KR100208968B1 - A novel saccharomyces cerevisiae ns9 which is used for producing yeast extract contained high density of nucleic acid - Google Patents

A novel saccharomyces cerevisiae ns9 which is used for producing yeast extract contained high density of nucleic acid Download PDF

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KR100208968B1
KR100208968B1 KR1019950007221A KR19950007221A KR100208968B1 KR 100208968 B1 KR100208968 B1 KR 100208968B1 KR 1019950007221 A KR1019950007221 A KR 1019950007221A KR 19950007221 A KR19950007221 A KR 19950007221A KR 100208968 B1 KR100208968 B1 KR 100208968B1
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yeast extract
nucleic acid
saccharomyces cerevisiae
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rna
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이형재
남희섭
김성룡
권오성
안종석
박찬선
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이상윤
주식회사농심
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Abstract

본 발명은 RNA를 다량 함유하고 있는 신규한 균주에 관한 것이며, 더욱 상세하게는 고핵산 효모엑기스 조미료 제조에 이용될 수 있는 사카로마이세스(Saccharomyces)속의 신규의 균주 Saccharomyces cerevisiae NS 9에 관한 것이다.The present invention relates to a novel strain containing a large amount of RNA, and more particularly to a novel strain Saccharomyces cerevisiae NS 9 of the genus Saccharomyces (Saccharomyces) that can be used for the preparation of high-nucleic acid yeast extract seasoning.

본 발명에 의하면 균체내에 RNA를 다량 함유하고 있어, 이미, 이취가 없고 정미성 및 감칠맛이 뛰어난 고핵산 효모엑기스 조미료 제조에 이용될 수 있는 신규한 균주를 얻을 수 있다.According to the present invention, it is possible to obtain a novel strain which contains a large amount of RNA in cells, and which can be used for preparing high-nucleic acid yeast extract seasonings that are free of odor and have excellent taste and rich taste.

Description

고핵산 효모 엑기스 제조에 이용되는 Saccharomyces cerevisiae NS 9 균주Saccharomyces cerevisiae NS 9 strain used for the preparation of high nucleic acid yeast extract

본 발명은 RNA를 다량 함유하고 있는 신규한 균주에 관한 것이며, 더욱 상세하게는 이미, 이취가 없고 정미성 및 감칠맛이 뛰어난 고핵산 효모엑기스 조미료 제조에 이용될 수 있는 사카로마이세스(Saccharomyces)속의 신규의 균주에 관한 것이다.The present invention relates to a novel strain containing a large amount of RNA, and more particularly, Saccharomyces genus that can be used to prepare a high-nucleic acid yeast extract seasoning that has no off-flavor and is excellent in taste and taste. It relates to a novel strain.

효모 엑기스는 천연 조미료로서 HAP(Hydrolized Animal Protein)와 HVP(Hydrolized Vegetable Protein)의 대체재로서 주목받고 있는 식품소재이며, 그 풍미가 고기향과 유사하므로 구미에서는 쇠고기 추출물의 대체재로서 널리 이용되고 있다. 또한, 효모 엑기스는 복잡한 정미성과 농후한 맛을 지니고, 이미, 이취를 마스킹하는 등 다른 조미료에서 볼 수 없는 기능을 가진다. 이는 아미노산만이 주요 정미성분인 다른 조미료와 달리, 효모 엑기스에는 아미노산 뿐만 아니라 펩티드, 헥산등의 성분이 더 함유되어 있기 때문에 나타나는 장점이다.Yeast extract is a natural food seasoning that is attracting attention as a substitute for HAP (Hydrolized Animal Protein) and HVP (Hydrolized Vegetable Protein), and because its flavor is similar to meat flavor, it is widely used as a substitute for beef extract in Gumi. In addition, yeast extract has a complex taste and rich taste, and already has a function not found in other seasonings, such as masking off-flavor. Unlike other seasonings in which only amino acids are the main seasoning ingredients, yeast extract is an advantage because it contains not only amino acids but also components such as peptides and hexane.

그러나 효모 엑기스에는 효모 고유의 효모취가 있어 사용량이 제한되므로, 최근에는 기존의 효모 엑기스 보다 정미성과 감칠맛이 더욱 강하면서, 효모취는 제거되어 있는 고핵산 효모 엑기스를 제조하여 사용하는 추세이다. 고핵산 효모엑기스를 제조하기 위해서는, 핵산 조미료의 성분이 되는 5'-뉴클레오티드를 생성하는 전구체 RNA가 균체내에 다량 함유되어 있어야 한다. 효모 중의 RNA 함량은 균종에 따라 차이가 있으나, 야생균주 상태로서 캔디다(Candida)속 효모는 9%, 제빵용 효모는 7∼8%, 맥주 제조용(brewer) 효모는 5∼6%의 RNA를 함유하고 있는 것으로 알려져 있다. 현재, 고핵산 효모엑기스 제조에 이용되고 있는 효모는 일반적으로 7∼8% 정도로 RNA를 함유하고 있다.However, since yeast extract has a yeast inherent yeast odor, the amount of use is limited, and in recent years, while the taste and rich taste are stronger than the existing yeast extract, yeast odor is a trend to manufacture and use the high-nucleic acid yeast extract is removed. In order to prepare a high nucleic acid yeast extract, a large amount of precursor RNA that generates 5'-nucleotides as a component of nucleic acid seasonings should be contained in the cells. The RNA content in yeast varies depending on the type of fungi, but it is a wild strain containing 9% RNA of Candida genus, 7% to 8% baker's yeast, and 5% to 6% brewer's yeast. It is known to do. Currently, yeast used for producing high-nucleic acid yeast extract generally contains RNA at about 7 to 8%.

일본공개특허 소 56-16824호에는 RNA 함량이 높은 균체를 얻기 위한 방법으로서, 저온감수성 변이주를 연속식 발효법으로 배양하여 RNA 함량을 증가시키는 기술을 기재하고 있다. 이러한 방법에 의할 경우 RNA 함량이 증가된 균체를 얻을 수 있기는 하지만, 야생균주에 대해서 변이주는 상대적으로 불안정하므로 균일성 있는 제품 생산면에서 문제가 있을 수 있고, 또한 변이제 처리에 따른 경비 증가의 문제도 경시할 수 없는 단점으로 지적되어 왔다. 생육배지 조성이나 배지 첨가물을 바꾸는 등 배양환경을 조절함으로써 RNA 함량을 증가시키는 방법도 연구되었는데, 예를들어, 일본공개특허 소40-15960호, 일본공개특허 소 50-16875호, 일본공개특허 소 45-15946호 등에서는 무기염이나 항생물질을 배지중에 첨가하여 RNA 함량을 증가시키는 방법을 기술한다. 이러한 배지 첨가물에 의한 방법은 균체당 RNA 함량을 증가시킬 수는 있으나, 균체생육이 저하하여 최종적으로 얻어지는 전체 RNA 함량이 감소하는 문제가 있었다. 따라서, 일본공개특허 평 5-176757호에는 유가식 배양 중 배지공급속도를 조절하여 효모균체의 생육속도를 0.18h-1미만으로 유지함으로써, 균체농도를 증가시키면서 RNA 함량도 높은 상태를 유지하는 방법을 기재하고 있다. 그러나, 종래 알려져 있는 인공적으로 변이 또는 특수배양하여 RNA 함량이 높은 균주를 얻는 것 보다는, 본래 자연 상태에서 균체 내부에 RNA를 다량 함유하고 있는 균주를 천연적으로 발견, 분리해 내어 이용하는 것이 안정성 있는 균주 확보는 물론 경제적인 측면에 있어서도 훨씬 바람직하다.Japanese Laid-Open Patent Publication No. 56-16824 describes a technique for increasing RNA content by culturing low-temperature susceptible mutants in a continuous fermentation method as a method for obtaining cells with high RNA content. Although this method can be used to obtain cells with increased RNA content, mutant strains are relatively unstable for wild strains, which may cause problems in the production of uniform products. Has also been pointed out as a disadvantage that cannot be neglected. Methods of increasing the RNA content by controlling the culture environment, such as changing the growth medium composition and medium additives, have also been studied. For example, Japanese Patent Application Laid-Open No. 40-15960, Japanese Patent Application Laid-open No. 50-16875, and Japanese Patent Laid-Open 45-15946 et al describe methods for increasing RNA content by adding inorganic salts or antibiotics to the medium. The method using such a medium additive can increase the RNA content per cell, but there is a problem that the total growth of the final RNA content is reduced due to the decrease in cell growth. Therefore, Japanese Laid-Open Patent Publication No. 5-176757 maintains a high RNA content while increasing the cell concentration by maintaining the growth rate of the yeast cells to less than 0.18h -1 by adjusting the medium feed rate during fed-batch culture. It is described. However, rather than obtaining a strain having a high RNA content by artificially mutating or specially culturing it, it is more stable to naturally find, isolate, and use a strain that contains a large amount of RNA in the cell in its natural state. It is much more desirable in terms of security as well as economics.

이에 본 발명자들은 전국의 토양과 과일, 야채 시료를 대상으로 광범위한 조사(screening)를 실시하여 유망한 균주를 분리해 내고, 분리된 균주에 대해서는 액체 배지 중에서 균체내 RNA 함량을 정량하는 등의 방법을 통하여 RNA 함량이 특이적으로 높은 사카로마이세스(Saccharomyces)속에 속하는 새로운 균주(Saccharomyces cerevisiae NS 9로 명명)를 찾아내어 본 발명을 완성하게 되었다.Accordingly, the present inventors have carried out extensive screening of soil, fruit and vegetable samples from all over the country to isolate promising strains and quantify RNA content in the cells in liquid medium. The present invention was completed by finding a new strain (named Saccharomyces cerevisiae NS 9) belonging to the genus Saccharomyces, which has a particularly high RNA content.

따라서, 본 발명의 목적은 균체내에 RNA 함량이 높은 새로운 균주를 제공하는 데에 있다.Accordingly, it is an object of the present invention to provide a new strain having a high RNA content in the cells.

EH한, 본 발명의 목적은 이미, 이취가 없고 정미성과 감칠맛이 뛰어난 고핵산 효모엑기스 조미료 제조에 이용될 수 있는 RNA 함량이 높은 신규한 균주를 제공하는 데에 있다.An object of the present invention is to provide a novel strain having a high RNA content, which can be used for preparing a high-nucleic acid yeast extract seasoning that has no off-flavor and is excellent in taste and taste.

본 발명의 내용을 보다 상세히 설명하면 다음과 같다.Hereinafter, the content of the present invention will be described in detail.

본 발명에 의한 새로운 균주(Saccharomyces cerevisiae NS 9)는 본 발명이 속하는 분야에서는 관용화 되어 있는 균주 스크리닝 방법을 이용하여 선발하였다. 예를 들어, 하기와 같은 방법으로 선발하였다.A new strain (Saccharomyces cerevisiae NS 9) according to the present invention was selected using a strain screening method that is tolerated in the field to which the present invention belongs. For example, it was selected by the following method.

효모를 분리하기 위한 분리 원시료는 일반적으로 효모가 잘 서식할 수 있는 채소류와 과일류 등을 선택하였다. 또한, 토양시료로는 과수원 토양으로서 지표 5∼10cm 깊이의 토양을 채취하였다.As a separate raw material for separating yeast, vegetables and fruits are generally selected in which yeast can inhabit well. In addition, soil samples of 5-10 cm depth were taken as orchard soil as soil samples.

채취한 토양시료는 실험실 내의 통풍이 잘 되는 곳에서 일주일정도 풍건하여 사용하였다.The collected soil samples were used for a week in a well-ventilated place in the laboratory.

효모균주 분리원 시료중 과채 및 발효 식품은 세절한 후 멸균생리식염수에 최종 10%되게 첨가하고 믹서에서 잘 갈고 거즈로 거른후, 10ml를 시험관에 옮겨 정치시켰다. 토양시료도 1g을 멸균 생리식염수 10ml에 넣고 진탕한 후 정치시켰다. 이렇게 한 정치 시료액의 상승액을 취하여 100∼10-5까지 희석한 후 0.2ml를 YM 평판 배지(테트라시 클린 20mg/L, 암피실린 20ml/L 함유)에 30도에서 도말하여 24∼48시간 배양하고, 발생된 콜리니를 순수분리하였다.Fruits and fermented foods in yeast strain isolate samples were chopped and added to sterile saline solution at a final 10%, finely ground in a mixer, filtered with gauze, and then placed in 10 ml of the test tube. Soil samples were also placed in 10 ml of sterile physiological saline and shaken. After taking the synergistic solution of the static sample solution and diluting it to 10 0 to 10 -5 , 0.2 ml was plated at 30 degrees in YM flat medium (containing Tetracycline 20 mg / L and Ampicillin 20 ml / L) and incubated for 24 to 48 hours. The generated colony was purified purely.

이렇게 하여 순수분리된 콜로니를 YM 평판배지에서 재배양하여 콜로니의 모양과 색깔, 광택 등을 관찰하여 콜로니를 1차 선발한 후, 현미경(400배) 관찰에서 단일 셀의 형태, 균사체 형태 등을 분석하여 최종적으로 300주의 효모를 분리하였다. 균체내 RNA 함량을 정량하는 등의 방법을 통하여 RNA 함량이 특이적으로 높은 사카로마이세스(Saccharomyces)속에 속하는 새로운 균주를 선발하였다.In this way, purely isolated colonies were grown in YM plate medium to observe colonies' shape, color, and gloss to select colonies first, and then analyze the shape of single cells and mycelium in a microscope (400 times). Finally, 300 strains of yeast were isolated. New strains belonging to Saccharomyces genus with high RNA content were selected by quantifying RNA content in cells.

본 발명자들은 이러한 방법으로 선발한 균주를 대한민국 균주기탁기관인 유전자 은행에 기탁하여(기탁일 : 1994년 11월 21일) KCTC 8635P의 수탁번호를 부여 받았다.The present inventors deposited the strain selected by this method to the Gene Bank, a strain depositing institution of Korea (deposit date: November 21, 1994) was given an accession number of KCTC 8635P.

본 발명에 따른 새로운 고핵산 균주의 특성은 다음과 같다.The characteristics of the novel high nucleic acid strain according to the present invention are as follows.

(1)균체내 RNA 함량(1) RNA content in cells

와이 엠 브로스(YM broth) 배지를 이용하여 플라스크 배양할때, 균체 생육이 활발한 대수기 중에 13%의 RNA를 균체내에 함유한다.When the flask was cultured using YM broth medium, 13% of the RNA was contained in the cells during the logarithmic growth period.

(2) 형태학적 특성(2) morphological characteristics

형태학적 특성은 사카로마이세스(Saccharomyces)속 효모(yeast)가 갖는 일반적인 특성과 유사하다. 즉, 배양접시(Plate)상에서 흰색 군락(colony)을 형성하고, 균주를 최적 배지조건에서 배양한 후 관찰하면 다변성 발아배(multilateral budding) 형태를 보인다. 또한, 섬유사(filament)와 균막(pellicle)을 지니지 않는 특징 등이 사카로마이세스(Sqaccharomyces)속과 일치한다.Morphological properties are similar to the general properties of yeasts of the genus Saccharomyces. That is, a white colony is formed on a plate, and the strain is observed after culturing in an optimal medium condition to show a multilateral budding form. In addition, the filament and the pellicle-free feature is consistent with the genus Sqaccharomyces.

(3) 생리학적 특성(3) physiological characteristics

분리된 미생물의 생리학적 특성도 사카로마이세스 세레비스(Saccharomyces cerevisiae)가 갖는 일반적 특징과 거의 같다. 즉, 표1 및 2에서 알 수 있는 바와 같이, 분리된 미생물의 당이용 특성은 사카로마이세스 세레비스(Saccharomyces cerevisiae)의 경우와 일치한다.The physiological characteristics of the isolated microorganisms are almost the same as those of Saccharomyces cerevisiae. That is, as can be seen in Tables 1 and 2, the sugar utilization characteristics of the isolated microorganisms are consistent with those of Saccharomyces cerevisiae.

(4) 배양학적 특성(4) culture characteristics

분리된 미생물을 25℃에서 42℃까지의 범위에서 생육 가능하고, 10% 글루코오스를 포함한 배지에서 6% 정도의 에탄올을 생산한다. 그리고 요소를 가수분해하지 못하며, 다아조늄 블루(diazonium blue) B 반응에서 음성을 보이고, 전자전달계에서 조효소 Q를 이용한다.The separated microorganisms can be grown in a range of 25 ° C to 42 ° C, and produce about 6% ethanol in a medium containing 10% glucose. In addition, it does not hydrolyze urea, it is negative in the diazonium blue B reaction, and coenzyme Q is used in the electron transfer system.

이상과 같이 본 발명에 의하여 분리된 미생물에 대한 형태학적, 생리학적, 배양학적 특성 분석 결과가 일반적인 사카로마이세스 세레비스(Saccharomyces cerevisiae)의 특성과 유사하므로, 동 균주가 사카로마이세스 세레비스(Saccharomyces cerevisiae)에 속하는 균주임을 알수 있다. 따라서, 본 발명자들은 동 신규의 균주를 Saccharomyces cerevisiae NS 9으로 명명하고, 대한민국 특허균주 기탁기관인 유전자은행에 1994년 11월 21일에 균주를 기탁하여 KCTC 8635P의 수탁번호를 부여받았다.As described above, the morphological, physiological and culture characteristics of the microorganisms isolated by the present invention are similar to those of Saccharomyces cerevisiae, so that the strain is Saccharomyces cerevisiae. It can be seen that the strain belongs to (Saccharomyces cerevisiae). Therefore, the present inventors named the novel strain Saccharomyces cerevisiae NS 9, and deposited the strain on November 21, 1994 to Genebank, a Korean patent strain depositing institution, was given an accession number of KCTC 8635P.

본 발명의 신규 균주를 본 발명이 속하는 분야에서 관용화된 균체배양 방법에 의하여 배양하면, RNA를 다량 함유한 효모를 수확할 수 있으며, 이를 이용하여 이미, 이취가 없고 정미성과 감칠맛이 뛰어난 고핵산 효모엑기스 조미료를 제조할 수 있다.When the new strain of the present invention is cultured by the conventional cell culture method in the field to which the present invention belongs, it is possible to harvest yeast containing a large amount of RNA, and by using this, there is no odor and excellent taste and taste. Yeast extract seasonings can be prepared.

이하 본 발명의 균주의 배양 방법과 이를 이용한 고핵산 효모엑기스의 제조방법을 하기의 실시예에 의하여 더욱 상세히 설명한다. 이들 실시예는 본 발명의 이해를 돕기 위하여 제공되는 것으로 본 발명의 범위가 이에 한정되는 것은 아니다.Hereinafter, the method for culturing the strain of the present invention and a method for preparing a high nucleic acid yeast extract using the same will be described in more detail by the following examples. These examples are provided to aid the understanding of the present invention, but the scope of the present invention is not limited thereto.

[실시예 1]Example 1

효모배양Yeast culture

효모추출물 3g/ℓ, 맥아추출물 3 g/ℓ, 펩톤 5g/ℓ, 글루코오스 10g/ℓ로 구성된 배양용 배지 2ℓ를 2.5ℓ들이 발효조에 넣어 살균한후 Saccharomyces cerevisiae NS 9 균주 100㎖를 배지에 접종하고, 회전속도 500rpm, 통기량 1vvm, 배양온도 30℃에서 회분식으로 20시간 배양하여 10%의 RNA를 함유하는 Saccharomyces cerevisiae NS 9 균체 15g/ℓ를 얻었다.Two liters of culture medium consisting of 3g / l yeast extract, 3g / l malt extract, 5g / l peptone and 10g / l glucose were sterilized in a 2.5l fermenter, and then 100ml of Saccharomyces cerevisiae NS 9 strain was inoculated into the medium. 15 g / L of Saccharomyces cerevisiae NS 9 cells containing 10% RNA were obtained by incubating batchwise for 20 hours at a rotational speed of 500 rpm, aeration rate of 1 vvm and a culture temperature of 30 ° C. for 20 hours.

얻어진 균체 중의 RNA 함량과 현재 사용되고 있는 고핵산 효모엑기스 제조용 효모(타사 제품)중의 RNA 함량을 비교하여 표 4에 나타내었다.The RNA content in the obtained cells and the RNA content in the yeast (manufactured by other companies) for preparing high nucleic acid yeast extract are shown in Table 4 below.

[실시예 2]Example 2

고핵산 효모 엑기스의 제조Preparation of High Nucleic Acid Yeast Extract

실시예 1에서 얻어진 균체를 가열 처리하고, 세포벽 분해 효소, 단백질 분해 효소, 핵산 분해 효소 등을 차례로 처리한 후 효모취를 제거하여, 5'-뉴클레오티드를 3% 함유한 고핵산 효모 엑기스를 제조하였다.The cells obtained in Example 1 were heat-treated, followed by treatment with cell wall enzymes, proteolytic enzymes, nucleic acid enzymes, and the like, and then the yeast odor was removed to prepare a high nucleic acid yeast extract containing 3% of 5'-nucleotides. .

얻어진 고핵산 효모 엑기스와 일반 자가소화법으로 제조된 표준효모 엑기스의 지미와 효모취에 대한 관능 평가를 실시하여, 그 결과를 표 5에 나타내었다. 결과는 각 점수를 매우 강하다(5점), 강하다(4점), 약하다(3점), 매우 약하다(2점), 느낄 수 없다(1점)로 하여, 관능 시험요원 20명의 점수를 모두 합하여 나타낸 것이다.The sensory evaluation of Jimmy and yeast odor of the obtained high-nucleic acid yeast extract and the standard yeast extract prepared by the general self-extinguishing method was performed, and the results are shown in Table 5. The result is that the scores are very strong (5 points), strong (4 points), weak (3 points), very weak (2 points), and insignificant (1 point). It is shown.

표 5로부터, 본 발명의 Saccharomyces cerevisiae NS 9 균주를 이용함으로써 지미가 강한 반면 효모취는 상당히 제거된 고핵산 효모 엑기스를 제조할 수 있음을 알 수 있다.From Table 5, it can be seen that by using the Saccharomyces cerevisiae NS 9 strain of the present invention, a high-nucleic acid yeast extract with strong Jimmy but yeast odor can be prepared.

Claims (2)

와이 엠 브로스(YM broth) 배지에서 배양시 대수기 중에 13%의 RNA를 균체 내에 함유하며, 대한민국 유전자은행에 수탁번호 KCTC 8635P로 기탁되어 있는 Saccharomyces cerevisiae NS 9 균주.Saccharomyces cerevisiae NS 9 strain which contains 13% of RNA in cells during incubation in YM broth medium and has been deposited with accession number KCTC 8635P in the Korean Gene Bank. 고핵산 효모 엑기스 제조에 사용되는 제1항에 따른 균주.The strain according to claim 1 used for producing a high nucleic acid yeast extract.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100442574B1 (en) * 2001-08-31 2004-08-02 주식회사농심 Novel yeast mutant containing increased RNA content and method of mass-producing of the same
KR101189888B1 (en) 2010-03-29 2012-10-10 샘표식품 주식회사 Novel yeast mutant containing increased RNA content, method for producing the same and use thereof
KR20170012693A (en) 2015-07-22 2017-02-03 영남대학교 산학협력단 Osmotolerant Saccharomyces cerevisiae strain isolated from Korean Grapes

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100442574B1 (en) * 2001-08-31 2004-08-02 주식회사농심 Novel yeast mutant containing increased RNA content and method of mass-producing of the same
KR101189888B1 (en) 2010-03-29 2012-10-10 샘표식품 주식회사 Novel yeast mutant containing increased RNA content, method for producing the same and use thereof
KR20170012693A (en) 2015-07-22 2017-02-03 영남대학교 산학협력단 Osmotolerant Saccharomyces cerevisiae strain isolated from Korean Grapes

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