KR100273950B1 - Corynebacterium glutamicum CJ31-0210 producing L-lysine and method for producing L-lysine using the same - Google Patents
Corynebacterium glutamicum CJ31-0210 producing L-lysine and method for producing L-lysine using the same Download PDFInfo
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Abstract
본 발명은 L-라이신을 생산하는 코리네박테리움 글루타미컴(Corynebacterium glutamicum) CJ31-0210(기탁번호 제 KFCC-11043 호) 및 그를 배양하여 그 배양물로부터 L-라이신을 생산하는 방법에 관한 것으로서, L-라이신의 발효농도 및 대당수율을 향상시켜 L-라이신의 생산에 유용하게 사용될 수 있다.The present invention relates to Corynebacterium glutamicum (Corynebacterium glutamicum) CJ31-0210 (Accession No. KFCC-11043) for producing L- lysine, and to a method for culturing the same to produce L- lysine from the culture. To improve the fermentation concentration and yield of L-lysine, it can be usefully used for the production of L-lysine.
Description
본 발명은 L-라이신을 생산하는 신규한 코리네박테리움 글루타미컴 (Corynebacterium glutamicum) CJ31-0210에 관한 것으로서, 더욱 상세하게는 대한민국 특허출원 제 97-68670 호(1997년 12월 15일 출원)에 따른 코리네박테리움 글루타미컴 KFCC 11001로부터 유래되고, L-라이신의 발효농도 및 대당수율을 획기적으로 증가시킨 코리네박테리움 글루타미컴 CJ31-0210(기탁번호 제 KFCC-11043 호) 및 그를 배양하여 그 배양물로부터 L-라이신을 생산하는 방법에 관한 것이다.The present invention relates to a novel Corynebacterium glutamicum CJ31-0210 producing L-lysine, and more particularly, Korean Patent Application No. 97-68670 (filed December 15, 1997) Corynebacterium glutamicum CJ31-0210 (Accession No. KFCC-11043) derived from Corynebacterium glutamicum KFCC 11001 and dramatically increasing the fermentation concentration and the yield of L-lysine and It relates to a method for producing L-lysine from the culture by culturing.
L-라이신은 필수 아미노산의 일종으로 가축의 사료첨가제, 식품첨가제, 의약원료 등으로 사용되고 있다. 특히 L-라이신의 사료첨가제로서의 시장규모는 1997년에 약 40여만톤에 이르고 있어 L-라이신은 그야말로 거대한 생물공학 제품이라 할 수 있다. 사료첨가제로서의 L-라이신은 현재까지 년 8∼10%의 수요 증대를 나타내어 왔을 뿐 아니라, 가축배설물에 대한 규제가 강화됨에 따라 향후 더욱 급격한 시장 확대가 예상되고 있다. 따라서 L-라이신 생산 균주의 개발 또는 발효공정의 개선에 의한 L-라이신의 생산성의 향상은 더욱 절실한 과제라 할 수 있을 것이다.L-lysine is an essential amino acid and is used as feed additives, food additives, and pharmaceutical raw materials for livestock. In particular, the market size of L-lysine as a feed additive amounted to about 400,000 tons in 1997, so L-lysine is a huge biotech product. L-lysine as a feed additive has not only increased demand by 8-10% a year so far, but also is expected to expand more rapidly in the future as the regulations on livestock excretion are tightened. Therefore, the improvement of the productivity of L-lysine by the development of the L- lysine production strain or the improvement of the fermentation process will be more urgent task.
현재까지 미생물을 이용한 L-라이신의 생산방법으로는 야생균주를 사용하는 직접발효법(direct fermentation), 생합성 전구물질(예: α-아미노아디프산, α-케토아디프산)을 가하여 대사시키는 방법, 변이주를 사용하는 2단법(디아미노피멜릭산(diaminopimelic acid)을 통한 생산법), 효소에 의한 생산법(DL-α-아미노카프로락탐의 전환) 및 변이주를 사용하는 직접발효법이 알려져 있으며, 그 중에서도 변이주를 사용하는 직접발효법이 주축을 이루고 있다.Until now, the production of L-lysine using microorganisms includes direct fermentation using wild strains and metabolism by adding biosynthetic precursors (eg, α-aminoadipic acid and α-ketoadipic acid). , Two-stage method using mutant strains (production method through diaminopimelic acid), enzyme production method (conversion of DL-α-aminocaprolactam) and direct fermentation method using mutant strains are known. Among them, direct fermentation method using mutant strain is the main axis.
L-라이신은 세균, 방선균에서는 하기하는 바와 같은 디아미노피멜릭산 경로(DAP pathway)를 통하여 합성되며, 또한 코리네박테리움 글루타미컴에서는 하기와 같은 조절하에 있게 된다.L-lysine is synthesized through the diaminopimelic acid pathway (DAP pathway) as described below in bacteria and actinomycetes, and under Corynebacterium glutamicum under the following control.
즉 아스파토키나제(aspartokinase)는 라이신과 트레오닌에 의한 피드백 저해(feedback inhibition)를 받으며, 호모세린 디하이드로게나제(homoserine dehydrogenase)는 메티오틴에 의한 억제(repression) 및 트레오닌에 의한 피드백 저해를 받는다.That is, aspartokinase receives feedback inhibition by lysine and threonine, and homoserine dehydrogenase receives inhibition by methionine and feedback inhibition by threonine. .
따라서 변이주를 사용하는 직접발효법에서는 L-라이신 생산 균주로서 일반적으로 대사 제어 물질의 균체내 농도를 인위적으로 조절하기 위한 영양요구성 변이주(auxotroph), 예를 들면, 호모세린 영양요구성, 트레오닌 및 메티오닌 영양요구성 균주나, 최소배지에서는 야생균주와 같이 왕성하게 생육하나 트레오닌, 메티오닌을 소량 첨가하면 생육이 저해되는 성질을 갖는 트레오닌 피드백 저해에 과민성인 호모세린 디하이드로게나제를 갖는 트레오닌 및 메티오닌 감수성 변이주(threonine-methionine sensitive mutant)나, L-라이신과 L-트레오닌의 피드백 저해가 해제되고 호모세린 디하이드로게나제에 대한 트레오닌의 저해만 지속되는 라이신 유사체(예: S(β-아미노에틸)-L-시스테인(AEC))내성 변이주 등을 사용하는 것이 바람직하다.Thus, in direct fermentation methods using mutant strains, L-lysine producing strains are generally auxotrophs, such as homoserine nutrients, threonine and methionine, for artificially controlling the intracellular concentrations of metabolic control substances. Threonine and methionine susceptible strains with homoserine dehydrogenase that are hyperactive in nutrient constitutive strains or minimal media, but are hyperactive to threonine feedback inhibition, which has a property of inhibiting growth when a small amount of threonine and methionine are inhibited. (threonine-methionine sensitive mutant) or lysine analogues (e.g. S (β-aminoethyl) -L which release the feedback inhibition of L-lysine and L-threonine and persist only the inhibition of threonine to homoserine dehydrogenase -Cysteine (AEC)) resistant strains and the like are preferably used.
상기한 바와 같은 원리로 직접발효법에 의하여 L-라이신을 생산하기 위한 종래의 방법으로는 예를 들면, 일본공개특허 제 평4-88991, 평4-91794, 평5-111386, 평5-30985, 평6-7182 및 평7-155184 호, 및 국제공개특허 제 WO96-17930 및 95-23864 호 등이 공지되어 있다.As a conventional method for producing L-lysine by the direct fermentation method on the principle as described above, for example, Japanese Patent Laid-Open No. Hei 4-88991, Hei 4-91794, Hei 5-111386, Hei 5-30985, JP-A 6-7182 and JP-A 7-155184, and WO 96-17930 and 95-23864, and the like are known.
한편 본 출원인은 L-루이신(L-leucine) 비영양요구성 및 초산(acetic acid) 자화성의 L-라이신 생산 균주인 코리네박테리움 글루타미컴 KFCC 11001을 대한민국 특허출원 제 97-68670 호로 출원한 바 있다.On the other hand, the applicant of the L-leucine non-nutrient composition and acetic acid magnetization L- lysine production strain Corynebacterium glutamicum KFCC 11001 to Korean Patent Application No. 97-68670 It has been filed.
그러나 상기 방법들은 미생물에 의한 L-라이신의 발효농도 및 대당수율에 있어서 개선의 여지를 여전히 갖고 있는 방법들이다.However, these methods still have room for improvement in fermentation concentration and yield of L-lysine by microorganisms.
이에 본 발명자들은 대한민국 특허출원 제 97-68670 호에 공지된 바 있는 코리네박테리움 글루타미컴 KFCC 11001을 모균주로 하여 이로부터 보다 우수한 L-라이신 발효농도 및 대당수율을 얻을 수 있는 신균주를 개발해내기 위하여 지속적인 연구를 수행하였으며, 그 결과 초산 비자화성을 갖는 본 발명의 균주가 상기 목적에 부합할 뿐 아니라 기존에 전혀 공지된 바 없는 신규한 균주임을 확인하고 본 발명을 완성하였다.Accordingly, the present inventors have identified a new strain capable of obtaining better L-lysine fermentation concentration and yield from the Corynebacterium glutamicum KFCC 11001, which is known from Korean Patent Application No. 97-68670, as a parent strain. Continuous research was carried out to develop, and as a result, the strain of the present invention having non-acetic acid non-magnetic properties was confirmed to be a novel strain that was not previously known as well as a novel strain and completed the present invention.
따라서 본 발명의 목적은 L-라이신을 생산하는 코리네박테리움 글루타미컴 CJ31-0210을 제공하기 위한 것이다.It is therefore an object of the present invention to provide Corynebacterium glutamicum CJ31-0210 to produce L-lysine.
본 발명의 또다른 목적은 상기 미생물을 배양하여 그 배양물로부터 L-라이신을 생산하는 방법을 제공하기 위한 것이다.It is another object of the present invention to provide a method of culturing the microorganism to produce L-lysine from the culture.
본 발명은 L-라이신을 생산하는 코리네박테리움 글루타미컴 CJ31-0210(기탁번호 제 KFCC-11043 호)을 제공한다.The present invention provides Corynebacterium glutamicum CJ31-0210 (Accession No. KFCC-11043) for producing L-lysine.
본 발명은 또한 코리네박테리움 글루타미컴 CJ31-0210을 배양하여 그 배양물로부터 L-라이신을 생산하는 방법을 제공한다.The present invention also provides a method of culturing Corynebacterium glutamicum CJ31-0210 to produce L-lysine from the culture.
이하 본 발명을 더욱 상세히 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명에 있어서, 모균주로 사용한 코리네박테리움 글루타미컴 KFCC 11001은 대한민국 특허출원 제 97-68670 호에 개시된 바 있는 균주로서, 하기 표 1과 같은 균학적 성질을 갖는 균주이다.In the present invention, Corynebacterium glutamicum KFCC 11001 used as a parent strain is a strain as disclosed in Korean Patent Application No. 97-68670, which has a bacteriological property as shown in Table 1 below.
상기 균주는 L-라이신의 생합성 경로에 있어서, L-라이신과 L-트레오닌의 피드백 저해가 해제되고 호모세린 디하이드로게나제에 대한 트레오닌의 저해만 지속되는, 라이신 유사체인 S(β-아미노에틸)-L-시스테인에 내성을 갖는 균주이므로 L-라이신 생산 균주로서 적합한 균주이다.The strain is a lysine analogue S (β-aminoethyl) in which L-lysine and L-threonine feedback inhibition of L-lysine and L-threonine are released and only inhibition of threonine to homoserine dehydrogenase is sustained. Since the strain is resistant to -L-cysteine, it is suitable as an L-lysine producing strain.
본 발명에서는 상기 코리네박테리움 글루타미컴 KFCC 11001을 모균주로 하고 이 균주를 인공돌연변이시켜 초산 자화성에서 초산 비자화성으로 변이된 변이주를 획득하고자 하였다.In the present invention, the parent strain of Corynebacterium glutamicum KFCC 11001 was used as a parent strain, and the strain was artificially mutated to obtain a mutant strain that was changed from acetic acid magnetization to nonacetic acid.
인공돌연변이 유발원으로는 알킬화제의 일종인 N-메틸-N'-니트로-N-니트로소구아니딘(N-methyl-N'-nitro-N-nitrosoguanidine: NTG)을 107∼108세포/㎖에 대하여 최종농도 1,000㎍/㎖으로 약 5분간 처리하였다. NTG는 생체내에서 분해되어 산성 조건에서는 아질산(nitrous acid)으로, 알칼리성 조건에서는 디아조메탄으로 각각 전환되며 DNA 단편의 부정확한 복제를 일으킴으로써 돌연변이를 유발하게 된다.As a source of the mutagenicity, N-methyl-N'-nitro-N-nitrosoguanidine (NTG), a kind of alkylating agent, was added to 10 7-10 8 cells / ml. It was treated for about 5 minutes at a final concentration of 1,000㎍ / ㎖. NTG is degraded in vivo and converted to nitrous acid in acidic conditions and to diazomethane in alkaline conditions, causing mutations by inaccurate replication of DNA fragments.
그런 후 포도당 함유 배지에서는 세포의 성장이 일어나나 초산 함유 배지에서는 세포의 성장이 일어나지 않는 초산 비자화성 균주를 선별한 후, 그 균주의 L-라이신 발효농도를 시험해 본 결과, A-16 균주(하기 실시예 2 참조)가 모균주인 KFCC 11001에 비하여 그 농도를 획기적으로 증가시킨다는 놀라운 사실을 확인하였다. 이에 상기 A-16 균주 및 모균주인 KFCC 11001를 7ℓ 발효조에서 L-라이신 발효농도에 대하여 재확인 실험을 행하였으며, 그 결과 역시 A-16 균주가 KFCC 11001에 비하여 L-라이신 발효농도를 증가시킨다는 사실을 확인할 수 있었다.Then, non-acetic acid-free strains were selected in which the growth of the cells occurred in the medium containing glucose but not the growth of the cells in the medium containing acetic acid, and the L-lysine fermentation concentration of the strain was tested. (See Example 2) was found to surprisingly increase the concentration significantly compared to the parent strain KFCC 11001. Thus, the A-16 strain and the parent strain KFCC 11001 were reconfirmed for the L-lysine fermentation concentration in a 7 L fermenter. As a result, the A-16 strain increased the L-lysine fermentation concentration compared to KFCC 11001. Could confirm.
따라서 상기 A-16 균주를 코리네박테리움 글루타미컴 CJ31-0210라 명명하고, 이 균주를 1998년 7월 14일자로 사단법인 한국종균협회에 기탁하고, 기탁번호 제 KFCC-11043 호를 부여받았다.Therefore, the strain A-16 was named Corynebacterium glutamicum CJ31-0210, and the strain was deposited on July 14, 1998 with the Korean spawn association, and was given accession number KFCC-11043. .
본 발명에 따른 코리네박테리움 글루타미컴 CJ31-0210은 모균주인 코리네박테리움 글루타미컴 KFCC 11001로부터 유래된 변이주이나, 초산 비자화성 균주라는 점에 있어서 명백히 구별되는 균주이다.Corynebacterium glutamicum CJ31-0210 according to the present invention is a mutant derived from the parent strain Corynebacterium glutamicum KFCC 11001, but is clearly distinguished in that it is a nonacetic acid-free strain.
본 발명에 따른 코리네박테리움 글루타미컴 CJ31-0210을 창제하기 위한 과정을 상술하면 하기 실시예와 같다.The process for creating Corynebacterium glutamicum CJ31-0210 according to the present invention will be described in detail below.
[실시예]EXAMPLE
이하의 실험에서 변이주 선별을 위한 최소배지로는 하기의 한천 최소배지를 사용하였고, 선별된 변이주의 평가를 위한 배지로는 하기의 플라스크 발효배지를 사용하였다. L-라이신의 농도는 HPLC(High Performance Liquid Chromatography)로 측정하였고, 당량은 필요한 경우 버트란트(Bertrand)법을 사용하여 정량하였다.In the following experiments, the following agar minimum medium was used as the medium for selection of mutant strains, and the following flask fermentation medium was used as a medium for evaluation of the selected strains. The concentration of L-lysine was measured by HPLC (High Performance Liquid Chromatography), and the equivalent was quantified using Bertrand method if necessary.
한천 최소배지Agar Minimum Badge
포도당 10g(필요한 경우 초산 10g 사용), (NH4)2SO42g, 우레아 2g, KH2PO41.0g, K2HPO43.0g, MgSO4·7H2O 0.5g, FeSO4·7H2O 10mg, MnSO4·5H2O 10mg, 바이오틴 100㎍, 티아민·HCl 100㎍, CaCl2·2H2O 0.1g, Na2B4O7·10H2O 80㎍, (NH4)6MoO27·4H2O 40㎍, ZnSO4·7H2O 10㎍, CuSO4·7H2O 300㎍, MnCl2·4H2O 10㎍, FeCl3·6H2O 1mg, 한천 20g, 증류수 1리터당 (pH 7.0(살균전)).10 g of glucose (if necessary, 10 g of acetic acid), (NH 4 ) 2 SO 4 2 g, urea 2 g, KH 2 PO 4 1.0 g, K 2 HPO 4 3.0 g, MgSO 4 H 7 O 0.5 g, FeSO 4 7H 2 O 10mg, MnSO 4 5H 2 O 10mg, Biotin 100μg, Thiamine HCl 100μg, CaCl 2 · 2H 2 O 0.1g, Na 2 B 4 O 7 · 10H 2 O 80μg, (NH 4 ) 6 MoO 27 40 μg of 4H 2 O, 10 μg of ZnSO 4 7H 2 O, 300 μg of CuSO 4 7H 2 O, 10 μg of MnCl 2 4H 2 O, 1 mg of FeCl 3 , 6H 2 O, 20 g of agar, per liter of distilled water (pH 7.0 (prior to sterilization)).
플라스크 발효배지Flask fermentation medium
당밀(환원당으로서) 100g, 효모엑기스 4g, (NH4)2SO440g, 우레아 4g, KH2PO41g, NaCl 2.5g, MgSO4·7H2O 0.5g, FeSO4·7H2O 10mg, MnSO4·5H2O 10mg, 바이오틴 100㎍, 티아민·HCl 200㎍, CaCO440g, 공정수 1리터당 (pH 7.0(살균후)).Molasses (as reduced sugar) 100 g, yeast extract 4 g, (NH 4 ) 2 SO 4 40 g, urea 4 g, KH 2 PO 4 1 g, NaCl 2.5 g, MgSO 4 7H 2 O 0.5 g, FeSO 4 7H 2 O 10 mg, 10 mg MnSO 4 5H 2 O, 100 μg biotin, 200 μg thiamine-HCl, 40 g CaCO 4 , per liter of process water (pH 7.0 (after sterilization)).
[실시예 1]Example 1
모균주인 KFCC 11001의 초산 자화성을 시험하기 위하여 루리아 버타니(LB) 액체배지에서 16시간 성장시킨 세포를 멸균 생리식염수로 2회 세척한 후 적당히 희석하여 탄소원으로써 포도당과 초산을 각각 첨가한 한천 최소배지에 도말하였다. 30℃ 배양기에서 4일간 배양한 후 그 성장도를 측정한 결과, KFCC 11001의 경우 초산을 자화할 수 있는 능력이 있음을 알 수 있었다(표 2).To test the acetic acid magnetization of the parent strain KFCC 11001, cells grown in Luria Bertani (LB) liquid medium for 16 hours were washed twice with sterile saline solution and diluted appropriately with agar containing glucose and acetic acid as carbon sources, respectively. Plated on minimal medium. After incubation for 4 days in a 30 ℃ incubator, the growth was measured, it can be seen that KFCC 11001 has the ability to magnetize acetic acid (Table 2).
+ : 세포 성장 일어남+: Cell growth occurs
[실시예 2]Example 2
초산을 자화하지 못하는 균을 창제하기 위하여 루리아 버타니 액체배지에서 대수기 중반까지 성장한 코리네박테리움 글루타미컴 KFCC 11001을 시트레이트 완충액(pH 5.5)에 107∼108세포/㎖로 현탁시킨 후, NTG의 최종농도가 1,000㎍/㎖이 되도록 한 후 30℃ 진탕기에서 5분간 처리하였다. 이어서 인산칼륨 완충액(pH 7.0)으로 3회 세척하고 포도당이 함유된 한천 최소배지에 도말하였다. 30℃의 항온기에서 7일간 배양시킨 후 성장한 미생물 군락(colony)을 포도당과 초산이 각각 함유된 한천 최소배지에 투스픽킹(toothpicking)하여 포도당이 함유된 배지에서는 세포의 성장이 일어나지만 초산이 함유된 배지에서는 세포의 성장이 일어나지 않는 초산 비자화성 균주를 선별하였다. 그 결과 5개의 초산 비자화성 미생물 군락을 최종적으로 획득할 수 있었다.In order to create a bacterium that does not magnetize acetic acid, Corynebacterium glutamicum KFCC 11001, grown in Luria bertani liquid medium to the middle of the log phase, was suspended in citrate buffer (pH 5.5) at 10 7-10 8 cells / ml. After that, the final concentration of NTG was 1,000 ㎍ / ㎖ and then treated for 5 minutes in a 30 ℃ shaker. It was then washed three times with potassium phosphate buffer (pH 7.0) and plated in agar medium containing glucose. After 7 days of incubation at 30 ° C, microorganism colonies grown after toothpicking were grown on agar medium containing glucose and acetic acid, respectively, and cell growth occurred in the medium containing glucose, but acetic acid was contained. In the medium, non-acetic acid-free strains were selected in which cell growth did not occur. As a result, five communities of nonacetic acid non-microbial microorganisms were finally obtained.
그 미생물 군락을 30℃, 230rpm(revolution per minute), 72시간의 배양조건의 진탕배양기에서 배플 플라스크를 사용하여 L-라이신의 발효농도를 측정하였다. 그 결과를 하기 표 3에 나타내었으며, A-16 균주만이 KFCC 11001에 비해 L-라이신의 발효농도를 향상시켰음을 알 수 있었다.The fermentation concentration of L-lysine was measured using a baffle flask in a shaker incubator at 30 ° C., 230 rpm (revolution per minute), 72 hours of culture conditions. The results are shown in Table 3 below, and it was found that only the A-16 strain improved the fermentation concentration of L-lysine compared to KFCC 11001.
* PCV : Packed Cell Volume* PCV: Packed Cell Volume
[실시예 3]Example 3
실시예 2에서 획득한 A-16 균주를 CJ31-0210 균주라 명명하고, CJ31-0210 및 모균주인 KFCC11001를 7ℓ 발효조에서 하기와 같은 방법으로 재확인 실험을 행하였다. 먼저 두 균주 모두 각각의 플라스크 종배지에서 20시간 동안 진탕배양기에서 종배양한 후 7ℓ 발효조에 직접 접종하였다. 추가당 및 추가물을 배양중에 첨가하는 유가식 발효(fed-batch fermentation)로 균주의 평가실험을 행하였으며 사용된 배지 및 배양조건은 다음과 같다.The A-16 strain obtained in Example 2 was named CJ31-0210 strain, and CJ31-0210 and the parent strain KFCC11001 were reconfirmed in a 7 L fermenter in the following manner. First, both strains were seeded in a shaker incubator for 20 hours in each flask species, and then directly inoculated directly into a 7 L fermenter. Evaluation of the strain was carried out by fed-batch fermentation in which additional sugars and additional products were added during the cultivation. The medium and culture conditions used were as follows.
종배양 배지성분 및 배양조건Species Culture and Culture Conditions
원당 50g(멸살), 펩톤 10g, 효모엑기스 10g, KH2PO41.5g, MgSO4·7H2O 0.5g, 우레아 5g, 바이오틴 50㎍, 공정수 1ℓ(pH 7.0, 멸균전)50g per raw (annihilated), peptone 10g, yeast extract 10g, KH 2 PO 4 1.5g, MgSO 4 · 7H 2 O 0.5g, urea 5g, biotin 50㎍, process water 1ℓ (pH 7.0, before sterilization)
30℃, 220 rpm, 20시간 배양30 ° C, 220 rpm, 20 hours incubation
7ℓ 발효조 배지성분 및 배양조건7ℓ fermenter medium components and culture conditions
당밀(환원당으로서) 280g/ℓ, 콘 스팁 리쿼(Corn steep liquor) 100㎖/ℓ, (NH4)2SO435g/ℓ, KH2PO41.0g/ℓ, MgSO4·7H2O 0.5 g/ℓ, FeSO4·7H2O 10mg/ℓ, 바이오틴 300㎍/ℓ, 티아민·HCl 500㎍/ℓ, 소포제 3㎖/ℓ)Molasses (as reduced sugar) 280 g / l, Corn steep liquor 100 ml / l, (NH 4 ) 2 SO 4 35 g / l, KH 2 PO 4 1.0 g / l, MgSO 4 · 7H 2 O 0.5 g / l, FeSO 4 · 7H 2 O 10mg / l, Biotin 300µg / l, Thiamine-HCl 500µg / l, Defoamer 3ml / l)
온도 30℃, pH 7.2, 교반속도 600rpm, 통기량 1vvm(air volume/medium volume/minute)Temperature 30 ℃, pH 7.2, Stirring Speed 600rpm, Aeration Volume 1vvm (air volume / medium volume / minute)
그 발효 결과, CJ31-0210은 모균주 KFCC11001에 비해 발효농도가 2.6% 가량 증가하고 대당수율 또한 증가한 우량 균주임을 확인하였다(표 4).As a result of the fermentation, it was confirmed that CJ31-0210 was a superior strain in which the fermentation concentration was increased by about 2.6% and the yield was also increased compared to the parent strain KFCC11001 (Table 4).
본 발명에 따른 코리네박테리움 글루타미컴 CJ31-0210은 모균주인 코리네박테리움 글루타미컴 KFCC 11001를 인공돌연변이시켜 초산 자화성을 초산 비자화성 으로 변이시킨 균주로서, 모균주에 비해 L-라이신의 발효농도 및 대당수율을 향상시켜 L-라이신의 생산에 유용하게 사용될 수 있다.Corynebacterium glutamicum CJ31-0210 according to the present invention is a strain that transformed acetic acid magnetization to non-acetic acid by artificial mutation of the parent strain Corynebacterium glutamicum KFCC 11001, L- compared to the parent strain By improving the fermentation concentration and yield of lysine can be useful for the production of L- lysine.
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| KR1019980039305A KR100273950B1 (en) | 1998-09-22 | 1998-09-22 | Corynebacterium glutamicum CJ31-0210 producing L-lysine and method for producing L-lysine using the same |
| PCT/KR1999/000563 WO2000017379A2 (en) | 1998-09-22 | 1999-09-17 | An l-lysine-producing microorganism and a method for producing l-lysine using said microorganism |
| AU57630/99A AU5763099A (en) | 1998-09-22 | 1999-09-17 | An l-lysine-producing microorganism and a method for producing l-lysine using said microorganism |
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| KR100614219B1 (en) * | 2004-07-05 | 2006-08-21 | 씨제이 주식회사 | Fermentation of L-lysine using acid-treated product of soybean meal |
| CN116496950B (en) * | 2022-09-27 | 2023-10-24 | 欧铭庄生物科技(天津)有限公司滨海新区分公司 | Lysine production strain and application thereof, and lysine production method |
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| KR940001307B1 (en) * | 1991-12-27 | 1994-02-19 | 제일제당 주식회사 | Microorganism for producing l-lysine |
| SK278468B6 (en) * | 1992-03-11 | 1997-06-04 | Jiri Plachy | Strain of microorganism corynebacterium glutamicum ccm 4265 producing l-lysine |
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