JP3859031B2 - New gonococci and their uses - Google Patents

Description

（上記形態は２５℃、７日間培養のものを測定した）
【００１０】
（Ｂ）生育状態
（２５℃及び３５℃、７日間培養）
（ａ）麹汁寒天培地
（Ｉ）２５℃培養
径が６０．０ｍｍ、平坦状の巨大集落を形成し、中央部分が深緑色、外周は黄緑からクリーム色を呈し、胞子を形成し、シャーレ裏面のヒダは無く、クリーム色を呈する。
（II) ３５℃培養
径が４７．０ｍｍ、平坦状の巨大集落を形成し、中央部分がクリーム色、その外側が深緑色、外周がクリーム色を呈し、胞子を形成し、シャーレ裏面に放射状のヒダがあり、淡黄色を呈する。
（ｂ）ツアペック寒天培地
（Ｉ）２５℃培養
径が３３．０ｍｍ、平坦状の集落を形成し、中央部分が深緑色、外周はほとんど白色を呈し、胞子を形成し、裏面にヒダは無く、中央部が褐色から黄色、周辺は白色を呈する。
（II) ３５℃培養
径が２５．０ｍｍ、平坦状の集落を形成し、中央のみわずかに深緑色でほとんどクリーム色を呈し、胞子を形成し、裏面にヒダはなく、中央部が黄褐色、周辺はクリーム色を呈する。
【００１１】
（Ｃ）生理的性質
（ａ）生育温度 ： １５〜４５℃
（ｂ）最適生育温度 ： ２５〜３５℃
（ｃ）生育ｐＨ ： ２〜１０
（ｄ）最適生育ｐＨ ： ３〜９
（ｅ）有機酸の生成
元株のＡ．オリーゼに比べて、クエン酸生産が顕著に増加している。
【００１２】
２．ＫＩ４９

（上記形態は２５℃、７日間培養のものを測定した）
【００１３】
（Ｂ）生育状態
（２５℃及び３５℃、７日間培養）
（ａ）麹汁寒天培地
（Ｉ）２５℃培養
径が５８．０ｍｍ、平坦状の巨大集落を形成し、中央部分が深緑色、外周は黄緑からクリーム色を呈し、胞子を形成し、シャーレ裏面のヒダは無く、クリーム色を呈する。
（II) ３５℃培養
径が４４．０ｍｍ、平坦状の巨大集落を形成し、中央部分がクリーム色、その外側が深緑色、外周がクリーム色を呈し、胞子を形成し、シャーレ裏面に放射状のヒダがあり、淡黄色を呈する。
（ｂ）ツアペック寒天培地
（Ｉ）２５℃培養
径が３２．０ｍｍ、平坦状の集落を形成し、中央部分が深緑色、外周はほとんど白色を呈し、胞子を形成し、裏面にヒダは無く、中央部が褐色から黄色、周辺は白色を呈する。
（II) ３５℃培養
径が２４．０ｍｍ、平坦状の集落を形成し、中央のみわずかに深緑色でほとんどクリーム色を呈し、胞子を形成し、裏面にヒダはなく、中央部が黄褐色、周辺はクリーム色を呈する。
【００１４】
（Ｃ）生理的性質
（ａ）生育温度 ： １５〜４５℃
（ｂ）最適生育温度 ： ２５〜３５℃
（ｃ）生育ｐＨ ： ２〜１０
（ｄ）最適生育ｐＨ ： ３〜９
（ｅ）有機酸の生成
元株のＡ．オリーゼに比べて、クエン酸生産が顕著に増加している。
【００１５】
これら変異処理して得られた本菌株は、明らかにＡ．オリーゼに属するものであるが、クエン酸高生産性の向上の形質から、変異株とするのが妥当であり、それぞれ、変異株Ａ．オリーゼＫＤ２５及び変異株Ａ．オリーゼＫＩ４９と命名したものである。
【００１６】
本発明による新変異株は、クエン酸生産能が非常に高く、極めて有用性が高いものであるが、これら新変異株の胞子を蒸米に接種して製麹した後、清酒の醸造に使用すると、クエン酸含量の多いそう快な酸味を有し、しかも香味バランスが良好で、清酒独特の風味を合せ持つ新しいタイプの清酒を製造できる点で特に有用性が高いものである。したがって、本発明による新変異株の産業上の利用性は極めて顕著である。
【００１７】
【実施例】
以下、実施例によって本発明を更に具体的に説明するが、本発明はこれらの実施例に限定されない。
【００１８】
実施例１
クエン酸高生産株の取得手順は、市販の清酒用種もやしのＡ．オリーゼの胞子を用い、麹汁〔米麹：井水＝１：５（重量）比率で混合し、５５℃で２４時間反応後のろ液〕寒天プレート上へ接種し、単一コロニーを生成させ、胞子生成前に別のスラント（麹汁寒天培地）へ移植し、スラント中で胞子を形成させる。この胞子塊は０．０１％ツイーン（Tween)８０を含む滅菌水溶液を加え、３０℃で３０分間振とうすることにより胞子を分散させ、それぞれ単一にした懸濁液を得る。この胞子懸濁液に２０ｃｍの距離から１５ＷＵＶランプを３分間照射した。このときの生存率は３％であった。胞子懸濁液０．１ｍｌを下記表１に示すｐＨ指示薬を含む麹汁培地に塗布して、３０℃で９６時間培養することによりコロニーを作らせ、コロニー周辺の培地の色を緑から黄色に変色させる株を有機酸高生産株として取得する。これらの取得株を殺菌した１０ｇ蒸米に植菌し、３０℃、１２０時間の培養により種麹を作成する。この種麹０．０５ｇを５０ｇ蒸米に種付けし、４２時間の製麹（通常条件、スタート３０℃、３６時間後３７．５℃、以後３７．５℃定温、相対湿度９０％以上）を行う。得られた清酒麹１０ｇを蒸留水５０ｍｌで２０℃で３時間抽出後、その抽出液について有機酸自動分析計にて有機酸を定量し、クエン酸含量の高い株を選択した。これらの株を用いて米麹を調製し、その酸度及びクエン酸含量の比較を後記表２に示した。
【００１９】
【表１】

【００２０】
【表２】

【００２１】
表２からクエン酸高生産麹とは、クエン酸含量が２３０ｍｇ／１００ｇ米麹以上であり、クエン酸が２３０〜９００ｍｇ／１００ｇ米麹のクエン酸高生産変異株Ａ．オリーゼが得られる。この選択により、クエン酸高生産の変異株であるＫＤ２５及びＫＩ４９を得た。
【００２２】
実施例２
酒造用精白米（精米歩合７０％）５００ｇを常法に従って洗米し、井水に１６時間浸漬した後、３時間水切りした後６０分間１００℃で蒸きょうした。この蒸米に、本発明による変異株Ｎｏ．１、２、３、４、５（ＫＤ２５）及び６（ＫＩ４９）株、並びに対照としてＫＤ２５株及びＫＩ４９株の元株であるＡ．オリーゼをそれぞれ胞子を蒸米に対し０．１％ｗ／ｗ種付けし、４２時間製麹（スタート３０℃、３９時間後４０．０℃、以後４０．０℃定温、相対湿度９０％以上）した。
得られた米麹１０ｇへ蒸留水５０ｍｌを加え、２０℃で３時間で有機酸を抽出し、この抽出液中の有機酸を有機酸自動分析計を用いて定量した。下記表３に結果を示す。表３からもわかるとおり、本菌株のクエン酸生産量が極めて高く、元株Ａ．オリーゼのクエン酸生産に対して、ＫＤ２５株の場合は６．３倍以上、及びＫＩ４９株の場合では７．３倍以上で顕著に向上していることがわかる。
【００２３】
【表３】

【００２４】
同様の製麹条件で得たＫＤ２５株及びＫＩ４９株の米麹を用い、対照を元株Ａ．オリーゼの米麹を用いる清酒の仕込を下記表４に示す仕込配合で行った。
【００２５】
【表４】

【００２６】
まず、酵母（協会酵母７０１号、０．６ｇ使用）、米麹、乳酸、汲水を１３℃で混合してかくはんし、３時間後蒸米をこの混合物中へ投入して初添を行った。１日後初添と同様の方法で仲添を行い、５日後留添を行い仕込を終了した。醪の温度は１５℃で管理し、留添後１８日で上槽し清酒を得た。また、元株のＡ．オリーゼ米麹を用いた清酒へクエン酸を添加して、ＫＤ２５株を用いた清酒と同水準の酸度となるように清酒（元株＋クエン酸）を調整した。
この方法によって得られた清酒の一般分析値を下記表５に示した。この結果から明らかなように、本発明によるＫＤ２５株及びＫＩ４９株は対照菌元株Ａ．オリーゼに比べ清酒中の総酸量を著しく増加させ、それぞれ１．２３倍及び１．２７倍であった。
【００２７】
【表５】

【００２８】
次に増加した有機酸について検討した。すなわち、得られた清酒を試料として有機酸自動分析計により、主要な有機酸について定量した。結果を下記表６に示すが、この結果からも明らかなように、本発明によるＫＤ２５株及びＫＩ４９株の米麹を用いた場合には、対照菌元株Ａ．オリーゼを用いた場合より、清酒中のクエン酸含量が顕著に増加し、それぞれ５．４５倍及び７．０７倍となった。
得られたそれぞれの清酒をパネラー１０名で、３点法（１良−３悪）で官能評価を行った。その結果を後記表７に示すように、ＫＤ２５株及びＫＩ４９株の米麹を用いた清酒は、元株のＡ．オリーゼ米麹を用いた清酒に比べ、酸味がそう快で、香味は元株とほぼ同様で香味バランスのとれた酒質となり、喉ごし感が良好であった。
元株のＡ．オリーゼ米麹を用いた清酒へクエン酸を添加して酸味を付与した清酒は、酸味と清酒本来の風味のバランスが悪く、酸味と調和せず、酸味が口に残り、味、香りとも低い評価であった。
【００２９】
【表６】

【００３０】
【表７】

【００３１】
実施例３
本発明Ａ．オリーゼＫＤ２５麹を用いて、新規な低アルコール清酒を調製した。
すなわち、実施例１で用いた表４の仕込製造方法で示した麹歩合２０％を３５％及び５０％に変更し、それ以外は同様の操作でそれぞれ上槽、精製して原酒に相当する清酒を得た。次に、それら原酒を井水で割水して、アルコール濃度１０．２ｖ／ｖ％に調製して低アルコール清酒を調製した。
それらの分析結果を下記表８に示す。
【００３２】
【表８】

【００３３】
これらそれぞれの清酒、及び対照として市販の清酒（アルコール濃度１５ｖ／ｖ％）をアルコール濃度１０．２ｖ／ｖ％になるように井水で希釈したものを、パネラー１０名で、３点法（１良−３悪）で官能評価を行った。その結果を下記表９に示す。
【００３４】
【表９】

【００３５】
表９より、対照である通常の市販清酒を加水してアルコール濃度を１０．２ｖ／ｖ％にした対照では水っぽく感じられたのに対し、麹歩合３５％及び５０％にしてＡ．オリーゼＫＤ２５の麹を用いた通常の清酒（原酒、アルコール濃度が１５．２ｖ／ｖ％）は酸味の強い濃厚な酒質に仕上がり、これに加水してアルコール濃度を１０．２ｖ／ｖ％にした本発明の低アルコール清酒（アルコール濃度１０．２ｖ／ｖ％）は本発明の麹株の酸味成分であるクエン酸や麹由来の成分が調和し、香味のバランスがよく、さわやかですっきりした飲みやすい新規な清酒となった。
【００３６】
【発明の効果】
以上のことからも明らかなように、本発明のクエン酸高生産Ａ．オリーゼを用いて清酒醸造を行う場合には、従来の黒カビや焼酎麹菌の使用を行わずとも、この麹菌変異株さえ供すれば従来の清酒香味を有する新規なタイプのそう快な酸味を有する清酒を得ることができるという顕著な効果が得られる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a high-citric acid-producing strain of Aspergillus oryzae and a method for producing sake using the bacterial strain.
[0002]
[Prior art]
Conventionally, citric acid has been added to the production of alcoholic beverages with a pleasant sour taste, but adding citric acid to alcoholic beverages has also led to a decline in product image, and the amount of citric acid added to alcoholic beverages There are limitations. In addition, aspergillus niger which is black mold, and Aspergillus awamori, Aspergillus kawachii and Aspergillus usamii, which have a high amount of citric acid, such as Aspergillus niger. For example, a method for brewing sake has been proposed. In this method, although the citric acid content in sake is increased, A. produced by Aspergillus oryzae (hereinafter abbreviated as A. oryzae). It is different from the flavor unique to sake based on Orise, and the balance of flavor unique to sake is lost, and the original flavor of sake cannot be obtained.
[0003]
[Problems to be solved by the invention]
Currently, sake with a high citric acid content is produced by the method using black mold or shochu in sake, for example, sake, but the balance of flavor in sake and the improvement of heterogeneous flavor remain as issues. Is the current situation.
An object of the present invention is to provide a conventional balance and sake original flavor flavor of sake, novel can be used in the production of sake exhibiting refreshing acidity A. Obtaining an Olyse, and A. The object is to provide a method for producing sake using orise.
[0004]
[Means for Solving the Problems]
To summarize the present invention, the first invention of the present invention is an Aspergillus oryzae KD25 (FERM P), which has a high citric acid-producing ability, does not impair the flavor balance of sake, and does not impair the original flavor of sake. -16122) or Aspergillus oryzae KI49 (FERM P-16123) , and the second invention is characterized by producing koji using the koji mold and producing sake using the koji obtained. The present invention relates to a method for producing sake .
[0005]
A. According to the present invention Oryze is a koji mold with significantly improved citric acid production. A. of the present invention. Without impairing the balance collapsing it and sake original flavor flavor in sake by using oryzae brewing Qing liquor, moreover citric acid content much, it becomes possible to produce a sake with refreshing acidity.
[0006]
DETAILED DESCRIPTION OF THE INVENTION
The present invention will be specifically described below.
The novel gonococcus of the present invention comprises A. Any citrate high-producing strain belonging to Olyse may be used, and the method for obtaining the citrate is not limited, but examples include mutation. Examples of the mutation treatment method include physical ultraviolet irradiation and the use of chemical mutation agents. For example, it is acquired as follows. A. After irradiating oryzae spores with UV irradiation, they are cultured on a plate and screened for mutants with improved organic acid production ability that significantly reduce the pH of the medium by the generated organic acid. Production A. A mutant of Orise is obtained.
[0007]
One strain of Aspergillus oryzae newly isolated by the above method has the following mycological properties. Although it belongs to Olyse, its ability to produce citric acid is remarkably high, so it was identified as a mutant strain, and the first strain was named and displayed as Aspergillus oryzae KD25 (hereinafter abbreviated as KD25). Deposited as FERMP-16122 at the Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology. The second strain is named and displayed as Aspergillus oryzae KI49 (hereinafter abbreviated as KI49), and is deposited as FERM P-16123 at the Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology.
[0008]
The respective mycological properties are as follows.
[0009]
1. KD25

(The above form was measured at 25 ° C. for 7 days)
[0010]
(B) Growth state (cultured at 25 ° C and 35 ° C for 7 days)
(A) Boiled agar medium (I) 25 ° C. Culture diameter is 60.0 mm, flat large colony is formed, the central part is dark green, the outer periphery is creamy from yellow green, forming spores, petri dish There is no crease on the back side, and it has a cream color.
(II) 35 ° C. culture diameter 47.0 mm, forming a large flat colony, cream in the center, dark green on the outside, cream on the periphery, forming spores, radial on the petri dish back There are folds and light yellow.
(B) Tuapec agar medium (I) 25 ° C culture diameter is 33.0mm, flat colony is formed, the central part is dark green, the outer periphery is almost white, spores are formed, there are no folds on the back, The central part is brown to yellow and the periphery is white.
(II) 35 ° C. culture diameter is 25.0 mm, forming a flat settlement, slightly dark green at the center and almost creamy, forming spores, no folds on the back, yellowish brown at the center, The surrounding area is creamy.
[0011]
(C) Physiological properties (a) Growth temperature: 15-45 ° C
(B) Optimal growth temperature: 25-35 ° C
(C) Growth pH: 2 to 10
(D) Optimal growth pH: 3-9
(E) A. of the source strain of the organic acid Citric acid production is significantly increased compared to Orise.
[0012]
2. KI49

(The above form was measured at 25 ° C. for 7 days)
[0013]
(B) Growth state (cultured at 25 ° C and 35 ° C for 7 days)
(A) Boiled agar medium (I) 25 ° C. Culture diameter is 58.0 mm, a large flat colony is formed, the central part is dark green, the outer periphery is creamy from yellow green, forming spores, petri dish There is no crease on the back side, and it has a cream color.
(II) 35 ° C. culture diameter is 44.0 mm, forming a flat giant colony, creamy in the center, dark green on the outside, creamy on the periphery, forming spores, radial on the petri dish back There are folds and light yellow.
(B) Tuapec agar medium (I) 25 ° C culture diameter is 32.0 mm, flat colony is formed, the central part is dark green, the outer periphery is almost white, spores are formed, there are no folds on the back, The central part is brown to yellow and the periphery is white.
(II) 35 ° C. culture diameter of 24.0 mm, forming a flat colony, slightly dark green at the center and almost creamy, forming spores, no folds on the back, yellowish brown at the center, The surrounding area is creamy.
[0014]
(C) Physiological properties (a) Growth temperature: 15-45 ° C
(B) Optimal growth temperature: 25-35 ° C
(C) Growth pH: 2 to 10
(D) Optimal growth pH: 3-9
(E) A. of the source strain of the organic acid Citric acid production is significantly increased compared to Orise.
[0015]
This strain obtained by the mutation treatment is clearly A. Although it belongs to Olyse, it is appropriate to make it a mutant strain from the character of improvement in high productivity of citric acid. Olyse KD25 and mutant A. It is named Olise KI49.
[0016]
New mutants according to the present invention, citric acid-producing ability is very high, very but those highly useful, after koji was inoculated with spores of these new mutants to steamed rice, use in brewing Qing liquor Then, it is particularly useful in that a new type of sake having a pleasant acidity with a high citric acid content, a good flavor balance, and a unique flavor of sake can be produced. Therefore, industrial applicability of the new mutants according to the present invention is Ru quite remarkable der.
[0017]
【Example】
EXAMPLES Hereinafter, although an Example demonstrates this invention further more concretely, this invention is not limited to these Examples.
[0018]
Example 1
The procedure for acquiring a high-citric acid production strain is as follows. Using oryzae spores, inoculate on the agar plate with the rice bran soup (mixed at a ratio of rice bran: well water = 1: 5 (weight) and reacted at 55 ° C. for 24 hours) to form a single colony. Before sporulation, transplant to another slant (boiled agar medium) to form spores in the slant. To this spore mass, a sterilized aqueous solution containing 0.01% Tween 80 is added, and the spore is dispersed by shaking at 30 ° C. for 30 minutes to obtain a single suspension. This spore suspension was irradiated with a 15 W UV lamp for 3 minutes from a distance of 20 cm. The survival rate at this time was 3%. Apply 0.1 ml of the spore suspension to a broth medium containing the pH indicator shown in Table 1 below, and incubate at 30 ° C. for 96 hours to form colonies. The color of the medium around the colonies changes from green to yellow. Acquire the color change stock as a high organic acid production strain. These acquired strains are inoculated into sterilized 10 g steamed rice, and seed meal is prepared by culturing at 30 ° C. for 120 hours. 0.05 g of this seed koji is seeded on 50 g steamed rice and subjected to koji making for 42 hours (normal conditions, start 30 ° C., 36 hours later 37.5 ° C., then 37.5 ° C. constant temperature, relative humidity 90% or more). After extracting 10 g of the obtained sake lees with 50 ml of distilled water at 20 ° C. for 3 hours, the organic acid was quantified with an organic acid automatic analyzer for the extract, and a strain having a high citric acid content was selected. Rice bran was prepared using these strains, and the acidity and citric acid content were compared in Table 2 below.
[0019]
[Table 1]

[0020]
[Table 2]

[0021]
From Table 2, a high citric acid-producing rice bran is a citric acid-rich mutant having a citric acid content of 230 mg / 100 g or more and a citric acid of 230-900 mg / 100 g rice koji. Olyse is obtained. By this selection, KD25 and KI49, which are mutants with high citrate production, were obtained.
[0022]
Example 2
500 g of polished rice for sake brewing (milled rice ratio 70%) was washed according to a conventional method, immersed in well water for 16 hours, drained for 3 hours, and steamed at 100 ° C. for 60 minutes. To this steamed rice, the mutant No. 1, 2, 3, 4, 5 (KD25) and 6 (KI49) strains, and A. which is a former strain of KD25 strain and KI49 strain as a control. Each spore was seeded with 0.1% w / w of steamed rice, and then smelted for 42 hours (start 30 ° C, 39 hours later 40.0 ° C, then 40.0 ° C constant temperature, relative humidity 90% or more).
Distilled water (50 ml) was added to 10 g of the obtained rice bran, the organic acid was extracted at 20 ° C. for 3 hours, and the organic acid in the extract was quantified using an organic acid automatic analyzer. The results are shown in Table 3 below. As can be seen from Table 3, the amount of citric acid produced by this strain is extremely high. It can be seen that the citrate production of Orise is significantly improved by 6.3 times or more in the case of the KD25 strain and 7.3 times or more in the case of the KI49 strain.
[0023]
[Table 3]

[0024]
KD25 strain and KI49 strain rice bran obtained under the same koji-making conditions were used, and the control was used for the former strain A. The preparation of sake using rice bran of Orise was performed with the preparation composition shown in Table 4 below.
[0025]
[Table 4]

[0026]
First, yeast (Association Yeast No. 701, 0.6 g used), rice bran, lactic acid and scooped water were mixed at 13 ° C., stirred, and steamed rice was added into this mixture after 3 hours for initial addition. One day later, mediation was conducted in the same manner as the first attendance. The temperature of the koji was controlled at 15 ° C., and the sake was obtained in the upper tank 18 days after the distillation. In addition, A. Citric acid was added to sake using rice bran, and sake (original stock + citric acid) was adjusted to have the same level of acidity as sake using KD25 strain.
The general analysis values of sake obtained by this method are shown in Table 5 below. As is apparent from the results, the KD25 strain and KI49 strain according to the present invention are the control strain A. Compared with Orise, the total acid amount in sake was remarkably increased, being 1.23 times and 1.27 times, respectively.
[0027]
[Table 5]

[0028]
Next, the increased organic acid was examined. That is, the major organic acids were quantified using the obtained sake as a sample by an organic acid automatic analyzer. The results are shown in Table 6 below. As is apparent from these results, when rice bran of the KD25 strain and KI49 strain according to the present invention was used, the control strain A. The citric acid content in sake was significantly increased from the case of using orise, which was 5.45 times and 7.07 times, respectively.
Each obtained sake was subjected to sensory evaluation by 10 panelists using a three-point method (1 good-3 bad). As shown in Table 7 below, the sake using rice bran of the KD25 and KI49 strains was obtained from the original strain A. Compared to sake using Olise rice bran, the sour taste was so pleasant, the flavor was almost the same as the original strain, and the quality of the sake was well balanced, and the feeling of squeezing was good.
A. of the former stock Sake made by adding citric acid to Sake using Orise rice bran has a bad balance between sourness and Sake's original flavor, does not match the sourness, sourness remains in the mouth, and taste and aroma are low. Met.
[0029]
[Table 6]

[0030]
[Table 7]

[0031]
Example 3
Invention A. A new low alcohol sake was prepared using Orise KD25 ゼ.
That is, 20% of the koji ratio shown in Table 4 used in Example 1 was changed to 35% and 50%, and the remaining tanks were refined in the same manner except that the sake was equivalent to the original sake. Got. Next, these raw sakes were divided with well water and adjusted to an alcohol concentration of 10.2 v / v% to prepare low alcohol sake.
The analysis results are shown in Table 8 below.
[0032]
[Table 8]

[0033]
Each of these sakes and a commercially available sake (alcohol concentration 15 v / v%) as a control diluted with well water so as to have an alcohol concentration of 10.2 v / v% were obtained using a three-point method (1 Sensory evaluation was carried out with a good-3 bad). The results are shown in Table 9 below.
[0034]
[Table 9]

[0035]
From Table 9, it was felt watery in the control in which the normal commercial sake as a control was added to make the alcohol concentration 10.2 v / v%, whereas it was found to be A.I. Ordinary refined sake using Orise KD25 (raw liquor, alcohol concentration 15.2 v / v%) is finished in a strong acidity and rich liquor, and water is added to make the alcohol concentration 10.2 v / v% The low alcohol sake (alcohol concentration of 10.2 v / v%) of the present invention is harmonized with citric acid, which is the sour component of the koji stock of the present invention, and components derived from koji, and has a good balance of flavor and is refreshing and easy to drink It became a new sake.
[0036]
【The invention's effect】
As is clear from the above, the citric acid high production A. of the present invention. In the case of performing the Qing sake brewing using oryzae it is, even without the use of conventional black mold and shochu koji mold, with a refreshing acidity of a novel type having a conventional sake flavor if Kyosure even this Aspergillus oryzae mutant strain The remarkable effect that sake can be obtained is acquired.

Claims (2)

Aspergillus oryzae KD25 (FERM P-16122) or Aspergillus oryzae KI49 (FERM P-16123), which has a high citric acid production ability and does not impair the flavor balance of sake. Neisseria gonorrhoeae. Method of manufacturing sake, characterized in that to koji preparation using koji mold according to claim 1, to produce a sake using the obtained koji.