KR101177358B1 - Endolysin lysb4 with peptidase activity, food composition and feed composition thereof - Google Patents
Endolysin lysb4 with peptidase activity, food composition and feed composition thereof Download PDFInfo
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Abstract
Description
본 발명은 박테리오파지 유래의 엔돌라이신에 관한 것으로, 더욱 상세하게는 펩티다아제 활성이 있는 엔돌라이신 LysB4에 관한 것이다.
The present invention relates to an endolisin derived from bacteriophage, and more particularly, to an endolisin LysB4 having peptidase activity.
바실러스 세레우스(Bacillus cereus)는 그람 양성균이며, 포자를 형성하고, 호기성 또는 통성호기성균으로 주로 토양에서 발견된다. 이것은 구토형과 설사형과 같은 두 가지 타입의 독소를 형성하며 식중독을 일으킨다. 바실러스 세레우스와 관련된 식중독에 의한 증상은 상대적으로 가벼우나, 그 경우가 빈번히 일어나며, 심각한 질병으로 발전될 가능성이 크다. 게다가 바실러스 세레우스는 넓은 온도 범위에서 살 수 있으며, 열악한 환경에서 내열성을 보이는 포자(spore)를 형성할 수 있기 때문에 식품 산업에서 바실러스 세레우스를 효과적으로 제거할 수 있는 기술 개발이 시급하다. Bacillus cereus is a Gram-positive bacterium that forms spores and is found in soil mainly as aerobic or aerobic aerobes. It forms two types of toxins, vomiting and diarrhea, and causes food poisoning. Symptoms of food poisoning associated with Bacillus cereus are relatively mild, but they are frequent and likely to develop serious diseases. In addition, because Bacillus cereus can live in a wide temperature range and can form spores that exhibit heat resistance in harsh environments, there is an urgent need to develop techniques to effectively remove Bacillus cereus from the food industry.
한편, 엔돌라이신(Endolysin)은 최근 유망한 항생물질로 주목받고 있다. 엔돌라이신은 박테리아 세포벽의 펩티도글리칸(peptidoglcan)을 분해하는 효소로 박테리오파지(bacteriophage)의 유전물질에 암호화되어 있다. 저항성 균의 형성이 잘 되지 않고, 수 초 혹은 수 분 이내에 특정 박테리아만 죽이는 특성이 있다.Endolysin, on the other hand, has recently attracted attention as a promising antibiotic. Endolicin is an enzyme that breaks down peptidoglcan on bacterial cell walls and is encoded in the genetic material of bacteriophage. It does not form resistant bacteria and kills only certain bacteria within a few seconds or minutes.
현재 엔돌라이신을 처리한 우유 혹은 바나나주스가 스타필로코커스 오리우스 (Staphylococcus aureus) 혹은 몇몇의 그람 음성균으로부터 오염되지 않은 사례가 보고되었지만, 아직까지 바실러스 세레우스를 제어한 실례는 없는 실정이다. Currently, there have been reported cases where endorcinin-treated milk or banana juice is not contaminated with Staphylococcus aureus or some Gram-negative bacteria, but there are no examples of controlling Bacillus cereus.
한편, 바실러스 세레우스 감염 박테리오파지로부터 보고된 엔돌라이신은 아직 세 개뿐인데, 이들 모두 한가지 유형인 엔-아세틸무라모일-엘-알라닌 아미다아제(N-acetylmuramoyl-L-alanine amidase)의 특성을 갖는다. On the other hand, there are still three reported endolicins from Bacillus cereus-infected bacteriophages, all of which have one type of N-acetylmuramoyl-L-alanine amidase. .
따라서, 바실러스 세레우스를 타겟으로 하여, 이를 효과적으로 제어하기 위한 바이오컨트롤(biocontrol) 제제의 개발을 위해서는 더 많은 박테리오파지 혹은 더 많은 엔돌라이신이 분리되고 그 특성 분석이 이루어져야 한다.
Therefore, more bacteriophages or more endolicin must be isolated and characterized for the development of biocontrol formulations that target Bacillus cereus and effectively control them.
본 발명에서는 기존의 엔-아세틸무라모일-엘-알라닌 아미다아제(N-acetylmuramoyl-L-alanine amidase) 특성을 갖는 바실러스 세레우스 감염 박테리오파지 유래의 엔돌라이신과는 다른, 새로운 작용 메커니즘을 갖는 신규의 바실러스 세레우스 감염 박테리오 파지 유래의 엔돌라이신을 개발하여 제공하고자 한다.
In the present invention, a novel novel mechanism having a new mechanism of action, which is different from the endolisin derived from Bacillus cereus infected bacteriophage, which has N-acetylmuramoyl-L-alanine amidase properties. Bacillus cereus infection of bacteriophage-derived endolisin derived from to provide.
상기 목적을 달성하기 위하여, 본 발명은 서열번호 2의 아미노산 서열을 갖는 엔돌라이신 LysB4을 제공한다. In order to achieve the above object, the present invention provides an endolisin LysB4 having the amino acid sequence of SEQ ID NO: 2.
본 발명에서 새롭게 분리한 엔돌라이신 LysB4는 식중독의 원인균 중 하나인 바실러스 속 및 리스테리아 모노사이토제네스에 속하는 균주들에 대해 특이적인 용균 작용을 보이며, EDTA와 같은 화학약품 처리시 이 콜라이 혹은 살모넬라 같은 그람 음성균의 제어에도 효과적이다. The newly isolated endolisin LysB4 shows a specific lytic action against strains belonging to the genus Bacillus and Listeria monocytogenes, one of the causative agents of food poisoning, and the grams such as E. coli or Salmonella during the treatment of chemicals such as EDTA. It is also effective for the control of negative bacteria.
또한, 본 발명의 엔돌라이신 LysB4는 비교적 넓은 온도와 넓은 pH 범위에서 안정적인 항균 활성을 보이는 것으로 확인되었다.In addition, it was confirmed that the endolisin LysB4 of the present invention exhibits stable antimicrobial activity at a relatively wide temperature and a wide pH range.
또한, 본 발명의 엔돌라이신 LysB4는 바실러스 박테리오파지 유래로서는 처음으로 엘-알라노일-디-글루타메이트 펩티다아제(L-alanoyl-D-glutamate peptidase) 활성을 갖는 효소임이 확인되었다. In addition, it was confirmed that the endolicin LysB4 of the present invention is an enzyme having L-alanoyl-D-glutamate peptidase activity as the first bacteriophage-derived bacterium.
한편, 본 발명에 있어서, 서열번호 2의 아미노산 서열은 서열번호 1의 핵산 서열로 암호화되는 것이 좋다. On the other hand, in the present invention, the amino acid sequence of SEQ ID NO: 2 is preferably encoded by the nucleic acid sequence of SEQ ID NO: 1.
한편, 본 발명의 엔돌라이신 LysB4는 바실러스 세레우스(Bacillus cereus) 감염 박테리오파지 B4 (KCTC 12013BP) 유래인 것일 수 있다. On the other hand, the endolicin LysB4 of the present invention is Bacillus cereus ( Bacillus cereus ) may be derived from infected bacteriophage B4 (KCTC 12013BP).
한편, 본 발명은 상기 엔돌라이신 LysB4를 함유하는 식품 첨가제 또는 식품 조성물을 제공한다. 상기 식품 첨가제 또는 식품 조성물은 엔돌라이신 LysB4를 함유하기만 하면 그 제형에 특별히 국한되지 않으나, 대상 식품은 바실러스 속이나 리스테리아 모노사이토제네스에 의해 오염되기 쉬운 식품이면 더욱 바람직하다. On the other hand, the present invention provides a food additive or food composition containing the endolacin LysB4. The food additive or food composition is not particularly limited to the formulation as long as it contains endolacin LysB4, but the target food is more preferably a food that is susceptible to contamination by Bacillus or Listeria monocytogenes.
또한, 본 발명은 상기 엔돌라이신 LysB4를 함유하는 사료 첨가제 또는 사료 조성물을 제공한다. 상기 사료 첨가제 또는 사료 조성물은 엔돌라이신 LysB4를 함유하기만 하면 그 제형에 국한되지는 않으나, 바실러스 속이나 리스테리아 모노사이토제네스에 감염되기 쉬운 가축을 대상으로 하는 사료 첨가제 또는 사료이면 더욱 바람직하다.
The present invention also provides a feed additive or feed composition containing the endolacin LysB4. The feed additive or feed composition is not limited to the formulation as long as it contains endolacin LysB4, but more preferably is a feed additive or feed for livestock susceptible to the genus Bacillus or Listeria monocytogenes.
본 발명에서 새롭게 분리한 엔돌라이신 LysB4는 식중독의 원인균 중 하나인 바실러스 속 및 리스테리아 모노사이토제네스에 속하는 균주들에 대해 특이적인 용균 작용을 보이며, EDTA와 같은 화학약품 처리시 이 콜라이 혹은 살모넬라 같은 그람 음성균의 제어에도 효과적이기 때문에 식품 및 사료 산업에 효과적으로 활용될 수 있다.The newly isolated endolisin LysB4 shows a specific lytic action against strains belonging to the genus Bacillus and Listeria monocytogenes, one of the causative agents of food poisoning, and the grams such as E. coli or Salmonella during the treatment of chemicals such as EDTA. It is also effective in the control of negative bacteria, so it can be effectively used in the food and feed industry.
도 1은 본 발명 엔돌라이신 LysB4에서 펩티다아제 활성을 나타내는 도메인과 호스트 바인딩 도메인을 보여준다.
도 2는 본 발명 엔돌라이신 LysB4의 발현 정도 및 순수 분리 결과 (a)와 LysB4 농도에 따른 바실러스 세레우스의 항균 효과 (b)를 보여준다.
도 3은 여러 가지 pH (a)와 온도 (b) 범위에서 본 발명 엔돌라이신 LysB4의 항균효과를 보여준다.
도 4는 바실러스 세레우스 펩티도클리칸에 엔돌라이신 LysB4 처리 후, 0분 (a), 60분 (b) 뒤의 고속액체크로마토그래피 결과를 보여주며, 본 발명의 엔돌라이신 LysB4가 바실러스 세레우스 펩티도글리칸에서 펩티드 결합(peptide bond) 중 자르는 위치를 보여준다(c에서 화살표로 표시).Figure 1 shows the host binding domains and domains showing peptidase activity in the endolisin LysB4 of the present invention.
Figure 2 shows the antimicrobial effect of Bacillus cereus according to the expression level and pure separation result (a) and LysB4 concentration of the present invention endolaly LysB4.
Figure 3 shows the antimicrobial effect of the present invention endola lysine LysB4 in various pH (a) and temperature (b) range.
Figure 4 shows the results of high-performance liquid chromatography after 0 minutes (a), 60 minutes (b) after treatment with Bacillus cereus peptidoglycan endorlysine LysB4, the endolisin LysB4 of the present invention is Bacillus sere The cleavage site of the peptide bond in the Us peptidoglycan is shown (indicated by the arrow in c).
이하, 본 발명의 내용을 하기 실시예를 통해 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.
Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.
[[ 실시예Example 1: 본 발명 1: present invention 엔돌라이신Endolicin LysB4LysB4 의 분리 및 확인]Separation and confirmation of
(1) LysB4 분리 및 확인 (1) LysB4 isolation and identification
크기가 789 bp인 엔돌라이신 유전자는 박테리오파 B4 (KCTC 12013BP)로부터, 두가지 프라이머인 lysB4F(5'-AGTGGAAGTCATATGGCAATGGCATTA-3')와 lysB4R(5'-TAAAAAAAGGATCCCCGAAGGACTTCC-3')를 이용하여 PCR되었고, pET15b 벡터에 클로닝되었다. 그 후 이 콜라이(E. coli) BL21에 형질전환되었다. The endolicin gene, 789 bp in size, was PCR from bacteriopa B4 (KCTC 12013BP) using two primers, lysB4F (5'-AGTGGAAGT CATATG GCAATGGCATTA-3 ') and lysB4R (5'-TAAAAAAAGGATCCCCGAAGGACTTCC-3'). cloned into pET15b vector. This E. coli BL21 was then transformed.
이어, LysB4는 0.1 mM IPTG의 첨가에 의해 발현이 유도되었다. 그 후, 균주의 세포벽을 제거하고 원심분리를 한 뒤, 상등액을 Ni-NTA Superflow 컬럼에 통과시킴으로써 순수한 LysB4를 얻을 수 있었다. LysB4 was then induced to expression by addition of 0.1 mM IPTG. Then, after removing the cell wall of the strain and centrifugation, the pure LysB4 was obtained by passing the supernatant through the Ni-NTA Superflow column.
분리한 LysB4의 아미노산 서열은 서열번호 2와 같았고, 이를 암호화하는 핵산서열은 서열번호 1과 같았다. The amino acid sequence of the isolated LysB4 was the same as SEQ ID NO: 2, and the nucleic acid sequence encoding the same was as SEQ ID NO: 1.
한편, 도 1은 LysB4 아미노산 서열에서 펩타이드 활성을 나타내는 도메인과 호스트 바인딩 도메인을 보여준다. On the other hand, Figure 1 shows a domain showing the peptide activity and the host binding domain in the LysB4 amino acid sequence.
분리된 LysB4는 크기가 약 28 KDa 정도이었으며, 순수하게 분리되었음을 확인할 수 있었다(도 2의 a)
LysB4 was about 28 KDa in size and was confirmed to be purely separated (FIG. 2A).
[[ 실시예Example 2: 2: 엔돌라이신Endolicin LysB4LysB4 의 특성 분석]Characteristic Analysis of
(1) 엔돌라이신 LysB4의 숙주 용해능 (1) Host Solubility of Endolysine LysB4
정체기 단계의 바실러스 세레우스 ATCC 10876 배양액에 실시예 1에서 분리된 엔돌라이신 LysB4를 서로 다른 농도로 처리하였는데, 버퍼만 첨가한 대조군에 비해 흡광도가 효과적으로 감소하였고, 이를 통해 실시예 1에서 분리된 엔돌라이신 LysB4가 바실러스 세레우스를 사멸시키는 것으로 확인할 수 있었다.The Bacillus cereus ATCC 10876 cultures at the stagnation stage were treated with different concentrations of endolisin LysB4 isolated in Example 1, and the absorbance was effectively reduced compared to the control group added with buffer only. Dolysine LysB4 was found to kill Bacillus cereus.
한편, 서로 다른 농도를 첨가하였을 때는 농도가 높아질수록 그 흡광도가 점점 더 많이 줄어들었다(도 2의 b).On the other hand, when different concentrations were added, the higher the concentration, the more the absorbance decreased (b of FIG. 2).
이상과 같은 결과로서, 본 발명의 엔돌라이신 LysB4는 감수성 균주의 성장을 단순 억제시키는 것이 아니라, 해당 균주를 용균시키는 것임을 알 수 있었으며, 적은 양의 엔돌라이신으로도 감수성 균주를 효과적으로 제어한다고 판단할 수 있었다.
As a result, it could be seen that the endolicin LysB4 of the present invention does not simply inhibit the growth of the susceptible strain, but rather lyses the strain, and judges that the susceptible strain is effectively controlled even with a small amount of endolicin. Could.
(2) 엔돌라이신 LysB4의 효과적인 항균 활성 조건 (2) Effective Antimicrobial Activity Conditions of Endolicin LysB4
분리한 엔돌라이신 LysB4의 pH 및 온도에 따른 항균활성을 비교하고자 하였다. 실험결과, 항균 활성은 pH 8.0에서 10.5까지의 염기 범위에서 안정하게 나타났으며(도 3의 a), 37℃에서 75℃까지 비교적 안정한 활성을 보였다(도 3의 b). 특히, 45~55℃에서 가장 효과적인 항균 활성을 나타냈다.
The antimicrobial activity of the isolated endolacin LysB4 with pH and temperature was compared. As a result, the antimicrobial activity was stable in the base range from pH 8.0 to 10.5 (Fig. 3 a), and showed a relatively stable activity from 37 ℃ to 75 ℃ (Fig. 3 b). In particular, the most effective antimicrobial activity was shown at 45 ~ 55 ℃.
(3) 엔돌라이신의 항균 활성 범위 (3) the antimicrobial activity range of endolicin
실시예 1에서 분리한 엔돌라이신 LysB3에 대해 감수성을 나타내는 숙주 스펙트럼을 확인하고자 하였다. 실험은 상기 '(1)'의 방법과 동일하게 수행하였다. The host spectrum showing the sensitivity to the endolicin LysB3 isolated in Example 1 was attempted. The experiment was carried out in the same manner as in the '(1)'.
실혐결과, 하기 표 1에 있는 것처럼, 여러 가지 바실러스 속의 균주에 대해 효과적인 항균 활성을 나타냈는데, 리스테리아 모노싸이토제네스 균주들에 대해서 항균 활성을 보였다. 다만, 스트렙토코커스 써모필로스(Streptococcus thermophilus)나 스타필로코커스 오리우스(Staphylococcus aureus), 스타필로코커스 에피덜미디스(Staphylococcus epidermis) 및 엔테로코커스 피칼리스(Enterococcus faecalis)와 같은 다른 그람 양성균에는 성장 억제 효과를 나타내지 않았다. As a result, as shown in Table 1, it showed an effective antimicrobial activity against various strains of the genus Bacillus, it showed an antimicrobial activity against Listeria monocytogenes strains. However, Streptococcus thermophilus or Staphylococcus aureus ), Staphylococcus Other Gram-positive bacteria such as epidermis and Enterococcus faecalis did not show growth inhibition.
한편, 그람 음성균에 대해서는, EDTA와 같은 세포벽 제거제를 함께 처리했을 때, 살모넬라, 대장균과 같은 그람 음성균에서 성장 억제 효과를 보였고, 그 효과 정도는 균주마다 다소 다르게 나타났다. On the other hand, when Gram-negative bacteria were treated with cell wall remover such as EDTA, they showed growth inhibitory effect in Gram-negative bacteria such as Salmonella and Escherichia coli, and the degree of the effect was somewhat different for each strain.
따라서, 본 발명의 엔돌라이신 LysB4는 그 자체만 처리했을 때는 바실러스 및 리스테리아 모노싸이토제네스에 대해서만 감수성이 나타났으나, 세포벽 제거제를 함께 처리하면, 그람 음성균의 성장까지 억제할 수 있는 넓은 범위의 항균 활성 을 나타낸다고 할 수 있었다. Therefore, the endolisin LysB4 of the present invention exhibited susceptibility only to Bacillus and Listeria monocytogenes when treated alone, but when treated with cell wall remover, a wide range of gram-negative bacteria can be inhibited. It can be said that it exhibits antimicrobial activity.
그람-음성 박테리아
(Gram-negative bacteria)
Gram-negative bacteria
Gram-negative bacteria
그람-양성 박테리아
(Gram-positive bacteria)
Gram-positive bacteria
Gram-positive bacteria
(3) 엔돌라이신 LysB4의 펩티다아제 활성 측정 (3) Determination of Peptidase Activity of Endolysine LysB4
실시예 1에서 분리한 엔돌라이신 LysB4의 작용 기전으로서, 펩티다아제(peptidase) 활성이 있는지를 확인하고자 하였다. As a mechanism of action of endolicin LysB4 isolated in Example 1, it was intended to confirm whether there is peptidase activity.
바실러스 세레우스의 펩티도글리칸을 분리해 낸 뒤, 엔돌라이신 LysB4를 15 μg 처리하고, 0 분(도 4a)과 60 분(도 4b) 후 FDNB를 처리하여 새로 생겨난 아미노기에 DNP가 표지되도록 하였다. 그 후, 염산으로 12시간 정도를 끓여 펩티도글리칸의 펩티드 결합과 당 결합을 끊었다. 이 후, 샘플을 채취하여 고속액체크로마토그래피를 통해서 60분 뒤에 새로 생긴 피크(peak)를 확인하였고, 그 피크의 분획(fraction)을 받아 분자량 분석을 실시하였다. Pepsitiglycans of Bacillus cereus were isolated, treated with 15 μg of endolicin LysB4, and treated with FDNB after 0 min (FIG. 4A) and 60 min (FIG. 4B) to label DNP with newly formed amino groups. It was. Thereafter, the mixture was boiled with hydrochloric acid for about 12 hours to break the peptide bond and the sugar bond of peptidoglycan. Thereafter, a sample was taken and confirmed a new peak after 60 minutes through high performance liquid chromatography.
분석 결과, 분자량 313 정도의 DNP가 라벨링된 D-글루타메이트(glutamate)가 확인되었다.As a result, D-glutamate labeled with DNP having a molecular weight of about 313 was identified.
이상의 결과로부터 본 발명의 엔돌라이신 LysB4는 펩티도글리칸의 L-알라닌(alanine)과 D-글루타메이트(glutamate) 사이를 자르는 펩티다아제 효소임을 알 수 있었다 (도 4c).
From the above results, it was found that the endolicin LysB4 of the present invention is a peptidase enzyme that cuts between L-alanine and D-glutamate of peptidoglycan (FIG. 4C).
[[ 실시예Example 3: 3: LysB4LysB4 를 함유하는 Containing 식품 첨가제의Of food additives 제조] Produce]
표고버섯 분말 50중량%, 새우 분말 30중량%, 다시마 분말 15 중량%, 본 발명의 엔돌라이신 LysB4 5중량%를 혼합하여 조미료로 사용될 수 있는 식품 첨가제 300g을 제조하였다.
50% by weight of shiitake powder, 30% by weight of shrimp powder, 15% by weight of kelp powder, and 5% by weight of endolisin LysB4 of the present invention were mixed to prepare 300 g of a food additive that can be used as a seasoning.
[[ 실시예Example 4: 4: LysB4LysB4 를 함유하는 식품의 제조] Preparation of food containing
<우유 제조> <Milk production>
시판 중인 S 제조의 우유 200 mL에 본 발명의 엔돌라이신 LysB4를 1%(w/v) 첨가하여 본 발명에 따른 우유 조성물을 제조하였다.
A milk composition according to the present invention was prepared by adding 1% (w / v) of the endorlicin LysB4 of the present invention to 200 mL of commercially available S-produced milk.
<바나나 쥬스 제조> Banana juice production
바나나를 믹서에 넣고 갈아 바나나 원액 180 mL를 준비한 후, 본 발명의 엔돌라이신 LysB4를 1%(w/v) 첨가하여 본 발명에 따른 바나나 쥬스 조성물을 제조하였다.
After the bananas were put into a mixer to prepare a 180 mL banana stock solution, 1% (w / v) of endolicin LysB4 of the present invention was added to prepare a banana juice composition according to the present invention.
[[ 실시예Example 5: 5: LysB4LysB4 를 함유하는 사료 첨가제 제조] Preparation of feed additives containing
α-토코페롤 5.0중량%, 오징어간유 0.75중량%, 효모 93.25중량%, 본 발명의 엔돌라이신 LysB4 1.0중량%를 혼합하여 사료 첨가제를 1 kg을 제조하였다.
1 kg of a feed additive was prepared by mixing 5.0% by weight of α-tocopherol, 0.75% by weight of squid liver oil, 93.25% by weight of yeast, and 1.0% by weight of endolisin LysB4 of the present invention.
[[ 실시예Example 6: 6: LysB4LysB4 를 함유하는 사료의 제조] Preparation of feed containing
< 육우 사료조성물 제조> <Beef feed composition manufacturing>
육우용 사료조성물을 제조하였다. 기본사료는 미가공옥수수 5 kg, 면실피 펠렛 1.5 kg, 생미강 0.5 kg, 소맥피1.5 kg, 면실박 0.45 kg, 대두박 1.0 kg, 소금 0.1 kg, 석회석 0.15 kg, 당밀 0.25 kg, 미량광물질 0.05 kg으로 조제되었다. 여기에 본 발명의 엔돌라이신 LysB4를 0.1 kg 첨가하였다.
A feed composition for beef cattle was prepared. The basic feed is 5 kg of raw corn, 1.5 kg of cotton thread pellets, 0.5 kg of fresh rice bran, 1.5 kg of wheat buckwheat, 0.45 kg of cotton bean paste, 1.0 kg of soybean meal, 0.1 kg of salt, 0.15 kg of limestone, 0.25 kg of molasses, and 0.05 kg of trace minerals. Prepared. To this was added 0.1 kg of the endolisin LysB4 of the present invention.
<산란계용 사료조성물 제조> <Manufacturing feed composition for laying hens>
본 발명의 엔돌라이신 LysB4 1.0%(w/w)를 시판 산란계용 사료(디럭스 산란(제품명), 부국사료, 한국)에 첨가하여 산란계용 사료조성물 1 kg을 제조하였다.
Endolysine LysB4 1.0% (w / w) of the present invention was added to a commercial laying hen's feed (deluxe egg laying (product name), Bukook feed, Korea) to prepare a feed composition for laying
<넙치 양식용 사료조성물 제조><Furniture composition for halibut farming>
어분 58중량%, 대두박 4중량%, 콘글루텐분 3중량%, 소맥분 28.7중량%, 오징어간유 4중량%, 비타민 혼합제 1.2중량% 및 미네랄 혼합제 1중량%를 포함한 기본 사료에 본 발명의 엔돌라이신 LysB4 1.0%중량%를 첨가하여 넙치 양식용 사료조성물 1 kg을 제조하였다.
Endolysine of the present invention in a basic feed comprising 58% by weight of fish meal, 4% by weight of soybean meal, 3% by weight of gluten meal, 28.7% by weight of wheat flour, 4% by weight of squid liver oil, 1.2% by weight of vitamin mixture and 1% by weight of mineral mixture 1.0 kg by weight of LysB4 was added to prepare 1 kg of aquaculture feed composition.
서열목록 전자파일 첨부Attach an electronic file to a sequence list
Claims (7)
Endolicin LysB4 having the amino acid sequence of SEQ ID NO: 2.
상기 서열번호 2의 아미노산 서열은,
서열번호 1의 핵산 서열로 암호화되는 것을 특징으로 하는 엔돌라이신 LysB4.
The method of claim 1,
The amino acid sequence of SEQ ID NO: 2,
Endolicin LysB4, which is encoded by the nucleic acid sequence of SEQ ID NO: 1.
상기 엔돌라이신 LysB4는,
바실러스 세레우스(Bacillus cereus) 감염 박테리오파지 B4 (KCTC 12013BP) 유래인 것을 특징으로 하는 엔돌라이신 LysB4.
The method of claim 1,
The endolicin LysB4 is,
Bacillus cereus (Bacillus cereus ) endolicin LysB4, characterized in that it is derived from bacteriophage B4 (KCTC 12013BP).
A food additive containing endolicin LysB4 having the amino acid sequence of SEQ ID NO: 2.
A food composition comprising endolacin LysB4 having the amino acid sequence of SEQ ID NO: 2.
A feed additive containing endolicin LysB4 having the amino acid sequence of SEQ ID NO: 2.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110205306A (en) * | 2019-06-10 | 2019-09-06 | 武汉轻工大学 | Bacillus cereus bacteriophage, bacteriophage composition and inhibiting-bacteria preparation |
| KR102035283B1 (en) | 2019-01-31 | 2019-10-22 | 서울대학교 산학협력단 | Thermal stable endolysin LysCPD2 with lytic activity against Clostridium perfringens |
| CN115427429A (en) * | 2020-03-09 | 2022-12-02 | 尹特荣生物科技株式会社 | Antibacterial protein with cracking activity on bacillus and preparation method thereof |
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| US20100310522A1 (en) | 2007-11-26 | 2010-12-09 | Plant Bioscience Limited | Novel Polypeptides Having Endolysin Activity and Uses Thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102035283B1 (en) | 2019-01-31 | 2019-10-22 | 서울대학교 산학협력단 | Thermal stable endolysin LysCPD2 with lytic activity against Clostridium perfringens |
| CN110205306A (en) * | 2019-06-10 | 2019-09-06 | 武汉轻工大学 | Bacillus cereus bacteriophage, bacteriophage composition and inhibiting-bacteria preparation |
| CN115427429A (en) * | 2020-03-09 | 2022-12-02 | 尹特荣生物科技株式会社 | Antibacterial protein with cracking activity on bacillus and preparation method thereof |
| CN115427429B (en) * | 2020-03-09 | 2023-09-12 | 尹特荣生物科技株式会社 | Antibacterial protein with cracking activity to bacillus and preparation method thereof |
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