KR100442574B1 - Novel yeast mutant containing increased RNA content and method of mass-producing of the same - Google Patents
Novel yeast mutant containing increased RNA content and method of mass-producing of the same Download PDFInfo
- Publication number
- KR100442574B1 KR100442574B1 KR10-2001-0053264A KR20010053264A KR100442574B1 KR 100442574 B1 KR100442574 B1 KR 100442574B1 KR 20010053264 A KR20010053264 A KR 20010053264A KR 100442574 B1 KR100442574 B1 KR 100442574B1
- Authority
- KR
- South Korea
- Prior art keywords
- yeast
- mutant
- nucleic acid
- rna content
- medium
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
본 발명은 식품산업에서 천연 풍미 및 향미 소재로 사용되는 효모(Sacchromyces cerevisiae)를 돌연변이제(EMS ; Ethyl Methanesulfonate)로 돌연변이 시켜 얻은 고핵산 효모 변이주 특히,S. cerevisiaeM40-10에 관한 것이다. 또한 본 발명은 당밀과 CSL(corn steep liquor) 등 산업용 배지에서 이 변이주를 배양하여 변이주를 대량 생산하는 방법에 관한 것이다.The present invention relates to a high nucleic acid yeast mutant strain obtained by mutating a yeast ( Sacchromyces cerevisiae ) used as a natural flavor and flavoring material in the food industry with a mutant (EMS; Ethyl Methanesulfonate), in particular, S. cerevisiae M40-10. The present invention also relates to a method for mass production of mutant strains by culturing the mutant strains in an industrial medium such as molasses and corn steep liquor (CSL).
Description
본 발명은 RNA 함량이 증대된 사카로마이세스 세레비지에(Saccharomyces cerevisiae) 변이주에 관한 것으로, 더욱 상세하게는 식품산업에서 천연 풍미 소재로 유용하게 사용될 수 있는 높은 세포농도 및 RNA농도를 갖는 고핵산 효모 변이주 특히, 사카로마이세스 세레비지에 M40-10(KCTC 1030BP) 및 그 대량 생산 방법에 관한 것이다.The present invention relates to Saccharomyces cerevisiae mutants with increased RNA content, and more particularly, high nucleic acid having high cell concentration and RNA concentration, which can be usefully used as a natural flavor material in the food industry. Yeast mutant strains, in particular, Saccharomyces cerevisiae M40-10 (KCTC 1030BP) and its mass production method.
식품 산업에서 널리 사용되는 풍미 소재의 사용량이 증가함에 따라, 다양한 소재개발이 이루어지고 있으며, 특히 최근 들어 화학적 합성으로 만든 제품에 대한 기호도가 떨어지면서 MSG (monosodium glutamic acid)를 대체할 수 있는 천연 풍미 소재에 대한 관심이 높아지고 있다. 이러한 천연 풍미 소재로 등장한 것이 바로효모세포 추출물이다.As the amount of flavoring materials widely used in the food industry increases, various materials are being developed. In particular, recently, natural flavors, which can replace MSG (monosodium glutamic acid), have become less palatable for products made by chemical synthesis. Interest in materials is increasing. It is the yeast cell extract that appeared as a natural flavor material.
일반적으로 효모는 GRAS(generally recognized as safe)의 범주에 들어가는 안전한 미생물로 식품 및 음료에 있어서 알콜 발효에 이용될 뿐만 아니라, 효모 세포 자체를 보강하거나 변형시켜 얻어낸 추출물 등이 다양하게 이용되고 있다.Generally, yeast is a safe microorganism that falls into the category of GRAS (generally recognized as safe), and is used not only for alcohol fermentation in foods and beverages, but also for extracts obtained by reinforcing or modifying yeast cells themselves.
효모는 핵산이 비교적 풍부한 미생물로써, 그 함량은 균종에 따라 차이가 있으나, 약 7 내지 12% 정도의 RNA 함유량을 갖는다. 이러한 효모 내의 핵산을 추출하여 화학적으로 또는 효소에 의해 분해하면, 뉴클레오타이드가 생성되며, 이러한 뉴클레오타이드는 다음과 같은 조건에서 정미성을 가지므로 일종의 핵산 조미료의 원료가 된다.Yeast is a microorganism that is relatively rich in nucleic acid, the content of which varies depending on the species, but has an RNA content of about 7 to 12%. When nucleic acids in such yeast are extracted and chemically or decomposed by enzymes, nucleotides are produced, and these nucleotides have a taste in the following conditions, and thus are a kind of nucleic acid seasoning.
일반적으로 뉴클레오타이드가 정미성을 나타내는 조건은, 첫째로, 고분자 뉴클레오타이드나 뉴클레오사이드가 아닌 모노뉴클레오타이드일 것, 둘째로, 염기가 퓨린계의 것일 것, 세째로, 퓨린 환의 6번 위치에 OH기를 가질 것, 넷째로, 리보오스의 5'위치에 인산기를 가질 것, 다섯째, 뉴클레오타이드의 당은 리보오스, 또는 데옥시리보오스 두 가지 다 정미성을 갖지만, 리보오스일 경우 더욱 바람직하다는 것 등이다.In general, the conditions in which the nucleotides show the taste are: first, the polymer nucleotide or the non-nucleoside mononucleotide, second, the base is purine-based, third, the OH group at the 6 position of the purine ring And, fourthly, to have a phosphate group at the 5 'position of the ribose. Fifth, both sugars of ribose or deoxyribose have taste, but are more preferable in the case of ribose.
따라서 효모로부터 상기와 같은 정미성 뉴클레오타이드를 높은 수율로 얻기 위하여는 RNA 함량이 높은 고핵산 효모 균주가 필요하다.Therefore, high nucleic acid yeast strains with high RNA content are required in order to obtain such nucleotides from yeast in high yield.
종래, RNA 함량을 높인 효모 균주를 얻기 위한 방법으로는, UV(1.9W/m2를 10초 동안 조사)와 γ-ray (45 Gy)를 40%의 세포가 살아남는 조사시간 동안 조사하여 돌연변이를 일으켜서 고핵산 효모 변이주를 선별하는 방법이 있었다. 이와 같이 선별된 균주는 건조 균체량당 RNA 함량이 모균주에 비하여 높기는 하나 그 효과가 현저하지는 못하였다. 이는, 효모 균주에 있어서 RNA 함량을 높이는 것은 다른 대사 산물의 수율을 높이는 것에 비하여 그 효과를 얻기가 매우 힘들기 때문이다.Conventionally, in order to obtain a yeast strain with a high RNA content, UV (1.9 W / m 2 irradiated for 10 seconds) and γ-ray (45 Gy) irradiated 40% of the cells during the irradiation time of the mutation and mutation There was a way to screen for high nucleic acid yeast mutants. The selected strains had higher RNA content per dry cell weight than the parent strain, but the effect was not remarkable. This is because increasing the RNA content in the yeast strain is very difficult to obtain the effect compared to increasing the yield of other metabolites.
본 발명의 목적은 정미성 뉴클레오타이드를 높은 수율로 생산할 수 있는, 높은 RNA 함량을 갖는 고핵산 효모 돌연변이주를 제공하는 것이다.It is an object of the present invention to provide a high nucleic acid yeast mutant strain having a high RNA content, which can produce a high yield of nucleotide nucleotides.
또한 본 발명의 목적은, 상기와 같은 고핵산 효모 돌연변이주를 대량 생산하는 방법을 제공하는 것이다.It is also an object of the present invention to provide a method for mass production of such high nucleic acid yeast mutant strains.
도 1은 본 발명의 고핵산 효모 변이주를 발효조 회분 배양했을 때의 세포 농도와 RNA농도를 나타낸 그래프이다.1 is a graph showing the cell concentration and RNA concentration when the cultured high nucleic acid yeast mutant strain of the present invention.
상기와 같은 목적을 달성하기 위하여 본 발명은,The present invention to achieve the above object,
모균주인 사카로마이세스 세레비지에(Saccharomyces cerevisiae)를 화학적 돌연변이제(EMS)로 처리한 후, 효모의 산성 혹은 알칼리성 RNase의 활성을 억제하는 금속염이 첨가된 배지에서 자라지 못하는 콜로니를 선별하여 분리한, 높은 RNA 함량을 갖는 고핵산 효모 돌연 변이주를 제공한다. 이 때, RNase의 활성을 억제하는 금속염은 KCl인 것이 바람직하다. 또한 바람직하게는 상기의 고핵산 효모 돌연변이주는,S. cerevisiaeM40-10(KCTC 1030BP)이다.After treatment with Saccharomyces cerevisiae , a parent strain, with a chemical mutant (EMS), colonies that do not grow in the medium containing metal salts that inhibit the activity of acidic or alkaline RNase in yeast are screened and isolated. One provides a high nucleic acid yeast mutant strain with a high RNA content. At this time, the metal salt that inhibits the activity of RNase is preferably KCl. Also preferably, the high nucleic acid yeast mutant strain is S. cerevisiae M40-10 (KCTC 1030BP).
또한 본 발명은, 상기의 고핵산 효모 돌연 변이주를 당밀과 CSL을 포함하는 산업용배지로 배양하여 대량으로 생산하는 방법을 제공한다.In another aspect, the present invention provides a method for producing a large amount by culturing the high nucleic acid yeast mutant strain in an industrial medium containing molasses and CSL.
이하, 본 발명을 더욱 상세히 설명한다.Hereinafter, the present invention will be described in more detail.
효모 균체의 RNA 함량은 합성 속도와 분해 속도의 평형에 의해 결정될 수 있으며, 분해를 억제한다면 균체내 RNA 함량은 자연적으로 증가하게 된다. 본 발명에서는 이러한 점에 착안하여, RNA 분해에 관여하는 효모의 산성 혹은 알카리성 RNase의 활성을 억제하는 금속염을 찾아 돌연변이주 선별 방법에 이용하고자 하였다.The RNA content of the yeast cells can be determined by the equilibrium of the synthesis rate and the degradation rate. If the degradation is inhibited, the RNA content in the cells increases naturally. In view of the above, the present invention was intended to find a metal salt that inhibits the activity of the acidic or alkaline RNase of the yeast involved in RNA degradation, and to use it in the mutant strain selection method.
즉, 산성 및 알카리성 RNase가 모두 억제된 효모 변이주는 치사 변이주 이며, 일반 배지에서 생육이 가능하나 알칼리성 RNase를 억제하는 배지에서 자라지 못하는 치사 변이주는, 산성 RNase 활성이 억제되거나 핵산 분해 대사에 이상이 생겨 효모의 정상 생육 pH인 산성에서 RNA를 세포내에 다량 축적하게 된다. Na+와 K+의 경우 RNase의 활성을 특이적으로 저해하며, 같은 농도에서 K+가 저해효과가 더 뛰어 나다. 캔디다 리폴리티카(Candida lipolytica)의 RNase 활성에 금속염이 미치는 영향에 대한 연구에서도 알칼리성 RNas의 활성이 Na+와 K+에 의해 저해되는 것으로 나타난 바 있다.In other words, yeast mutants that inhibit both acidic and alkaline RNases are lethal mutants, and lethal mutants that can be grown in normal medium but do not grow in medium that inhibits alkaline RNases are inhibited in acidic RNase activity or have abnormalities in nucleic acid metabolism. At acidic, normal growth pH of yeast, RNA is accumulated in cells. Na + and K + specifically inhibit the activity of RNase, and at the same concentration K + is more effective. Studies on the effect of metal salts on the RNase activity of Candida lipolytica have been shown to inhibit the activity of alkaline RNas by Na + and K + .
본 발명에서는, 이러한 사실을 기초로,S. cerevisie, 특히S.cerevisieMTY62를 EMS로 처리하여 돌연변이 시켜서 얻은 변이주를 YPD 배지 및 1.5 M KCl이 첨가된 YPD 배지에 도말하여, YPD 배지에서는 잘자라고, 1.5 M KCl이 첨가된 YPD 배지에서는 자라지 못하는 콜로니를 선별하여 이를 고핵산 효모 변이주로 선별하였다. 상기에서 KCl 첨가 이유는 고농도의 KCl이 알카리성 RNase를 저해하므로 RNA를 축적하는 효모를 효율적으로 선별하기 위해서이다.In the present invention, on the basis of this fact, S. cerevisie, in particular to smear the mutant obtained by mutagenesis with EMS to handle S.cerevisie MTY62 in YPD medium with the YPD culture medium, and 1.5 M KCl is added, in the YPD medium good night, Colonies that did not grow in YPD medium added with 1.5 M KCl were selected as high nucleic acid yeast mutants. The reason for adding KCl is to efficiently select yeasts that accumulate RNA because KCl inhibits alkaline RNase.
상기와 같은 방법으로 선별한 균주는 대한민국 균주기탁기관인 유전자 은행에 기탁하여 2001년 6월 13일 KCTC 1030BP의 기탁번호를 부여받았다.The strains selected in the above manner were deposited with the Gene Bank, a strain depositing institution of the Republic of Korea, and received the accession number of KCTC 1030BP on June 13, 2001.
본 발명에 따라 선별된 효모 변이주는 세포내 RNA 함량 및 RNA 농도가 모균주에 비하여 높게 나타나는데, 이는 RNA 함량은 단 1%도 증가시키기 어려운 점을 감안할 때 매우 현저한 효과라고 볼 수 있다.The yeast mutant strain selected according to the present invention appears to have a high intracellular RNA content and RNA concentration compared to the parent strain, which can be seen as a very significant effect given that the RNA content is difficult to increase by only 1%.
이하, 실시예를 통하여 본 발명의 효모 변이주 및 그 유도 방법에 대하여 더욱 상세히 설명한다.Hereinafter, the yeast mutant strain of the present invention and a method of inducing the same will be described in more detail with reference to Examples.
하기하는 실시예는 본 발명을 설명하기 위한 것이며, 본 발명을 한정하고자 하는 의도로 이해되어서는 안 될 것이다.The following examples are intended to illustrate the invention and should not be understood as intended to limit the invention.
실시예 1- 고핵산효모변이주의 분리 Example 1 Isolation of High Nucleic Acid Yeast Mutants
고핵산 돌연변이 균주는 다음과 같이 분리하였다.The high nucleic acid mutant strain was isolated as follows.
먼저, 돌연변이 시키고자 하는 효모를 24시간 배양하여 세포농도가 약 2×108cells/mL일 때 원심 분리하여 효모 균체를 얻은 후, 1 mL(50 mM) 인산완충액 (pH 7.0)으로 두 번 세척한 다음 4 mL(50 mM) 인산완충액 (pH 7.0)에 현탁하였다.First, the yeast to be mutated is incubated for 24 hours and centrifuged when the cell concentration is about 2 × 10 8 cells / mL to obtain yeast cells, and then washed twice with 1 mL (50 mM) phosphate buffer (pH 7.0). It was then suspended in 4 mL (50 mM) phosphate buffer (pH 7.0).
그리고 나서, 대조구로서는, EMS를 첨가하지 않고, 100∼200개의 콜로니가 형성되도록 YPD 배지에 적절한 배율로 희석·도말하였다. Then, as a control, without adding EMS, dilution and coating were carried out at an appropriate magnification ratio in the YPD medium so that 100 to 200 colonies were formed.
실험구로서는, 돌연변이시키기 위해 현탁한 배양액에 120 ㎕의 EMS (0.3%)를 첨가하고 30℃에서 반응시켜서, 반응 0분부터 10분 간격으로 반응액을 채취하여 5% 소듐 티오설페이트 (돌연변이 정지제)를 반응액과 동량을 넣은 후, YPD 배지 (1% 효모추출물, 2% 펩톤, 2% 덱스트로스)에 적절히 희석, 도말하였다. 상기와 같이 희석·도말한 후 생성된 콜로니 수를 대조구와 비교해 생존율이 40∼60% 일 때의 콜로니를 선별하였다.As a test group, 120 μl of EMS (0.3%) was added to the culture suspension to be mutated and reacted at 30 ° C., and the reaction solution was collected every 10 minutes from 0 minutes to 5% sodium thiosulfate (mutant stopper). ) Was added to the reaction solution, and then diluted and plated in YPD medium (1% yeast extract, 2% peptone, 2% dextrose) as appropriate. After dilution and plating as described above, the number of colonies produced was compared with the control group, and colonies with a survival rate of 40 to 60% were selected.
이렇게 해서 얻은 변이주를 YPD 배지 및 1.5 M KCl이 첨가된 YPD 배지에 도말하여 YPD 배지에서는 잘자라고, 1.5 M KCl이 첨가된 YPD 배지에서는 자라지 못하는 콜로니를 선별하여 이를 고핵산 효모 변이주로 선별하였다.The mutants obtained in this way were plated on YPD medium and YPD medium to which 1.5 M KCl was added, and colonies that grew well in YPD medium and did not grow on YPD medium to which 1.5 M KCl was added were selected as high nucleic acid yeast strains.
이와 같이 선별한 고핵산 변이주를 YPD 배지 및 산업용배지 (10% 당밀, 5% CSL)에서 배양하여, 최종적으로 고핵산 변이주 M40-10을 선별하였다.The high nucleic acid mutant strains thus selected were cultured in YPD medium and industrial medium (10% molasses, 5% CSL) to finally select the high nucleic acid mutant strain M40-10.
이 고핵산 변이주 M40-10은 2001년 6월 13일에 한국 유전자 은행에 KCTC 1030BP의 기탁번호로 기탁하였다.The high nucleic acid mutant strain M40-10 was deposited on June 13, 2001, with a deposit number of KCTC 1030BP to the Korea Gene Bank.
상기의 방법에 따라 선별된 고핵산 효모 변이주 M40-10( )을 10% 당밀과 5% CSL이 첨가된 산업용배지를 이용하여 발효조 회분배양하여 대량 생산하였다.The high-nucleic acid yeast mutant strain M40-10 () selected according to the above method was mass-produced by batch fermentation using an industrial medium to which 10% molasses and 5% CSL were added.
실시예 2- 효모변이주의 균학적 성질 Example 2 -Mycological Properties of Yeast Mutants
본 발명의 변이주의 균학적 성질은 모균주인S. cerevisiaeMTY62와 대체로 유사하며, 다음과 같다.The mycological properties of the mutant strains of the present invention are generally similar to that of the parent strain S. cerevisiae MTY62.
실시예 3- 세포농도, RNA 농도 및 RNA 함량 측정 Example 3 Measurement of Cell Concentration, RNA Concentration and RNA Content
모균주S. cerevisiaeMTY62와 본 발명의 변이주S. cerevisiaeM40-10의 세포농도 및 RNA 농도와 함량을 측정하였다.The cell concentration and the RNA concentration and the amount of the parental strain S. cerevisiae mutants of the present invention MTY62 and S. cerevisiae M40-10 were measured.
1차 종배지로는 YPD 배지(1% Yeast Extract, 2% Peptone, 2% Dextrose)를 이용하였고, 2차 종배지로는 산업용 배지 (5% 당밀, 2% CSL)를 이용하였다.YPD medium (1% Yeast Extract, 2% Peptone, 2% Dextrose) was used as the primary seed medium, and industrial medium (5% molasses, 2% CSL) was used as the secondary seed medium.
발효배지로는 산업용배지 (10% 당밀, 5% CSL), 0.03% 우레아, 0.004% KH2PO4, 0.02% MgSO4, 0.02% ZnSO4, 1 ppm 비오틴을 이용하였다. 산업용배지의 사용시 당밀은 105℃에서 30분 열처리 후 7,000 rpm에서 7분 원심 분리하여 얻은 상등액을 사용하였고, CSL은 7,000 rpm에서 7분간 원심분리하여 사용하였다.As fermentation medium, industrial medium (10% molasses, 5% CSL), 0.03% urea, 0.004% KH 2 PO 4 , 0.02% MgSO 4 , 0.02% ZnSO 4 , 1 ppm biotin was used. The molasses of the industrial medium was used as a supernatant obtained by centrifugation at 7,000 rpm for 7 minutes after heat treatment at 105 ° C. for 30 minutes, and CSL was used by centrifugation at 7,000 rpm for 7 minutes.
발효 배양방법은 다음과 같다.Fermentation culture method is as follows.
먼저, 전배양으로 고핵산 효모 변이주를 1차 종배지 함유 시험관에서 18시간 배양한 다음, 2차 종배지 50 mL 함유 플라스크 두 개에 15 시간 배양 후, 발효조에 접종하였다. 배양 조건은 작업 부피 1.0 L, pH 5.5, 30 ℃, 교반 속도 300∼600 rpm, DO 2ppm을 공기포화의 30% 이상으로 유지하였다. 이와 같은 조건으로 30 시간까지 배양했을 때, 시간에 따른, RNA 농도와 균체 농도의 변화를 도 1의 그래프에 나타내었다.First, the high-nucleic acid yeast mutant strains were cultured for 18 hours in a test tube containing a primary seed medium as a preculture, and then cultured in two flasks containing 50 mL of a secondary seed medium for 15 hours, and then inoculated into a fermenter. Culture conditions were 1.0 L working volume, pH 5.5, 30 ° C., stirring speed 300-600 rpm, DO 2 ppm was maintained above 30% of air saturation. When cultured up to 30 hours under these conditions, changes in RNA concentration and cell concentration over time are shown in the graph of FIG. 1.
또한 상기와 같이 모균주S. cerevisiaeMTY62와 본 발명의 변이주S. cerevisiaeM40-10를 산업용배지를 이용하여 발효조 회분배양했을 때의 각 균주의 세포농도, RNA농도, 및 RNA 함량을 비교한 결과를 표 1에 나타내었다.In addition, the result of comparing the cell concentration, RNA concentration, and the RNA content of each of the strain when cultured using the batch fermenters industrial medium a parent strain S. cerevisiae mutants of the present invention MTY62 and S. cerevisiae as the M40-10 Table 1 shows.
본 발명에 따라 얻어진 효모 변이주는 산업용배지로 발효조 회분 배양했을 때, 모균주에 비해 세포농도는 약 10∼20%, RNA농도는 약 20% 이상, RNA 함량은 약 10% 증가한 결과를 나타내어, 고핵산 효모를 얻을 수 있었다. 이러한 효과는 세포 내 RNA 함량을 증가시키는 것이 매우 어려운 점임을 감안할 때, 매우 현저한 효과라고 할 수 있다. 이와 같이 얻은 고핵산 효모는 천연 풍미 소재로서 안전할 뿐만 아니라 정미성이 뛰어나므로, 천연 조미료의 원료로서 유용하게 사용될 수 있다.Yeast mutant strain obtained according to the present invention when cultured in a batch of fermenter in an industrial medium, cell concentration is about 10 to 20%, RNA concentration is about 20% or more, RNA content is increased by about 10% compared to the parent strain, Nucleic acid yeast could be obtained. This effect is very significant considering that it is very difficult to increase the RNA content in the cell. The high-nucleic acid yeast thus obtained is not only safe as a natural flavor material but also excellent in taste, and thus can be usefully used as a raw material for natural seasonings.
Claims (4)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2001-0053264A KR100442574B1 (en) | 2001-08-31 | 2001-08-31 | Novel yeast mutant containing increased RNA content and method of mass-producing of the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2001-0053264A KR100442574B1 (en) | 2001-08-31 | 2001-08-31 | Novel yeast mutant containing increased RNA content and method of mass-producing of the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20030018736A KR20030018736A (en) | 2003-03-06 |
| KR100442574B1 true KR100442574B1 (en) | 2004-08-02 |
Family
ID=27721844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR10-2001-0053264A KR100442574B1 (en) | 2001-08-31 | 2001-08-31 | Novel yeast mutant containing increased RNA content and method of mass-producing of the same |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR100442574B1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5102076B2 (en) * | 2008-03-06 | 2012-12-19 | アサヒグループホールディングス株式会社 | A Saccharomyces cerevisiae mutant and a method for producing a yeast with a high RNA content using the mutant. |
| CN116286939B (en) * | 2023-02-03 | 2023-09-01 | 齐鲁工业大学(山东省科学院) | Method for improving nucleic acid yield of saccharomyces cerevisiae and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05176757A (en) * | 1990-02-15 | 1993-07-20 | Daicel Chem Ind Ltd | Production of yeast cell |
| KR0141404B1 (en) * | 1995-03-31 | 1998-06-15 | 이상윤 | Novel saccharomyces cerevisiae ns53 which produces high density nucleic acid |
| KR100208968B1 (en) * | 1995-03-31 | 1999-07-15 | 이상윤 | A novel saccharomyces cerevisiae ns9 which is used for producing yeast extract contained high density of nucleic acid |
-
2001
- 2001-08-31 KR KR10-2001-0053264A patent/KR100442574B1/en not_active IP Right Cessation
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05176757A (en) * | 1990-02-15 | 1993-07-20 | Daicel Chem Ind Ltd | Production of yeast cell |
| KR0141404B1 (en) * | 1995-03-31 | 1998-06-15 | 이상윤 | Novel saccharomyces cerevisiae ns53 which produces high density nucleic acid |
| KR100208968B1 (en) * | 1995-03-31 | 1999-07-15 | 이상윤 | A novel saccharomyces cerevisiae ns9 which is used for producing yeast extract contained high density of nucleic acid |
Non-Patent Citations (3)
| Title |
|---|
| Korean journal of food science & technology vol 31 (2)" and the pp.465-474 (1999. * |
| Korean journal of food science & technology vol 31 (2)" and the pp.465-474 (1999.) * |
| Vol. 28, No. 1, pp. 122~126, 1996 * |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20030018736A (en) | 2003-03-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US9556444B2 (en) | Non-recombinant saccharomyces strains that grow on xylose | |
| Khurshid et al. | Mutation of Aspergillus niger for hyperproduction of citric acid from black strap molasses | |
| WO1993001299A1 (en) | Novel yeast strains for the production of xylitol | |
| EP2256184B1 (en) | Saccharomyces cerevisiae mutants and a production process of RNA-rich yeast utilizing the mutants | |
| KR100442574B1 (en) | Novel yeast mutant containing increased RNA content and method of mass-producing of the same | |
| US5998181A (en) | Fermentation process for preparing xylitol using Candida tropicalis | |
| Gasent-Ramírez et al. | Characterization of genetically transformed Saccharomyces cerevisiae baker's yeasts able to metabolize melibiose | |
| JP3023326B2 (en) | Trichosporonoides mutant strain and method for producing erythritol using the same | |
| US6287830B1 (en) | Fermentation process for preparing erythritol by a high salt tolerant mutant of Candida sp. | |
| Ogawa et al. | Hybridization and breeding of the benomyl resistant mutant, Trichoderma harzianum antagonized to phytopathogenic fungi by protoplast fusion | |
| JPH11196859A (en) | Mutant | |
| US6916639B2 (en) | Erythritol-producing moniliella strains | |
| US20040214308A1 (en) | Gamma-glutamylcystein-producing yeast and method of screening the same | |
| EP1092781B1 (en) | Fermentation process for preparing erythritol by a high salt tolerant mutant of candida sp. | |
| Christensen | Cross-breeding of distillers' yeast by hybridization of spore derived clones | |
| Fiedurek et al. | Direct methods for selecting mutants with increased production of invertase from mutagenized cultures of Aspergillus fumigatus | |
| KR100434518B1 (en) | Erythritol fermentation method by a novel strain, Pseudozyma tsukubaensis | |
| KR100314903B1 (en) | Novel yeast strain with high content of nucleic acid, Candida norvegica | |
| WO2005054442A2 (en) | Yeast strains resistant to purine base analogues, process for the production thereof, yeast extracts obtainable from said yeast strains and use of said yeast strains or yeast extracts in food, drink and beverabes | |
| KR101189888B1 (en) | Novel yeast mutant containing increased RNA content, method for producing the same and use thereof | |
| KR100262249B1 (en) | Yeast strain containing organic germanium at high concentration in cells and its manufacturing method | |
| KR100955325B1 (en) | Novel Brevibacterium sp. and Preparation Method of L-Ornithine using Thereof | |
| KR101233667B1 (en) | Mutants of Candida Utilis Containing High-Concentration Ribonucleic Acid | |
| Yoshida et al. | Production of lactase by mutants of Kluyveromyces lactis | |
| CS252991B1 (en) | Method of yeasts and strain saccharomyces cerevisiae ccy 22-15-11 and ccy 22-15-12 bredding won like this |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant | ||
| FPAY | Annual fee payment |
Payment date: 20130530 Year of fee payment: 10 |
|
| FPAY | Annual fee payment |
Payment date: 20140630 Year of fee payment: 11 |
|
| LAPS | Lapse due to unpaid annual fee |