JP3527661B2 - Culture medium for lactic acid bacteria separated from barley shochu distillation residue - Google Patents
Culture medium for lactic acid bacteria separated from barley shochu distillation residueInfo
- Publication number
- JP3527661B2 JP3527661B2 JP15732699A JP15732699A JP3527661B2 JP 3527661 B2 JP3527661 B2 JP 3527661B2 JP 15732699 A JP15732699 A JP 15732699A JP 15732699 A JP15732699 A JP 15732699A JP 3527661 B2 JP3527661 B2 JP 3527661B2
- Authority
- JP
- Japan
- Prior art keywords
- medium
- culture
- liquid
- lactic acid
- shochu
- 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
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【０００１】[0001]
【発明の属する技術分野】本発明は焼酎製造で副成する
焼酎蒸留残液から得られる微生物用培地、およびその製
造方法に関する。より詳しくは本発明は，大麦を原料と
する焼酎製造において副成する焼酎蒸留残液を固液分離
して液体分を得、該液体分をろ過して清澄液を得、該清
澄液を濃縮して濃縮液を得、該濃縮液を合成吸着剤を用
いる吸着処理に付して非吸着性画分を得、該非吸着性画
分を乾燥することにより得られる乾燥物を有効成分とし
て含有することを特徴とする微生物用培地およびその製
造方法に関する。TECHNICAL FIELD The present invention relates to a culture medium for microorganisms obtained from shochu distillation residual liquid by-produced in the production of shochu, and a method for producing the same. More specifically, the present invention is to obtain a liquid component by solid-liquid separation of a shochu distillation residual liquid by-produced in the production of shochu using barley as a raw material, obtain a clear liquid by filtering the liquid content, and concentrate the clear liquid. To obtain a concentrated liquid, and subject the concentrated liquid to an adsorption treatment using a synthetic adsorbent to obtain a non-adsorptive fraction, which contains a dried product obtained by drying the non-adsorptive fraction as an active ingredient. The present invention relates to a microorganism culture medium and a method for producing the same.
【０００２】[0002]
【従来の技術】焼酎製造において副成する焼酎蒸留残液
は，家畜用飼料あるいは肥料として一部で利用される場
合があるが、大部分は海洋投棄，大地還元，焼却処分な
どにより廃棄処分されるのが一般的である。海洋投棄は
焼酎蒸留残液の安価な処分方法であるが，世界的な環境
問題の高まりにより，法的に規制され全面禁止されるこ
とになっている。また，大地還元も地下水や河川を汚染
する問題があり，焼却処理はコストおよびダイオキシン
発生等の問題がある。このようなことから、現在、多方
面において焼酎蒸留残液の有効利用が検討されている。
これまでに、上述したように焼酎蒸留残液を飼料あるい
は肥料として利用する方法等が提案されている。2. Description of the Related Art The shochu distillation residual liquid by-produced in the production of shochu may be used partially as livestock feed or fertilizer, but most of it is discarded by ocean dumping, land return, incineration, etc. It is common to Ocean dumping is an inexpensive way to dispose of shochu distillation residue, but due to rising global environmental problems, it is legally regulated and banned altogether. In addition, ground return also has the problem of contaminating groundwater and rivers, and incineration has problems such as cost and dioxin generation. Therefore, at present, effective utilization of the shochu distillation residual liquid is being studied in various fields.
So far, there have been proposed methods for utilizing the shochu distillation residual liquid as feed or fertilizer as described above.
【０００３】また、上述の提案の他に、焼酎蒸留残液の
微生物用培地への利用が提案されている。例えば、特開
平８−３０８５９０号公報（以下、文献１と言う。）に
は、ポリ−γ−グルタミン酸を製造する際に、ポリ−γ
−グルタミン酸産生土壌細菌の培地として焼酎蒸留残液
を用いることが記載されている。一般に、焼酎蒸留残液
は０．１〜２ｇ／Ｌのグルタミン酸を含むことに鑑み
て、文献１においては、ポリ−γ−グルタミン酸生産の
ためのグルタミン酸供給源として焼酎蒸留残液を利用し
ている。具体的には、固液分離を行って得られた焼酎蒸
留残液を固液分離に付して得られた上清（ｐＨ３．８）
を、水酸化ナトリウム水溶液でｐＨ６．５に調整し、こ
れをそのまま培地として使用している。しかしながら、
焼酎蒸留残液は、水分を９０％近く含み、しかもそのＢ
ＯＤが７００００〜１０００００ｍｇ／Ｌと極めて高い
ため腐敗の進行が早く、文献１に記されているように、
焼酎蒸留残液の上清を中和後そのまま培地として用いた
場合には保存性が極めて悪い。また当該焼酎蒸留残液の
上清は水不溶性成分を多く含み、さらにＳＳ（懸濁物
質：水中に分散している固形粒子分のなかで、２ｍｍ以
下の径でろ紙の目にとどまるもの）を含有するため、こ
れを微生物用培地として用いた場合には、培養中の菌体
濃度の測定や培養終了後の菌体の回収が困難である。そ
のため培地としての価値を著しく低める原因となってし
まうことから好ましくない。この他に当該焼酎蒸留残液
の上清は、培地成分としては望ましくない着色成分も多
く含んでいるため、着色成分を含んだ微生物用培地をそ
のまま用いる場合には、培養中の菌体濃度の測定が困難
になるだけでなく、培養終了後に得られる菌体が着色さ
れてしまう。このことは、培地としての価値を著しく低
めてしまう。In addition to the above-mentioned proposals, it has been proposed to use the shochu distillation residue liquid as a culture medium for microorganisms. For example, in Japanese Unexamined Patent Publication No. 8-308590 (hereinafter referred to as Document 1), when poly-γ-glutamic acid is produced, poly-γ is used.
-The use of shochu distillation residue as a medium for glutamate producing soil bacteria is described. In general, in view of the fact that the shochu distilled residue contains 0.1 to 2 g / L of glutamic acid, Reference 1 uses the shochu distilled residue as a glutamic acid supply source for poly-γ-glutamic acid production. . Specifically, the supernatant (pH 3.8) obtained by subjecting the shochu distillation residual liquid obtained by solid-liquid separation to solid-liquid separation
Was adjusted to pH 6.5 with an aqueous solution of sodium hydroxide and used as it was as a medium. However,
The shochu distillation residue contains nearly 90% water, and its B
Since the OD is extremely high at 70,000 to 100,000 mg / L, the decay progresses quickly, and as described in Document 1,
When the supernatant of the distillation residue of shochu is neutralized and used as a medium as it is, the preservability is extremely poor. In addition, the supernatant of the distilled spirits distillation residual liquid contains a large amount of water-insoluble components, and further contains SS (suspended substance: solid particles dispersed in water that remain in the eyes of filter paper with a diameter of 2 mm or less). Therefore, when this is used as a culture medium for microorganisms, it is difficult to measure the bacterial cell concentration during the culture and collect the bacterial cells after the culture is completed. Therefore, it is not preferable because it causes the value of the medium to be significantly reduced. In addition to this, the supernatant of the shochu distillation residual liquid contains a large amount of coloring components which are not desirable as medium components, so when using a microbial medium containing coloring components as it is, Not only is the measurement difficult, but the cells obtained after the culture are colored. This significantly reduces its value as a medium.
【０００４】ところで、本発明者らの一人は、焼酎蒸留
残液を有効利用する観点から、特開平８−５６５８４号
公報（以下、文献２と言う。）において、焼酎蒸留残液
から飼料を製造する方法を提案している。文献２では、
焼酎蒸留残液を固液分離して液体分と固体分に分け，該
液体分のＳＳ（懸濁物質：水中に分散している固形粒子
分のなかで、２ｍｍ以下の径でろ紙の目にとどまるも
の）の量を１００ｇ／Ｌ以下に調整後に所定の水分まで
乾燥させた前記液体分の乾燥物と、別の方法で乾燥させ
た前記固体分の乾燥物を混合することにより飼料を製造
する方法が記載されている。By the way, one of the inventors of the present invention, from the viewpoint of effectively utilizing the distilled spirits distillation residual liquid, in Japanese Patent Application Laid-Open No. 8-56584 (hereinafter referred to as Document 2), produces a feed from the distilled spirits distillation residual liquid. Suggesting a way to do it. In reference 2,
The shochu distillation residual liquid is separated into solid and liquid to separate it into liquid and solid, and SS (suspended substance: solid particles dispersed in water) of the liquid has a diameter of 2 mm or less The amount of the (retaining substance) is adjusted to 100 g / L or less and then the dried product of the liquid component dried to a predetermined water content is mixed with the dried product of the solid component dried by another method to produce a feed. The method is described.
【０００５】本発明者らは，文献２において得られる飼
料を、そのまま酵母等の微生物用培地として用いたとこ
ろ、以下のような問題があることが判明した。すなわ
ち、当該飼料は固体分乾燥物を混合しているために水不
溶性成分が多く，さらに着色成分も多いため、そのまま
では微生物用培地として使用に適していないことが判っ
た。また、前記液体分の乾燥物をそのまま酵母等の微生
物用培地として用いたところ、同じく水不溶性成分と着
色成分が多く，そのままでは微生物用培地として使用に
適していないことが判った。The present inventors have found that when the feed obtained in Document 2 is used as it is as a culture medium for microorganisms such as yeast, there are the following problems. That is, it was found that the feed is not suitable for use as a culture medium for microorganisms as it is because the feed contains a large amount of water-insoluble components due to the mixture of dried solids and also contains a large amount of coloring components. Further, when the dried product of the liquid was used as it was as a culture medium for microorganisms such as yeast, it was found that it was not suitable for use as a culture medium for microorganisms as it was since it contained a large amount of water-insoluble components and coloring components.
【０００６】[0006]
【発明が解決しようとする課題】以上述べたように，文
献１においては、固液分離を行った焼酎蒸留残液の上清
を水酸化ナトリウム水溶液でｐＨ６．５に調整して培地
として用いるが、この場合、当該培地は液体であるため
に保存性が悪く、さらに水不溶性成分およびＳＳ、さら
に培地成分としては望ましくない着色成分を含有し、こ
れらは培地としての価値を著しく低めてしまうという問
題がある。また、文献２に記載された飼料、あるいは液
体分の乾燥物をそのまま微生物用培地として用いた場合
には、上述したように水不溶性成分およびＳＳ、さらに
培地成分としては望ましくない着色成分を含有するため
に、そのままでは微生物用培地として使用に適していな
い。これらの問題は焼酎蒸留残液の微生物用培地への有
効利用を進める上で早急に解決を要する問題である。As described above, in Reference 1, the supernatant of the shochu distillation residue after solid-liquid separation is adjusted to pH 6.5 with an aqueous sodium hydroxide solution and used as a medium. , In this case, since the medium is liquid, it is poor in storability, and further contains a water-insoluble component and SS, and a coloring component which is not desirable as a medium component, and these significantly reduce the value of the medium. There is. Further, when the feed described in Reference 2 or the dried product of the liquid content is used as it is as a culture medium for microorganisms, it contains a water-insoluble component and SS, and further, a coloring component which is not desirable as a culture medium component, as described above. Therefore, as it is, it is not suitable for use as a culture medium for microorganisms. These problems are problems that need to be solved as soon as possible in order to effectively utilize the distillation residue of shochu in a culture medium for microorganisms.
【０００７】本発明は，上述した従来技術における問題
点に鑑みて、さらなる研究の結果完成に至ったものであ
る。本発明の主たる目的は，焼酎蒸留残液からの微生物
用培地製造に係る従来技術における上記問題点を解決
し，焼酎蒸留残液から、水不溶性成分および着色成分が
極めて少なく、これを微生物用培地として用いた場合
に、得られる培養菌体の量が著しく増加する微生物用培
地およびその製造方法を提供することにある。The present invention has been completed as a result of further research in view of the above-mentioned problems in the prior art. The main object of the present invention is to solve the above-mentioned problems in the prior art relating to the production of a culture medium for microorganisms from a shochu-distilling residual liquid, and to obtain a very small amount of a water-insoluble component and a coloring component from the shochu-distilling residual liquid. It is intended to provide a microorganism medium and a method for producing the same, in which the amount of the obtained cultured cells is remarkably increased when used as the above.
【０００８】[0008]
【課題を解決するための手段】本発明者らは，焼酎蒸留
残液からの微生物用培地製造における上述した問題を解
決し，焼酎蒸留残液をより有効に利用する観点から，水
不溶性成分および着色成分が極めて少なく，微生物用培
地として用いた場合に、得られる培養菌体の量が著しく
増加する微生物用培地を得ることを目的として，実験を
介して鋭意研究を重ねた。その結果，大麦を原料とする
焼酎製造において副成する焼酎蒸留残液を固液分離して
液体分を得、該液体分をろ過して清澄液を得、該清澄液
を濃縮して濃縮液を得、該濃縮液を合成吸着剤を用いる
吸着処理に付して非吸着性画分を得、該非吸着性画分を
乾燥することにより、水不溶性成分および着色成分が極
めて少なく，これを微生物用培地として用いた場合、得
られる培養菌体の量が著しく増加する微生物用培地が得
られ，上記課題が解決されることが判明した。本発明は
この判明した事実に基づくものである。[Means for Solving the Problems] From the viewpoint of solving the above-mentioned problems in producing a culture medium for microorganisms from a distilled spirits distillation residual liquid and more effectively utilizing the distilled spirits distillation residual liquid, the present inventors We have conducted intensive studies through experiments for the purpose of obtaining a microbial medium in which the amount of the cultured cells obtained is significantly increased when used as a microbial medium with very few coloring components. As a result, a shochu distillation residual liquid by-produced in the production of shochu using barley as a raw material is subjected to solid-liquid separation to obtain a liquid component, the liquid component is filtered to obtain a clear liquid, and the clear liquid is concentrated to form a concentrated liquid. To obtain a non-adsorptive fraction by subjecting the concentrated liquid to an adsorption treatment using a synthetic adsorbent, and drying the non-adsorptive fraction, whereby the water-insoluble component and the coloring component are extremely small. It was found that when used as a culture medium, a culture medium for microorganisms in which the amount of the obtained cultured cells is remarkably increased is obtained, and the above-mentioned problems are solved. The present invention is based on this discovered fact.
【０００９】以下本発明の好ましい態様について述べる
が，これらによって何ら本発明が制限されるものではな
い。本発明の微生物用培地は、図１に示すように、大麦
を原料とする焼酎製造において副成する焼酎蒸留残液を
固液分離して液体分を得る第１の工程、該液体分をろ過
して清澄液を得る第２の工程、該清澄液を濃縮して濃縮
液を得る第３の工程、該濃縮液を合成吸着剤を用いる吸
着処理に付して非吸着性画分を得る第４の工程、得られ
た該非吸着性画分を乾燥する第５の工程により得られる
ものである。以下に本発明の微生物用培地を製造する際
に原料として用いる焼酎蒸留残液、および各工程につい
て詳述する。The preferred embodiments of the present invention will be described below, but the present invention is not limited thereto. The microbial medium of the present invention is, as shown in FIG. 1, a first step of solid-liquid separating a shochu distillation residual liquid by-produced in the production of shochu using barley as a raw material to obtain a liquid content, and filtering the liquid content. To obtain a clarified liquid, a third step of concentrating the clarified liquid to obtain a concentrated solution, a third step of subjecting the concentrated solution to an adsorption treatment using a synthetic adsorbent to obtain a non-adsorbable fraction It is obtained by the step 4 and the fifth step of drying the obtained non-adsorptive fraction. The shochu distillation residual liquid used as a raw material when producing the microbial medium of the present invention, and each step will be described in detail below.
【００１０】本発明において言う焼酎蒸留残液は、大麦
又は精白大麦を原料として大麦麹および蒸麦を製造し、
得られた大麦麹および蒸麦中に含まれるでんぷんを麹お
よび／又は酵素剤を使用して糖化し、さらに酵母による
アルコール発酵を行い焼酎熟成もろみを得、得られた焼
酎熟成もろみを減圧蒸留または常圧蒸留等の単式蒸留装
置を用いて蒸留する際に蒸留残さとして副成するものを
意味し、代表的には例えば大麦焼酎の蒸留残液が挙げら
れる。さらに米焼酎、甘藷焼酎、そば焼酎の製造におい
ても、これらの焼酎製造において原料の一部として大麦
を使用する場合に副成する焼酎蒸留残液も本発明におい
て言う焼酎蒸留残液に包含される。The shochu distillation residual liquid as used in the present invention is obtained by producing barley koji and steamed barley using barley or refined barley as a raw material,
The starch contained in the obtained barley koji and steamed barley was saccharified using koji and / or an enzymatic agent, and shochu aged moromi was obtained by alcohol fermentation with yeast, and the obtained shochu aged moromi was distilled under reduced pressure or It means a by-product as a distillation residue when distilling using a single-distillation apparatus such as atmospheric distillation, and typically, for example, a distillation residual liquid of barley shochu is mentioned. Further, also in the production of rice shochu, sweet potato shochu, and buckwheat shochu, a shochu distillation residual liquid by-produced when barley is used as a part of the raw material in the production of these shochu is also included in the shochu distillation residual liquid referred to in the present invention. .
【００１１】本発明において、焼酎製造の蒸留工程で得
られた焼酎蒸留残液を固液分離して液体分を得る第１の
工程は、焼酎蒸留残液から原料大麦あるいは麹由来の水
不溶性の発酵残渣や酵母菌体を除去し、液体分を得るこ
とを目的として行うものである。この第１の工程におい
ては、一般的にスクリュープレス方式やローラープレス
方式、あるいはろ過圧搾式の固液分離機を用いて行うこ
とが出来る。こうして、第１の工程で得られる該液体分
をろ過して清澄液を得る第２の工程は、該液体分に依然
として多く含まれているＳＳを除去して清澄液を得るこ
とを目的として行うものである。第２の工程のろ過処理
においては、各種の遠心分離機、ケイソウ土ろ過装置、
セラミックろ過装置、あるいはろ過圧搾機等を用いて行
うことができる。In the present invention, the first step for solid-liquid separation of the shochu distillation residual liquid obtained in the distillation step for producing shochu to obtain a liquid component is a water-insoluble material derived from barley or koji derived from the shochu distillation residual liquid. The purpose is to remove the fermentation residue and yeast cells to obtain a liquid content. The first step can be generally performed using a screw press system, a roller press system, or a filtration / compression type solid-liquid separator. In this way, the second step of filtering the liquid component obtained in the first step to obtain a clear liquid is carried out for the purpose of removing SS still contained in a large amount in the liquid component to obtain a clear liquid. It is a thing. In the filtration process of the second step, various centrifuges, diatomaceous earth filtration devices,
It can be performed using a ceramic filtration device or a filtration press.
【００１２】また、本発明において、第２の工程で得ら
れる清澄液を濃縮して濃縮液を得る第３の工程は、次の
第４の工程において吸着処理に付す清澄液の濃度を高め
る目的のために実施するものである。この第３の工程の
清澄液の濃縮方法には公知の方法を用いることができ、
具体的には、減圧濃縮装置、真空蒸発装置等を用いて行
うことができる。Further, in the present invention, the third step of concentrating the clarified liquid obtained in the second step to obtain a concentrated liquid is to increase the concentration of the clarified liquid subjected to the adsorption treatment in the next fourth step. It is to be carried out for. A known method can be used for the method of concentrating the clarified liquid in the third step,
Specifically, it can be performed using a vacuum concentration device, a vacuum evaporation device, or the like.
【００１３】第３の工程で得られる該濃縮液を合成吸着
剤を用いる吸着処理に付して非吸着性画分を得る第４の
工程は、該濃縮液に含まれる培地成分としては望ましく
ない着色成分を取り除くことを目的として行うものであ
る。なお、着色成分が培養液中に多量に存在すると、培
養中の菌体濃度の測定が困難になるだけでなく、培養終
了後に得られる菌体が着色されてしまうことから、培地
としての価値を著しく低めてしまう。第４の工程で使用
する合成吸着剤としては，芳香族系、芳香族系修飾型、
メタクリル系の合成吸着剤を用いることができる。例え
ば、合成吸着剤として好適なものとしては、三菱化学社
製のセパビーズＳＰ８５０およびダイヤイオンＨＰ２
０、さらにオルガノ社製のアンバーライトＸＡＤ１６等
を使用することができる。The fourth step of subjecting the concentrated solution obtained in the third step to an adsorption treatment with a synthetic adsorbent to obtain a non-adsorbable fraction is not desirable as a medium component contained in the concentrated solution. The purpose is to remove the coloring component. It should be noted that if a large amount of a coloring component is present in the culture solution, not only it becomes difficult to measure the bacterial cell concentration during culture, but also the bacterial cells obtained after the completion of the culture will be colored, which makes it a valuable medium. It significantly lowers. The synthetic adsorbent used in the fourth step is an aromatic type, an aromatic type modified type,
A methacrylic synthetic adsorbent can be used. For example, as a suitable synthetic adsorbent, Sepabeads SP850 and DIAION HP2 manufactured by Mitsubishi Chemical Corporation are used.
0, and Amberlite XAD16 or the like manufactured by Organo can be used.
【００１４】また第４の工程で得られる非吸着性画分を
乾燥する第５の工程は、該非吸着性画分を乾燥する方法
として，ディスク型ドライヤー、ドラム型ドライヤー、
スプレー型ドライヤー等の乾燥装置、あるいは凍結乾燥
機を用いて行うことができる。The fifth step of drying the non-adsorptive fraction obtained in the fourth step is as a method for drying the non-adsorptive fraction, which includes a disk dryer, a drum dryer,
It can be performed using a drying device such as a spray dryer or a freeze dryer.
【００１５】ところで、微生物の生育用培地について
は、それぞれの微生物に最適のｐＨがあり、細菌用の培
地はｐＨ６．８〜７．２、カビ・酵母用の培地はｐＨ
５．０〜６．０に調整するのが一般的である。焼酎蒸留
残液は麹菌由来のクエン酸を含むため、そのｐＨは３〜
４と低く、これをそのまま培地原料として用いた場合に
は、得られた培地のｐＨ値が一般的に微生物の培養に最
適とされるｐＨ値よりもかなり低くなり好ましくない。
従って、本発明においても、第２の工程で得られた清澄
液、あるいは第３の工程で得られた濃縮液を適当な中和
剤を用いて中和処理することが望ましい。こうした中和
剤としては，水酸化ナトリウム／水酸化カリウム等を使
用することができる。By the way, regarding the growth medium for the microorganisms, there is an optimum pH for each microorganism, the medium for bacteria has a pH of 6.8 to 7.2, and the medium for molds and yeasts has a pH.
Generally, it is adjusted to 5.0 to 6.0. Since the distilled spirits residual liquid contains citric acid derived from Aspergillus oryzae, its pH is 3 ~.
It is as low as 4, and when it is used as it is as a raw material for the medium, the pH value of the obtained medium is considerably lower than the pH value generally optimized for culturing microorganisms, which is not preferable.
Therefore, also in the present invention, it is desirable to neutralize the clarified liquid obtained in the second step or the concentrated liquid obtained in the third step with an appropriate neutralizing agent. As such a neutralizing agent, sodium hydroxide / potassium hydroxide or the like can be used.
【００１６】さらに、第４の工程で得られる該非吸着性
画分を乾燥する第５の工程においては、微生物用培地乾
燥物の粒子の均一化を図るために、乾燥前の該非吸着性
画分に賦形剤を添加することもでき、そうした賦形剤と
しては、噴霧乾燥により粉末製品を得る際に一般的に用
いられる賦形剤を使用することができる。好ましい具体
例としては、ばい焼デキストリンや酵素変性デキストリ
ン等のデキストリン類、あるいはエーテル化デンプンや
酸処理デンプン等のデンプン類を挙げることができる。Further, in the fifth step of drying the non-adsorptive fraction obtained in the fourth step, the non-adsorptive fraction before drying is used in order to homogenize the particles of the dried product of the culture medium for microorganisms. It is also possible to add an excipient to the above, and as such an excipient, an excipient that is generally used in obtaining a powder product by spray drying can be used. Preferred specific examples include dextrins such as roasted dextrin and enzyme-modified dextrin, and starches such as etherified starch and acid-treated starch.
【００１７】なお、本発明の微生物用培地は、特に、酵
母、乳酸菌、及びビフィズス菌の菌体を培養する際に、
酵母エキス、カゼインペプトン、肉エキス等の窒素源の
代替として使用することができる。この場合、本発明の
微生物用培地が窒素源として利用されるだけではなく、
培養菌体の増殖速度を高め、その結果、得られる培養菌
体の量が著しく増大する。この他、本発明の微生物用培
地は、特に、酵母、乳酸菌、及びビフィズス菌の菌体を
培養する際に、窒素源を十分に含有する培地に増殖促進
因子として添加することができる。この場合、本発明の
微生物用培地により菌体の増殖が一層促進されると共
に、菌体の増殖速度が一層高められ、そしてまた、前記
の場合と同様に、得られる培養菌体の量が著しく増大す
る。前記酵母としてはSaccharomyces属の酵母を挙げる
ことができ、前記乳酸菌としてはLactobacillus属のLac
tobacillus acidophilus、Lactobacillus plantarum、
Lactobacillus fermentum等を挙げることができ、さら
に前記ビフィズス菌としてはBifidobacterium属のBifid
obacterium bifidum、Bifidobacterium longum等を挙げ
ることができる。The microbial medium of the present invention is particularly suitable for culturing yeast, lactic acid bacterium and bifidobacteria cells.
It can be used as a substitute for nitrogen sources such as yeast extract, casein peptone, and meat extract. In this case, the microbial medium of the present invention is not only used as a nitrogen source,
The growth rate of the cultured cells is increased, and as a result, the amount of the obtained cultured cells is significantly increased. In addition, the culture medium for microorganisms of the present invention can be added as a growth-promoting factor to a culture medium containing a sufficient nitrogen source, particularly when culturing yeast, lactic acid bacteria, and bifidobacteria. In this case, the microbial medium of the present invention further promotes the growth of the bacterial cells, further increases the growth rate of the bacterial cells, and, similarly to the above case, the amount of the obtained cultured bacterial cells is significantly increased. Increase. Examples of the yeast include yeasts of the genus Saccharomyces, and the lactic acid bacteria include Lac of the genus Lactobacillus.
tobacillus acidophilus, Lactobacillus plantarum,
Lactobacillus fermentum and the like, and further, the Bifidobacteria include Bifid of the genus Bifidobacterium.
Examples include obacterium bifidum and Bifidobacterium longum.
【００１８】[0018]
【実施例】以下に実施例を挙げて本発明を具体的に説明
するが，本発明はこれらの実施例によって何ら限定され
るものではない。EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.
【００１９】[0019]
【大麦製焼酎蒸留残液の取得】以下の実施例に供する目
的で大麦製焼酎の製造を行った。仕込みの割合は表１に
示す通りとした。原料としては、大麦（７０％精白）を
用いた。麹の製造は大麦を４０％（Ｗ／Ｗ）吸水させ、
４０分間蒸した後、４０℃まで放冷し、大麦トンあたり
１ｋｇの種麹（白麹菌）を接種し、３８℃、ＲＨ９５％
で２４時間、３２℃、ＲＨ９２％で２０時間で行った。
蒸麦は大麦を４０％（Ｗ／Ｗ）吸水させ４０分間蒸した
後、４０℃まで放冷後、１次仕込みに加えた。１次仕込
みでは前述の方法で製造した大麦麹（大麦として３ト
ン）に、水３．６キロリットル及び酵母として焼酎酵母
の培養菌体１ｋｇ（湿重量）を加えて１次もろみを得、
得られた１次もろみを５日間の発酵（１段目の発酵）に
付した。次いで、２次仕込みでは、上記１段目の発酵を
終えた１次もろみに、水１１．４キロリットル、前述の
方法で製造した蒸麦（大麦として７トン）を加えて１１
日間の発酵（２段目の発酵）に付した。発酵温度は１次
仕込み、２次仕込みとも２５℃とした。上記２段目の発
酵を終えた２次もろみを常法により単式蒸留に付し、大
麦製焼酎１０キロリットルと大麦製焼酎蒸留残液１５キ
ロリットルを得た。得られた大麦製焼酎蒸留残液を以下
の実施例に用いた。[Acquisition of Barley Shochu Distillation Residue] Barley shochu was produced for the purpose of the following examples. The charging ratio was as shown in Table 1. Barley (70% polished) was used as a raw material. Koji is made to absorb 40% (W / W) of barley,
After steaming for 40 minutes, let cool to 40 ° C, inoculate 1 kg of seed koji (white koji mold) per ton of barley, 38 ° C, RH 95%
24 hours, 32 ° C., RH 92%, 20 hours.
The steamed barley was prepared by absorbing 40% (W / W) of barley and steaming it for 40 minutes, allowing it to cool to 40 ° C., and adding it to the primary charge. In the primary charging, barley koji (3 tons as barley) produced by the above-mentioned method was added with 3.6 kiloliters of water and 1 kg (wet weight) of cultured bacterial cells of shochu yeast as yeast to obtain primary mash.
The obtained primary moromi was subjected to fermentation for 5 days (first stage fermentation). Next, in the secondary charging, 11.4 kiloliters of water and steamed barley (7 tons as barley) produced by the above-mentioned method were added to the primary mash after the above first fermentation to produce 11
It was subjected to fermentation for the day (second stage fermentation). The fermentation temperature was 25 ° C. for both the primary charge and the secondary charge. The secondary moromi that had undergone the second-stage fermentation was subjected to simple distillation by a conventional method to obtain 10 kiloliters of barley shochu and 15 kiloliters of barley shochu distillation residual liquid. The obtained barley shochu distillation residual liquid was used in the following examples.
【００２０】[0020]
【実施例１】大麦製焼酎製造の蒸留工程で得られた前記
大麦製焼酎蒸留残液１キロリットルを信和エンジニアリ
ング（株）製のスクリュープレス方式の固液分離機で固
液分離し，得られた液体分をさらに藪田産業（株）社製
のろ過圧搾機を用いてさらにＳＳ分を分離し，約０．８
５キロリットルの清澄液を得た。次に、該清澄液を
（株）大河原製作所製の真空蒸発装置を用いて約１／３
に濃縮して濃縮液を得た。得られた該濃縮液を三菱化学
社製の合成吸着剤セパビーズＳＰ８５０を充填したカラ
ムに接触させ、当該カラムから溶出してきた当該合成吸
着剤に対して非吸着性を示す非吸着性画分を得た。得ら
れた非吸着性画分を（株）大河原化工機製スプレー型乾
燥機を用いて、入口温度 150℃，出口温度80℃，熱風量
0.45m3 / min，および噴霧空気量 1.0 kgf /cm2の条件
で噴霧乾燥し、微生物用培地に使用する約１０ｋｇの粉
末状の培地用乾燥物を得た。Example 1 1 kg of the barley shochu distillation residual liquid obtained in the distillation step of barley shochu production was solid-liquid separated by a screw press type solid-liquid separator manufactured by Shinwa Engineering Co., Ltd. The liquid component was further separated into SS components using a filtration press manufactured by Yabuta Sangyo Co., Ltd.
5 kiloliters of clear liquid was obtained. Next, the clarified liquid was used for about 1/3 using a vacuum evaporator manufactured by Okawara Seisakusho.
Concentrated to give a concentrated solution. The obtained concentrated liquid is brought into contact with a column packed with Mitsubishi Chemical's synthetic adsorbent SepaBeads SP850 to obtain a non-adsorptive fraction showing non-adsorptivity to the synthetic adsorbent eluted from the column. It was The resulting non-adsorbable fraction was applied to a spray dryer made by Okawara Kakoki Co., Ltd. at an inlet temperature of 150 ° C, an outlet temperature of 80 ° C, and a hot air flow rate.
Spray-drying was performed under the conditions of 0.45 m 3 / min and the amount of sprayed air of 1.0 kgf / cm 2 to obtain about 10 kg of a powdery dried medium for medium used as a medium for microorganisms.
【００２１】[0021]
【比較例１】大麦製焼酎製造の蒸留工程で得られた前記
大麦製焼酎蒸留残液１キロリットルを信和エンジニアリ
ング（株）製のスクリュープレス方式の固液分離機で固
液分離し，約０．８５キロリットルの液体分を得た。該
液体分を（株）西村鉄鋼所製ディスク型乾燥機を用いて
乾燥し、約７０ｋｇの乾燥物を得た。得られた該乾燥物
を（株）西村鉄鋼所製の粉砕機により粉砕し，微生物用
培地に使用する粉末状の培地用乾燥物を得た。Comparative Example 1 1 kiloliter of the barley shochu distillation residual liquid obtained in the distillation step of barley shochu production was subjected to solid-liquid separation with a screw press type solid-liquid separator manufactured by Shinwa Engineering Co., Ltd. A liquid content of 0.85 kiloliters was obtained. The liquid content was dried using a disk type dryer manufactured by Nishimura Steel Co., Ltd. to obtain about 70 kg of a dried product. The obtained dried product was crushed by a crusher manufactured by Nishimura Steel Co., Ltd. to obtain a powdery dried medium for medium used as a microorganism medium.
【００２２】[0022]
【評価】前記実施例１および前記比較例１で得られた培
地用乾燥物について、以下の実験を介して、それらの有
用性を評価した。[Evaluation] The usefulness of the dried medium product obtained in Example 1 and Comparative Example 1 was evaluated through the following experiments.
【００２３】[0023]
【実験１】微生物用培地に求められる基本的な条件とし
て、水に溶解した場合の水不溶性成分量が少ないことが
挙げられる。このような水不溶性成分が培養液中に多量
に存在すると、培養中の菌体濃度の測定や培養終了後の
菌体の回収が困難になり、培地としての価値を著しく低
めてしまう。そこで前記実施例１および前記比較例１で
得られた培地用乾燥物について、以下の実験１におい
て、それらを水に溶解した場合の水不溶性成分量の比較
をおこなった。[Experiment 1] A basic condition required for a microbial medium is that the amount of water-insoluble components when dissolved in water is small. If such a water-insoluble component is present in a large amount in the culture solution, it will be difficult to measure the concentration of the bacterial cells during the culture and to recover the bacterial cells after the completion of the culture, and the value of the medium will be significantly reduced. Therefore, with respect to the dried medium products obtained in Example 1 and Comparative Example 1, the amounts of water-insoluble components when dissolved in water were compared in Experiment 1 below.
【００２４】実施例１で得られた培地用乾燥物１０ｇを
２０℃の水１リットルに溶解し攪拌後、得られた溶解液
を８０００ｒｐｍで１５分間遠心分離し，湿重量で０．
０５ｇの水不溶性成分を得た。これと同様に、比較例１
で得られた培地用乾燥物１０ｇを２０℃の水１リットル
に溶解し攪拌後、得られた溶解液を８０００ｒｐｍで１
５分間遠心分離し，湿重量で３．２ｇの水不溶性成分を
得た。10 g of the dried culture medium obtained in Example 1 was dissolved in 1 liter of water at 20 ° C. and stirred, and the resulting solution was centrifuged at 8000 rpm for 15 minutes to give a wet weight of 0.
05 g of water-insoluble component was obtained. Similarly to this, Comparative Example 1
10 g of the dried medium for medium obtained in 1 was dissolved in 1 liter of water at 20 ° C. and stirred, and the resulting solution was stirred at 8000 rpm for 1 hour.
After centrifugation for 5 minutes, 3.2 g of a water-insoluble component was obtained by wet weight.
【００２５】上記実験１において得られた水不溶性成分
量の結果を表２に示す。表２に示した結果から明らかな
ように、比較例１で得られた微生物用培地の場合には、
水不溶性成分の割合が使用した培地用乾燥物の３２％に
達した。一方、本発明の実施例１で得られた前記微生物
用培地の場合には、水不溶性成分の割合は使用した培地
用乾燥物のわずか０．５％以下であった。以上のことか
ら、本発明の実施例１で得られた培地用乾燥物に含まれ
る水不溶性成分は、比較例１で得られた培地用乾燥物に
含まれる水不溶性成分の１／６０以下の量であった。す
なわち、本発明で得られた培地用乾燥物は、培養中の菌
体濃度の測定や培養終了後の菌体の回収の際に何ら問題
を生じない、微生物培養に極めて適した性質を有してい
ることが明らかとなった。Table 2 shows the results of the amounts of the water-insoluble components obtained in Experiment 1 above. As is clear from the results shown in Table 2, in the case of the microorganism culture medium obtained in Comparative Example 1,
The proportion of water-insoluble components reached 32% of the dry medium used. On the other hand, in the case of the culture medium for microorganisms obtained in Example 1 of the present invention, the proportion of the water-insoluble component was only 0.5% or less of the dry medium used. From the above, the water-insoluble component contained in the dried medium product obtained in Example 1 of the present invention was 1/60 or less of the water-insoluble component contained in the dried medium product obtained in Comparative Example 1. Was the amount. That is, the dried product for a medium obtained in the present invention does not cause any problem in measuring the bacterial cell concentration during the culture or collecting the bacterial cells after the culture, and has a property extremely suitable for microbial culture. It became clear.
【００２６】[0026]
【実験２】微生物用培地に求められる基本的な条件とし
て、水に溶解した場合の着色成分量が少ないことが挙げ
られる。着色成分は培地成分として不要であり、これら
の着色成分が培地中に多量に存在すると、培養中の菌体
濃度の測定が困難になるだけでなく、培養終了後に得ら
れる菌体が着色されてしまう。このため、培地としての
価値を著しく低めてしまう。そこで以下の実験２におい
て、前記実施例１および前記比較例１で得られた培地用
乾燥物の着色成分についての評価を行った。[Experiment 2] A basic condition required for a microbial medium is that the amount of coloring components when dissolved in water is small. Coloring components are unnecessary as medium components, and the presence of a large amount of these coloring components in the medium not only makes it difficult to measure the bacterial cell concentration during culture, but also causes the bacterial cells obtained after the culture to be colored. I will end up. Therefore, the value of the medium is significantly reduced. Therefore, in Experiment 2 below, the coloring components of the dried culture medium obtained in Example 1 and Comparative Example 1 were evaluated.
【００２７】上記実験１で得られたそれぞれの溶解液を
遠心分離して上清を得、得られた上清について、該上清
の色を目視観察し、さらに吸光光度計を用いて該上清の
４３０ｎｍ、および４８０ｎｍにおける吸光度を測定し
た。実験２において調べたそれぞれの溶解液の結果を表
３に示す。表３に示した結果から明らかなように、それ
ぞれの溶解液の上清についてその色を目視観察した結
果、本発明の実施例１で得られた溶解液の上清は無色を
呈し、着色は認められなかった。一方、比較例１で得ら
れた溶解液の上清は濃褐色を呈し、着色成分が多量に含
まれていることが明らかであった。また、表３に示した
結果から明らかなように、それぞれの溶解液の上清につ
いてその４３０ｎｍおよび４８０ｎｍの吸光度を測定し
た結果、本発明の実施例１で得られた溶解液の上清は、
比較例１で得られた溶解液の上清と比較して、極めて低
い吸光度を示した。The respective lysates obtained in the above Experiment 1 were centrifuged to obtain a supernatant, and the color of the obtained supernatant was visually observed and further measured using an absorptiometer. Absorbances at 430 nm and 480 nm of Qing were measured. Table 3 shows the results of the respective lysates examined in Experiment 2. As is clear from the results shown in Table 3, as a result of visually observing the color of the supernatant of each lysate, the supernatant of the lysate obtained in Example 1 of the present invention was colorless and showed no coloration. I was not able to admit. On the other hand, the supernatant of the solution obtained in Comparative Example 1 had a dark brown color, and it was clear that the coloring component was contained in a large amount. Further, as is clear from the results shown in Table 3, as a result of measuring the absorbance at 430 nm and 480 nm of the supernatants of the respective lysates, the lysate supernatants obtained in Example 1 of the present invention were:
Compared with the supernatant of the lysate obtained in Comparative Example 1, the absorbance was extremely low.
【００２８】さらに以下の実験３および実験４におい
て、前記実施例１において得た清澄液と、該清澄液を合
成吸着剤を用いる吸着処理に付して得た非吸着性画分の
それぞれの凍結乾燥物について、それらの着色成分につ
いての評価を行った。Further, in the following Experiments 3 and 4, freezing of the clarified liquid obtained in Example 1 and non-adsorbable fractions obtained by subjecting the clarified liquid to an adsorption treatment using a synthetic adsorbent The dried product was evaluated for those coloring components.
【実験３】上記実施例１において得た清澄液と該清澄液
を合成吸着剤を用いる吸着処理に付して得た非吸着性画
分のそれぞれ１リットルを凍結乾燥し、得られたそれぞ
れの凍結乾燥物の重量を測定した。その結果、該清澄液
１リットルからは２８．１ｇの凍結乾燥物が得られたの
に対して、非吸着性画分１リットルからは、２５．７ｇ
の凍結乾燥物が得られた。このことから清澄液を合成吸
着剤を用いる吸着処理に付して非吸着性画分を得ること
により、該清澄液１リットル当たり２．４ｇの不要な着
色成分を取り除くことができることがわかった。[Experiment 3] 1 liter of each of the clarified liquid obtained in Example 1 above and the non-adsorbed fraction obtained by subjecting the clarified liquid to an adsorption treatment using a synthetic adsorbent was freeze-dried. The weight of the freeze-dried product was measured. As a result, 28.1 g of a freeze-dried product was obtained from 1 liter of the clarified liquid, while 25.7 g was obtained from 1 liter of the non-adsorbable fraction.
Lyophilized product was obtained. From this, it was found that 2.4 g of unnecessary coloring components can be removed per liter of the clear solution by subjecting the clear solution to an adsorption treatment using a synthetic adsorbent to obtain a non-adsorbable fraction.
【００２９】[0029]
【実験４】さらに別に以下の実験４を行った。すなわ
ち、上記実施例１において得た培地用乾燥物と、上記実
施例１における該濃縮液を合成吸着剤を用いる吸着処理
に付さずにそのまま噴霧乾燥して得た培地用乾燥物を用
いて、酵母の培養試験を行い、得られる培養酵母菌体の
色の評価を行った。[Experiment 4] The following Experiment 4 was conducted separately. That is, using the dried medium product obtained in the above Example 1 and the dried medium product obtained by directly spray-drying the concentrated solution in the above Example 1 without being subjected to an adsorption treatment using a synthetic adsorbent, The yeast culture test was performed and the color of the obtained cultured yeast cells was evaluated.
【本発明の培地による培養試験】上記実施例１において
得た培地用乾燥物３０ｇ、ブドウ糖３２．５ｇ，炭酸ア
ンモニウム８．５ｇ，リン酸アンモニウム２ｇ，硫酸マ
グネシウム０．３ｇ，及び５０％乳酸２７．５ｇを水１
リットルに溶解し，ｐＨ４．５に調整後，市販の焼酎酵
母を接種し，３０℃で４０時間好気培養し、得られた培
養液を１００００ｒｐｍで１５分間遠心分離した。その
結果，該酵母菌体本来の白色を呈する湿重量３６．２ｇ
の酵母菌体が得られた。[Cultivation test using the medium of the present invention] 30 g of the dried product for medium obtained in Example 1 above, 32.5 g of glucose, 8.5 g of ammonium carbonate, 2 g of ammonium phosphate, 0.3 g of magnesium sulfate, and 27.50% lactic acid. 5 g of water
After dissolving in liter and adjusting to pH 4.5, commercially available shochu yeast was inoculated, aerobically cultured at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes. As a result, the wet weight of the yeast cells was 36.2 g, which was originally white.
Yeast cells were obtained.
【対照の培地による培養試験】上記実施例１における該
濃縮液を合成吸着剤を用いる吸着処理に付さずにそのま
ま噴霧乾燥して得た培地用乾燥物３０ｇ，ブドウ糖３
２．５ｇ，炭酸アンモニウム８．５ｇ，リン酸アンモニ
ウム２ｇ，硫酸マグネシウム０．３ｇ，及び５０％乳酸
２７．５ｇを水１リットルに溶解し，ｐＨ４．５に調整
後，市販の焼酎酵母を接種し，３０℃で４０時間好気培
養し、得られた培養液を１００００ｒｐｍで１５分間遠
心分離した。その結果，該酵母菌体本来の白色とは異な
る褐色を呈した湿重量３２．５ｇの酵母菌体が得られ
た。以上のことから、本発明による微生物用培地は、着
色成分の含有量が極めて低いため、培養中の菌体濃度の
測定の際に何ら問題を生じず、得られる培養菌体におい
ても着色がほとんど認められない微生物培養に極めて適
した性質を有していることが明らかとなった。[Cultivation test using a control medium] 30 g of a dried product for medium obtained by spray-drying the concentrated liquid as it is without subjecting it to an adsorption treatment using a synthetic adsorbent, glucose 3
2.5 g, ammonium carbonate 8.5 g, ammonium phosphate 2 g, magnesium sulfate 0.3 g, and 50% lactic acid 27.5 g were dissolved in 1 liter of water, adjusted to pH 4.5, and inoculated with commercially available shochu yeast. Aerobic culture was performed at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes. As a result, a yeast cell having a wet weight of 32.5 g, which had a brown color different from the original white color of the yeast cell, was obtained. From the above, the culture medium for microorganisms according to the present invention has an extremely low content of coloring components, and therefore does not cause any problem when measuring the concentration of bacterial cells in the culture, and the resulting cultured bacterial cells show almost no coloring. It has been revealed that it has extremely suitable properties for culturing microorganisms that are not recognized.
【００３０】[0030]
【実験５】前記実施例１で得られた培地用乾燥物、およ
び従来から一般的に使用されている廃糖蜜を用いて、酵
母の培養試験を行い、本発明により得られた微生物用培
地の評価を行った。[Experiment 5] A yeast culture test was carried out using the dried medium for culture medium obtained in Example 1 above and the molasses that has been generally used in the past, to obtain the medium for microorganisms obtained by the present invention. An evaluation was made.
【００３１】[0031]
【本発明の培地による培養試験】実施例１で得られた培
地用乾燥物３０ｇ，ブドウ糖３２．５ｇ，炭酸アンモニ
ウム８．５ｇ，リン酸アンモニウム２ｇ，硫酸マグネシ
ウム０．３ｇ，及び５０％乳酸２７．５ｇを水１リット
ルに溶解し，ｐＨ４．５に調整後，市販の焼酎酵母を接
種し，３０℃で４０時間好気培養し、得られた培養液を
１００００ｒｐｍで１５分間遠心分離した。その結果，
湿重量で３７．５ｇの菌体が得られた。[Cultivation test using the medium of the present invention] 30 g of the dried medium for medium obtained in Example 1, 32.5 g of glucose, 8.5 g of ammonium carbonate, 2 g of ammonium phosphate, 0.3 g of magnesium sulfate, and 27.50% lactic acid. After dissolving 5 g in 1 liter of water and adjusting the pH to 4.5, commercially available shochu yeast was inoculated, aerobically cultured at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes. as a result,
As a result, 37.5 g of wet cells was obtained.
【対照の培地による培養試験】廃糖蜜５０ｇ、ブドウ糖
３２．５ｇ，炭酸アンモニウム８．５ｇ，リン酸アンモ
ニウム２ｇ，硫酸マグネシウム０．３ｇ，及び５０％乳
酸２７．５ｇを水１リットルに溶解し，ｐＨ４．５に調
整後，市販の焼酎酵母を接種し，３０℃で４０時間好気
培養し、得られた培養液を１００００ｒｐｍで１５分間
遠心分離し，湿重量で２８．０ｇの菌体が得られた。[Cultivation test using a control medium] 50 g of molasses, 32.5 g of glucose, 8.5 g of ammonium carbonate, 2 g of ammonium phosphate, 0.3 g of magnesium sulfate, and 27.5 g of 50% lactic acid were dissolved in 1 liter of water to have a pH of 4. After adjusting to 0.5, commercially available shochu yeast was inoculated, aerobically cultivated at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes to obtain 28.0 g of wet cells. It was
【００３２】実験５において調べた酵母培養試験の結果
を表４に示す。表４に示した結果から明らかなように、
本発明の実施例１で得られた培地用乾燥物を用いた場合
の酵母菌体の湿重量は、対照の廃糖蜜を用いた場合の
１．３４倍に達した。さらに，この結果を培地添加量あ
たりで比較すると、本発明の実施例１で得られた培地用
乾燥物を用いた場合の酵母菌体の湿重量は、対照の廃糖
蜜を用いた場合の２．２３倍に達することが判った。さ
らに、培養中の酵母菌体の湿重量を経時的に調べたとこ
ろ、本発明の実施例１で得られた培地用乾燥物を用いた
場合には、培養開始後３０時間目において、すでに対照
の廃糖蜜を用いた場合の培養終了時（４０時間目）とほ
ぼ同じ菌体湿重量に達していることが判った。このこと
から、本発明の焼酎蒸留残液から得られる微生物用培地
を用いることにより、従来よりも短い培養日数で所望の
酵母菌体が得られることが明らかとなった。The results of the yeast culture test examined in Experiment 5 are shown in Table 4. As is clear from the results shown in Table 4,
The wet weight of the yeast cells when using the dried medium for culture medium obtained in Example 1 of the present invention reached 1.34 times that when using the control molasses. Furthermore, comparing these results per medium addition amount, the wet weight of the yeast cells when using the dried medium for culture obtained in Example 1 of the present invention was 2 when the control molasses was used. It turned out to reach .23 times. Furthermore, when the wet weight of the yeast cells in the culture was examined over time, when the dried medium for medium obtained in Example 1 of the present invention was used, the control was already performed at 30 hours after the start of the culture. It was found that when the molasses was used, the wet weight of the cells reached almost the same as that at the end of the culture (40th hour). From this, it was clarified that the desired yeast cells can be obtained in a shorter number of culture days than before by using the microorganism culture medium obtained from the shochu distillation residue of the present invention.
【００３３】[0033]
【実験６】前記実施例１で得られた培地用乾燥物、およ
び従来から一般的に使用されている培地用酵母エキスを
用いて、乳酸菌の培養試験を行い、本発明により得られ
た微生物用培地の評価を行った。[Experiment 6] A culture test of lactic acid bacteria was carried out using the dried medium for culture medium obtained in Example 1 and the yeast extract for culture medium which has been generally used from the past. The culture medium was evaluated.
【００３４】[0034]
【本発明の培地による培養試験】実施例１で得られた培
地用乾燥物５ｇ，ブドウ糖１０ｇ，ポリペプトン５ｇ，
及び塩化ナトリウム５ｇを水１リットルに溶解し，ｐＨ
７に調整後，ラクトバチルス アシドフィラスＩＦＯ１
３９５１Tを接種し，３０℃で４０時間培養し、得られ
た培養液を１００００ｒｐｍで１５分間遠心分離した。
その結果，湿重量で４１．５ｇの菌体が得られた。[Cultivation test using the medium of the present invention] 5 g of the dried medium for medium obtained in Example 1, 10 g of glucose, 5 g of polypeptone,
And 5 g of sodium chloride are dissolved in 1 liter of water, and the pH is
After adjusting to 7, Lactobacillus acidophilus IFO1
3951 T was inoculated and cultured at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes.
As a result, 41.5 g of wet cells were obtained.
【対照の培地による培養試験】培地用酵母エキス５ｇ，
ブドウ糖１０ｇ，ポリペプトン５ｇ，及び塩化ナトリウ
ム５ｇを水１リットルに溶解し，ｐＨ７に調整後，ラク
トバチルス アシドフィラスＩＦＯ１３９５１Tを接種
し，３０度で４０時間培養し、得られた培養液を１００
００ｒｐｍで１５分間遠心分離し，湿重量で２９．４ｇ
の菌体が得られた。[Culture test using control medium] Yeast extract 5 g for medium,
Glucose 10 g, polypeptone 5 g, and sodium chloride 5 g were dissolved in water 1 liter, and after adjusting the pH to 7, Lactobacillus acidophilus IFO13951 T was inoculated and cultured at 30 ° C. for 40 hours.
Centrifuge at 00 rpm for 15 minutes, wet weight 29.4g
Was obtained.
【００３５】実験６において調べた乳酸菌培養試験の結
果を表５に示す。表５に示した結果から明らかなよう
に、本発明の実施例１で得られた培地用乾燥物を用いた
場合の乳酸菌菌体の湿重量は、培地用酵母エキスを用い
た対照の場合の１．４１倍に達した。また、培養中の乳
酸菌菌体の湿重量を経時的に調べたところ、本発明の実
施例１で得られた培地用乾燥物を用いた場合には、培養
開始後２８時間目において、すでに培地用酵母エキスを
用いた対照の場合の培養終了時（４０時間目）とほぼ同
じ菌体湿重量に達していることが判った。このことか
ら、本発明の焼酎蒸留残液から得られる微生物用培地を
用いることにより、従来よりも短い培養日数で所望の乳
酸菌菌体が得られることが明らかとなった。The results of the lactic acid bacterium culture test examined in Experiment 6 are shown in Table 5. As is clear from the results shown in Table 5, the wet weight of the lactic acid bacterium cells when the dried medium product obtained in Example 1 of the present invention was used was the same as when the yeast extract for the medium was used as a control. It reached 1.41 times. Further, when the wet weight of the lactic acid bacterial cells in the culture was examined with time, when the dried medium for culture obtained in Example 1 of the present invention was used, the medium was already cultured at 28 hours after the start of the culture. It was found that the wet weight of the cells reached almost the same as that at the end of the culture (40th hour) in the case of the control using the yeast extract. From this, it was clarified that the desired lactic acid bacterium can be obtained in a shorter number of culture days than before by using the microorganism culture medium obtained from the shochu distillation residue of the present invention.
【００３６】[0036]
【実験７】前記実施例１で得られた培地用乾燥物を、従
来の乳酸菌培養の際に用いる乳酸菌培養用の対照培地に
１％添加して、乳酸菌の培養試験を行い、本発明により
得られた培地用乾燥物の乳酸菌増殖促進因子としての評
価を行った。[Experiment 7] The dried medium product obtained in Example 1 was added to a control medium for lactic acid bacterium culture used in conventional lactic acid bacterium culture in an amount of 1% to perform a lactic acid bacterium culture test. The dried product for medium was evaluated as a lactic acid bacterium growth promoting factor.
【００３７】[0037]
【対照培地による培養試験】培地用酵母エキス５ｇ，ブ
ドウ糖１０ｇ，ポリペプトン５ｇ，及び塩化ナトリウム
５ｇを水１リットルに溶解し，ｐＨ７に調整する手法で
３個の培地を作製した。ラクトバチルス プランタラム
ＩＦＯ３０７０，ラクトバチルス アシドフィラスＩＦ
Ｏ１３９５１T，及びラクトバチルス ファーメンタム
ＩＦＯ３０７１のそれぞれを個別に前記培地の１つに接
種し，３０℃で４０時間培養した。これにより３個の培
養液を得た。得られた３個の培養液のそれぞれを１００
００ｒｐｍで１５分間遠心分離した。このようにして前
記３種類の乳酸菌のそれぞれについて培養菌体を得た。[Cultivation test using control medium] 3 types of medium were prepared by dissolving 5 g of yeast extract for medium, 10 g of glucose, 5 g of polypeptone, and 5 g of sodium chloride in 1 liter of water and adjusting the pH to 7. Lactobacillus plantarum IFO3070, Lactobacillus acidophilus IF
Each of O13951 T and Lactobacillus fermentum IFO3071 was individually inoculated into one of the above-mentioned medium and cultured at 30 ° C. for 40 hours. As a result, three culture solutions were obtained. 100 of each of the 3 cultures obtained
Centrifuge at 00 rpm for 15 minutes. In this way, cultured cells were obtained for each of the above three types of lactic acid bacteria.
【対照培地に本発明の培地を１％添加した培養試験】実
施例１で得られた培地用乾燥物１０ｇ，培地用酵母エキ
ス５ｇ，ブドウ糖１０ｇ，ポリペプトン５ｇ，及び塩化
ナトリウム５ｇを水１リットルに溶解し，ｐＨ７に調整
する手法で３個の培地を作製した。ラクトバチルス プ
ランタラムＩＦＯ３０７０，ラクトバチルス アシドフ
ィラスＩＦＯ１３９５１T，及びラクトバチルス ファ
ーメンタムＩＦＯ３０７１のそれぞれを個別に前記培地
の１つに接種し，３０℃で４０時間培養した。これによ
り３個の培養液を得た。得られた３個の培養液のそれぞ
れを１００００ｒｐｍで１５分間遠心分離した。このよ
うにして前記３種類の乳酸菌のそれぞれについて培養菌
体を得た。[Cultivation test in which 1% of the medium of the present invention was added to the control medium] 10 g of the dried product for medium obtained in Example 1, 5 g of yeast extract for medium, 10 g of glucose, 5 g of polypeptone, and 5 g of sodium chloride were added to 1 liter of water. Three media were prepared by the method of dissolving and adjusting the pH to 7. Each of Lactobacillus plantarum IFO 3070, Lactobacillus acidophilus IFO 13951 T , and Lactobacillus fermentum IFO 3071 was individually inoculated into one of the above-mentioned medium, and cultured at 30 ° C. for 40 hours. As a result, three culture solutions were obtained. Each of the three cultures obtained was centrifuged at 10,000 rpm for 15 minutes. In this way, cultured cells were obtained for each of the above three types of lactic acid bacteria.
【００３８】その結果、前記対照培地に本発明の実施例
１で得られた培地用乾燥物を１％添加することにより、
上述の乳酸菌それぞれの増殖が著しく促進されることが
判った。具体的には、前記対照培地に本発明の実施例１
で得られた培地用乾燥物を１％添加することにより、乳
酸菌の菌体湿重量が、ラクトバチルス プランタラムＩ
ＦＯ３０７０では対照培地の３．１３倍、ラクトバチル
ス アシドフィラスＩＦＯ１３９５１Tでは対照培地の
２．５８倍、ラクトバチルス ファーメンタムＩＦＯ３
０７１では対照培地の２．６７倍に達した。このことか
ら本発明により得られる微生物用培地は優れた乳酸菌の
増殖促進効果も有することが判った。As a result, by adding 1% of the dried product for a medium obtained in Example 1 of the present invention to the control medium,
It was found that the growth of each of the above-mentioned lactic acid bacteria was significantly promoted. Specifically, the control medium described above was used in Example 1 of the present invention.
By adding 1% of the dried product for medium obtained in the above, the wet weight of lactic acid bacteria was determined to be Lactobacillus plantarum I.
FO3070 was 3.13 times that of the control medium, Lactobacillus acidophilus IFO13951 T was 2.58 times that of the control medium, and Lactobacillus fermentum IFO3.
At 071, it reached 2.67 times that of the control medium. From this, it was found that the microbial medium obtained by the present invention also has an excellent growth promoting effect on lactic acid bacteria.
【００３９】[0039]
【実験８】前記実施例１で得られた培地用乾燥物、およ
び従来から一般的に使用されている培地用酵母エキスを
用いて、ビフィズス菌の培養試験を行い、本発明により
得られた微生物用培地の評価を行った。[Experiment 8] A culture test of Bifidobacterium was performed using the dried medium for culture obtained in Example 1 above and the yeast extract for medium generally used conventionally, and the microorganism obtained by the present invention was tested. The culture medium was evaluated.
【００４０】[0040]
【本発明の培地による培養試験】実施例１で得られた培
地用乾燥物１０ｇ，ブドウ糖１０ｇ，カゼインペプトン
１０ｇ，肉エキス５g，リン酸水素２カリウム３ｇ，L-
システイン塩酸塩０．５ｇ，アスコルビン酸ナトリウム
１０g ，及び１mlの界面活性剤Ｔｗｅｅｎ８０(商標名)
を水１リットルに溶解し，ｐＨ７に調整後，ビフィドバ
クテリウム ビフィダムJCM１２５４を接種し，３７℃
で４８時間培養し、得られた培養液を１００００ｒｐｍ
で１５分間遠心分離し，ビフィズス菌菌体を得た。[Culture test using medium of the present invention] 10 g of dried medium for medium obtained in Example 1, 10 g of glucose, 10 g of casein peptone, 5 g of meat extract, 3 g of dipotassium hydrogen phosphate, 3 g of L-
Cysteine hydrochloride 0.5 g, sodium ascorbate 10 g, and 1 ml of surfactant Tween 80 (trade name)
Dissolved in 1 liter of water and adjusted to pH 7, inoculated with Bifidobacterium bifidum JCM1254, 37 ℃
Cultivated for 48 hours at 10,000 rpm
After centrifugation for 15 minutes, bifidobacteria were obtained.
【対照の培地による培養試験】培地用酵母エキス５ｇ，
ブドウ糖１０ｇ，カゼインペプトン１０ｇ，肉エキス５
g，リン酸水素２カリウム３ｇ，L-システイン塩酸塩
０．５ｇ，アスコルビン酸ナトリウム１０g ，及び１ml
の界面活性剤Ｔｗｅｅｎ８０(商標名)を水１リットルに
溶解し，ｐＨ７に調整後，ビフィドバクテリウム ビフ
ィダムJCM１２５４を接種し，３７℃で４８時間培養
し、得られた培養液を１００００ｒｐｍで１５分間遠心
分離し，ビフィズス菌菌体を得た。[Culture test using control medium] Yeast extract 5 g for medium,
Glucose 10g, casein peptone 10g, meat extract 5
g, dipotassium hydrogen phosphate 3 g, L-cysteine hydrochloride 0.5 g, sodium ascorbate 10 g, and 1 ml
The surfactant Tween 80 (trademark) of 1 is dissolved in 1 liter of water, adjusted to pH 7, and inoculated with Bifidobacterium bifidum JCM1254, incubated at 37 ° C. for 48 hours, and the obtained culture broth at 10,000 rpm for 15 minutes. Centrifugation gave bifidobacteria.
【００４１】その結果、それぞれの培養試験において得
られたビフィズス菌菌体の湿重量を比較したところ、本
発明の実施例１で得られた培地用乾燥物を用いた場合の
ビフィズス菌菌体の湿重量は、培地用酵母エキスを用い
た対照の場合の１．６３倍に達した。また、培養中のビ
フィズス菌菌体の湿重量を経時的に調べたところ、本発
明の実施例１で得られた培地用乾燥物を用いた場合に
は、培養開始後３５時間目において、すでに培地用酵母
エキスを用いた対照の場合の培養終了時（４８時間目）
とほぼ同じ菌体湿重量に達していることが判った。この
ことから、本発明の焼酎蒸留残液から得られる微生物用
培地を用いることにより、従来よりも短い培養日数で所
望のビフィズス菌菌体が得られることが明らかとなっ
た。As a result, the wet weights of the bifidobacteria obtained in the respective culture tests were compared. As a result, the bifidobacteria of the bifidobacteria obtained when the dried medium for culture obtained in Example 1 of the present invention was used. The wet weight reached 1.63 times that of the control using the yeast extract for the medium. In addition, when the wet weight of the Bifidobacteria cells in the culture was examined with time, when the dried medium for medium obtained in Example 1 of the present invention was used, at 35 hours after the start of the culture, At the end of culture (48 hours) in the case of control using yeast extract for medium
It was found that the wet weight of the cells was almost the same as the above. From this, it was clarified that the desired bifidobacteria can be obtained in a shorter number of culture days than before by using the microbial medium obtained from the shochu distilled residue of the present invention.
【００４２】[0042]
【実施例２】大麦製焼酎製造の蒸留工程で得られた前記
大麦製焼酎蒸留残液１キロリットルを信和エンジニアリ
ング（株）製のスクリュープレス方式の固液分離機で固
液を分離し，得られた液体分をさらに巴工業（株）社製
のデカンタ型遠心分離器を用いて固液を分離し，得られ
た液体分をさらに日本シューマッハー（株）社製のセラ
ミック濾過装置を用いてさらにＳＳ分を分離し，約０．
８５キロリットルの清澄液を得、当該清澄液を水酸化ナ
トリウムで中和し，約０．９キロリットルの中和液を得
た。次に、当該中和液を（株）大河原製作所製の真空蒸
発装置を用いて約３倍まで濃縮して濃縮液を得、得られ
た当該濃縮液を三菱化学社製の合成吸着剤セパビーズＳ
Ｐ８５０を充填したカラムに接触させ、当該充填カラム
から溶出してきた当該合成吸着剤に対して非吸着性を示
す非吸着性画分を得、得られた非吸着性画分を（株）大
河原化工機製スプレー型乾燥機を用いて、入口温度 150
℃，出口温度80℃，熱風量0.45m3 / min，および噴霧空
気量 1.0 kgf /cm2の条件で噴霧乾燥し、微生物用培地
に使用する約８ｋｇの培地用乾燥物を得た。[Example 2] 1 kiloliter of the barley shochu distillation residual liquid obtained in the distillation step of barley shochu production was separated by a screw press type solid-liquid separator manufactured by Shinwa Engineering Co., Ltd. The obtained liquid content is further separated into solid and liquid using a decanter type centrifuge manufactured by Tomoe Kogyo Co., Ltd., and the obtained liquid content is further processed using a ceramic filtration device manufactured by Nippon Schumacher Co., Ltd. Separate the SS content to about 0.
Eighty-five kiloliters of clear liquid was obtained, and the clear liquid was neutralized with sodium hydroxide to obtain about 0.9 kiloliter of neutralized liquid. Next, the neutralized solution is concentrated to about 3 times by using a vacuum evaporator manufactured by Okawara Seisakusho Co., Ltd. to obtain a concentrated solution, and the obtained concentrated solution is a synthetic adsorbent SepaBeads S manufactured by Mitsubishi Chemical Corporation.
By contacting with a column packed with P850, a non-adsorptive fraction showing non-adsorptivity to the synthetic adsorbent eluted from the packed column was obtained, and the obtained non-adsorptive fraction was collected from Okawara Kako Co., Ltd. Inlet temperature of 150 using a mechanical spray dryer
° C., an outlet temperature of 80 ° C., hot-air quantity 0.45 m 3 / min, and spray dried under the conditions of the spray air volume 1.0 kgf / cm 2, to obtain a medium for drying of approximately 8kg use in microbiological media.
【００４３】[0043]
【実験９】前記実施例２で得られた培地用乾燥物、およ
び従来から一般的に使用されている培地用酵母エキスを
用いて、乳酸菌の培養試験を行い、本発明により得られ
た微生物用培地の評価を行った。[Experiment 9] A lactic acid bacterium culture test was carried out using the dried medium for culture medium obtained in Example 2 and the yeast extract for medium which has been generally used from the past, for the microorganism obtained by the present invention. The culture medium was evaluated.
【００４４】[0044]
【本発明の培地による培養試験】実施例２で得られた培
地用乾燥物５ｇ，ブドウ糖１０ｇ，ポリペプトン５ｇ，
及び塩化ナトリウム５ｇを水１リットルに溶解し，ｐＨ
７に調整後，ラクトバチルス アシドフィラスＩＦＯ１
３９５１Tを接種し，３０℃で４０時間培養し、得られ
た培養液を１００００ｒｐｍで１５分間遠心分離した。
その結果，湿重量で４５．３ｇの菌体が得られた。[Cultivation test using the medium of the present invention] 5 g of the dried product for medium obtained in Example 2, 10 g of glucose, 5 g of polypeptone,
And 5 g of sodium chloride are dissolved in 1 liter of water, and the pH is
After adjusting to 7, Lactobacillus acidophilus IFO1
3951 T was inoculated and cultured at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes.
As a result, 45.3 g of wet cells were obtained.
【対照の培地による培養試験】培地用酵母エキス５ｇ，
ブドウ糖１０ｇ，ポリペプトン５ｇ，及び塩化ナトリウ
ム５ｇを水１リットルに溶解し，ｐＨ７に調整後，ラク
トバチルス アシドフィラスＩＦＯ１３９５１Tを接種
し，３０度で４０時間培養し、得られた培養液を１００
００ｒｐｍで１５分間遠心分離し，湿重量で２９．７ｇ
の菌体が得られた。[Culture test using control medium] Yeast extract 5 g for medium,
Glucose 10 g, polypeptone 5 g, and sodium chloride 5 g were dissolved in water 1 liter, and after adjusting the pH to 7, Lactobacillus acidophilus IFO13951 T was inoculated and cultured at 30 ° C. for 40 hours.
Centrifuge at 00 rpm for 15 minutes, wet weight 29.7g
Was obtained.
【００４５】実験９において調べた乳酸菌培養試験の結
果を表６に示す。表６に示した結果から明らかなよう
に、本発明の実施例２で得られた培地用乾燥物を用いた
場合の乳酸菌菌体の湿重量は、培地用酵母エキスを用い
た対照の場合の１．５３倍に達した。また、培養中の乳
酸菌菌体の湿重量を経時的に調べたところ、本発明の実
施例２で得られた培地用乾燥物を用いた場合には、培養
開始後２８時間目において、すでに培地用酵母エキスを
用いた対照の場合の培養終了時（４０時間目）とほぼ同
じ菌体湿重量に達していることが判った。このことか
ら、本発明の焼酎蒸留残液から得られる微生物用培地を
用いることにより、従来よりも短い培養日数で所望の乳
酸菌菌体が得られることが明らかとなった。The results of the lactic acid bacterium culture test examined in Experiment 9 are shown in Table 6. As is clear from the results shown in Table 6, the wet weight of the lactic acid bacterium cells when the dried medium product obtained in Example 2 of the present invention was used was the same as when the control using the yeast extract for the medium was used. It reached 1.53 times. Further, when the wet weight of the lactic acid bacterium in the culture was examined with time, when the dried medium for culture obtained in Example 2 of the present invention was used, it was found that the medium was already cultured at 28 hours after the start of culture. It was found that the wet weight of the cells reached almost the same as that at the end of the culture (40th hour) in the case of the control using the yeast extract. From this, it was clarified that the desired lactic acid bacterium can be obtained in a shorter number of culture days than before by using the microorganism culture medium obtained from the shochu distillation residue of the present invention.
【００４６】[0046]
【実験１０】前記実施例２で得られた培地用乾燥物、お
よび従来から一般的に使用されている培地用酵母エキス
を用いて、ビフィズス菌の培養試験を行い、本発明によ
り得られた微生物用培地の評価を行った。[Experiment 10] A microorganism obtained by the present invention was subjected to a culture test of Bifidobacterium using the dried medium for culture obtained in Example 2 and the yeast extract for medium generally used conventionally. The culture medium was evaluated.
【００４７】[0047]
【本発明の培地による培養試験】実施例２で得られた培
地用乾燥物１０ｇ，ブドウ糖１０ｇ，カゼインペプトン
１０ｇ，肉エキス５g，リン酸水素２カリウム３ｇ，L-
システイン塩酸塩０．５ｇ，アスコルビン酸ナトリウム
１０g ，及び１mlの界面活性剤Ｔｗｅｅｎ８０(商標名)
を水１リットルに溶解し，ｐＨ７に調整後，ビフィドバ
クテリウム ビフィダムJCM１２５４を接種し，３７℃
で４８時間培養し、得られた培養液を１００００ｒｐｍ
で１５分間遠心分離し，ビフィズス菌菌体を得た。[Cultivation test using the medium of the present invention] 10 g of the dried medium for medium obtained in Example 2, 10 g of glucose, 10 g of casein peptone, 5 g of meat extract, 3 g of dipotassium hydrogen phosphate, 3 g of L-
Cysteine hydrochloride 0.5 g, sodium ascorbate 10 g, and 1 ml of surfactant Tween 80 (trade name)
Dissolved in 1 liter of water and adjusted to pH 7, inoculated with Bifidobacterium bifidum JCM1254, 37 ℃
Cultivated for 48 hours at 10,000 rpm
After centrifugation for 15 minutes, bifidobacteria were obtained.
【対照の培地による培養試験】培地用酵母エキス５ｇ，
ブドウ糖１０ｇ，カゼインペプトン１０ｇ，肉エキス５
g，リン酸水素２カリウム３ｇ，L-システイン塩酸塩
０．５ｇ，アスコルビン酸ナトリウム１０g ，及び１ml
の界面活性剤Ｔｗｅｅｎ８０(商標名)を水１リットルに
溶解し，ｐＨ７に調整後，ビフィドバクテリウム ビフ
ィダムJCM１２５４を接種し，３７℃で４８時間培養
し、得られた培養液を１００００ｒｐｍで１５分間遠心
分離し，ビフィズス菌菌体を得た。[Culture test using control medium] Yeast extract 5 g for medium,
Glucose 10g, casein peptone 10g, meat extract 5
g, dipotassium hydrogen phosphate 3 g, L-cysteine hydrochloride 0.5 g, sodium ascorbate 10 g, and 1 ml
The surfactant Tween 80 (trademark) of 1 is dissolved in 1 liter of water, adjusted to pH 7, and inoculated with Bifidobacterium bifidum JCM1254, incubated at 37 ° C. for 48 hours, and the obtained culture broth at 10,000 rpm for 15 minutes. Centrifugation gave bifidobacteria.
【００４８】その結果、それぞれの培養試験において得
られたビフィズス菌菌体の湿重量を比較したところ、本
発明の実施例２で得られた培地用乾燥物を用いた場合の
ビフィズス菌菌体の湿重量は、培地用酵母エキスを用い
た対照の場合の１．８１倍に達した。また、培養中のビ
フィズス菌菌体の湿重量を経時的に調べたところ、本発
明の実施例２で得られた培地用乾燥物を用いた場合に
は、培養開始後３５時間目において、すでに培地用酵母
エキスを用いた対照の場合の培養終了時（４８時間目）
とほぼ同じ菌体湿重量に達していることが判った。この
ことから、本発明の焼酎蒸留残液から得られる微生物用
培地を用いることにより、従来よりも短い培養日数で所
望のビフィズス菌菌体が得られることが明らかとなっ
た。As a result, when the wet weights of the bifidobacteria obtained in the respective culture tests were compared, it was found that the bifidobacteria of the bifidobacteria obtained when the dried medium for culture obtained in Example 2 of the present invention was used. The wet weight reached 1.81 times that of the control using the yeast extract for the medium. Further, when the wet weight of the bifidobacteria cells in the culture was examined with time, it was found that when the dried medium product obtained in Example 2 of the present invention was used, it was already observed at 35 hours after the start of the culture. At the end of culture (48 hours) in the case of control using yeast extract for medium
It was found that the wet weight of the cells was almost the same as the above. From this, it was clarified that the desired bifidobacteria can be obtained in a shorter number of culture days than before by using the microbial medium obtained from the shochu distilled residue of the present invention.
【００４９】[0049]
【実施例３】大麦製焼酎製造の蒸留工程で得られた前記
大麦製焼酎蒸留残液１キロリットルを信和エンジニアリ
ング（株）製のスクリュープレス方式の固液分離機で固
液を分離し，得られた液体分をさらに巴工業（株）社製
のデカンタ型遠心分離器を用いて固液を分離し，約０．
８５キロリットルの液体分を得た。次に、当該液体分を
（株）大河原製作所製の真空蒸発装置を用いて約１／３
に濃縮して濃縮液を得、得られた当該濃縮液を三菱化学
社製の合成吸着剤セパビーズＳＰ８５０を充填したカラ
ムに接触させ、当該充填カラムから溶出してきた当該合
成吸着剤に対して非吸着性を示す非吸着性画分を得、得
られた非吸着性画分に可溶分の 1.5倍量の（株）日澱化
学製食品添加用デキストリン・アミコール 6-Lを添加
後，（株）大河原化工機製スプレー型乾燥機を用いて、
入口温度 150℃，出口温度80℃，熱風量0.45m3 / min，
および噴霧空気量 1.0 kgf /cm2の条件で噴霧乾燥し、
微生物用培地に使用する約１２ｋｇの乾燥物を得た。Example 3 1 kiloliter of the barley shochu distillation residual liquid obtained in the distillation step of barley shochu production was separated by a screw press type solid-liquid separator manufactured by Shinwa Engineering Co., Ltd. to obtain a solid-liquid separator. The obtained liquid component is further separated into solid and liquid by using a decanter type centrifugal separator manufactured by Tomoe Kogyo Co., Ltd.
Obtained 85 kiloliters of liquid. Next, about 1/3 of the liquid content was applied using a vacuum evaporator manufactured by Okawara Seisakusho.
Is concentrated to obtain a concentrated liquid, and the obtained concentrated liquid is brought into contact with a column packed with a synthetic adsorbent SepaBeads SP850 manufactured by Mitsubishi Chemical Co., Inc., and the synthetic adsorbent eluted from the packed column is not adsorbed. Non-adsorptive fraction exhibiting the property was added, and after adding 1.5 times the soluble amount of dextrin / amicol 6-L for food addition manufactured by Nitto Chemical Co., Ltd. to the non-adsorptive fraction ) Using a spray type dryer manufactured by Okawara Kakoki,
Inlet temperature 150 ℃, outlet temperature 80 ℃, hot air flow 0.45m 3 / min,
And spray-dried under the condition of spray air volume 1.0 kgf / cm 2 ,
About 12 kg of dried product to be used as a microorganism culture medium was obtained.
【００５０】[0050]
【実験１１】前記実施例３で得られた培地用乾燥物、お
よび従来から一般的に使用されている培地用酵母エキス
を用いて、乳酸菌の培養試験を行い、本発明により得ら
れた微生物用培地の評価を行った。[Experiment 11] A culture test of lactic acid bacteria was carried out using the dried medium for culture medium obtained in Example 3 and yeast extract for culture medium which has been generally used from the past. The culture medium was evaluated.
【００５１】[0051]
【本発明の培地による培養試験】実施例３で得られた培
地用乾燥物５ｇ，ブドウ糖１０ｇ，ポリペプトン５ｇ，
及び塩化ナトリウム５ｇを水１リットルに溶解し，ｐＨ
７に調整後，ラクトバチルス アシドフィラスＩＦＯ１
３９５１Tを接種し，３０℃で４０時間培養し、得られ
た培養液を１００００ｒｐｍで１５分間遠心分離した。
その結果，湿重量で４３．１ｇの菌体が得られた。[Culture test using the medium of the present invention] 5 g of the dried product for medium obtained in Example 3, 10 g of glucose, 5 g of polypeptone,
And 5 g of sodium chloride are dissolved in 1 liter of water, and the pH is
After adjusting to 7, Lactobacillus acidophilus IFO1
3951 T was inoculated and cultured at 30 ° C. for 40 hours, and the obtained culture solution was centrifuged at 10,000 rpm for 15 minutes.
As a result, 43.1 g of wet cells were obtained.
【対照の培地による培養試験】培地用酵母エキス５ｇ，
ブドウ糖１０ｇ，ポリペプトン５ｇ，及び塩化ナトリウ
ム５ｇを水１リットルに溶解し，ｐＨ７に調整後，ラク
トバチルス アシドフィラスＩＦＯ１３９５１Tを接種
し，３０度で４０時間培養し、得られた培養液を１００
００ｒｐｍで１５分間遠心分離し，湿重量で２９．２ｇ
の菌体が得られた。[Culture test using control medium] Yeast extract 5 g for medium,
Glucose 10 g, polypeptone 5 g, and sodium chloride 5 g were dissolved in water 1 liter, and after adjusting the pH to 7, Lactobacillus acidophilus IFO13951 T was inoculated and cultured at 30 ° C. for 40 hours.
Centrifuge at 00 rpm for 15 minutes, wet weight 29.2g
Was obtained.
【００５２】実験１１において調べた乳酸菌培養試験の
結果を表７に示す。表７に示した結果から明らかなよう
に、本発明の実施例３で得られた培地用乾燥物を用いた
場合の乳酸菌菌体の湿重量は、培地用酵母エキスを用い
た対照の場合の１．４８倍に達した。また、培養中の乳
酸菌菌体の湿重量を経時的に調べたところ、本発明の実
施例３で得られた培地用乾燥物を用いた場合には、培養
開始後２８時間目において、すでに培地用酵母エキスを
用いた対照の場合の培養終了時（４０時間目）とほぼ同
じ菌体湿重量に達していることが判った。このことか
ら、本発明の焼酎蒸留残液から得られる微生物用培地を
用いることにより、従来よりも短い培養日数で所望の乳
酸菌菌体が得られることが明らかとなった。The results of the lactic acid bacterium culture test investigated in Experiment 11 are shown in Table 7. As is clear from the results shown in Table 7, the wet weight of the lactic acid bacterium cells when the dried medium product obtained in Example 3 of the present invention was used was the same as when the yeast extract for medium was used as a control. It reached 1.48 times. In addition, when the wet weight of the lactic acid bacterial cells in the culture was examined with time, when the dried medium for culture obtained in Example 3 of the present invention was used, it was found that at 28 hours after the start of the culture, the medium was already cultured. It was found that the wet weight of the cells reached almost the same as that at the end of the culture (40th hour) in the case of the control using the yeast extract. From this, it was clarified that the desired lactic acid bacterium can be obtained in a shorter number of culture days than before by using the microorganism culture medium obtained from the shochu distillation residue of the present invention.
【００５３】[0053]
【実験１２】前記実施例３で得られた培地用乾燥物、お
よび従来から一般的に使用されている培地用酵母エキス
を用いて、ビフィズス菌の培養試験を行い、本発明によ
り得られた微生物用培地の評価を行った。[Experiment 12] A culture test of Bifidobacterium was carried out using the dried medium for culture medium obtained in Example 3 and the yeast extract for culture medium which has been generally used conventionally, and the microorganism obtained by the present invention was tested. The culture medium was evaluated.
【００５４】[0054]
【本発明の培地による培養試験】実施例３で得られた培
地用乾燥物１０ｇ，ブドウ糖１０ｇ，カゼインペプトン
１０ｇ，肉エキス５g，リン酸水素２カリウム３ｇ，L-
システイン塩酸塩０．５ｇ，アスコルビン酸ナトリウム
１０g ，及び１mlの界面活性剤Ｔｗｅｅｎ８０(商標名)
を水１リットルに溶解し，ｐＨ７に調整後，ビフィドバ
クテリウム ビフィダムJCM１２５４を接種し，３７℃
で４８時間培養し、得られた培養液を１００００ｒｐｍ
で１５分間遠心分離し，ビフィズス菌菌体を得た。[Cultivation test using the medium of the present invention] 10 g of the dried product for medium obtained in Example 3, 10 g of glucose, 10 g of casein peptone, 5 g of meat extract, 3 g of dipotassium hydrogen phosphate, 3 g of L-
Cysteine hydrochloride 0.5 g, sodium ascorbate 10 g, and 1 ml of surfactant Tween 80 (trade name)
Dissolved in 1 liter of water and adjusted to pH 7, inoculated with Bifidobacterium bifidum JCM1254, 37 ℃
Cultivated for 48 hours at 10,000 rpm
After centrifugation for 15 minutes, bifidobacteria were obtained.
【対照の培地による培養試験】培地用酵母エキス５ｇ，
ブドウ糖１０ｇ，カゼインペプトン１０ｇ，肉エキス５
g，リン酸水素２カリウム３ｇ，L-システイン塩酸塩
０．５ｇ，アスコルビン酸ナトリウム１０g ，及び１ml
の界面活性剤Ｔｗｅｅｎ８０(商標名)を水１リットルに
溶解し，ｐＨ７に調整後，ビフィドバクテリウム ビフ
ィダムJCM１２５４を接種し，３７℃で４８時間培養
し、得られた培養液を１００００ｒｐｍで１５分間遠心
分離し，ビフィズス菌菌体を得た。[Culture test using control medium] Yeast extract 5 g for medium,
Glucose 10g, casein peptone 10g, meat extract 5
g, dipotassium hydrogen phosphate 3 g, L-cysteine hydrochloride 0.5 g, sodium ascorbate 10 g, and 1 ml
The surfactant Tween 80 (trademark) of 1 is dissolved in 1 liter of water, adjusted to pH 7, and inoculated with Bifidobacterium bifidum JCM1254, incubated at 37 ° C. for 48 hours, and the obtained culture broth at 10,000 rpm for 15 minutes. Centrifugation gave bifidobacteria.
【００５５】その結果、それぞれの培養試験において得
られたビフィズス菌菌体の湿重量を比較したところ、本
発明の実施例３で得られた培地用乾燥物を用いた場合の
ビフィズス菌菌体の湿重量は、培地用酵母エキスを用い
た対照の場合の１．７３倍に達した。また、培養中のビ
フィズス菌菌体の湿重量を経時的に調べたところ、本発
明の実施例３で得られた培地用乾燥物を用いた場合に
は、培養開始後３５時間目において、すでに培地用酵母
エキスを用いた対照の場合の培養終了時（４８時間目）
とほぼ同じ菌体湿重量に達していることが判った。この
ことから、本発明の焼酎蒸留残液から得られる微生物用
培地を用いることにより、従来よりも短い培養日数で所
望のビフィズス菌菌体が得られることが明らかとなっ
た。 As a result, when the wet weights of the bifidobacteria obtained in the respective culture tests were compared, it was found that the bifidobacteria of the bifidobacteria obtained when the dried medium for culture obtained in Example 3 of the present invention was used. The wet weight reached 1.73 times that of the control using the yeast extract for the medium. Further, when the wet weight of the bifidobacteria cells in the culture was examined with time, when the dried medium for medium obtained in Example 3 of the present invention was used, at 35 hours after the start of the culture, At the end of culture (48 hours) in the case of control using yeast extract for medium
It was found that the wet weight of the cells was almost the same as the above. From this, it was clarified that the desired bifidobacteria can be obtained in a shorter number of culture days than before by using the microbial medium obtained from the shochu distilled residue of the present invention.
【００５６】[0056]
【表１】 [Table 1]
【００５７】[0057]
【表２】 [Table 2]
【００５８】[0058]
【表３】 [Table 3]
【００５９】[0059]
【表４】 [Table 4]
【００６０】[0060]
【表５】 [Table 5]
【００６１】[0061]
【表６】 [Table 6]
【００６２】[0062]
【表７】 [Table 7]
【００６３】[0063]
【発明の効果】以上詳述したように、本発明の焼酎蒸留
残液から得られる微生物用培地の製造方法によれば、以
下の効果を奏することができる。すなわち、大麦を原料
とする焼酎製造において副成する焼酎蒸留残液を固液分
離して液体分を得、該液体分をろ過して清澄液を得、該
清澄液を濃縮して濃縮液を得、該濃縮液を合成吸着剤を
用いる吸着処理に付して非吸着性画分を得、該非吸着性
画分を乾燥することにより、これを微生物用培地として
用いた場合、水不溶性成分および着色成分が極めて少な
くなり，得られる培養菌体の量が著しく増加する。As described in detail above, according to the method for producing a culture medium for microorganisms obtained from the shochu distillation residue of the present invention, the following effects can be obtained. That is, the shochu distillation residual liquid by-produced in the production of shochu using barley as a raw material is subjected to solid-liquid separation to obtain a liquid component, the liquid component is filtered to obtain a clear liquid, and the clear liquid is concentrated to obtain a concentrated liquid. Then, the concentrated solution is subjected to an adsorption treatment using a synthetic adsorbent to obtain a non-adsorptive fraction, and the non-adsorptive fraction is dried to obtain a water-insoluble component and The amount of coloring components becomes extremely small, and the amount of cultured cells obtained increases significantly.
【図１】 本発明の微生物用培地の製造工程を示す製造
工程図である。FIG. 1 is a manufacturing process diagram showing a manufacturing process of a culture medium for a microorganism of the present invention.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 梅本 泰史 大分県宇佐市大字山本2231−１ 三和酒 類株式会社内 (72)発明者 下田 雅彦 大分県宇佐市大字山本2231−１ 三和酒 類株式会社内 (56)参考文献 熊本県工業技術センター研究報告第33 号（平成７年11月）第58−61頁 リサイクル技術百科「プラスチック ス」２月号別冊（1994年２月10日発行) (58)調査した分野(Int.Cl.7，ＤＢ名) C12N 1/20 C12N 1/00 ＢＩＯＳＩＳ／ＷＰＩ（ＤＩＡＬＯＧ) ＰｕｂＭｅｄ ＪＳＴＰｌｕｓ（ＪＯＩＳ)─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Yasushi Umemoto 2331-1, Yamamoto, Usa-shi, Oita Sanwa Sake Co., Ltd. (72) Masahiko Shimoda 2231-1, Yamamoto, Usa-shi, Oita Sanwa Sake Co., Ltd. (56) References Kumamoto Prefectural Industrial Technology Center, Research Report No. 33 (November 1995) pp. 58-61 Recycle Technology Encyclopedia "Plastics" February issue, separate volume (published on February 10, 1994) (58) Fields investigated (Int.Cl. 7 , DB name) C12N 1/20 C12N 1/00 BIOSIS / WPI (DIALOG) PubMed JSTPlus (JOIS)
Claims (2)
する焼酎蒸留残液を固液分離して液体分を得、該液体分
をろ過して清澄液を得、該清澄液を濃縮して濃縮液を
得、該濃縮液を芳香族系合成吸着剤を用いる吸着処理に
付して前記合成吸着剤に非吸着の画分を分取し、分取し
た前記画分を乾燥することにより得られる乾燥物を有効
成分として含有することを特徴とする乳酸菌培養用培
地。1. A shochu distillation residual liquid by-produced in the production of shochu using barley as a raw material is subjected to solid-liquid separation to obtain a liquid content, the liquid content is filtered to obtain a clear solution, and the clear solution is concentrated. to obtain a concentrated solution, it was collected fractions of the non-adsorbed on the synthetic adsorbent is subjected to adsorption treatment using an aromatic synthetic adsorbent the concentrate min, Shi prep
A lactic acid bacterium culture medium, which comprises a dried product obtained by drying the above-mentioned fraction as an active ingredient.
して乳酸菌を培養することを特徴とする乳酸菌の培養方
法。2. Use of the culture medium for lactic acid bacterium culture according to claim 1.
Method for lactic acid bacterium characterized by culturing lactic acid bacterium
Law .
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| JP4921499B2 (en) * | 2008-02-05 | 2012-04-25 | 株式会社キティー | Antiallergic composition using new strains Lactobacillus crispatas KT-11, KT-23, and KT-25 |
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| JP6698581B2 (en) * | 2017-04-25 | 2020-05-27 | 三和酒類株式会社 | Animal cell culture medium and method of using the same |
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| Title |
|---|
| リサイクル技術百科「プラスチックス」２月号別冊（１９９４年２月１０日発行） |
| 熊本県工業技術センター研究報告第３３号（平成７年１１月）第５８−６１頁 |
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