KR890005265A - New Yeast Strains Providing Increased Glucose Efficacy, Methods of Making Such Yeasts, and Use of These Yeasts - Google Patents

New Yeast Strains Providing Increased Glucose Efficacy, Methods of Making Such Yeasts, and Use of These Yeasts Download PDF

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KR890005265A
KR890005265A KR1019880011377A KR880011377A KR890005265A KR 890005265 A KR890005265 A KR 890005265A KR 1019880011377 A KR1019880011377 A KR 1019880011377A KR 880011377 A KR880011377 A KR 880011377A KR 890005265 A KR890005265 A KR 890005265A
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안네 오싱가 클라아스
프란시스쿠스 보이데케르 로베르트
베르투스 반 데르 플라아트 요하네스
아브라함 데 홀란데르 요하네스
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안스 월터 라벤
기스트브로카데스 나암로제 베노오트하프
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Abstract

New yeast strains providing for an enhanced rate of the fermentation of sugars, and a process to obtain such yeasts and the use of these yeasts. Yeasts capable of improved fermentation of sugars, a process to obtain these yeasts and the use of these yeasts are provided. The yeasts show higher rates of metabolism resulting in for example higher carbon dioxide and ethanol production in media containing sugars, such as maltose, as main carbon and energy source. The fermentation rate of sugars is improved by the introduction into a yeast of one or more DNA constructs comprising at least one gene encoding a protein promoting the uptake and/or initial metabolic conversion of a transported sugar substrate.

Description

증가된 당 발효율을 제공하는 신규한 효모균주, 그러한 효모의 제조방법 및 이들 효모의 사용New Yeast Strains Providing Increased Glucose Efficacy, Methods of Making Such Yeasts, and Use of These Yeasts

본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음As this is a public information case, the full text was not included.

제1도는 플라스미드 pGb-eMAL69의 제조과정을 도시한다. 굵은 화살표는 표시된 유전자의 전사의 방향을 가리킨다. 플라스미드는 일정비례로 하지않고 대략적으로 도시되었다.1 shows the preparation of plasmid pGb-eMAL69. The thick arrows indicate the direction of transcription of the indicated genes. Plasmids are shown roughly and not in proportion.

제2도는 pGb-eMAL61의 제조과정을 도시한다. 플라스미드는 일정비례로 하지 않고 대략적으로 도시되었다.2 shows the preparation of pGb-eMAL61. Plasmids are shown roughly, not in proportion.

제3도는 플라스미드 pGb-eMAL63의 제조과정을 도시한다. 플라스미드는 일정비례로 하지 않고 대략적으로 도시되었다.3 shows the preparation of plasmid pGb-eMAL63. Plasmids are shown roughly, not in proportion.

Claims (41)

형질전환되지 않은 어버이 효모와 비교하여 증가된 당 발효율을 제공하는 형질전환된 효모에 있어서, 상기 형질전환된 효모는 : 상기 효모에 존재하는 적어도 하나의, 바람직하게는 상동 DNA구조를 포함하며, 상기 DNA구조는 운반된 당기질의 흡수 및/또는 초기 대사 전환을 촉진하는 단백질을 코드화하고, 상기 효모에서 발현되는 유전자를 적어도 하나 포함하며, 유전자는 상기 효모에서 발현할 수 있게되는 것을 특징으로 하는 효모.In a transformed yeast that provides increased sugar developmental efficiency compared to an untransformed parental yeast, the transformed yeast comprises: at least one, preferably homologous DNA structure present in the yeast, The DNA structure encodes a protein that promotes uptake and / or early metabolic conversion of the transported pulley, and includes at least one gene expressed in the yeast, wherein the gene is capable of being expressed in the yeast. . 제1항에 있어서, 상기 구조는 말토오스 투과효소 활성, 말타아제 활성 또는 말토오스 조절 단백질 활성, 또는 그의 어느 조합을 가지는 효소를 코드화하는 유전자를 적어도 하나 포함하는 것을 특징으로 하는 효모.The yeast of claim 1, wherein the structure comprises at least one gene encoding an enzyme having maltose permease activity, maltase activity or maltose regulatory protein activity, or any combination thereof. 제2항에 있어서, 상기 유전자 또는 그의 조합은 글루코스 억제에 민감하지 않고 말토오스 유도에 지배되지 않는 전사단계 제어하에 놓이게 되는 것을 특징으로 하는 효모.3. The yeast of claim 2, wherein said gene or combinations thereof are placed under transcriptional stage control that is not sensitive to glucose inhibition and is not governed by maltose induction. 제2항에 있어서, 상기 유전자는 알코올데히드로게나제 I(ADHI) 및/또는 번역 연장인자(EFlαA)촉진유전자의 전사단계 제어하에 있음을 특징으로 하는 효모.3. The yeast of claim 2, wherein the gene is under the control of the transcriptional stage of the alcohol dehydrogenase I (ADHI) and / or translational prolongation factor (EFlαA) promoter. 제4항에 있어서, 상기 촉진유전자는 시카로마이세스 속에 속하는 효모, 바람직하게는 맥주효모균으로 부터 유도되는 것을 특징으로 하는 효모.5. The yeast of claim 4, wherein said promoter is derived from yeast, preferably brewer's yeast, belonging to the genus Cicaromyces. 제1항 내지 제5항중 어느 하나에 있어서, 상기 구조는 에피솜성 성분의 일부인 것을 특징으로 하는 효모.The yeast according to any one of claims 1 to 5, wherein the structure is part of an episomal component. 제1항 내지 제6항중 어느 하나에 있어서, 상기 구조는 상기 효모의 염색체안으로 함입되는 것을 특징으로 하는 효모.The yeast according to any one of claims 1 to 6, wherein the structure is incorporated into the chromosome of the yeast. 제1항 내지 제7항중 어느 한 항에 있어서, 적어도 2개의 상기 구조를 포함하는 것을 특징으로 하는 효모.8. Yeast according to any one of claims 1 to 7, comprising at least two of said structures. 제1항 내지 제8항중 어느 한 항에 있어서, 상기 효모에는 이종 DNA가 없음을 특징으로 하는 효모.9. The yeast according to claim 1, wherein said yeast is free of heterologous DNA. 에탄올과 이산화탄소로 되는 말토오스의 발효율을 향상시킬 수 있는 형질전환된 효모에 있어서, 실질적으로 원핵성 DNA가 없는 DNA구조를 포함하며, 상기 DNA구조는 다음의 유전자 : 말타아제 활성, 말토오스 투과효소 활성 또는 말토오스 조절 단백질 활성을 가지는 효소를 코드화하는 유전자를 적어도 하나 포함하는 것을 특징으로 하는 효모.In a transformed yeast capable of improving the fermentation efficiency of maltose consisting of ethanol and carbon dioxide, it comprises a DNA structure substantially free of prokaryotic DNA, the DNA structure comprising the following genes: maltase activity, maltose permease activity A yeast comprising at least one gene encoding an enzyme having maltose regulatory protein activity. 제10항에 있어서, 말타아제는 번역 연장 인자(EFlαA)촉진 유전자의 전사단계 유전자 발현 제어하에 놓이게 되는 것을 특징으로 하는 효모.The yeast of claim 10, wherein the maltase is placed under the control of transcriptional gene expression of a translational extension factor (EF1αA) promoting gene. 제10 또는 제11항에 있어서, 말토오스 투과효소는 알코올데히드로게나제 I(ADHI)촉진유전자의 전사단계 유전자 발현 제어하에 놓이게 되는 것을 특징으로 하는 효모.12. The yeast of claim 10 or 11, wherein the maltose permease is placed under the control of transcriptional gene expression of the alcohol dehydrogenase I (ADHI) promoter. 제10항 내지 제12항중 어느 한 항에 있어서, 상기 촉진유전자는 사카로마이세스 속에 속하는 효모로 부터 유도되는 것을 특징으로 하는 효모.The yeast of any one of claims 10 to 12, wherein the promoter is derived from yeast belonging to the genus Saccharomyces. 제1항 내지 제13항의 어느 하나를 따라 얻어지는 효모를 건조시킴으로써 얻어지는 습기 함량이 3 내지 8%인 효모.A yeast having a moisture content of 3 to 8% obtained by drying the yeast obtained according to any one of claims 1 to 13. 제14항에 있어서, 상기 효모는 사카로마이세스속에 속하고, 바람직하게는 상기 효모는 맥주 효모균인 것을 특징으로 하는 효모.15. The yeast of claim 14, wherein said yeast belongs to the genus Saccharomyces, preferably said yeast is brewer's yeast. 제1항 내지 제15항의 어느 하나를 따르는 효모를 사용하는 DNA중개된 형질전환 이외의 균주 개선과정에 의해 얻을 수 있는 효모.A yeast obtained by a strain improvement process other than DNA mediated transformation using a yeast according to any one of claims 1 to 15. 맥주효모균주 ApGb-iA32/G418, 맥주효모균주 ApGb-iRRo1, 맥주효모균주 ApGb-eMAL6g, 맥주효모균주 ApGb-eMAL61, 맥주효모균주 ApGb--eMAL63, 맥주효모균주 CpGb-eMAL6g, 맥주효모균주 CpGb-eMAL61 및 맥주효모균주 ApGb-p2RBRRO1#1로 이루어지는 군으로 부터 선택되는 효모.Beer Yeast strain ApGb-iA32 / G418, Beer yeast strain ApGb-iRRo1, Beer yeast strain ApGb-eMAL6g, Beer yeast strain ApGb-eMAL61, Beer yeast strain ApGb--eMAL63, Beer yeast strain CpGb-eMAL6g, Beer yeast strain CpGb-CpGb- Yeast selected from the group consisting of eMAL61 and brewer's yeast strain ApGb-p2RBRRO1 # 1. 제1항 내지 제17항중 어느 하나의 효모로 부터 얻을 수 잇는 압착, 즉석건조 또는 활성건조 효모.A compressed, instant dried or active dried yeast obtainable from the yeast of any one of claims 1 to 17. 제1항의 형질전환된 효모로 부터 얻을 수 있고, 시험 B에서는 165분후에 효모의 건조중량 285mg당 적어도 340ml의 기체생성을 보이고 시험 B'에서는 165분후에 효모의 건조중량 285mg당 적어도 170ml의 기체생성을 보이며, 바람직하게는 시험 B에서는 165분후에 효모의 건조 중량 285mg당 380 내지 450ml의 기체생성을 보이고 시험 B'에서는 165분후에 효모의 건조중량 285mg당 180 내지 240ml의 기체생성을 보이며, 더욱 바람직하게는 시험 B에서는 165분후에 효모의 건조중량 285mg당 적어도 400ml의 기체생성을 보이고 시험 B'에서는 165분후에 효모의 건조중량285mg당 적어도 190ml의 기체생성을 보이는 압착 효모.Obtained from the transformed yeast of claim 1, and in test B, at least 340 ml of gas is produced per 285 mg of dry weight of yeast after 165 minutes and at least 170 ml of gas per 285 mg of dry weight of yeast after 165 minutes of test B '. Preferably, in test B, 380 to 450 ml of gas is produced per 285 mg of dry weight of yeast after 165 minutes, and in test B ', 180 to 240 ml of gas is produced per 285 mg of dry weight of yeast after 165 minutes, more preferably. Preferably, the test yeast produced at least 400 ml per 285 mg of dry weight of yeast after 165 minutes in test B and at least 190 ml of gas per 285 mg of dry weight of yeast after 165 minutes in test B '. 제1항의 형질전환된 효모로 부터 얻을 수 있고, 시험 B에서 165분후에 효모의 건조중량 285mg당 400 내지 500ml의 기체생성을 보이고, 바람직하게는 시험 B에서 165분후에 효모의 건조중량 285mg당 440ml의 기체생성을 보이는 압착효모.It can be obtained from the transformed yeast of claim 1 and shows gas formation of 400 to 500 ml per 285 mg of dry weight of yeast after 165 minutes in Test B, preferably 440 ml per 285 mg of dry weight of yeast after 165 minutes in Test B. Pressurized yeast showing gas formation. 제19항 또는 제20항의 압착 효모를 건조시킴으로써 얻을 수 있는 즉석건조 또는 활성건조 효모.Instant dry or active dry yeast obtained by drying the compressed yeast of claim 19 or 20. 제1항의 효모로 부터 얻을 수 잇고, 시험 C에서는 165분후에 효모의 건조중량 285mg당 310 내지 360ml의 기체생성을 시험 C'에서는 165분후에 효모의 건조중량 285mg당 145 내지 195ml의 기체생성을 각각 보이며, 바람직하게는 시험 C에서 165분후에 효모의 건조중량 285mg당 적어도 330ml의 기체생성을 보이고 시험 C'에서는 165분후에 효모의 건조중량 285mg당 적어도 155ml의 기체생성을 보이며, 또는 시험 C에서 165분후에 효모의 건조중량 285mg당 320 내지 400ml의 기체생성을 보이고, 바람직하게는 시험 C에서 165분후에 효모의 건조중량 285mg당 320ml 내지 400ml의 기체생성을 보이고, 바람직하게는 시험 C에서 165분후에 효모의 건조중량 285mg당 적어도 350ml의 기체생성을 보이는 건조된 효모.It can be obtained from the yeast of claim 1, and in test C, the production of 310 to 360ml per 285mg of dry weight of yeast after 165 minutes, in test C ', the production of 145 to 195ml per 285mg of dry weight of yeast after 165 minutes, respectively. Preferably at least 330 ml per 285 mg of dry weight of yeast after 165 minutes in test C and at least 155 ml per 285 mg of dry weight of yeast after 165 minutes in test C, or 165 in test C After minutes, it shows 320 to 400 ml of gas production per 285 mg of dry weight of yeast, preferably after 165 minutes in test C, it shows 320 to 400 ml of gas per 285 mg of dry weight of yeast, preferably after 165 minutes in test C Dried yeast showing at least 350 ml of gas production per 285 mg dry weight of yeast. pGb-eMAL6g, pGb-eMAL61, pGb-eMAL63, pGb-iA32/G418, pGb-iRRo1, pGb-M6g(Δ-9), pGb-RBRR01, pGb-RBN3 및 pGb-RBREGO1로 이루어지는 군으로 부터 선택되는 벡터.pGb-eMAL6g, pGb-eMAL61, pGb-eMAL63, pGb-iA32 / G418, pGb-iRRo1, pGb-M6g (Δ-9), pGb-RBRR01, pGb-RBN3 and pGb-RBREGO1 . 제1항 내지 제22항중 어느 하나의 효모를 포함하는, 반죽 또는 유사한 방법.23. Dough or similar method comprising yeast of any one of the preceding claims. 제1항 내지 제22항중 어느 하나의 효모를 사용하는 것을 포함하는, 발효된 밀가루제품, 알코올성 음료 및 다른 알코올성 제품을 제조하는 방법.23. A method of making fermented wheat flour products, alcoholic beverages and other alcoholic products, comprising using the yeast of any one of claims 1 to 22. 운반된 기질의 흡수 및/또는 초기 대사전환을 촉진하는 단백질을 코드화하는 유전자를 적어도 하나 포함하는 상동 DNA를 적어도 하나 효모안으로 도입시키는 것을 포함하는 당의 발효율을 향상시킬 수 있는 형질전환된 효모의 제조방법.Preparation of a transformed yeast capable of improving sugar developmental efficiency comprising introducing into the yeast into at least one homologous DNA comprising at least one gene encoding a protein that promotes uptake and / or early metabolic transformation of the transported substrate. Way. 제26항에 있어서, 구조는 말타아제 활성, 말토오스 투과효소 활성 또는 말토오스 조절단백질 활성을 가지는 효소를 코드화하는 유전자를 적어도 하나 포함하는 것을 특징으로 하는 방법.27. The method of claim 26, wherein the structure comprises at least one gene encoding an enzyme having maltase activity, maltose permease activity or maltose regulatory protein activity. 제27항에 있어서, 구조가 상기 유전자를 적어도 2개 포함하는 것을 특징으로 하는 방법.The method of claim 27, wherein the structure comprises at least two of said genes. 제27항에 있어서, 상기 유전자는 각각 알코올데히드로게나제 I(ADHI) 및/또는 번역 연장인자(EFlαA)촉진유전자의 전사단계 제어하에 있음을 특징으로 하는 방법.28. The method of claim 27, wherein said gene is under the control of the transcriptional stage of the alcohol dehydrogenase I (ADHI) and / or translational prolongation factor (EF1αA) promoter. 제26항 내지 제29항중 어느 하나에 있어서, 상기 유전자 또는 그의 조합이 글루코스 억제에 민감하지 않고 말토오스 유도에 지배되지 않는 전사단계 제어하에 놓이게 되는 것을 특징으로 하는 방법.30. The method of any one of claims 26-29, wherein said gene or combinations thereof are placed under transcriptional stage control that is not sensitive to glucose inhibition and is not governed by maltose induction. 제26항 내지 제30항중 어느 하나에 있어서, 상기 구조가 에피솜성 성분의 일부인 것을 특징으로 하는 방법.31. The method of any one of claims 26-30, wherein the structure is part of an episomal component. 제26항 내지 제30항중 어느 하나에 있어서, 상기 구조는 상기 효모의 염색체 안으로 함입되는 것을 특징으로 하는 방법.31. The method of any of claims 26-30, wherein the structure is incorporated into the chromosome of the yeast. 제27항 내지 제32항중 어느 하나에 있어서, 적어도 2개의 상기 DNA구조의 도입을 포함하는 것을 특징으로 하는 방법.33. The method of any one of claims 27-32, comprising introducing at least two of said DNA structures. 제26항 내지 30항 또는 제32항 또는 제33항중 어느 하나에 있어서, 상기 효모에는 이종 DNA가 없고 유전자 대치기술을 사용하여 유전자의 염색체 안으로의 함입을 포함하는 것을 특징으로 하는 방법.34. The method of any one of claims 26-30 or 32 or 33, wherein said yeast is free of heterologous DNA and comprises incorporation of genes into the chromosome using gene replacement techniques. 제34항에 있어서, 염색체의 포자형성-특이적 유전자가 제26항 또는 제32항에 기재되어 있는 유전자가 삽입되어 있는 동일한 포자형성-특이적 유전자를 포함하는 DNA절편에 의해 대치되는 것을 특징으로 하는 방법.The method of claim 34, wherein the sporulation-specific gene of the chromosome is replaced by a DNA fragment comprising the same sporulation-specific gene into which the gene described in claim 26 or 32 is inserted. How to. 제29항에 있어서, 상기 촉진유전자는 사카로마이세스 속에 속하는 효모로 부터 유도되는 것을 특징으로 하는 방법.30. The method of claim 29, wherein said promoter is derived from yeast belonging to the genus Saccharomyces. 제26항 내지 제36항중 어느 하나에 있어서, 상기 효모는 사카로마이세스 속에 속하며 바람직하게는 맥주효모균인 것을 특징으로 하는 방법.37. The method according to any one of claims 26 to 36, wherein said yeast belongs to Saccharomyces and preferably is brewer's yeast. 제26항 내지 제37항중 어느 하나에 따라 얻어진 효모의 응용을 포함하는 반죽 또는 유사한 제품을 제조하는 방법.38. A method of making a dough or similar product comprising the application of yeast obtained according to any one of claims 26-37. 제26항 내지 제37항중 어느 하나에 따라 얻어진 효모의 사용을 포함하는 알코올성 음료 및 알코올성 제품의 제조방법.38. A process for the preparation of alcoholic beverages and alcoholic products comprising the use of yeast obtained according to any one of claims 26-37. 제26항 내지 제37항중 어느 하나에 따라 얻어진 효모의 사용을 포함하는 빵 및 관련제품의 제조방법.38. A method of making bread and related products comprising the use of yeast obtained according to any one of claims 26-37. 제26항 내지 제37항중 어느 하나에 따라 얻어진 효모의 사용을 포함하는 말타아제 또는 말토오스 투과효소 활성이 있는 효소의 제조방법.38. A method for preparing an enzyme having maltase or maltose permease activity comprising the use of yeast obtained according to any of claims 26-37. ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.※ Note: The disclosure is based on the initial application.
KR1019880011377A 1987-09-03 1988-09-03 New Yeast Strains Providing Increased Glucose Efficacy, Methods of Making Such Yeasts, and Use of These Yeasts KR890005265A (en)

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