was heat-treated at 110 ° C. for 20 minutes to inactivate the bacteria and enzymes, then dried with a ventilator until the water content became 10% or less, and powdered and prepared with a pulverizer. Thereafter, the productivity of monacolin K is evaluated by the following method.
モナコリンＫ含量は、調製した紅麹粉末に２０〜１００倍量の６０％エタノールを加え、攪拌しながら３０分間抽出し、その抽出液を高速液体クロマトグラフィーにかけて定量し、紫外線処理前の菌株の１．５倍以上、好ましくは２倍以上、より好ましくは２．５倍以上となる菌株をモナコリンＫ高生産性菌の候補株として選抜する。上述の製麹方法であれば、通常、紫外線処理前の菌株のモナコリンＫ生産量（紅麹100g中のモナコリンK含有量）は、50〜300mg/100gである。 Monacolin K content is determined by adding 20 to 100 times the amount of 60% ethanol to the prepared red koji powder, extracting with stirring for 30 minutes, and quantifying the extract by high performance liquid chromatography. A strain that is 5 times or more, preferably 2 times or more, and more preferably 2.5 times or more is selected as a candidate strain for a highly productive monacolin K-producing bacterium. In the case of the above-mentioned koji making method, the amount of monacolin K produced by the strain before ultraviolet treatment (monacolin K content in 100 g of red yeast rice) is usually 50 to 300 mg / 100 g.
以上のようにして得られる具体的な菌株としては、例えば独立行政法人製品評価技術基盤機構特許微生物寄託センター(NMPD)に、平成19年9月4日に受託番号：NITE P-412として寄託されている紅麹菌株が上げられる。NITE P-412は、モナスカス・ピローサス(Monascus pilosus) NBRC4520に生存率約０．５％の割合となる紫外線照射を行い、上記のスクリーニング法で取得したものである。 As a specific strain obtained as described above, for example, it was deposited at the National Institute of Technology and Evaluation (MICPD) on September 4, 2007 as the deposit number: NITE P-412. The strain of red yeast rice is raised. NITE P-412 was obtained by the above-described screening method by irradiating Monascus pilosus NBRC4520 with ultraviolet radiation at a survival rate of about 0.5%.
本発明のNITE P-412菌株と親菌株であるNBRC4520菌株の培養的性質を下記表１に示す。 The culture properties of the NITE P-412 strain of the present invention and the parent strain NBRC4520 strain are shown in Table 1 below.
NITE P-412菌株の形態的性質を表２に示す。 Table 2 shows the morphological properties of NITE P-412 strain.
NITE P-412菌株の生理的性質を表３に示す。 Table 3 shows the physiological properties of the NITE P-412 strain.
上記表１〜３に示されるように、NITE P-412菌株とNBRC4520菌株では、モナコリンＫ産生能以外の培養的性質、形態的性質及び生理的性質は同等であることが確認された。 As shown in Tables 1 to 3 above, it was confirmed that the NITE P-412 strain and the NBRC4520 strain had the same culture properties, morphological properties, and physiological properties other than the ability to produce monacolin K.
さらに、28S rDNAD1/D2およびITS-5.8S rDNA塩基配列の解析結果からも、NITE P-412菌株はモナスカス属の１種と推定され、親菌株であるM. pilosusの塩基配列と98.7〜100％の高い相同率を示した。 Furthermore, from the analysis results of 28S rDNAD1 / D2 and ITS-5.8S rDNA nucleotide sequences, NITE P-412 strain is estimated to be one species of the genus Monascus, and 98.7-100% of the nucleotide sequence of the parent strain M. pilosus. It showed a high homology rate.
本発明のモナスカス属菌株を用いて紅麹を製造し、食品素材等に用いることができる。製麹方法としては、従来公知の方法を採用すればよく、例えば特許第2591635号（米を製造原料とする紅麹の製造法）に記載される方法に従って紅麹を製造することができる。製麹は所望量のモナコリンＫを含有する紅麹が得られるまで行えばよく、製麹期間は適宜設定され得るが、本発明の紅麹菌株はモナコリンＫ高生産性であることから従来の紅麹菌株を用いる場合よりも製麹期間が短縮される。 Red yeast rice can be produced using the Monascus strain of the present invention and used as a food material or the like. As a koji-making method, a conventionally known method may be employed, and for example, koji can be produced according to a method described in Japanese Patent No. 2591635 (a method for producing koji rice using rice as a production raw material). The koji making may be performed until a koji mold containing a desired amount of monacolin K is obtained, and the koji making period can be appropriately set. However, since the koji mold strain of the present invention has high productivity of monakolin K, The ironmaking period is shortened compared with the case of using a koji strain.
本発明の紅麹菌株を用いて得られる紅麹には、血中コレステロール値の低減作用に有効とされるモナコリンＫが通常（紫外線処理前）の紅麹菌株に比べて好ましくは約２倍以上も含有されており、血中コレステロール値改善のための健康食品素材、特にサプリメント用途として有用である。 In the red yeast rice obtained using the red yeast rice strain of the present invention, monacolin K, which is effective in reducing blood cholesterol level, is preferably about twice or more that of the normal (before UV treatment). Is also contained, and is useful as a health food material for blood cholesterol level improvement, particularly as a supplement.
本発明の菌株を用いて得られた紅麹について、モナコリンＫの生産性を分析した試験例を挙げて説明するが、これに限定されるものではない。 The red yeast rice obtained using the strain of the present invention will be described with reference to a test example in which the productivity of monacolin K is analyzed, but is not limited thereto.
試験例１
精白米を一晩水に浸漬し約1時間水切りした後、オートクレーブで125℃、20分間蒸気滅菌し蒸米を得た。これをフラスコ当たり３０ｇ入れた培地に本発明菌株(Monascus pilosus NITE P-412)、または親菌株(Monascus pilosus NBRC4520)を植菌し、最初の４日間は30〜35℃、４日目以後は２３〜２５℃で、計１７日間培養し、培養途上７、１０、１４日目にサンプリングを行った。なお、培養期間中の水分率は４０〜５０％に調整した。こうして得た培養物および培養終了後の紅麹を１１０℃、２０分間の熱処理により紅麹菌と酵素を失活させた後、通風60℃で水分率を約10％まで乾燥し、さらに粉砕して紅麹粉末とし、モナコリンＫ含量を定量した。その結果を表４に示す。
Test example 1
The polished rice was immersed in water overnight and drained for about 1 hour, and then steam sterilized in an autoclave at 125 ° C. for 20 minutes to obtain steamed rice. The strain of the present invention (Monascus pilosus NITE P-412) or the parent strain (Monascus pilosus NBRC4520) is inoculated into a medium containing 30 g of flask per flask. The culture was performed at ˜25 ° C. for a total of 17 days, and sampling was performed on the 7th, 10th, and 14th days of the culture. The moisture content during the culture period was adjusted to 40 to 50%. The culture thus obtained and the red yeast rice after completion of the culture are inactivated by heat treatment at 110 ° C. for 20 minutes, dried at 60 ° C. to a moisture content of about 10%, and further pulverized. Monacholine K content was quantified using red yeast rice powder. The results are shown in Table 4.
これによると、本発明菌株を用いることにより、培養１７日間でモナコリンＫ含量を212mg/100gから634mg/100gへと約３倍に高めることができた。またモナコリンＫ含量が0．2％の紅麹を製造するのに、親菌株を用いた場合は１７日間を要するのに対し、本発明菌株を用いれば１０日間で達成することができ、培養期間を約1週間短縮できることがわかった。 According to this, by using the strain of the present invention, the monacolin K content could be increased about 3 times from 212 mg / 100 g to 634 mg / 100 g in 17 days of culture. In addition, it takes 17 days to produce red yeast rice with a monacholine K content of 0.2% when the parent strain is used, whereas it can be achieved in 10 days using the strain of the present invention. It was found that can be shortened by about one week.
試験例２
精白米を一晩水に浸漬し約1時間水切りした後、オートクレーブで125℃、20分間蒸気滅菌し蒸米を得た。これをフラスコ当たり３０ｇ入れた培地に本発明菌株(Monascus pilosus NITE P-412)、または親菌株(Monascus pilosus NBRC4520)を植菌し、最初の４日間は30〜35℃、４日目以後は２３〜２５℃で、計８週間培養し、培養途上１週間目毎にサンプリングを行った。なお、培養期間中の水分率は４０〜６８％に調整した。こうして得た培養物および培養終了後の紅麹を１１０℃、２０分間の熱処理により紅麹菌と酵素を失活させた後、通風60℃で水分率を約10％まで乾燥し、さらに粉砕して紅麹粉末とし、モナコリンＫ含量を定量した。その結果を図１に示す。
Test example 2
The polished rice was immersed in water overnight and drained for about 1 hour, and then steam sterilized in an autoclave at 125 ° C. for 20 minutes to obtain steamed rice. The strain of the present invention (Monascus pilosus NITE P-412) or the parent strain (Monascus pilosus NBRC4520) is inoculated into a medium containing 30 g of flask per flask. The culture was performed at ˜25 ° C. for a total of 8 weeks, and sampling was performed every week during the culture. The water content during the culture period was adjusted to 40 to 68%. The culture thus obtained and the red yeast rice after completion of the culture are inactivated by heat treatment at 110 ° C. for 20 minutes, dried at 60 ° C. to a moisture content of about 10%, and further pulverized. Monacholine K content was quantified using red yeast rice powder. The result is shown in FIG.
これによると、本発明菌株を用いることにより、８週間の培養でモナコリンＫ含量はほぼ2.5％に達するのに対し、親菌株では0．9％程度までしか達しなかった。 According to this, by using the strain of the present invention, the monacolin K content reached approximately 2.5% after 8 weeks of culture, whereas the parent strain reached only about 0.9%.
Claims (3)
Red potato obtained using the red potato strain according to claim 1.
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| CN108865897A (en) * | 2018-06-29 | 2018-11-23 | 夏放军 | A kind of high yield type monascus strain culture medium and preparation method thereof |
| CN108865898A (en) * | 2018-06-29 | 2018-11-23 | 夏放军 | A kind of breeding method of monascus strain |
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| CN108641968B (en) * | 2018-05-24 | 2021-08-31 | 富乐顿生物工程科技（北京）有限公司 | Spatial monascus purpureus FuH-23-4 screening and application thereof in producing monascus pigment |
| CN111534443B (en) * | 2020-06-12 | 2022-04-01 | 山东省农业科学院农产品研究所 | High-yield MonacolinK purple monascus strain and method for producing functional red yeast rice by fermenting MonacolinK purple monascus strain |
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| JP2000106835A (en) * | 1998-09-30 | 2000-04-18 | Gunze Ltd | Anka and its production |
| JP2000106834A (en) * | 1998-09-30 | 2000-04-18 | Gunze Ltd | Embryo anka |
| JP2004298027A (en) * | 2003-03-28 | 2004-10-28 | Iwata Kagaku Kogyo Kk | Ang-khak culture product containing physiologically active substance and method for producing the same |
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| JP2000106835A (en) * | 1998-09-30 | 2000-04-18 | Gunze Ltd | Anka and its production |
| JP2000106834A (en) * | 1998-09-30 | 2000-04-18 | Gunze Ltd | Embryo anka |
| JP2004298027A (en) * | 2003-03-28 | 2004-10-28 | Iwata Kagaku Kogyo Kk | Ang-khak culture product containing physiologically active substance and method for producing the same |
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| CN108865897A (en) * | 2018-06-29 | 2018-11-23 | 夏放军 | A kind of high yield type monascus strain culture medium and preparation method thereof |
| CN108865898A (en) * | 2018-06-29 | 2018-11-23 | 夏放军 | A kind of breeding method of monascus strain |
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