JP5934458B2 - Eukaryotic microorganism culture method - Google Patents
Eukaryotic microorganism culture method Download PDFInfo
- Publication number
- JP5934458B2 JP5934458B2 JP2009074595A JP2009074595A JP5934458B2 JP 5934458 B2 JP5934458 B2 JP 5934458B2 JP 2009074595 A JP2009074595 A JP 2009074595A JP 2009074595 A JP2009074595 A JP 2009074595A JP 5934458 B2 JP5934458 B2 JP 5934458B2
- Authority
- JP
- Japan
- Prior art keywords
- light
- eukaryotic microorganism
- yeast
- eukaryotic
- growth
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 244000005700 microbiome Species 0.000 title claims description 22
- 238000012136 culture method Methods 0.000 title description 5
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 18
- 230000012010 growth Effects 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 11
- 230000001678 irradiating effect Effects 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 6
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 17
- 241000228212 Aspergillus Species 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 230000001737 promoting effect Effects 0.000 description 4
- 241000235070 Saccharomyces Species 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 240000006439 Aspergillus oryzae Species 0.000 description 2
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000221198 Basidiomycota Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 238000000018 DNA microarray Methods 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000004378 air conditioning Methods 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
この発明は、簡便な手法により酵母やカビ等の真核微生物の増殖速度を容易に制御することができる培養方法に関するものである。 The present invention relates to a culture method capable of easily controlling the growth rate of eukaryotic microorganisms such as yeast and mold by a simple technique.
清酒は、米を原料とする醸造酒であるが、米にはブドウ糖が含まれないため、酵母（主にSaccharomyces cereviseae）による発酵に先立って、コウジカビ（Aspregillus）による糖化（デンプンのブドウ糖への分解）が必要である。そして、糖化と発酵とが並行して行われる（並行複発酵）ことが日本酒の特徴である。 Sake is a brewed liquor made from rice. However, since rice does not contain glucose, saccharification (degradation of starch into glucose) by Aspergillus precedes fermentation by yeast (mainly Saccharomyces cereviseae). )is necessary. A characteristic of Japanese sake is that saccharification and fermentation are performed in parallel (parallel double fermentation).
従来、コウジカビによる糖化や、酵母による発酵の制御は、主に温度を管理することによりなされており、コウジカビによる糖化は約３０℃で行われ、酵母による発酵は８〜１８℃程度で行われている。発酵時の温度がこれより高く酵母の活動が活発であると、雑味成分が増えるので、好ましくなく、このため、清酒造りは一般的に冬季に行われる。 Conventionally, saccharification by Aspergillus or yeast is controlled mainly by controlling the temperature, saccharification by Aspergillus oryzae is performed at about 30 ° C., and fermentation by yeast is performed at about 8 to 18 ° C. Yes. When the temperature at the time of fermentation is higher than this and the activity of yeast is active, miscellaneous components increase, which is not preferable. For this reason, sake brewing is generally performed in winter.
このように温度管理は清酒の出来栄えを左右する大きな要因であるが、温度を厳密に管理することは容易ではなく、温度を厳密に管理しようとすれば大がかりな空調設備等が必要となる。 As described above, temperature management is a major factor that influences the quality of sake. However, it is not easy to strictly control the temperature, and if air temperature is to be strictly controlled, a large-scale air conditioning facility is required.
ところで、従来、特定のスペクトルの光線を照射することにより、微生物による物質生産能を向上させる方法が知られているが（例えば、特許文献１等参照）、光照射により微生物の増殖速度を制御する方法は知られていない。 By the way, conventionally, there is known a method for improving the substance production ability by microorganisms by irradiating with light of a specific spectrum (for example, see Patent Document 1), but the growth rate of microorganisms is controlled by light irradiation. The method is not known.
そこで本発明は、温度管理に代わる簡便な手法により酵母やカビ等の真核微生物の増殖速度を容易に制御することができる培養方法を提供すべく図ったものである。 Therefore, the present invention is intended to provide a culture method capable of easily controlling the growth rate of eukaryotic microorganisms such as yeast and mold by a simple technique instead of temperature control.
本発明者は、鋭意検討の結果、酵母やカビ等の真核微生物を培養する際に、光を照射しながら培養すると、特定の波長の光を照射することにより、その増殖が促進されることを見出し、この知見に基づき本発明を完成させるに至った。 As a result of intensive studies, the present inventor, when cultivating eukaryotic microorganisms such as yeast and mold, while irradiating with light, the proliferation is promoted by irradiating with light of a specific wavelength. Based on this finding, the present invention has been completed.
すなわち本発明に係る真核微生物の培養方法は、真核微生物を培養する方法であって、前記真核微生物に５２０ｎｍ以上の波長の可視光線を照射して、前記真核微生物の増殖を促進する工程を有していることを特徴とする。 That is, the eukaryotic microorganism culturing method according to the present invention is a method for culturing a eukaryotic microorganism, and irradiates the eukaryotic microorganism with visible light having a wavelength of 520 nm or more to promote the growth of the eukaryotic microorganism. It has the process.
前記真核微生物としては、例えば、酵母、カビ等が挙げられる。 Examples of the eukaryotic microorganism include yeast, mold and the like.
前記５２０ｎｍ以上の波長の可視光線としては、黄色光が好適である。 As the visible light having a wavelength of 520 nm or more, yellow light is preferable.
このように本発明によれば、特定の波長の光を照射するという極めて簡便な手法により、酵母やカビ等の真核微生物の増殖を促進して、その増殖速度を制御することができるので、清酒を始めとする種々の醸造製品等を製造する際に、発酵等の速度を容易に制御することができる。 Thus, according to the present invention, by a very simple technique of irradiating light of a specific wavelength, it is possible to promote the growth of eukaryotic microorganisms such as yeast and mold, and to control the growth rate, When producing various brewed products such as sake, the rate of fermentation and the like can be easily controlled.
以下に本発明を詳述する。 The present invention is described in detail below.
本発明は、真核微生物を培養する方法である。前記真核微生物としては特に限定されず、例えば、酵母類、藻類、カビ類、担子菌類等が挙げられる。なかでも、醸造等に利用される酵母（Saccharomyces cereviseae）やコウジカビ（Aspregillus）を培養する際に、本発明に係る培養方法を好適に用いることができる。 The present invention is a method for culturing eukaryotic microorganisms. The eukaryotic microorganism is not particularly limited, and examples thereof include yeasts, algae, molds, basidiomycetes and the like. Among these, when culturing yeast (Saccharomyces cereviseae) and Aspergillus used for brewing or the like, the culture method according to the present invention can be suitably used.
本発明に係る培養方法は、前記真核微生物に５２０ｎｍ以上の波長の可視光線を照射して、前記真核微生物の増殖を促進する工程を有しているものである。 The culture method according to the present invention includes a step of irradiating the eukaryotic microorganism with visible light having a wavelength of 520 nm or more to promote the growth of the eukaryotic microorganism.
前記５２０ｎｍ以上の波長の可視光線としては、例えば、黄色光、橙色光、赤色光等が挙げられるが、なかでも、黄色光を照射したときに、顕著な増殖促進効果が見られる。 Examples of the visible light having a wavelength of 520 nm or more include yellow light, orange light, and red light. Among them, a remarkable proliferation promoting effect is observed when yellow light is irradiated.
前記５２０ｎｍ以上の波長の可視光線の光強度としては、１〜１００μｍｏｌｍ−２ｓ−１であることが好ましく、より好ましくは１０〜６５μｍｏｌｍ−２ｓ−１である。１μｍｏｌｍ−２ｓ−１未満であると、光強度が不充分で、充分な増殖促進効果が見られないことがあり、１００μｍｏｌｍ−２ｓ−１を超えると、光強度が高すぎて、光照射対象の真核微生物に悪影響が及ぶことがある。 The light intensity of visible light having a wavelength of 520 nm or more is preferably 1 to 100 μmol −2 s −1 , more preferably 10 to 65 μmol −2 s −1 . If it is less than 1 μmolm −2 s −1 , the light intensity is insufficient and a sufficient growth promoting effect may not be seen. If it exceeds 100 μmolm −2 s −1 , the light intensity is too high and light irradiation is performed. The target eukaryotic microorganism may be adversely affected.
以下に実施例を掲げて本発明を更に詳細に説明するが、本発明はこれら実施
例のみに限定されるものではない。
The present invention will be described in more detail with reference to the following examples. However, the present invention is not limited to these examples.
＜実験１＞黄色光照射下における酵母の培養
独立行政法人酒類総合研究所より入手した下記の酵母（Saccharomyces cereviseae）を、試験用の株として使用した。
<Experiment 1> Culture of yeast under yellow light irradiation The following yeast (Saccharomyces cereviseae) obtained from the National Research Institute for Liquors was used as a test strain.
・ＲＩＢ １０２８株（ワイン酵母）
・ＲＩＢ ６００１株（清酒酵母、協会１号）
・ＲＩＢ ２０１０株（ビール酵母）
・ RIB 1028 strain (wine yeast)
-RIB 6001 strain (Sake Yeast, Association No. 1)
・ RIB 2010 strain (beer yeast)
これらの株の酵母を、ＹＭ液体培地（グルコース１０ｇ／Ｌ、ペプトン５ｇ／Ｌ、酵母エキス３ｇ／Ｌ、麦芽エキス３ｇ／Ｌ、以上を蒸留水に溶解）中において、３０℃で培養した。この際、条件１として、暗黒下で培養し、条件２として、３０μｍｏｌｍ−２ｓ−１の光強度の黄色光（ピーク波長５５６ｎｍ）を照射しながら培養し、条件３として、３０μｍｏｌｍ−２ｓ−１の光強度の琥珀色光（ピーク波長５９３ｎｍ）を照射しながら培養した。 Yeasts of these strains were cultured at 30 ° C. in YM liquid medium (glucose 10 g / L, peptone 5 g / L, yeast extract 3 g / L, malt extract 3 g / L, dissolved in distilled water). At this time, as condition 1, the cells were cultured in the dark, and as condition 2, the cells were cultured while irradiating with yellow light (peak wavelength 556 nm) having a light intensity of 30 μmolm −2 s −1 , and as condition 3, 30 μmolm −2 s − The cells were cultured while being irradiated with amber light having a light intensity of 1 (peak wavelength: 593 nm).
なお、黄色光の光源としては、ラムシード社製の白色ＬＥＤに黄色フィルタを装着したものを使用し、琥珀色光の光源としては、ラムシード社製の琥珀色ＬＥＤを使用した。なお、各照射光のスペクトルは図１に示した。 In addition, as a light source of yellow light, a white LED manufactured by Ramseed was used and a yellow filter was mounted, and as a light source of amber light, an amber LED manufactured by Ramseed was used. The spectrum of each irradiation light is shown in FIG.
培養開始後、０、２、４、６、１２、２４時間後の培地をサンプリングし、適宜希釈してヘマトサイトメータで菌数を計数し、生菌数を算出した。 After the start of culture, the medium after 0, 2, 4, 6, 12, and 24 hours was sampled, diluted as appropriate, and the number of bacteria was counted with a hematocytometer to calculate the number of viable bacteria.
結果を表１〜３に示し、図２〜４に増殖グラフ（対数）を示した。 The results are shown in Tables 1 to 3, and the proliferation graph (logarithm) is shown in FIGS.
表１〜３及び図２〜４に示すように、黄色光照射下における生菌数は暗黒下と比べて、ＲＩＢ １０２８株で培養開始６時間後に１．６倍、ＲＩＢ ６００１株で培養開始４時間後及び１０時間後に２．１倍になり、ＲＩＢ ２０１０株で培養開始４〜１２時間後に約２倍になったことが観察され、黄色光に増殖促進作用があり、特に対数増殖期にこの促進効果が顕著であることが認められた。 As shown in Tables 1 to 3 and FIGS. 2 to 4, the number of viable cells under yellow light irradiation is 1.6 times higher in the RIB 1028 strain 6 hours after the start of culture than in the dark, and the start of culture in the RIB 6001 strain 4 It was observed that it doubled 2.1 hours later and 10 hours later, and approximately doubled 4 to 12 hours after the start of culture in the RIB 2010 strain. Yellow light has a growth promoting action, especially in the logarithmic growth phase. It was recognized that the promoting effect was remarkable.
＜実験２＞黄色光照射下におけるコウジカビの培養
塚本鑛吉商店より入手したコウジカビ（ひかみ吟醸用コウジ）を試験用の株として使用して、コウジ１０ｇを６０℃２０ｍＬの湯に溶かし、マグネットスターラにて回転数２５０ｒｐｍで攪拌しながら３０℃にて３０分間放置した。この際、３０μｍｏｌｍ−２ｓ−１又は６０μｍｏｌｍ−２ｓ−１の光強度の黄色光（ピーク波長５５６ｎｍ、図１参照）を照射しながら培養した。また、コントロールとして、暗黒下でも培養した。各条件とも３サンプルずつ培養した。
<Experiment 2> Cultivation of Aspergillus under Yellow Light Using Aspergillus oryzae obtained from Tsukamoto Yukichi Shoten as a test strain, 10 g of Aspergillus is dissolved in 20 mL of water at 60 ° C., and a magnetic stirrer is used. The mixture was allowed to stand at 30 ° C. for 30 minutes with stirring at 250 rpm. Under the present circumstances, it culture | cultivated irradiating yellow light (peak wavelength 556 nm, refer FIG. 1) of the light intensity of 30 micromol - 2s - 1 or 60 micromol - 2s - 1 . As a control, the cells were cultured in the dark. Three samples were cultured for each condition.
なお、黄色光の光源としては、ラムシード社製の白色ＬＥＤに黄色フィルタを装着したものを使用した。 In addition, as a light source of yellow light, a white LED manufactured by Lambseed and a yellow filter attached thereto was used.
その後、上澄み液を５００ｍＬ採取し、更に遠心分離して得られた上澄み液を希釈し、攪拌した後、バイオチップ上に滴下して、塚本鑛吉商店製グルコース計ＧＭ−１０００を使用してグルコース濃度を測定した。結果を表４（３０μｍｏｌｍ−２ｓ−１黄色光照射）及び表５（６０μｍｏｌｍ−２ｓ−１黄色光照射）に示した。 Thereafter, 500 mL of the supernatant is collected, and the supernatant obtained by further centrifuging is diluted, stirred, dropped onto the biochip, and glucose is added using a glucose meter GM-1000 manufactured by Tsukamoto Yukichi Shoten. Concentration was measured. The results are shown in Table 4 (30 μmolm −2 s −1 yellow light irradiation) and Table 5 (60 μmolm −2 s −1 yellow light irradiation).
得られた結果より、黄色光の照射によりコウジカビのグルコース生成能が増強されていることが確認された。このため、黄色光照射によりコウジカビの活動が活性化されたものと考えられる。 From the obtained results, it was confirmed that the glucose producing ability of Aspergillus was enhanced by irradiation with yellow light. For this reason, it is thought that the activity of Aspergillus was activated by the yellow light irradiation.
本発明は、真核微生物を利用して、清酒を始めとする種々の醸造製品等を製造する際に、有用である。 The present invention is useful when producing various brewed products such as sake using eukaryotic microorganisms.
Claims (1)
前記真核微生物に黄色光を照射して、前記真核微生物の増殖を促進する工程を有しており、
前記工程において、前記可視光線の光強度が、１〜１００μｍｏｌｍ −２ ｓ −１ の範囲であることを特徴とする真核微生物の培養方法。
A method of culturing yeast, which is a eukaryotic microorganism, when producing a brewed product ,
Irradiating the eukaryotic microorganism with yellow light to promote the growth of the eukaryotic microorganism,
In the above step, the eukaryotic microorganism culturing method, wherein the light intensity of the visible light is in the range of 1 to 100 μmolm −2 s −1 .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2009074595A JP5934458B2 (en) | 2009-03-25 | 2009-03-25 | Eukaryotic microorganism culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2009074595A JP5934458B2 (en) | 2009-03-25 | 2009-03-25 | Eukaryotic microorganism culture method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2010220587A JP2010220587A (en) | 2010-10-07 |
| JP5934458B2 true JP5934458B2 (en) | 2016-06-15 |
Family
ID=43038480
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2009074595A Expired - Fee Related JP5934458B2 (en) | 2009-03-25 | 2009-03-25 | Eukaryotic microorganism culture method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP5934458B2 (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4691307B2 (en) * | 2002-03-24 | 2011-06-01 | 孝昭 石井 | Microbial growth control method |
| JP4941925B2 (en) * | 2006-08-01 | 2012-05-30 | 学校法人立命館 | Method for purifying soil or water contaminated with hydrocarbons |
-
2009
- 2009-03-25 JP JP2009074595A patent/JP5934458B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JP2010220587A (en) | 2010-10-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6329940B2 (en) | Euglena microalgae, polysaccharide manufacturing method, and organic compound manufacturing method | |
| JP5775862B2 (en) | Method for culturing microalgae and method for using microalgae | |
| Mathew et al. | Isolation of acetic acid bacteria and preparation of starter culture for apple cider vinegar fermentation | |
| CN104312834A (en) | A kind of preparation method of sugarcane fruit wine | |
| US20160230192A1 (en) | Triterpenoids high yielding strain of antrodia cinnamomea and use thereof | |
| KR101563148B1 (en) | Microalgae Chlamydomonas reinhardtii mutant with enhanced biomass, starch and lipid content by gamma ray irradiation and uses thereof | |
| JP2010233517A (en) | Fucoxanthin-containing microalgae culture and production method thereof | |
| US10364446B2 (en) | Streptomyces psammoticus and methods of using the same for vanillin production | |
| CN118556550A (en) | A method for cultivating edible fungus mycelium using bottom pot water of liquor byproduct | |
| CN102899236A (en) | Process method for brewing ginseng vinegar by immobilized fermentation | |
| CN110564580A (en) | A method for microbial co-cultivation and fermentation to produce vinegar containing pyrroloquinoline quinone | |
| CN103130550B (en) | Culture medium and culture method of male agaric mycelium | |
| JP5934458B2 (en) | Eukaryotic microorganism culture method | |
| CN102021212A (en) | Preparation method of ganoderma polysaccharide | |
| CN105199928A (en) | Method for producing flavor blending liquid from pichia kudriavzevii | |
| JP2005295871A (en) | Enzyme productivity adjustment method | |
| CN109321510B (en) | Application of strigolactone in promoting the accumulation of lipids in the algae | |
| CN101928699A (en) | Fermentation process for improving yield of lucid ganoderma laccase | |
| CN109601254A (en) | A kind of Antrodia camphorata cultural method that cultured products are kept completely separate with matrix | |
| US12618029B2 (en) | Method for producing agave cultures for tequila | |
| CN102634494B (en) | A kind of manganese peroxidase solid-state fermentation substrate and preparation and application thereof | |
| KR101633031B1 (en) | Method for producing mochi-koji | |
| CN109964727B (en) | Method for improving selenium enrichment rate of Cordyceps militaris by utilizing light | |
| JP6014700B2 (en) | Method for culturing microalgae and method for using microalgae | |
| CN105754863B (en) | Laccase-producing bidens macrophylla and method for preparing laccase by using same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20120302 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20131121 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20140120 |
|
| A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20140805 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20141104 |
|
| A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20141111 |
|
| A912 | Re-examination (zenchi) completed and case transferred to appeal board |
Free format text: JAPANESE INTERMEDIATE CODE: A912 Effective date: 20141226 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20160303 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20160509 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 5934458 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| LAPS | Cancellation because of no payment of annual fees |