JP2004000167A - Process for preparing fermentation product containing significant amount of nisin by using barley shochu distillation residual liquid as medium - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a process for preparing a fermentation product having an excellent taste which may be used as a food product. <P>SOLUTION: A medium used here is obtained by adding sugar to a liquid which comprises a liquid component (A) obtained through solid-liquid separation of a barley Shochu (distilled liquor) distillation residual liquid and whose Brix concentration is adjusted to 0.5-6.0. Nisin-producing lactic bacteria are added to the medium and incubated while controlling the sugar concentration of the culture broth to ≥0.1 g/l and its pH to 4.0-7.0. <P>COPYRIGHT: (C)2004,JPO

Description

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【発明の効果】
以上詳述したように、本発明のナイシンを含有する発酵生産物の製造方法は、大麦焼酎蒸留残液を固液分離することにより得られる液体分（Ａ）からなり、Ｂｒｉｘ濃度を０．５乃至６．０の範囲に調整した液体に糖を添加したものを培地として使用し、該培地にナイシン生産能を有する乳酸菌を加えて培養を行い、該培養は、培養液の糖濃度を少なくとも０．１ｇ／Ｌ以上に制御し且つそのｐＨ値を４．０乃至７．０の範囲に制御しながら行うことを特徴とし、ナイシンを著量含有する発酵生産物の効率的製造を可能にする。そして、得られるナイシンを著量含有する発酵生産物は、優れた呈味性を有し、食品として好適に使用することができる。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention is a method for producing a nisin-containing fermentation product for culturing lactic acid bacteria having nisin-producing ability using a liquid component obtained by solid-liquid separation of a shochu distillation residue, as a medium, As the liquid component (A) obtained by subjecting the barley shochu distillation residue to solid-liquid separation, a liquid prepared by adding a sugar to a liquid having a Brix concentration adjusted to a range of 0.5 to 6.0 is used, A lactic acid bacterium having a nisin-producing ability is added to the medium for culturing, and the culturing is performed by controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and adjusting the pH value thereof to a value in the range of 4.0 to 7.0. The present invention relates to a method for producing a fermentation product containing a significant amount of nisin, characterized in that the production is performed while controlling the amount of nisin. The method for producing a fermentation product containing a significant amount of nisin of the present invention includes an embodiment in which the culture is performed by a batch culture method and / or a continuous culture method. In addition, the method for producing a fermentation product containing a significant amount of nisin of the present invention comprises: converting a portion of the liquid component (A) obtained by solid-liquid separation of the barley shochu distillation residue into a rice shochu distillation residue. It includes a mode in which the liquid component (B) obtained by solid-liquid separation is replaced. The fermented product containing a significant amount of nisin obtained by the present invention has excellent taste and can be used as food.
[0002]
[Prior art]
What is generally known as nisin is a polypeptide consisting of 34 amino acids, a histidine at position 27 of the amino acid sequence is called nisin A, and a polypeptide at position 27 of the amino acid sequence is asparagine. Call it Nisin Z. Such nisin is available from Lactococcus lactis subsp. Lactis is a type of bacteriocin produced by Lactis, and has antibacterial activity against Gram-positive bacteria such as Streptococcus, Bacillus, Clostridium, and Staphylococcus. Has been recognized. The main such foods include cheese, canned milk, cream, mayonnaise and the like.
[0003]
For the production of nisin, the following method has been proposed. That is, in Japanese Patent Application Laid-Open No. 4-75596 [hereinafter referred to as "Document 1"], in fermentative production of nisin using lactic acid bacteria, the growth rate of the bacterium produced by nisin-producing bacteria is increased by metabolites other than nisin. Before decreasing or stopping the growth, the culture solution containing the cells is continuously filtered using a membrane, and separated into a solution containing the cells and a solution containing nisin without the cells, The liquid containing the cells is returned to the culture tank, a liquid containing nisin without the cells is extracted, and a fresh liquid medium having the same volume as the amount of the extracted liquid is used as a nisin-producing bacterial culture in the culture tank. The dilution rate is 0.1 to 10 hours ^-1 The filtration culture is continued while supplying the nisin, and the nisin is separated from the liquid containing nisin without the cells, the culture solution in the culture tank, or the cells themselves contained in the culture solution. A manufacturing method is described. More specifically, Document 1 describes that an MRS medium is used as a culture medium, Streptococcus lactis IFO12007 is used as a lactic acid bacterium strain having a nisin-producing ability, and fermentation is performed using the above-described filtration culture method. The nisin concentration of the culture obtained by the fermentation was 1.72 × 10 ⁵ U / L.
[0004]
Japanese Patent Application Laid-Open No. 4-126093 (hereinafter referred to as “Reference 2”) discloses that, when culturing a strain of Streptococcus lactis SBT1212 having a nisin-producing ability to produce nisin, serine or phenylalanine is added to the culture solution. There is described a method for producing nisin, which comprises adding 0.1 to 10 mg / ml and culturing to enhance nisin production. Document 2 describes that a nisin production amount of a fermentation product obtained by the production method using a medium consisting of whole milk to which serine is added is 1000 (IU / ml). .
JP-A-6-9690 [hereinafter referred to as “Reference 3”] discloses Lactococcus lactis subsp. LactisNRRL-B-18809 as an MRS medium supplemented with a 1% yeast extract, or a 0.5% yeast extract. Describes a method for producing bacteriocin (Nisin), which is cultured using a medium consisting of 7% whey to which nisin has been added, and that the nisin titer of the fermented product obtained in the production method is 1600 AU / ml. Has been described.
[0005]
However, since the nisin concentration in the fermentation product achieved by the nisin production methods described in Literatures 1 to 3 is extremely low as described above, these nisin production methods are not suitable for industrially producing large amounts of nisin. It is not suitable. Incidentally, at present, these nisin production methods have not been industrially practiced.
[0006]
By the way, it has been proposed to use a shochu distillation residue as a culture medium for lactic acid bacteria. That is, Japanese Patent Application Laid-Open No. 2000-236891 [hereinafter referred to as “Document 4”. ] Describes a method for producing lactic acid, characterized in that lactic acid bacteria are added to a medium obtained from solids by solid-liquid separation of distilled spirits of rice shochu, followed by fermentation. JP-A-2000-245491 [hereinafter referred to as “Reference 5”] discloses a lactic acid bacterium that produces L-lactic acid in a medium obtained from a solid obtained by solid-liquid separation of potato shochu distilled cake in an open system without steam sterilization. A method for producing high-purity L-lactic acid, characterized in that it is added and subjected to fermentation, is described.
JP-A-2000-342247 [hereinafter referred to as “Reference 6”] discloses that a shochu distillation residue, which is a by-product in the production of shochu using barley as a raw material, is subjected to solid-liquid separation to obtain a liquid component. Filtration to obtain a clarified solution, concentration of the clarified solution to obtain a concentrated solution, and a microorganism containing a non-adsorbable fraction obtained by subjecting the concentrated solution to an adsorption treatment using a synthetic adsorbent as an active ingredient. The medium is excellent as a culture medium for culturing yeast, lactic acid bacteria, and bifidobacteria.It is described that when the medium for microorganisms is used for culturing yeast, lactic acid bacteria, and bifidobacteria, the amount of cultured cells significantly increases. .
[0007]
Also, Japanese Patent Application Laid-Open No. 2002-369672 [hereinafter referred to as Document 7]. ] Describes a method for sterilizing foods using a lactic acid fermentation solution containing a bacteriocin such as nisin. It is described that the antibacterial action of the lactic acid fermentation solution can be maintained for a long time by adding the lactic acid fermentation solution and calcium ions to food.
[0008]
[Patent Document 1]
JP-A-4-75596
[Patent Document 2]
JP-A-4-126093
[Patent Document 3]
JP-A-6-9690
[Patent Document 4]
JP 2000-236891 A
[Patent Document 5]
JP-A-2000-245491
[Patent Document 6]
JP 2000-342247 A
[Patent Document 7]
JP-A-2002-369672
[0009]
However, the solids of the rice shochu distillation residue and the solids of the potato shochu distillation residue used as the culture medium raw materials in References 4 and 5 are used in the production of barley-based shochu used as the culture medium raw material in the present invention. The liquid component obtained by solid-liquid separation of the remaining barley shochu distillation residue and the liquid component obtained by solid-liquid separation of the rice shochu distillation residue by-produced in the production of shochu from rice are objective. Are clearly different. In addition, the inventions described in Documents 4 and 5 aim at producing lactic acid by culturing lactic acid bacteria using a medium consisting of solids obtained from rice shochu distillation residue or potato shochu distillation residue. There is no suggestion about a method for producing a fermented lactic acid bacterium solution containing nisin and having excellent taste.
[0010]
Document 6 describes that the microorganism culture medium is effective for the growth of lactic acid bacteria belonging to the genus Lactobacillus, that is, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus fermentum, and the like, but Lactococcus having nisin-producing ability. No mention is made of the bacterial cell growth effect of lactic acid bacteria belonging to Lactis. Document 6 describes that the culture medium for microorganisms is effective for the growth of cells such as lactic acid bacteria of the genus Lactobacillus, that is, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus fermentum. The Lactococcus lactis subsp. It may be easy to guess that when lactic acid bacteria belonging to Lactis are cultured, similar cell growth effects are exhibited. However, the amount of nisin produced in the culture by culturing the lactic acid bacterium having the nisin-producing ability cannot be increased at all only by an increase in the amount of cells in the culture. This point is the same even when using the microorganism culture medium having the above-mentioned bacterial cell growth effect. By the way, the present inventors have used the microorganism culture medium described in Document 6 to prepare Lactococcus lactis subsp. Each strain of several lactic acid bacteria belonging to Lactis was cultured under different culturing conditions, and the bacterial cell amount and nisin concentration in the obtained fermentation product were examined. No universal correlation was observed between the amount and the nisin concentration.
[0011]
Document 7 describes a method for sterilizing foods using a lactic acid fermentation solution containing a bacteriocin such as nisin. In addition, it is described that shochu lees can be used as a nitrogen source for a medium in obtaining the lactic acid fermentation liquor. However, Document 7 does not disclose that the productivity of nisin is remarkably increased when the shochu lees is used as a nitrogen source, and that the resulting lactic acid fermentation liquor has excellent taste.
[0012]
As is clear from the above description, even if the above-mentioned literature is examined closely, the liquid content obtained by solid-liquid separation of the barley shochu distillation residue or the rice shochu distillation residue is used as the medium. When lactic acid bacteria having nisin-producing ability are cultured, nisin production is remarkably increased, and that the obtained fermentation product contains a significant amount of nisin and has excellent taste is completely unpredictable. .
[0013]
[Problems to be solved by the invention]
As described above, in the conventional nisin production technology, the various media described above are used.However, using any of these conventional media, the nisin concentration in the obtained fermentation product is extremely low. . For these reasons, in order to industrially produce large amounts of nisin, there is a strong demand for early provision of means for further increasing the amount of nisin produced in the fermentation product.
Apart from this, as described above, Literature 7 discloses that a lactic acid fermented liquor containing nisin obtained by culturing lactic acid bacteria having nisin-producing ability using a medium containing shochu lees as a nitrogen source has an antibacterial effect. A method for sterilizing food by adding the lactic acid fermentation liquid to food is described. However, the lactic acid fermentation solution containing nisin obtained by culturing lactic acid bacteria having nisin-producing ability using such a conventional medium has a low nisin concentration as described above, and its taste is satisfactory. There is no way to go. When such a lactic acid fermentation liquid is used by adding it to food, it is desired that the lactic acid fermentation liquid contains a significant amount of nisin and has excellent taste.
[0014]
The present invention has been completed as a result of further research in view of the above-mentioned problems in the prior art. An object of the present invention is to solve the above-mentioned problems in the conventional nisin production technology and to provide an nisin industrial production method which enables mass production of nisin. Another object of the present invention is to provide a method that contains a significant amount of nisin, has excellent taste, and enables efficient production of a fermentation product extremely suitable for use in foods. is there.
[0015]
[Means for Solving the Problems]
The present inventors solve the above-mentioned problems in the conventional nisin production technology by more effectively utilizing the shochu distillation residue, that is, it is possible to increase nisin production by using the shochu distillation residue in a medium. We conducted intensive investigations through experiments to develop a method to make the data.
As a result, a liquid obtained by solid-liquid separation of the residual liquid from the distillation of barley shochu (A) and having a Brix concentration adjusted to a range of 0.5 to 6.0 and a sugar added to a liquid is used as a medium. A lactic acid bacterium having nisin-producing ability is added to the medium for culturing, and the culturing is performed by controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and adjusting the pH value to 4.0 to 7.0. It was found that, when the method of controlling while controlling the ratio to 0 was adopted, the production amount of nisin was significantly increased, and a fermentation product containing a significant amount of nisin was obtained. When the fermented product was subjected to a sensory test, it was found that the fermented product had an excellent taste as compared with a fermented product obtained by using a conventional medium, and was extremely suitable for use as a food. understood. The present invention has been completed based on the above findings.
[0016]
The present invention makes it possible to produce a large amount of nisin by using a barley shochu distillation residue, which has never been used as a medium in the conventional nisin production technology, as a medium. That is, the present invention comprises a liquid component (A) obtained by subjecting a barley shochu distillation residue to solid-liquid separation, wherein sugar is added to a liquid having a Brix concentration adjusted to a range of 0.5 to 6.0. Is used as a culture medium, and a lactic acid bacterium having nisin-producing ability is added to the medium for culturing. The culturing is performed by controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and adjusting the pH value to 4.0. The present invention provides a method for producing a fermentation product containing a significant amount of nisin, characterized in that the fermentation product is controlled while being controlled in the range of from 7.0 to 7.0. The culture in the production method of the present invention may be performed by a batch culture method and / or a continuous culture method. The fermented product containing a significant amount of nisin obtained by the present invention has excellent taste and can be suitably used as food.
[0017]
In view of the fact that when the lactic acid bacterium of the genus Lactobacillus is cultured using the medium for microorganisms described in Document 6, the amount of the cultured cells obtained is significantly increased. Using a medium, Lactococcus lactissubsp. There is no mention of culturing lactic acid bacteria of the genus Lactis, but when lactic acid bacteria having the nisin-producing ability are cultured using the above-mentioned medium for microorganisms, the degree of increase in the amount of cultured cells is not likely, We imagined that remarkable production of nisin would be brought about, and the present inventors conducted intensive studies through experiments. That is, the present inventors solid-liquid-separate the shochu distillation residue (hereinafter abbreviated as “barley shochu distillation residue”) by-produced in the production of shochu using barley as a raw material described in Document 6. To obtain a clarified liquid by concentrating the clarified liquid to obtain a concentrated liquid, and subjecting the concentrated liquid to an adsorption treatment using a synthetic adsorbent. After adjusting the Brix concentration of each of the fraction and the liquid to 8.0, the same amount of glucose was added to each, and then Lactococcus lactis subsp. Lactic acid bacteria having the ability to produce nisin belonging to Lactis were inoculated and separately subjected to batch culture. As a control, an MRS medium supplemented with the same amount of glucose as described above was used, and all others were subjected to batch culture by the same method as described above.
[0018]
The amount of lactic acid bacteria and the nisin concentration of the fermentation product obtained in each batch culture were measured. As a result, when the medium consisting of the liquid component described in Document 6 was used and when the medium consisting of the non-adsorbable fraction was used, the amount of lactic acid bacteria in the fermentation product obtained in these two cases and nisin The concentrations were found to be substantially the same. Furthermore, no significant difference was observed in the amount of the lactic acid bacteria cells and the nisin concentration as compared with the case where the control MRS medium was used, and it was found that they were substantially equivalent. That is, it was revealed that nisin productivity significantly higher than that of the control MRS medium could not be achieved simply by using the medium for microorganisms described in Reference 6.
[0019]
Therefore, the present inventors conducted an experiment on the production of nisin through experiments using a medium consisting of a liquid component obtained by solid-liquid separation of a barley shochu distillation residue.
That is, in producing nisin using the medium consisting of the liquid component, it is presumed that the Brix concentration of the liquid component may have a detrimental effect on the production of nisin. The study was conducted through experiments for the purpose of determining the concentration. That is, the Brix concentration of the liquid is 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10 0.0, 11.0, 12.0, 13.0, 14.0, and 15.0, and the same amount of glucose was added to each of them to produce the following nisin-producing lactic acid bacteria. A liquid medium adjusted to such an optimum pH value was obtained, and Lactococcus lactis subsp. Lactic acid bacteria capable of producing nisin belonging to Lactis were inoculated and separately subjected to batch culture. As a control, an MRS medium adjusted to the same pH value by adding the same amount of glucose as described above was used, and all other components were subjected to batch culture by the same method as described above. As a result, when the liquid medium obtained by adjusting the Brix concentration of the liquid to be in the range of 0.5 to 6.0 is used, the obtained fermentation production obtained is higher than when the control MRS medium is used. There was a tendency for the nisin concentration in the product to increase.
[0020]
Therefore, when culturing for the production of nisin using the medium composed of the above-mentioned liquid components, an experiment was conducted through experiments for the purpose of determining the optimal pH conditions for mass production of nisin. That is, a predetermined amount of glucose was added to the above-mentioned liquid whose Brix concentration was adjusted to 4.0 to obtain a plurality of liquid culture media whose pH was adjusted to 3.0 to 8.0. Lactococcus lactis subsp. Was added to each of the obtained liquid media having different pH values. Lactic acid-producing lactic acid bacteria belonging to Lactis were inoculated respectively and separately subjected to batch culture while maintaining the adjusted pH value. As a control, an MRS medium adjusted to pH 6.8 by adding the same amount of glucose as described above was used, and all other conditions were subjected to batch culture by the same method as described above. As a result, when the culture was performed while maintaining the pH value of the liquid medium in the range of 4.0 to 7.0, and when the culture was performed using the control MRS medium (maintained at pH 6.8). It was found that the concentration of nisin in the obtained fermentation product was significantly increased as compared with.
[0021]
From the above, the Brix concentration of the liquid obtained by subjecting the barley shochu distillation residue to solid-liquid separation is adjusted to the range of 0.5 to 6.0, and sugar is added thereto to adjust the pH value to 4. 0 to 7.0 and kept in the range of Lactococcus lactis subsp. By inoculating a lactic acid bacterium having nisin-producing ability belonging to Lactis and subjecting it to batch culture, the nisin concentration in the obtained fermentation product is remarkable as compared with the case where the culture is performed using a conventional MRS medium. It became clear that it would increase.
[0022]
the above
And
In the same manner as in the case of the liquid component (A) obtained by solid-liquid separation of the barley shochu distillation residue, the experiment described in As for the component (B), the same result as that of the liquid component (A) obtained by solid-liquid separation of the above-mentioned distillation residue of barley shochu was obtained. That is, the Brix concentration of the liquid component (B) obtained by solid-liquid separation of the rice shochu distillation residue is adjusted to a range of 0.5 to 6.0, and sugar is added thereto to adjust the pH value to 4. 0 to 7.0 and kept in the range of Lactococcus lactis subsp. By inoculating a lactic acid bacterium having nisin-producing ability belonging to Lactis and subjecting it to batch culture, the nisin concentration in the obtained fermentation product is remarkable as compared with the case where the culture is performed using a conventional MRS medium. It became clear that it would increase.
[0023]
Next, nisin production by a continuous culture method was examined through experiments using a medium composed of the above liquid components. That is, the Brix concentration of the liquid was adjusted to 4.0, glucose was added to the mixture at 3% by weight to adjust the pH value to 5.5, and a liquid medium was obtained, and Lactococcus lactis subsp. A lactic acid bacterium having the ability to produce nisin belonging to Lactis is inoculated, a continuous culturing device comprising a jar fermenter is used, the pH value is maintained at 5.5 with a pH controller, and the filtration culture method described in Document 1 is used. A liquid containing nisin without cells was extracted, and continuous culture was performed in which a fresh liquid medium having the same volume as the extracted liquid was supplied to a jar fermenter. As a control, continuous culture was carried out in the same manner as described above except that an MRS medium containing 3% by weight of glucose was used instead of using the fresh liquid medium in the filtration culture. As a result, by using the liquid medium as the fresh medium to be supplied to the jar fermenter in the filtration culture, the nisin concentration in the resulting fermentation product is significantly higher than when the MRS medium is used as the fresh medium. It turned out to be growing.
[0024]
Furthermore, Table 1 of Document 1 describes that the higher the glucose content of the MRS medium used as a fresh medium, the higher the nisin concentration in the culture solution is. As shown in FIGS. 4 and 5 of Document 1, As is evident, the glucose concentration in the culture solution after the shift to the continuous culture is substantially infinitely close to 0 g / L in any case regardless of the glucose concentration of the supplied MRS medium. Therefore, the present inventors speculated that the glucose concentration in the culture solution during continuous culture might affect nisin productivity, and studied through experiments.
[0025]
That is, the Brix concentration of the liquid was adjusted to 4.0, glucose was added at 3% by weight to adjust the pH value to 5.5, and a liquid medium was obtained, and Lactococcus lactis subsp. A lactic acid bacterium belonging to Lactis, which has the ability to produce nisin, is inoculated, a continuous culturing device comprising a jar fermenter is used, the pH value is maintained at 5.5 with a pH controller, and the filtration culture method described in Document 1 is used. A liquid containing nisin without the body was extracted, and continuous culture was performed in which a fresh liquid medium having the same volume as the extracted liquid medium was supplied to a jar fermenter. At that time, the glucose concentration of the culture solution was measured using an online biochemical controller equipped with a glucose electrode, and after shifting to continuous culture, the liquid medium was supplied to a jar fermenter to reduce the glucose concentration of the culture solution to 0.01. , 0.1, 1.0, 5.0, and 10.0 g / L, and continuous culture was separately performed. As a result, when the glucose concentration in the culture solution after the shift to the continuous culture was maintained at 0.1, 1.0, 5.0, and 10.0 g / L, the glucose concentration in the culture solution was 0.1 g / L. It was revealed that the nisin concentration in the culture solution was dramatically increased as compared with the case where the concentration was maintained at 01 g / L. From this, it is possible to continuously produce a culture solution containing an extremely high concentration of nisin by maintaining the glucose concentration in the culture solution after transition to continuous culture at 0.1 g / L or more. It has been found.
[0026]
Therefore, a liquid containing a liquid component (A) obtained by solid-liquid separation of the residual liquid of barley shochu distilled water and having a Brix concentration adjusted to a range of 0.5 to 6.0 and added with sugar is used as a medium. Then, a lactic acid bacterium having a nisin-producing ability is added to the medium for culturing, and the culturing is performed by controlling the sugar concentration of the culture solution to at least 0.1 g / L and adjusting the pH value to 4.0 to 7.0. A fermentation product was obtained by a method performed while controlling the temperature within the range. When the nisin concentration of the obtained fermentation product was examined, it was found that the nisin concentration was significantly higher than the nisin concentration of the fermentation product obtained by using the conventional MRS medium. When these fermented products were subjected to a sensory test by 10 panelists, the latter had a rough taste and an irritating taste and had a problem in taste, whereas the former had a rounded and harmonious taste. It has an extremely excellent taste consisting of a good flavor and a rich umami, and has been found to be extremely suitable as a food material.
[0027]
Also, using the liquid component (B) of the above-mentioned rice shochu distillation residue,
When the experiment was carried out by the method described above, the glucose concentration in the culture solution after the shift to the continuous culture was 0.1 g / L or more, as in the case where the liquid content of the barley shochu distillation residue was used. It has been found that the maintenance of the culture medium enables continuous production of a culture solution containing a very high concentration of nisin. Therefore, a liquid containing a liquid component (B) obtained by solid-liquid separation of a rice shochu distillation residue and having a Brix concentration adjusted to a range of 0.5 to 6.0 and added with sugar is used as a medium. Then, a lactic acid bacterium having a nisin-producing ability is added to the medium for culturing, and the culturing is performed by controlling the sugar concentration of the culture solution to at least 0.1 g / L and adjusting the pH value to 4.0 to 7.0. A fermentation product was obtained by a method performed while controlling the temperature within the range. When the nisin concentration of the obtained fermentation product was examined, it was found that the nisin concentration was significantly higher than the nisin concentration of the fermentation product obtained by using the conventional MRS medium. When these fermented products were subjected to a sensory test by 10 panelists, the latter had a rough taste and an irritating taste and had a problem in taste, whereas the former had a rounded and harmonious taste. It has an extremely excellent taste consisting of a good flavor and a rich umami, and has been found to be extremely suitable as a food material.
[0028]
Further, as the medium, a part of the liquid component (A) obtained by solid-liquid separation of the barley shochu distillation residue is converted into a liquid component (B) obtained by solid-liquid separation of the rice shochu distillation residue. A liquid obtained by adding a sugar to a liquid whose Brix concentration has been adjusted to a range of 0.5 to 6.0 is used, and a lactic acid bacterium having a nisin-producing ability is added to the medium, followed by culturing. The fermentation product was obtained by a method of controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and controlling the pH value in the range of 4.0 to 7.0. As a result, it was revealed that the nisin concentration of the fermentation product was significantly increased as compared with the nisin concentration of the fermentation product obtained by using the conventional MRS medium. When these fermented products were subjected to a sensory test by 10 panelists, the latter had a rough taste and an irritating taste and had a problem in taste, whereas the former had a rounded and harmonious taste. It has an extremely excellent taste consisting of a good flavor and a rich umami, and has been found to be extremely suitable as a food material.
[0029]
In addition, the non-adsorbed fraction obtained by subjecting at least a part of the liquid component (A) obtained by subjecting the barley shochu distillation residue to solid-liquid separation to an adsorption treatment using a synthetic adsorbent, At least a part of the liquid component (A ′) obtained by converting (A) to contain the non-adsorbed fraction and the liquid component (B) obtained by solid-liquid separation of rice shochu distillation residue is used as a synthetic adsorbent. A liquid component (B ') is prepared as a non-adsorbed fraction obtained by subjecting the liquid component (B) to a non-adsorbed fraction containing the non-adsorbed fraction. ), The sugar having been added to a liquid having a Brix concentration adjusted to a range of 0.5 to 6.0, and a Brix concentration of 0 using the liquid component (A ′) and the liquid component (B ′). The medium prepared by adding sugar to the liquid adjusted to the range of 0.5 to 6.0 was used as the medium. A lactic acid bacterium having nisin-producing ability is added to the medium for culturing, and the culturing is performed by controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and adjusting the pH value to 4.0 to 7.0. A fermentation product was obtained by a method performed while controlling to a range of 0. When the nisin concentration of the obtained fermentation product was examined, it was found that the nisin concentration of each fermentation product was significantly higher than the nisin concentration of the fermentation product obtained by using the conventional MRS medium. understood. When these fermented products were subjected to a sensory test by 10 panelists, the latter had a rough taste and an irritating taste and had a problem in taste, whereas the former had a rounded and harmonious taste. It has an extremely excellent taste consisting of a good flavor and a rich umami, and has been found to be extremely suitable as a food material.
[0030]
[Example of embodiment]
Hereinafter, preferred embodiments of the present invention will be described, but the present invention is not limited to these embodiments. In the present invention, a step (a) of obtaining a liquid component (A) by solid-liquid separation of a residual liquid of barley shochu distillation, and diluting the liquid component (A) with water to obtain a Brix concentration of 0.5 to 6. A step (b) of adjusting to a range of 0, a step (c) of adding a sugar to the product obtained in the step (b) to obtain a medium, and culturing by adding a lactic acid bacterium capable of producing nisin to the medium. The culturing is performed while controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and controlling the pH value in the range of 4.0 to 7.0. The present invention provides a method for producing a fermented product containing a significant amount of nisin. In the step (a), a part of the liquid component (A) obtained by solid-liquid separation of the barley shochu distillation residue is converted into a rice shochu distillation residue produced as a by-product in the production of shochu from rice. The liquid component (B) obtained by liquid separation may be replaced. Further, at least a part of the liquid component (A) and / or the liquid component (B) may be a non-adsorbed fraction obtained by performing an adsorption separation treatment using a synthetic adsorbent. The culture in the step (d) can be performed by a batch culture method and / or a continuous culture method.
[0031]
The barley shochu distillation residue used in the present invention is produced from barley or refined barley to produce barley koji and steamed barley, and the resulting barley koji and starch contained in steamed barley are koji, and / or enzyme agents. Saccharified using, and subjected to alcohol fermentation by yeast to obtain aged moromi, which is by-produced as a distillation residue when distilling the aged mash using a distillation apparatus such as vacuum distillation or atmospheric distillation, that is, , Means the residue of distillation of barley shochu. Further, in the production of rice shochu, sweet potato shochu, buckwheat shochu, the shochu distillation residue by-produced as a by-product when barley is used as a part of the raw materials in the production of these shochu is also the barley shochu distillation residue used in the present invention. Included.
In obtaining the barley shochu distillation residue, barley koji used in the production of barley shochu may be produced under the koji making conditions used in ordinary barley shochu production, and as a koji strain used, barley shochu is generally used. Aspergillus kawachii used in the production is preferred. Alternatively, Aspergillus strains such as Aspergillus awamori used in awamori production and Aspergillus oryzae used in sake production can be used. As the yeast used for producing barley shochu, various types of yeast for brewing shochu generally used for producing shochu can be used.
[0032]
The rice shochu distillation residue used in the present invention is produced by producing rice koji and steamed rice using rice as a raw material, and saccharifying the obtained rice koji and starch contained in steamed rice using koji and / or an enzyme agent. Then, it is further subjected to alcohol fermentation by yeast to obtain an aged moromi, which is by-produced as a distillation residue when distilling the aged moromi using a distillation apparatus such as vacuum distillation or atmospheric distillation, that is, rice shochu. It means the distillation residue. Further, in the production of barley shochu, sweet potato shochu, buckwheat shochu, the shochu distillation residue by-produced as a by-product when using rice as a part of the raw material in the production of these shochu is also the rice shochu distillation residue used in the present invention. Included.
In obtaining the rice shochu distillation residue, the rice koji used in the production of rice shochu may be produced under the koji-making conditions used in ordinary rice shochu production. Aspergillus kawachii used in the production is preferred. Alternatively, Aspergillus strains such as Aspergillus awamori used in awamori production and Aspergillus oryzae used in sake production can be used. As the yeast used for the production of rice shochu, various types of yeast for brewing shochu generally used in the production of shochu can be used.
[0033]
In step (a), the reason for obtaining the liquid component by solid-liquid separation of the barley shochu distillation residue is that the water-insoluble fermentation residue derived from the raw barley and barley koji is removed from the barley shochu distillation residue to obtain only the liquid component. Is to use. The solid-liquid separation is performed through a solid-liquid separation method such as a screw press method or a roller press method, or a preliminary separation is performed using a solid-liquid separator of a filtration and pressing type, and then a centrifugal separator, a diatomaceous earth filtration device, a ceramic A solid-liquid separation process is performed using a filtration device or a filtration press to obtain the liquid component.
[0034]
In the step (a), the reason for obtaining the liquid component by solid-liquid separation of the rice shochu distillation residue is that the water-insoluble fermentation residue derived from the raw rice and rice koji is removed from the rice shochu distillation residue to obtain only the liquid component. Is to use. The solid-liquid separation is performed through a solid-liquid separation method such as a screw press method or a roller press method, or a preliminary separation is performed using a solid-liquid separator of a filtration and pressing type, and then a centrifugal separator, a diatomaceous earth filtration device, a ceramic A solid-liquid separation process is performed using a filtration device or a filtration press to obtain the liquid component.
[0035]
The liquid component (A) and / or the liquid component (B) is subjected to an adsorption / separation treatment using a synthetic adsorbent to make a non-adsorbed fraction by removing a component exhibiting unpleasant taste contained in the liquid component. This is done for the purpose of doing so. Preferable specific examples of the synthetic adsorbent include Amberlite XAD-4, Amberlite XAD-16, Amberlite XAD-1180 and Amberlite XAD-2000 manufactured by Organo Corporation, Sepabeads manufactured by Mitsubishi Chemical Corporation. Aromatic (or styrene-based) synthetic adsorbents such as SP850 and Diaion HP20, Amberlite XAD-7 manufactured by Organo Co., Ltd., and methacryl-based (or Diaion HP2MG manufactured by Mitsubishi Chemical Corporation) (or (Also referred to as acrylic). In addition to the above, an aromatic-modified synthetic adsorbent such as Sepapise SP207 manufactured by Mitsubishi Chemical Corporation may be used in some cases. The non-adsorbed fraction thus obtained can be used in place of at least a part of the liquid component.
[0036]
In the step (b), the liquid component (A) (including the non-adsorbed fraction of the liquid component (A)) and the liquid component (B) supplied from the step (a) Of the above-mentioned non-adsorbed fraction) is adjusted to a range of 0.5 to 6.0 by adjusting the concentration of a medium component suitable for nisin productivity of lactic acid bacteria having nisin-producing ability. Therefore, the productivity of nisin can be increased. More preferably, the Brix concentration is adjusted in the range of 2.0 to 4.0. That is, in this case, nisin productivity can be significantly improved.
[0037]
Preferred specific examples of the sugar added in step (c) include glucose, xylose, sucrose, fructose, maltose, ribose, galactose, mannose, mannitol, cellobiose, saccharose, and trehalose. The sugar concentration of the liquid medium varies depending on the type of sugar used, but is generally in the range of 5 to 70 g / L. Among these sugars, glucose is particularly preferred, and in that case, the glucose concentration is preferably set in the range of 5 to 70 g / L.
[0038]
The lactic acid bacterium having nisin-producing ability used in step (d) may be any lactic acid-producing lactic acid bacterium, as long as it is a strain having nisin-producing ability, in particular, Lactococcus lactis subsp. Lactic acid bacteria belonging to Lactis are preferred. Specifically, when the nisin to be produced is nisin A, Lactococcus lactis NCDO497, Lactococcus lactis NIZO R5, Lactococcus lactis ATCC 7962 and Lactococcus lactis ATCC 11454 can be preferably mentioned. Further, as the strain when nisin producing is nisin Z, Lactococcus lactis NIZO 22186, Lactococcus lactis NRRL-B-18583, Lactococcus lactis NCFB2118, Lactococcus lactis NCFB2054, Lactococcus lactis NIZO N9, Lactococcus lactis NIZO 221186, Lactococcus lactis IO- 1 (JCM7638), Lactococcus lactis subsp. Lactis A. Ishizaki Chizuka (JCM11180), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 5B (JCM11181), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 7B (JCM11182), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 8B (JCM11183) and Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 9B (JCM11184) can be mentioned as a preferable example.
[0039]
The step of performing the culture in the step (d) while controlling the sugar concentration of the culture solution to at least 0.1 g / L and controlling the pH value of the culture solution to a range of 4.0 to 7.0 includes batch culture. It can be performed by a system or a continuous culture system. In addition, a fed-batch culture method can be adopted. When controlling the pH value of the culture solution to be in the range of 4.0 to 7.0, as described above, a composite glass electrode for pH measurement was attached to the culture tank, measured with a pH meter, and the actual pH value was measured. When the pH becomes lower than the set pH value, an alkaline solution such as a sodium hydroxide solution is supplied to the culture tank using a pH controller or the like, whereby the pH can be maintained at the set pH value. Adjust to a range of 0 to 6.5. It is preferable that the fluctuation range of the predetermined pH value is 0.1 or less. In the step (d), when controlling the sugar concentration (glucose concentration) of the culture solution to at least 0.1 g / L or more, if the culture is performed by a continuous culture method or a fed-batch culture method, a glucose analyzer or the like is used. Can be controlled using In this step, a substrate solution containing a nitrogen source consisting of yeast extract or polypeptone in addition to glucose or a substrate solution consisting of the above liquid medium can be supplied to the culture tank, whereby the liquid in the culture step can be obtained. The glucose concentration of the medium can be maintained at a desired value. The glucose concentration is at least 0.1 g / L or more, preferably 1.0 g / L or more, so that the activity of lactic acid bacteria having nisin-producing ability and the specific nisin production rate can be maintained at high values. . The culturing temperature at the time of performing the culturing may be set to a temperature optimal for the growth of the lactic acid bacterium to be used, and is generally in the range of 20 to 40 ° C, preferably 25 to 37 ° C. The stirring speed is suitably from 10 to 1000 rpm, preferably from 50 to 300 rpm.
[0040]
When the culture in the step (d) is performed by the continuous culture method, the culture is started according to the above-described culture conditions, the concentration of the bacterial cells is increased, and the culture is shifted to the continuous culture at the latter stage of the logarithmic growth phase. In the continuous culture method, a composite glass electrode for pH measurement is attached to a culture tank, measured with a pH meter, and when an actual pH value is lower than a set pH value, an alkaline solution such as a sodium hydroxide solution is cultured. By supplying to the tank, the set pH value can be maintained. At this time, by removing the same amount of sterilizing solution as the liquid medium or the alkaline solution supplied to the culture tank from the culture tank through an ultrafiltration membrane or a microfiltration membrane, etc., the liquid volume in the culture tank is kept constant. Can be held. Separately from the above pH control, a process online turbidimeter probe is attached to the culture tank, the output of which is input to the DDC controller, and the same amount of sterilized water or liquid medium supplied to the culture tank (cell culture) ) Is removed from the culture tank, thereby controlling the turbidity of the culture tank and keeping the concentration of the bacterial cells in the culture solution and the amount of the culture solution constant. The cell concentration is set in the range of 4.0 to 18.0 g / L, preferably in the range of 8.0 to 15.0 g / L. The dilution rate of the medium can be set to an arbitrary value depending on the cell concentration of the culture solution, the nisin production rate, and the like. ^-1 But preferably in the range of 0.2 to 1.2 hr ^-1 Range.
[0041]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.
[0042]
Barley shochu was produced for the purpose of the following examples. Barley (70% refined) was used as a raw material.
[Production of barley koji]
Barley is allowed to absorb 40% (w / w) water, steamed for 40 minutes, allowed to cool to 40 ° C., inoculated with 1 kg of koji (white koji mold) per ton of barley, and incubated at 38 ° C. and RH 95% for 24 hours for 32 hours. Barley koji was produced by keeping the mixture at RH 92% for 20 hours.
[Manufacture of steamed wheat]
Barley was allowed to absorb 40% (w / w) water, steamed for 40 minutes, and then allowed to cool to 40 ° C. to produce steamed barley.
[Barley shochu production and production of barley shochu distillation residue]
In the primary preparation, 3.6 kiloliters of water and 1 kg (wet weight) of cultured cells of shochu yeast as yeast were added to the barley koji (3 tons as barley) produced by the above-mentioned method to obtain a primary moromi. The obtained primary moromi was subjected to 5 days of fermentation (first stage fermentation). Next, in the secondary preparation, 11.4 kiloliters of water and steamed barley (7 tons as barley) produced by the above-mentioned method are added to the primary moromi after the first stage fermentation, and fermentation for 11 days ( (Second stage fermentation). The fermentation temperature was 25 ° C. for both the primary charge and the secondary charge. The secondary mash after the second-stage fermentation was subjected to simple distillation in a conventional manner to obtain 10 kiloliters of barley shochu and 15 kiloliters of barley shochu distillation residue. The obtained barley shochu distillation residue was used in the following Examples.
[0043]
Production of rice shochu was performed for the purpose of the following examples. As a raw material, milled rice (70% milled) was used.
[Production of rice koji]
Rice koji is produced by absorbing 35% (w / w) of rice, steaming for 40 minutes, allowing to cool to 40 ° C., inoculating 1 g of white koji mold per kg of rice, and 24 hours at 38 ° C. and 95% RH. At 32 ° C. and 92% RH for 20 hours.
[Manufacture of steamed rice]
Steamed rice was prepared by absorbing 35% (w / w) of rice, steaming for 40 minutes, and then allowing the rice to cool to 40 ° C.
[Production of rice shochu and production of rice shochu distillation residue]
In the primary preparation, rice koji (3 tons as rice) produced by the above-mentioned method was added with 3.6 kiloliters of water and 1 kg (wet weight) of cultured cells of shochu yeast as yeast to obtain primary moromi. The obtained primary moromi was subjected to 5 days of fermentation (first stage fermentation). Next, in the secondary preparation, 11.4 kiloliters of water and steamed rice (7 tons as rice) produced by the above-described method were added to the primary moromi after the first stage fermentation, and fermentation was performed for 15 days (2 days). Stage fermentation). The fermentation temperature was 25 ° C. for both the primary charge and the secondary charge. The secondary mash after the second-stage fermentation was subjected to simple distillation by a conventional method to obtain 10 kiloliters of rice shochu and 15 kiloliters of rice shochu distillation residue. The obtained rice shochu distillation residue was used in the following Examples.
[0044]
Embodiment 1
1. Preparation of lactic acid bacteria culture medium from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation of barley shochu distillation residue] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid content of the barley shochu distillation residue. The liquid was diluted with water to adjust its Brix concentration to 4, glucose was added at 3.6% by weight, the pH was adjusted to 5.5 using sodium hydroxide, and then sterilized at 121 ° C for 15 minutes. Was performed to obtain a culture medium for lactic acid bacteria culture.
2. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour.
3. Main culture of lactic acid bacteria capable of producing nisin
In a 2L jar fermenter, add 1. 500 ml of the culture medium for lactic acid bacteria obtained in 25 ml of the lactic acid bacterium preculture liquid obtained in the above was introduced, and batch culture was performed under the conditions of a stirring speed of 250 rpm, a culture temperature of 30 ° C., a culture time of 24 hours, and a pH of 5.5.
[0045]
[Comparative Example 1]
1. Preparation of MRS medium
MRS medium (peptone 1% by weight, meat extract 1% by weight, yeast extract 0.5% by weight, K ₂ HPO ₄ 0.2% by weight, 0.2% by weight of diammonium citrate, 3.6% by weight of glucose, 0.1% by weight of Tween 80, 0.5% by weight of sodium acetate, MGSO ₄ ・ 7H ₂ O 0.058% by weight, MnSO ₄ ・ 4H ₂ O (0.028% by weight) was adjusted to pH 5.5 using sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to obtain a lactic acid bacteria culture medium.
2. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour.
3. Main culture of lactic acid bacteria capable of producing nisin
In a 2L jar fermenter, add 1. And 500 ml of the MRS medium prepared in 25 ml of the lactic acid bacterium preculture liquid obtained in the above was introduced, and batch culture was performed under the conditions of a stirring speed of 250 rpm, a culture temperature of 30 ° C., a culture time of 24 hours, and a pH of 5.5.
[0046]
Embodiment 2
1. Preparation of lactic acid bacteria culture medium from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation of barley shochu distillation residue] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid content of the barley shochu distillation residue. The liquid was diluted with water to adjust its Brix concentration to 4, added 3.6% by weight of glucose, adjusted to pH 5.5 with sodium hydroxide, and then sterilized at 121 ° C for 15 minutes. The treatment was performed to obtain a lactic acid bacteria culture medium.
2. Preparation of substrate solution for adjusting glucose concentration from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation of barley shochu distillation residue] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid content of the barley shochu distillation residue. The liquid was diluted with water to adjust its Brix concentration to 4, added 1.0% by weight of glucose, adjusted to pH 5.5 with sodium hydroxide, and then sterilized at 121 ° C for 15 minutes. The treatment was performed to obtain a glucose concentration adjusting substrate solution.
[0047]
3. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour.
4. Main culture of lactic acid bacteria capable of producing nisin
The main culture of lactic acid bacteria having a nisin-producing ability was performed by a continuous culture method using a continuous culture device comprising a 3 L jar fermenter.
That is, 1800 ml of the lactic acid bacterium culture medium obtained in 1 above and 90 ml of the lactic acid bacterium preculture liquid obtained in 3 above were introduced into the 3 L jar fermenter, and the mixture was batched under the conditions of a stirring speed of 250 rpm, a culture temperature of 30 ° C., and a pH of 5.5. Eight hours after the start of the cultivation, when the glucose concentration of the culture solution reached 4 g / L, the culture was shifted to continuous culture with pH control, glucose concentration control, and turbidity control. The continuous culture was performed at a concentration of 4 g / L until 42 hours after the start of the culture.
The pH control was performed by using a pH sensor and supplying a 5N sodium hydroxide solution to the culture tank to maintain the pH at 5.5. The glucose concentration was controlled by measuring the glucose concentration of the culture using an online biochemical controller BF-410 (manufactured by Able Corporation) equipped with a glucose electrode, and measuring the glucose concentration obtained in step 2 during continuous culture. The adjustment was performed by supplying the substrate solution for adjustment to the culture tank and maintaining the glucose concentration of the culture solution at 4 g / L. Further, in the pH control and the glucose concentration control, a sterilizing solution having the same amount as the total amount of the 5N sodium hydroxide solution supplied to the culture tank and the amount of the glucose concentration adjusting substrate solution was subjected to precision filtration. The liquid volume in the culture tank was kept constant by extracting from the culture tank through the membrane. The turbidity control is performed by using a laser turbidity meter and using the same amount of culture solution (bacterium) as the turbidity control supply solution (yeast extract 0.5% by weight, polypeptone 0.5% by weight) supplied to the culture tank. Was removed from the culture tank, whereby the cell concentration and liquid volume in the culture tank were kept constant.
[0048]
[Comparative Example 2]
1. Preparation of MRS medium
MRS medium (peptone 1% by weight, meat extract 1% by weight, yeast extract 0.5% by weight, K ₂ HPO ₄ 0.2% by weight, 0.2% by weight of diammonium citrate, 3.6% by weight of glucose, 0.1% by weight of Tween 80, 0.5% by weight of sodium acetate, MGSO ₄ ・ 7H ₂ O 0.058% by weight, MnSO ₄ ・ 4H ₂ O (0.028% by weight) was adjusted to pH 5.5 with sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to obtain an MRS medium for lactic acid bacteria culture.
2. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand still at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, and 3 ml at 37 ° C. A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour. 3. Main culture of lactic acid bacteria capable of producing nisin
The main culture of lactic acid bacteria having a nisin-producing ability was performed by a continuous culture method using a continuous culture device comprising a 3 L jar fermenter.
That is, the above 1. 1800 ml of the lactic acid bacteria culture medium obtained in the above and 90 ml of the lactic acid bacteria preculture solution obtained in the above 3 were introduced, and subjected to batch culture at a stirring speed of 250 rpm, a culture temperature of 30 ° C. and a pH of 5.5, and 12 hours after the start of the culture. When the glucose concentration of the culture solution reaches less than 0.1 g / L, the process is shifted to continuous culture with pH control, glucose concentration control, and turbidity control, and the glucose concentration of the culture solution is reduced to less than 0.1 g / L. And the continuous culture was carried out up to 42 hours after the start of the culture.
The pH control was performed by using a pH sensor and supplying a 5N sodium hydroxide solution to the culture tank to maintain the pH at 5.5. The glucose concentration is controlled by measuring the glucose concentration of the culture solution using an online biochemical controller BF-410 (manufactured by Able Corporation) equipped with a glucose electrode. , Yeast extract 0.5% by weight, polypeptone 0.5% by weight, and NaCl 0.5% by weight) were supplied to the culture tank to maintain the glucose concentration of the culture solution at less than 0.1 g / L. Further, at the time of the pH control and the glucose concentration control, by removing the same amount of sterilizing solution as the 5N sodium hydroxide solution and the substrate solution supplied to the culture tank from the culture tank through a microfiltration membrane, The liquid volume in the culture tank was kept constant. The turbidity control is performed by using a laser turbidity meter and using the same amount of culture solution (bacterium) as the turbidity control supply solution (yeast extract 0.5% by weight, polypeptone 0.5% by weight) supplied to the culture tank. Was removed from the culture tank, whereby the cell concentration and liquid volume in the culture tank were kept constant.
[0049]
Embodiment 3
Instead of Lactococcus lactis IO-1 used in Example 2, Lactococcus lactisNIZO 22186, Lactococcus lactis NRRL-B-18583, Lactococcus lactis NCFB2118, Lactococcus lactis NCFB2054, Lactococcus lactis NIZO N9, Lactococcus lactis NIZO 221186, Lactococcus lactis subsp. Lactis A. Ishizaki Chizuka (JCM11180), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 5B (JCM11181), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 7B (JCM11182), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 8B (JCM11183) and Lactococcus lactis subsp. Lactis A. Continuous culture was performed in the same manner as in Example 2 except that Ishizaki Yasaka 9B (JCM11184) was used.
[0050]
[Comparative Example 3]
Instead of Lactococcus lactis IO-1 used in Comparative Example 2, Lactococcus lactis NIZO 22186, Lactococcus lactis NRRL-B-18583, Lactococcus lactis NCFB2118, Lactococcus lactis NCFB2054, Lactococcus lactis NIZO N9, Lactococcus lactis NIZO 221186, Lactococcus lactis subsp. Lactis A. Ishizaki Chizuka (JCM11180), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 5B (JCM11181), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 7B (JCM11182), Lactococcus lactis subsp. Lactis A. Ishizaki Yasaka 8B (JCM11183) and Lactococcus lactis subsp. Lactis A. Continuous culture was carried out in the same manner as in Comparative Example 2 except that Ishizaki Yasaka 9B (JCM11184) was used.
[0051]
Embodiment 4
1. Preparation of lactic acid bacteria culture medium from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation of barley shochu distillation residue] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid content of the barley shochu distillation residue. The liquid fraction was subjected to a ceramic filtration device to obtain a clarified solution, the clarified solution was subjected to a vacuum evaporator and concentrated to about 3 times to obtain a concentrated solution, and the concentrated solution was packed with a column filled with a synthetic adsorbent. To obtain a non-adsorbable fraction solution showing non-adsorbability to the synthetic adsorbent eluted from the packed column, and diluting the non-adsorbable fraction solution with water to adjust its Brix concentration to 4. After adding 3.6% by weight of glucose and adjusting the pH to 5.5 with sodium hydroxide, the mixture was sterilized at 121 ° C. for 15 minutes to obtain a lactic acid bacteria culture medium.
2. Preculture of lactic acid bacteria
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour.
3. Main culture of lactic acid bacteria
500 ml of the lactic acid bacterium culture medium obtained in 1 above and 25 ml of the lactic acid bacterium preculture liquid obtained in 2 above were introduced into a 2 L jar fermenter, and the stirring speed was 250 rpm, the culturing temperature was 30 ° C., the culturing time was 24 hours, and the pH was 5.5. Batch culture was performed under the conditions.
[0052]
Embodiment 5
1. Preparation of lactic acid bacteria culture medium from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation residue of barley shochu] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid component, and the liquid component is subjected to ceramic filtration. A clarified solution was obtained by subjecting the clarified solution to a vacuum evaporator, and the clarified solution was concentrated to about 3 times to obtain a concentrated solution. The concentrated solution was brought into contact with a column filled with a synthetic adsorbent, A non-adsorbable fraction solution showing non-adsorbability to the synthetic adsorbent eluted from the above is obtained, and the Brix concentration is adjusted to 4 by diluting the non-adsorbable fraction solution with water, and glucose is adjusted to 3. After adding 6% by weight and adjusting the pH to 5.5 using sodium hydroxide, sterilization was performed at 121 ° C. for 15 minutes to obtain a lactic acid bacteria culture medium.
2. Preparation of substrate solution for adjusting glucose concentration from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation residue of barley shochu] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid component, and the liquid component is subjected to ceramic filtration. A clarified solution was obtained by subjecting the clarified solution to a vacuum evaporator, and the clarified solution was concentrated to about 3 times to obtain a concentrated solution. The concentrated solution was brought into contact with a column filled with a synthetic adsorbent, A non-adsorbable fraction solution showing non-adsorbability to the synthetic adsorbent eluted from the above is obtained, and the Brix concentration is adjusted to 4 by diluting the non-adsorbable fraction solution with water. After adding 0% by weight and adjusting the pH to 5.5 using sodium hydroxide, a sterilization treatment was performed at 121 ° C. for 15 minutes to obtain a glucose concentration adjusting substrate solution.
3. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour.
[0053]
4. Main culture of lactic acid bacteria capable of producing nisin
The main culture of lactic acid bacteria having a nisin-producing ability was performed by a continuous culture method using a continuous culture device comprising a 3 L jar fermenter.
That is, the above 1. 1800 ml of the lactic acid bacterium culture medium obtained in the above and 90 ml of the lactic acid bacterium preculture liquid obtained in the above 3 were introduced, and subjected to batch culture under the conditions of a stirring speed of 250 rpm, a culture temperature of 30 ° C. and a pH of 5.5, and 8 hours after the start of the culture. When the glucose concentration of the culture solution reached 4 g / L, the culture was shifted to continuous culture with pH control, glucose concentration control, and turbidity control, and the culture was started while maintaining the glucose concentration of the culture solution at 4 g / L. The continuous culture was performed until 42 hours thereafter.
The pH control was performed by using a pH sensor and supplying a 5N sodium hydroxide solution to the culture tank to maintain the pH at 5.5. The glucose concentration was controlled by measuring the glucose concentration of the culture solution using an online biochemical controller BF-410 (manufactured by Able Co., Ltd.) equipped with a glucose electrode. The adjustment was performed by supplying the substrate solution for adjustment to the culture tank and maintaining the glucose concentration of the culture solution at 4 g / L. Further, at the time of the pH control and the glucose concentration control, the same amount of sterilizing solution as the 5N sodium hydroxide solution and the glucose concentration adjusting substrate solution supplied to the culture tank was passed through the microfiltration membrane from the culture tank. By withdrawing, the liquid volume in the culture tank was kept constant. The turbidity control is performed by using a laser turbidity meter and using the same amount of culture solution (bacterium) as the turbidity control supply liquid (yeast extract 0.5% by weight, polypeptone 0.5% by weight) supplied to the culture tank. Was removed from the culture tank to maintain the cell concentration and liquid volume in the culture tank constant.
[0054]
Embodiment 6
1. Preparation of lactic acid bacteria culture medium from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation of barley shochu distillation residue] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid content of the barley shochu distillation residue. The liquid was diluted with water to adjust its Brix concentration to 4, added 3.6% by weight of glucose, adjusted to pH 5.5 with sodium hydroxide, and then sterilized at 121 ° C for 15 minutes. The treatment was performed to obtain a lactic acid bacteria culture medium.
2. Preparation of lactic acid bacteria culture medium from rice shochu distillation residue
the above
The rice shochu distillation residue obtained in [Production of rice shochu and production of distillation residue of rice shochu] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid component, and the liquid component is washed with water. After dilution, the Brix concentration was adjusted to 4, glucose was added at 3.6% by weight, the pH was adjusted to 5.5 using sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to culture lactic acid bacteria. A medium was obtained.
3. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand still at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, and incubated at 37 ° C. for 3 hours. A preculture of lactic acid bacteria was obtained by culturing with shaking at 100 rpm for an hour.
4. Main culture of lactic acid bacteria capable of producing nisin
250 ml of the lactic acid bacteria culture medium obtained in 1 above, 250 ml of the lactic acid bacteria culture medium obtained in 2 above, and 25 ml of the lactic acid bacteria preculture liquid obtained in 3 above were introduced into a 2 L jar fermenter. Batch culture was carried out at 30 ° C., a culture time of 24 hours, and a pH of 5.5.
[0055]
Embodiment 7
1. Preparation of lactic acid bacteria culture medium from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation residue of barley shochu] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid component, and the liquid component is washed with water. After dilution, the Brix concentration was adjusted to 4, glucose was added at 3.6% by weight, the pH was adjusted to 5.5 using sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to culture lactic acid bacteria. A medium was obtained.
2. Preparation of substrate solution for adjusting glucose concentration from barley shochu distillation residue
the above
The barley shochu distillation residue obtained in [Production of barley shochu and distillation residue of barley shochu] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid component, and the liquid component is washed with water. After dilution, the Brix concentration was adjusted to 4, glucose was added at 1.0% by weight, the pH was adjusted to 5.5 using sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to adjust the glucose concentration. A substrate solution was obtained.
3. Preparation of lactic acid bacteria culture medium from rice shochu distillation residue
The rice shochu distillation residue obtained in the distillation process of rice shochu production is centrifuged at 8000 rpm and 10 min to obtain a liquid component of the rice shochu distillation residue, and the liquid component is adjusted to Brix 4 to give glucose of 3%. Then, the mixture was adjusted to pH 5.5 with sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to obtain a lactic acid bacteria culture medium.
4. Preparation of substrate solution for adjusting glucose concentration from distillation residue of rice shochu
the above
The rice shochu distillation residue obtained in [Production of rice shochu and production of distillation residue of rice shochu] described above is centrifuged at 8000 rpm for 10 minutes to obtain a liquid component, and the liquid component is washed with water. After dilution, the Brix concentration was adjusted to 4, glucose was added at 1.0% by weight, the pH was adjusted to 5.5 using sodium hydroxide, and then sterilized at 121 ° C. for 15 minutes to adjust the glucose concentration. A substrate solution was obtained.
[0056]
5. Preculture of lactic acid bacteria capable of producing nisin
50 μl of the stock strain of Lactococcus lactis IO-1 was inoculated into 10 ml of TGC medium, and allowed to stand still at 37 ° C. for 18 hours to obtain a culture solution. 10 ml of the culture solution was inoculated into 100 ml of CMG medium, and 3 ml at 37 ° C. A preculture of lactic acid bacteria was obtained by shaking culture at 100 rpm for an hour.
6. Main culture of lactic acid bacteria capable of producing nisin
The main culture of lactic acid bacteria having a nisin-producing ability was performed by a continuous culture method using a continuous culture device comprising a 3 L jar fermenter.
That is, 900 ml of the lactic acid bacteria culture medium obtained in 1 above, 900 ml of the lactic acid bacteria culture medium obtained in 3 above, and 90 ml of the lactic acid bacteria preculture liquid obtained in 5 above were introduced into the 3 L jar fermenter, and the stirring speed was 250 rpm. Batch culture was performed at a culture temperature of 30 ° C. and a pH of 5.5, and when the glucose concentration of the culture solution reached 4 g / L 8 hours after the start of the culture, pH control, glucose concentration control, and turbidity were performed. The culture was shifted to continuous culture with control, and the continuous culture was performed until 42 hours after the start of the culture while maintaining the glucose concentration of the culture solution at 4 g / L.
The pH control was performed by using a pH sensor and supplying a 5N sodium hydroxide solution to the culture tank to maintain the pH at 5.5. The glucose concentration was controlled by measuring the glucose concentration of the culture using an online biochemical controller BF-410 (manufactured by Able Corporation) equipped with a glucose electrode, and measuring the glucose concentration obtained in step 2 during continuous culture. The mixture obtained by mixing the substrate solution for adjustment and the substrate solution for glucose concentration adjustment obtained in 4 above at a ratio of 1: 1 is supplied to a culture tank to maintain the glucose concentration of the culture solution at 4 g / L. It was done by doing. Further, at the time of the pH control and the glucose concentration control, the same amount of sterilizing solution as the 5N sodium hydroxide solution and the glucose concentration adjusting substrate solution supplied to the culture tank was passed through the microfiltration membrane from the culture tank. By withdrawing, the liquid volume in the culture tank was kept constant. The turbidity control is performed by using a laser turbidity meter and using the same amount of culture solution (bacterium) as the turbidity control supply solution (yeast extract 0.5% by weight, polypeptone 0.5% by weight) supplied to the culture tank. Was removed from the culture tank, whereby the cell concentration and liquid volume in the culture tank were kept constant.
[0057]
Each of the fermentation products obtained in Example 1, Example 2, Examples 4 to 7, Comparative Example 1 and Comparative Example 2 was subjected to the following Test Example 1, and the nisin concentration of the fermentation product was measured. .
[0058]
[Test Example 1]
Each of the fermented products obtained in Example 1, Example 2, Examples 4 to 7, Comparative Example 1 and Comparative Example 2 was subjected to centrifugation at 9000 rpm for 30 minutes to obtain a top product. The supernatant 1 of each fermentation product was adjusted to pH 3.0 by adding concentrated hydrochloric acid to the supernatant 1 of each fermentation product, left at 4 ° C. overnight, and again subjected to centrifugation at 9000 rpm and 30 min. The supernatant 2 of the fermentation product was obtained, and the nisin concentration of the culture solution was measured using the obtained supernatant 2 of each fermentation product as a sample according to the following method. That is, Micrococcus luteus (IFO N0.3333) was used as an indicator bacterium, and nisin (manufactured by ICN) dissolved in 0.01 N HCl at 3000 IU / ml was used as a nisin A standard solution. One loopful of the indicator bacterium was inoculated into 10 ml of LB liquid medium, and shake culture was performed under the conditions of a culture temperature of 30 ° C., a stirring speed of 100 rpm, and a culture time of 18 hours to obtain a culture solution, and 100 μl of the culture solution was cooled to an appropriate temperature. The suspension was suspended in 2.0 ml of soft agar to obtain a suspension. The suspension was overlaid on an LB agar medium, allowed to stand for 1 hour, and then diluted with 0.01N HCl to an appropriate concentration to obtain the supernatant of the fermentation product. 2 was spotted on the LB agar medium in an amount of 10 μl each, and cultured at 30 ° C. for 24 hours. Then, the presence or absence of a growth inhibition circle around the spot was confirmed. The dilution rate D was determined, and the nisin antibacterial activity value was calculated from the relational expression of nisin antibacterial activity (AU / ml) = 1 / D × sample addition amount (ml).
[0059]
[Evaluation 1]
Table 1 shows the nisin antibacterial activity values of the respective fermentation products obtained 24 hours after the start of the culture in Example 1 and Comparative Example 1. From the results shown in Table 1, the following was found. That is, the nisin antibacterial activity 24 hours after the start of the culture was as low as 1200 IU / ml for the fermentation product obtained in Comparative Example 1, whereas the fermentation product obtained in Example 1 was 4500 IU. / Ml and an extremely high value. That is, the nisin antibacterial activity value of the fermentation product obtained by the method for producing nisin of the present invention using the lactic acid bacteria culture medium obtained from the barley shochu distillation residue is determined by the conventional method for producing nisin using the MRS medium. It was found that the obtained fermented product reached about 3.8 times the nisin antibacterial activity value.
Table 2 shows the nisin antibacterial activity values in the continuous culture of Example 2 and Comparative Example 2. From the results shown in Table 2, the following was found. That is, in Comparative Example 2, the nisin antibacterial activity value showed a maximum value at 12 hours after the start of culture, in which the batch culture was shifted to continuous culture, and then gradually decreased, and then, at 42 hours after the start of culture, 1000 IU / ml. On the other hand, in Example 2, the maximum value was shown at 12 hours after the start of culture, which shifted from batch culture to continuous culture, and then decreased until 24 hours, but after 24 hours, It rose again and reached 6300 IU / ml 42 hours after the start of the culture. That is, the lactic acid bacterium culture medium obtained from the barley shochu distillation residue is used, and the nisin production method of the present invention in which culturing is performed while maintaining the glucose concentration of the culture solution during continuous culture at 4 g / L. The nisin antibacterial activity value of the fermented product is obtained by a conventionally known method for producing nisin in which culture is performed using an MRS medium and maintaining the glucose concentration of the culture solution during continuous culture at less than 0.1 g / L. It was found to reach 6.3 times the nisin antibacterial activity value of the fermented product.
The following was found from the results of Example 3 and Comparative Example 3. That is, the nisin antibacterial activity value of each fermentation product obtained by subjecting various lactic acid bacteria strains having a nisin-producing ability to continuous culture in Example 3 was the same as in Comparative Example 3 regardless of which strain was used. The value was significantly higher than that in the case of employing the known nisin production method shown in FIG.
[0060]
The results of Example 4 and Example 5 revealed the following. That is, instead of the liquid component obtained by solid-liquid separation of the barley shochu distillation residue used in the present invention, the liquid component is filtered to obtain a clarified liquid, and the clarified liquid is concentrated to obtain a concentrated liquid. When a solution comprising the non-adsorbable fraction described in Document 6 obtained by subjecting the concentrated solution to an adsorption treatment using a synthetic adsorbent is used, the fermentation product obtained in Examples 4 and 5 is used. The nisin antibacterial activity value was found to be equivalent to the nisin antibacterial activity value of the fermented products obtained in each of Example 1 and Example 2.
[0061]
The results of Example 6 and Example 7 revealed the following. That is, Example 6 and Example in which a mixture of a liquid component obtained by solid-liquid separation of a barley shochu distillation residue and a liquid component obtained by solid-liquid separation of a rice shochu distillation residue was used as a medium. The nisin antibacterial activity value of the fermented product obtained in Example 7 was found to be equivalent to the nisin antibacterial activity value of the fermented product obtained in each of Example 1 and Example 2.
[0062]
[Test Example 2]
Each of the fermentation products obtained in Example 1, Example 2, Examples 4 to 7, Comparative Example 1 and Comparative Example 2 was subjected to a sensory test for taste. That is, each of the fermentation products obtained in Example 1, Example 2, Examples 4 to 7, Comparative Example 1 and Comparative Example 2 was subjected to a sensory test by 10 panelists, and the fermentation products were obtained. The taste was evaluated.
[0063]
[Evaluation 2]
Table 3 shows the sensory test results of the fermentation products obtained in Example 1, Example 2, Examples 4 to 7, Comparative Example 1 and Comparative Example 2. From the results shown in Table 3, the following was found. That is, the fermentation products obtained in Comparative Example 1 and Comparative Example 2 had extremely low monotonous flavor and unpleasant unpleasant taste, and thus had very low sensory evaluation values. 2, and all of the fermented products obtained in Examples 4 to 7 have a mellow and harmonious flavor and a rich umami, and thus have a significantly high sensory evaluation value, and thus are seasonings or food materials. It turned out to be suitable. From the comments of the panelists, among the fermentation products obtained in Examples 1, 2, and 4 to 7, the fermentation products obtained in Examples 4 and 5 are particularly The fermented products obtained in Example 6 and Example 7 are characterized by exhibiting a distinctly mellow flavor, and each of the fermented products obtained in Example 6 and Example 7 is more palatable than the fermented products obtained in Example 1 and Example 2. It has been revealed that it has the characteristic of exhibiting a rich umami.
[0064]
From the above results, it is understood that according to the present invention, nisin productivity can be drastically increased as compared with a conventionally known method for producing nisin. In addition, the fermented product obtained by the present invention contains a significant amount of nisin and, in addition to excellent antibacterial activity, exhibits a mellow, harmonious flavor and rich umami, and thus can be used very suitably as food. .
[0065]
[Table 1]

[0066]
[Table 2]

[0067]
[Table 3]

[0068]
【The invention's effect】
As described above in detail, the method for producing a fermentation product containing nisin of the present invention comprises a liquid component (A) obtained by solid-liquid separation of a barley shochu distillation residue, and has a Brix concentration of 0.5. A medium prepared by adding a saccharide to a liquid adjusted to a range of 6.0 to 6.0 and using a lactic acid bacterium having a nisin-producing ability is cultured. 0.1 g / L or more and the pH value is controlled in the range of 4.0 to 7.0, and enables efficient production of a fermentation product containing a significant amount of nisin. The resulting fermented product containing a significant amount of nisin has excellent taste and can be suitably used as food.

Claims (15)

A method for producing a fermentation product containing nisin for culturing a lactic acid bacterium having nisin-producing ability using a liquid component obtained by solid-liquid separation of a shochu distillation residue as a medium, wherein the medium is barley shochu A liquid comprising a liquid component (A) obtained by subjecting a distillation residue to solid-liquid separation and having a Brix concentration adjusted to a range of 0.5 to 6.0, to which sugar has been added. A lactic acid bacterium having a productivity is added to perform culturing, and the culturing is performed while controlling the sugar concentration of the culture solution to at least 0.1 g / L or more and controlling the pH value in the range of 4.0 to 7.0. A method for producing a fermentation product containing a significant amount of nisin. The fermentation production containing a significant amount of nisin according to claim 1, wherein the adjustment so that the Brix concentration of the liquid component (A) is within the above range is performed by diluting the liquid component with water. Method of manufacturing a product. The method for producing a fermentation product containing a significant amount of nisin according to claim 1 or 2, wherein the nisin is nisin A and / or nisin Z. The nisin according to any one of claims 1 to 3, wherein at least a part of the liquid component (A) is subjected to an adsorption treatment using a synthetic adsorbent to obtain a non-adsorbed fraction. Method for producing fermented products. The method according to claim 4, wherein the synthetic adsorbent is selected from aromatic, modified aromatic, and methacrylic synthetic adsorbents. The method for producing a fermentation product containing a significant amount of nisin according to any one of claims 1 to 5, wherein the culturing is performed by a batch culture method and / or a continuous culture method. The culturing is carried out by the continuous culturing method. At this time, the liquid is obtained by solid-liquid separation of the residual liquid of barley shochu distillation, and the sugar is adjusted to a liquid whose Brix concentration is adjusted to a range of 0.5 to 6.0. 7. The method for producing a fermentation product containing a significant amount of nisin according to claim 6, wherein the solution to which nisin is added is supplied as a substrate solution. 2. The nisin according to claim 1, wherein the liquid component (A) is partially replaced by a liquid component (B) obtained by solid-liquid separation of a residual liquid of rice shochu distillation. 3. A method for producing a fermentation product containing a significant amount. The liquid composed of the liquid component (A) and the liquid component (B) is diluted with water and adjusted to have a Brix concentration in the range of 0.5 to 6.0. Item 10. A method for producing a fermented product containing a significant amount of nisin according to Item 8. The method for producing a fermentation product containing a significant amount of nisin according to claim 8 or 9, wherein the nisin is nisin A and / or nisin Z. 11. The liquid fraction (A) and / or at least a part of the liquid fraction (B) is subjected to an adsorption treatment using a synthetic adsorbent to make a non-adsorbed fraction. 3. A method for producing a fermentation product containing a significant amount of nisin according to item 1. The method for producing a fermented product containing a significant amount of nisin according to claim 11, wherein the synthetic adsorbent is selected from aromatic, modified aromatic, and methacrylic synthetic adsorbents. The method for producing a fermentation product containing a significant amount of nisin according to any one of claims 8 to 12, wherein the culture is performed by a batch culture method and / or a continuous culture method. The culturing is carried out by the continuous culturing method. At this time, the liquid component (A) obtained by solid-liquid separation of the barley shochu distillation residue and a part of the liquid component (A) are converted to the rice shochu distillation residue. Supplying a liquid obtained by replacing the liquid component (B) obtained by solid-liquid separation with a Brix concentration adjusted to a range of 0.5 to 6.0 and adding a sugar as a substrate solution; The method for producing a fermentation product containing a significant amount of nisin according to claim 13. A fermented product containing a significant amount of nisin produced by the production method according to any one of claims 1 to 14, which has excellent taste.