JP5340607B2 - Yeast β-phenethyl alcohol production promoter and method for producing yeast fermented product containing β-phenethyl alcohol - Google Patents

Description

  The present invention relates to an accelerator for obtaining a fermented yeast product such as beer, shochu, and bread having a characteristic aroma component without using a brewed yeast having a special mutation, and a method for using the agent.

  Higher alcohols are known as one of the flavor components of alcoholic beverages such as liquor, shochu, and beer. Among them, β-phenethyl alcohol having a rose-like scent is known. Further, β-phenethyl alcohol is known to have a yeast odor masking effect in bread.

It is known that β-phenethyl alcohol is mainly produced by yeast and imparts flavor to foods using yeast such as miso, soy sauce, alcoholic beverages and breads.
For example, Patent Document 1 (Japanese Patent Laid-Open No. 3-98577) discloses a yeast (Saccharomyces cerevisiae) having an ability to produce phenethyl alcohol.
Patent Document 2 (Japanese Patent Laid-Open No. 3-94670) discloses β-phenethyl alcohol, β-phenethyl acetate high-producing yeast strains, and a method for producing alcoholic beverages using the same.
Patent Document 3 (Japanese Patent Application Laid-Open No. 3-98577) discloses phenylalanine analog-resistant mutant yeasts and breads using the same.

By the way, it is also known that β-phenethyl alcohol is produced by the metabolism of the amino acid phenylalanine. Yeast obtains this derived substance, phenylalanine, either in the form of peptides or free amino acids from the outside of the cell, or by the yeast's own amino acid biosynthetic pathway. If there are many, β-phenethyl alcohol is considered to be derived mainly from phenylalanine taken from outside the cell because feedback inhibition is applied and there is a system in which excess amino acids are not biosynthesized.
In sake yeast, a β-phenethyl alcohol high-producing strain in which an enzyme involved in this feedback inhibition is deficient is known.

  However, no oligopeptide having a β-phenethyl alcohol-producing ability superior to free amino acids, particularly phenylalanine, is known.

(References)
Japanese Unexamined Patent Publication No. 3-98577 Japanese Patent Laid-Open No. 3-94670 Japanese Unexamined Patent Publication No. 3-98577

  The present invention provides a yeast fermented product such as beer, shochu, and bread having a characteristic aroma component without using a brewed yeast having a special mutation, and the production of such aroma component β- The object was to obtain a phenethyl alcohol production promoter.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that an oligopeptide containing phenylalanine in the sequence has an excellent effect on the production of phenethyl alcohol than free phenylalanine, and have completed the present invention. .
That is, the present invention is a yeast β-phenethyl alcohol production promoter mainly composed of one or more oligopeptides containing phenylalanine in the sequence. The oligopeptide containing phenylalanine in the sequence is preferably selected from one or more of dipeptides or tripeptides.
The present invention also provides a yeast containing β-phenethyl alcohol, characterized in that a β-phenethyl alcohol production promoter of yeast mainly comprising one or more oligopeptides containing phenylalanine in the sequence is used as a medium. This is a method for producing a fermented product. The oligopeptide containing phenylalanine in the sequence is preferably selected from one or more of dipeptides or tripeptides.

  By using the β-phenethyl alcohol production promoter of the yeast of the present invention, beer, shochu, bread having a deep fragrance rich in β-phenethyl alcohol without using a brewed yeast or the like that has been subjected to a special mutation Yeast fermented products such as can be obtained.

First, a yeast β-phenethyl alcohol production promoter mainly composed of one or more dipeptides containing phenylalanine in the sequence will be described.
The oligopeptide containing phenylalanine in the sequence may be any of those containing phenylalanine at the N-terminus, those containing phenylalanine at the C-terminus, and those containing other sites.
In the present invention, the oligopeptide containing phenylalanine in the sequence may be produced by any production method. For example, it may be chemically synthesized by the tert-butyloxycarbonyl method or 9-fluorenylmethyloxycarbonyl method, etc., or the plant protein or animal protein rich in phenylalanine is decomposed with an enzyme or acid and fractionated. It may be obtained.
Even if these animal and vegetable proteins are simply enzymatically decomposed to obtain peptides containing oligopeptides as the main component, the nitrogen content of oligopeptides containing phenylalanine in the sequence per total nitrogen content is extremely rare.
On the other hand, the β-phenethyl alcohol production promoter of yeast of the present invention has a nitrogen content of oligopeptide containing phenylalanine in the sequence per total nitrogen content of 10% by weight or more, preferably 30% by weight or more, more preferably 50%. Those containing at least% by weight are suitable.
As the remaining components of the yeast β-phenethyl alcohol production promoter, other components useful for fermentation, such as nitrogen sources such as other amino acids and peptides, carbon sources, minerals, excipients, and sugars can be used.
As described above, when a yeast β-phenethyl alcohol production promoter having a high oligopeptide content containing phenylalanine in the sequence is used, a fermented product containing a large amount of β-phenethyl alcohol can be obtained with a small amount of the agent. If the ratio of the oligopeptide containing phenylalanine in the sequence is low, it is necessary to use a large amount of yeast β-phenethyl alcohol production promoter in order to obtain the same effect, which is not preferable.

Next, a yeast fermented product containing β-phenethyl alcohol will be described.
Examples of the fermented product include beer-based beverages such as beer, happoshu, and miscellaneous sake, and yeast fermented products such as sake, shochu, wine, bread, miso, and soy sauce.
The present invention provides a yeast fermentation product containing β-phenethyl alcohol by using a yeast β-phenethyl alcohol production promoter mainly composed of one or more oligopeptides containing phenylalanine in the sequence as a main component. Can be manufactured.
Hereinafter, a method for producing a yeast fermented product containing β-phenethyl alcohol will be described.

The amount to be used in the medium of the β-phenethyl alcohol production promoter of yeast mainly composed of one or more of the oligopeptides containing phenylalanine in the sequence and the more preferable sequence differ depending on the type of yeast fermentation product.
As an example of the addition ratio of β-phenethyl alcohol production promoter, in the case of a beer-based beverage, the appropriate amount (% by weight) of β-phenethyl alcohol production promoter is 0.0008, depending on the oligopeptide composition. % To 0.66%, preferably 0.0016% to 0.33%, and more preferably 0.004% to 0.165%.
If it is too small, the effect is small, and if it is too large, it will reach a peak and the cost will increase. Regarding the sequence, for example, in the case of a beer-based beverage, it is preferable in the order of Phe-Phe≈Phe-Asp ≧ Asp-Phe> Phe-Leu in which the amount of β-phenethyl alcohol produced is large.
The oligopeptide as used herein refers to a peptide having a size that can be taken in by yeast via a peptide transporter from a dipeptide consisting of two amino acids, and is preferably a dipeptide or a tripeptide.

  In addition, in the case of yeast fermented products such as shochu and bread, the medium is different, such as the amount of nitrogen source to be added is different from beer, so the appropriate range can be determined experimentally unlike the above ratio.

As described above, yeast obtains phenylalanine, which is a derived substance, either from the outside in the form of peptides or free amino acids, or is produced by the yeast's own amino acid biosynthesis pathway. If there is a large amount, there is a system in which feedback inhibition occurs and excess amino acids are not biosynthesized. Incorporation of peptides and free amino acids is a separate system where peptides are incorporated via peptide transporters and free amino acids are incorporated via amino acid transporters.
In brewer's yeast, there is a difference in the uptake speed of each free amino acid, and phenylalanine is known to be relatively slow in uptake. However, when phenylalanine is present in the state of an oligopeptide, the incorporated system is different, and it is thought that it is incorporated faster than it exists in the state of a free amino acid. In fact, when the total amount of phenylalanine outside the cell is measured in the form of amino acids obtained by acid degradation of proteins and peptides, it is added as an oligopeptide when added with free phenylalanine or when added with an oligopeptide of the above sequence. The amount of phenylalanine had decreased since the earlier case. Although the amount of β-phenethyl alcohol produced varies depending on the oligopeptide sequence, it is thought that this is due to the ease of incorporation and the difference in amino acid degradation rate after incorporation.

  Hereinafter, embodiments of the present invention will be described specifically by way of examples.

(Examples 1 to 3, Comparative Example 1)
A culture solution in which dipeptides and various free amino acids are added in the proportions of each example shown in Table 1 (upper table:% conversion, lower table: mmol / 100 ml conversion) as a nitrogen source, and the components in Table 2 are added as other nutrient sources Each was adjusted to pH 5.0 and autoclaved at 121 ° C. for 20 minutes to obtain a culture solution.
Yeast Bank Weihenstephan 34/70 was used as the yeast for the fermentation test. Previously shake cultured in 200 ml of YPD medium at 25 ° C. for 24 hours (120 rpm), further scaled up to 900 ml of the same medium and cultured with shaking at 25 ° C. for 72 hours, and then collected by centrifugation (1690 g, 10 minutes) . After washing once with sterilized water, a suspension of yeast obtained by centrifuging again and adding suspending sterilized water 3 times the weight of the slurry to obtain an added yeast solution.
To 2200 ml of the culture solution, add 10 ml of the added yeast solution, transfer to a glass fermentation tube (inner diameter: 55 mm, length: 1135 mm), and in order to vent the culture solution, hold the mouth of the fermentation tube and shake well. For 11 days. During the culture, the culture solution was sampled at regular intervals, and the amount of β-phenethyl alcohol produced was measured.
As a result, as shown in Table 3, when the content of β-phenethyl alcohol on the 11th day of culture was examined, it was about 100 ppm in Example 1 (Phe-ASP addition group) and about 2 in Example 2 (Phe-Phe addition group). In 80 ppm of Example 3 (Phe-Leu addition group), a high content of β-phenethyl alcohol of about 60 ppm was observed. On the other hand, in Comparative Example 1 (free Phe added group), only about 40 ppm of β-phenethyl alcohol was observed.

(Table 1)

(Table 2)

(Table 3)

(Comparative Example 2)
As a nitrogen source, “High New S” (manufactured by Fuji Oil Co., Ltd.), which is a soybean oligopeptide mixture, was used in a culture solution of 0.3%. As shown in Table 3, only about 30 ppm β-phenethyl alcohol was observed on the 10th day of culture.

  The present invention promotes the production of β-phenethyl alcohol in products that are fermented with yeast, such as beer, shochu, and sake, or breads, miso, and soy sauce, and enables fermented yeast products with excellent aromas. It has become.

Claims (2)

A β-phenethyl alcohol production promoter for beer bottom yeast , comprising as a main component one or more of three kinds of dipeptides of Phe-Asp, Phe-Phe, and Phe-Leu . It is characterized in that a β-phenethyl alcohol production promoter for beer bottom yeast , comprising as a main component one or more of three kinds of dipeptides of Phe-Asp, Phe-Phe, and Phe-Leu, is used as a medium. A method for producing a beer bottom yeast fermented product containing β-phenethyl alcohol.