• 생명과학회지
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• 제14권2호
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• pp.225-228
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• 2004

나타마이신 배양액을 4$^{\circ}C$와 실온에서 14일 동안 보관하였을 때, 4$^{\circ}C$에서는 나타마이신의 활성이 80% 이상 유지되었으나 실온에서는 27%로 급격하게 감소하였다. 이러한 사실은 나타마이신의 손실을 최소화하기 위해서는 최단 시간 내에 배양액으로부터의 나타마이신을 회수할 필요성과 장기간 보관 시에는 4$^{\circ}C$이하에서 보관할 필요성을 제시하고 있다. 효율적인 나타마이신 정제과정을 개발하기 위해 배양액으로부터의 나타마이신의 추출 용매 및 적정 사용량에 대해 조사한 결과 2g의 나타마이신을 추출하는데 1 l의 methanol을 사용하는 것이 가장 효율적이었다. 확립된 나타마이신 추출에 필요한 methanol의 양과 Diaion HP-20을 이용한 column chromatography를 적용하여 4.2 g의 나타마이신이 함유된 1,800 ml의 배양액으로부터 2.9 g의 나타마이신을 획득하였다. 본 연구에서 개발된 정제과정을 통해 순도가 96.6%이고 회수율이 69.1%인 나타마이신을 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.241-247
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• 2016

Natamycin is a widely used antifungal antibiotic. For natamycin biosynthesis, the gene pimE encodes cholesterol oxidase, which acts as a signalling protein. To confirm the positive effect of the gene pimE on natamycin biosynthesis, an additional copy of the gene pimE was inserted into the genome of Streptomyces gilvosporeus 712 under the control of the ermE* promoter (p_ermE*) using intergeneric conjugation. Overexpression of the target protein engendered 72% and 81% increases in the natamycin production and cell productivity, respectively, compared with the control strain. Further improvement in the antibiotic production was achieved in a 1 L fermenter to 7.0 g/l, which was a 153% improvement after 120 h cultivation. Exconjugants highly expressing pimE and pimM were constructed to investigate the effects of both genes on the increase of natamycin production. However, the co-effect of pimE and pimM did not enhance the antibiotic production obviously, compared with the exconjugants highly expressing pimE only. These results suggest not only a new application of cholesterol oxidase but also a useful strategy to genetically engineer natamycin production.

• 한국축산식품학회지
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• 제36권4호
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• pp.547-557
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• 2016

This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use.

• Journal of Dairy Science and Biotechnology
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• 제37권2호
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• pp.83-93
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• 2019

Food spoilage by fungi is responsible for considerable food waste and economical losses. Among the food products, fermented dairy products are susceptible to deterioration due to the growth of fungi, which are resistant to low pH and can proliferate at low storage temperatures. For controlling fungal growth in dairy products, potassium sorbate and natamycin are the main preservatives used, and natamycin is approved by most countries for use in cheese surface treatment. However, a strong societal demand for less processed and preservative-free food has emerged. In the dairy products, lactic acid bacteria (LAB) are naturally present or used as cultures and play a key role in the fermentation process. Fermentation is a natural preservation technique that improves food safety, nutritional value, and specific organoleptic features. Production of organic acids is one of the main features of the LAB used for outcompeting organisms that cause spoilage, although other mechanisms such as antifungal peptides obtained from the cleavage of food proteins and competition for nutrients also play a role. More studies for better understanding these mechanisms are required to increase antifungal LAB available in the market.

• 미생물학회지
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• 제41권2호
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• pp.140-145
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• 2005

상업적으로 중요한 macrolide계 항진균 학생물질인 natamycin을 생산하는 Streptomyces natalensis ATCC27448의 환자 유전학적인 연구를 위해 대장균으로부터 S. natalensis로 plasmid DNA를 직접 도입하는 형질전환법을 확립하였다. 이러한 S. natalensis의 형질전환은 oriT와 attP 단편을 가지고 있는, ${\Phi}C31$ 유래의 integration 벡터인 pSET152를 이용하여 Escherichia coli ET12567/pUZ28002을 DNA 공여체(donor)로 이용한 접합전달법(conjugal transfer)을 사용하여 확립하였다. 접합전달의 가장 높은 효율은 10 mM의 $MgCl_2$를 포함한 MS 배지에서, $6.25\times10^8$의 E. coli 공여체와 열처리를 하지 않은 S. natalensis의 포자를 사용하여 얻어졌다. 또 얻어진 접합전달체 (exconjugant)에 대하여 southern blot hybridization과 벡터가 삽입된 염색체부분의 염기서열분석을 통해 attB site와 pseudo-attB site를 확인하다. attB site의 경우에는 다른 방선균들처럼 S. natalensis 염색체의 pirin 상동체를 코드하는 ORF내에 존재하였으나 pseudo-attB site는 염색체내 다른 site (GenBank accession no. $YP\_117731$)에 존재하였고 그 염기서열은 attB 염기서열과 차이를 나타내었다.

• 생명과학회지
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• 제21권1호
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• pp.96-101
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• 2011

ATP와 L-methionine으로부터 SAM synthetase (MetK)에 의해 생합성 되는 S-adenosylmethionine (SAM)은 세포내 메틸화에 필요한 메틸기를 제공하는 중심적인 공급체의 역할을 할뿐만 아니라 방선균에서는 일차 및 이차대사산물의 생산 조절에 관여하고 있다는 사실이 밝혀졌다. 이에 논 연구에서는 산업적으로 매우 중요한 항진균성 항생물질인 natamycin을 생산하는 S. natalensis로부터 SAM synthetase 코드하는 metK 유전자를 클로닝하고 동정하였다. S. natalensis에서 클로닝된 metK는 1,209 bp의 염기를 가진 유전자로써 아미노산서열에서 S. pristinaespiralis ATCC 25486과 S. peucetius ATCC 27952의 MetK와 96%, S. violaceusniger Tu 4113과 95% 일치하는 매우 높은 상동성을 보였다. 또 pSET152ET 벡터를 이용해 구축한 metK 고발현용 재조합 플라스미드 pCD1를 S. lividans TK24의 genomic DNA에 도입하여 actinorhodin 생산 유도를 시도해 본 결과 R5 고체배지에서 pCD1이 도입되지 않은 균주에서는 actinorhodin 생산을 전혀 확인할 수 없었지만 pCD1이 도입된 형질전환체에서는 actinorhodin 생산이 강하게 유도되었으며 R4 액체배지에서는 actinorhodin 생산량이 10배 증가되었다. 따라서 본 연구를 통해 클로닝된 S. natalensis 유래 metK 유전자는 방선균에서 이차대사산물의 생산을 유도할 수 있음을 확인할 수 있었다.

• Journal of Dairy Science and Biotechnology
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• 제30권2호
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• pp.75-81
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• 2012

Molds cause severe cheese deterioration, even though some white and blue molds are used for the manufacture of Camembert and Blue cheese, respectively. The species of Geotrichum, Moniliella, Aspergillus, Penicillium, Mucor, Fusarium, Phoma, and Cladosporium are the main fungi that affect contamination during cheese ripening. Once deteriorated by fungal spoilage, cheese becomes toxic and inedible. Fungal deterioration of cheese decreases the nutritional value, flavor profiles, physicochemical and organoleptic properties, and increases toxicity and infectious disease. Fungal contamination during cheese ripening is highly damaging to cheese production in Korean farmstead milk processing companies. Therefore, these companies hesitate to develop natural and ripened cheese varieties. This article discusses the recent and ongoing developments in the removal techniques of fungal contamination during cheese ripening. There are 2 categories of antifungal agents: chemical and natural. Major chemical agents are preservatives (propionic acid, sodium propionate, and calcium propionate) and ethanol. Among the natural agents, grapefruit seed extract, phytoncide, essential oils, and garlic have been investigated as natural antifungal agents. Additionally, some studies have shown that antibiotics such as natamycin and Delvocid$^{(R)}$, have antifungal activities for cheese contaminated with fungi. Microbial resources such as probiotic lactic acid bacteria, Propionibacterium, lactic acid bacteria from Kimchi, and bacteriocin are well known as antifungal agents. In addition, ozonization treatment has been reported to inhibit the growth activity of cheese-contaminating fungi.