• Journal of Life Science
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• v.14 no.2
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• pp.225-228
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• 2004

Natamycin, produced by Streptomyces natalensis ATCC27448, is a polyene macrolide antibiotic, is widely used in the food industry in order to prevent mould contamination. This study carried out to develop an efficient purification process of natamycin from fermentation broth. The stability of natamycin in fermentation broth during storage period was investigated at 4$^{\circ}C$ and room temperature. After the storage of fermentation broth for 14 days at 4$^{\circ}C$, residual activity of natamycin was about 80% but decreased by 27% at room temperature. As solvent to extract natamycin from fermentation broth, methanol was the most efficient. A developed purification procedure includes methanol extraction and Diaion HP-20 column chromatography. Approximately 2.9 g of natamycin was obtained with a final yield of 69.1% and purity of 96.6% from 1.8 l of fermentation broth by this developed purification procedure.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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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.

• Food Science of Animal Resources
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• v.36 no.4
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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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• v.37 no.2
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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.

• Korean Journal of Microbiology
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• v.41 no.2
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• pp.140-145
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• 2005

Streptomyces natalensis ATCC27448 produces natamycin, a commercially important macrolide antifungal antibiotic. For molecular genetic study of S. natalensis, we have developed a system for introducing DNA into S. natalensis via conjugal transfer from Escherichia coli. An effective transformation procedure for S. natalensis was established based on transconjugation from E, coli ET12567/pUZ8002 using a ${\Phi}C31$-derived integration vector, pSET152, containing oriT and attP fragments. The high frequency was obtained on MS medium containing 10 mM $MgCl_2$ using $6.25\times10^8$ of E.coli donor cells without heat treatment of spores. In addition, southern blot analysis of exconjugants and the sequence of plasmids containing DNA flanking the insertion sites from the chromosome revealed that S. natalensis contains a single ${\Phi}C31$ attB site and at least a secondary or pseudo attB site. Similar to the case of various Streptomyces species, a single ${\Phi}C31$ attB site of S. natalensis is present within an ORF encoding a pirin-homolog, but a pseudo-attB site is present within a distinct site (GenBank accession no. $YP\_117731$) and also its sequence deviates from the consensus sequences of attB sequence.

• Journal of Life Science
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• v.21 no.1
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• pp.96-101
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• 2011

S-Adenosyl-L-methionine synthtase (SAM-s) catalyzes the biosynthesis of SAM from ATP and L-methionine. SAM plays important roles in the primary and secondary metabolism of cells. A metK encoding a SAM-s was searched from Streptomyces natalensis producing natamycin, a predominantly a strong antifungal agent, inhibiting the growth of both yeasts and molds and preventing the formation of aflatoxin in filamentous fungi. To obtain the metK of S. natalensis, PCR using primers designed from the two highly conserved regions for metK genes of Streptomyces strains was carried out, and an intact 1.2-kb metK gene of S. natalensis was cloned by genomic Southern hybridization with PCR product as a probe. To identify the function of the cloned metK gene, it was inserted into pSET152ET for its high expression in the Streptomyces strain, and then introduced into S. lividans TK24 as a host by transconjugation using E. coli ET12567(pUZ8002). The high expression of metK in S. lividans TK24 induced actinorhodin production on R5 solid medium, and its amount in R4 liquid medium was 10-fold higher than that by exconjugant including only pSET152ET.

• Journal of Dairy Science and Biotechnology
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• v.30 no.2
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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.