• 한국동물위생학회지
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• 제47권1호
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• pp.19-26
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• 2024

Canine atopic dermatitis (CAD) is an inflammatory and pruritic skin disease with a genetic predisposition, characterized by allergic sensitivity. It is known for its distinctive clinical features, including a high recurrence rate and chronic progression. To manage CAD, medications such as steroids and immunosuppressants are commonly used, but consideration should be given to the potential resistance and side effects associated with long-term use. In order to reduce these risks, various adjunctive factors are currently under consideration. One of these adjunctive agents, probiotics have shown effectiveness in regulating atopic dermatitis by modulating immune responses, as demonstrated in several recent studies. In this study, a substance combining three probiotics-L. plantarum, L. reuteri, and Ped. Acidilactici-was used in patients diagnosed with CAD, and its clinical effects and safety were evaluated. The trial involved four groups: a group receiving conventional treatment for atopic dermatitis (A), a group prescribed low-dose probiotics (B), a group prescribed high-dose probiotics (C), and a group prescribed topical probiotics (D). For assessment, the Canine Atopic Dermatitis Extent and Severity Index (CADESI), Trans-Epidermal Water Loss (TEWL) test, gut microbiome, and serum IgE test were conducted. As a result, the CAD severity index (CADESI-4) significantly decreased in the probiotics groups (B & C). In the serum total IgE test, the groups consuming probiotics showed a significant difference, while the group using topical probiotics (D) did not exhibit a significant change. Also, the TEWL test showed improved scores in the probiotics groups (B & C). Therefore, L. plantarum, L. reuteri, and Ped. Acidilactici probiotic combination could be considered as an effective adjunctive treatment, especially for atopic patients with moderate to severe skin lesions.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.403-411
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• 2005

The relationship between plasmid (~9.5 kb) and pediocin PA-1 in P. acidilactici K10 was confirmed by plasmid curing. The pediocin operon of P. acidilactici K10 was amplified by PCR (polymerase chain reaction), and the nucleotide sequence was analyzed. The sequence of the pediocin operon of P. acidilactici K10 was similar to those of P. acidilactici strains producing pediocin PA-1/ AcH. For the expression of pediocin PA-1 in E. coli, a pQEPED (pQE-30 Xa::mature pedA) was constructed. His-tagged recombinant pediocin PA-1 (-6.5 kDa) was translated by cell-free in vitro transcription and translation using pQEPED as a DNA template. Theresult of slot blotting assay showed that transcription of recombinant pedA in E. coli M15 was induced by the addition of isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) at the final concentration of 1 mM. Although the recombinant pediocin PA-1 inhibited the growth of E. coli, it was expressed in the host strain and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography under denaturing condition. This is the first report for the production and one-step purification of biologically active recombinant pediocin PA-1 in E. coli.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.149-153
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• 2001

To investigate the mode of bactericidal action for antimicrobial peptide, pediocin, synthetic and mutant pediocins were prepared by direct chemical synthesis. Native pediocin was purified from Pedio-coccus acidilactici M and its conformational structure and bactericidal functions were analyzed and compared to synthetic pediocin. Schematic mode of pediocin actions, how pediocin binds on the target cell membrane, penetrates and makes tunnel are proposed. For these purposes, primary and secondary structures of pediocin was analyzed and disulfide bond assignment was also done. The pediocin purified from P. acidilactici M had high effective bactericidal ability against gram positive bacteria, especially Listeria monocytogenes and was very stable at extreme pHs and even at high temperatures such as autoclaving temperature (121$^{\circ}C$). Pediocin was consisted of 44 amino acids with four cysteines. Novel synthetic peptides were achieved by solid phase peptide synthesis(SPPS) method. To explain the function of cysteine in C-terminal region, mutant pediocin, Ped[C24A＋C44A], was synthesized and their structural and biological functions were analyzed. Second mutant pediocin, Ped[KllE], was prepared to explain the function of lysine at 11 of N-terminal part of pediocin, especially loop of $\beta$-sheet, and to predict the initial binding site of pediocin. The native and synthetic pediocins was showed random coil conformation by spectropolarimetry in moderate conditions. This conformation was observed in extreme conditions such as high temperature and low and high pHs, also. Circular dichroism(CD) data also showed the existence of $\beta$-turn structure in N-terminal part both native and synthetic pediocins. A structural model for pediocin predicts that 18 amino acids in the N-terminal part of the peptide assume a three-strand $\beta$-sheet conformation. This random coil in C-terminal part of pediocin was converted to folding structure, helix structure, in nonpolar solvents such as alcohol and TFE. The disulfide bond between $^{9}$ Cys and $^{14}$ Cys was concrete and inevitable, however, evidences of disulfide bond between $^{24}$ Cys and $^{44}$ Cys was not. Data of Ped[C24A＋C44A], pediocin mutant showed that $^{44}$ Cys was required during killing the target cells but not inevitable, since Ped[C24A＋C44A] still have bactericidal activity but much less than native pediocin. Another pediocin mutant, Ped[KllE], had still bactericidal activity, was controversial to propose that positive charge like as $^{11}$ Lys in loop or hinge in bacteriocin bound or helped to binding to microorganism with electrostatic interaction between cell membrane especially teichoic acid and positive amino acid nonspecifically. The conformation of pediocin among native, synthetic and mutant pediocins did not show big difference. The conformations between oxidized and reduced pediocin were almost similar regardless of native or synthetic.

• 한국식품조리과학회지
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• 제7권1호
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• pp.15-25
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• 1991

김치 발효 중에 주로 나타나는 Lac. plantarum, Leu. mesenteroides, Ped. acidilactici, Lac. brevis 등의 젖산균들이 김치 발효에 미치는 영향을 규명하고자, 이 젖산균들을 순수배양하여 염도 2.31%의 김치즙에 접종한 후 $30^{\circ}C$, $21^{\circ}C$, $14^{\circ}C$, $7^{\circ}C$에서 발효시키면서 pH및 총산도의 변화, 휘발성 유기산과 비휘발성 유기산의 변화를 관찰하였다. Control group과 Leu. mesenteroides를 접종한 시료, 4종의 젖산균을 혼합 접종한 시효들의 발효온도에 따른 pH 및 산도 변화 경향은 서로 매우 유사하며, 특히 1$14^{\circ}C$에서 이러한 경향은 더 뚜렷하게 나타났다. Lac. plantarum이나 Ped. acidilactici 그리고 Lac. brevis를 접종한 시료들은 control group이나 Leu. mesenteroides, 4종의 젖산균을 혼합접종한 시료보다 더 느리게 pH 및 총산도가 변화하였으며 특히 $14^{\circ}C$에서는 control group과 Leu. mesenteroides 그리고 4종의 젖산균을 혼합 접종한 시료에서만이 발효가 진행되었다. 휘발성산과 비휘발성산의 변화 양상도 control group, Leu. mesenteroides를 접종한 시료, 4종의 젖산균을 혼합 접종한 시료에 있어서 서로 매우 유사하였다. 검출한 휘발성 유기산으로는 formfic, acetic acid뿐이었으며, 이들은 Leu. mesenteroides나 Lac. brevis 등에 의해 주로 생성됨을 알 수 있다. Leu. mesenteroides를 접종한 시료에서 다른 발효온도에서보다 $14^{\circ}C$에서 acetic acid의 함량이 높았다. 비휘발성 유기산은 모든 시료에서 lactic acid가 검출되었고, Leu. mesenteroides를 접종한 시료에서 $21^{\circ}C$와 $14^{\circ}C$에 citric aicd가 검출되었으며 succinic acid는 controlgroup과 Lac. brevis를 접종한 시료에서 검출되었다.

• 한국식품조리과학회지
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• 제7권2호
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• pp.89-95
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• 1991

김치발효 중에 주로 나타나는 Lac. plantarum, Leu. mesenteoides, Ped. acidilactici, Lac. brevis 등의 젖산균들이 김치발효에 미치는 영향을 규명하고자, 이 젖산균들을 순수 배양하여 염도 2.31%의 김치즙에 접종한 후 $30^{\circ}C$, $21^{\circ}C$, $7^{\circ}C$에서 발효시키면서 알코올 당의 변화 및 생균수의 변화를 관찰하고 관능검사를 하였다. Ethyl alcohol은 control group과 Leu. mesenteoides를 접종한 시료, 4종의 젖산균을 혼합 접종한 시료에서 검출되었으며 특히 $14^{\circ}C$에서 더 많은 양이 검출되었다. 당은 발효가 진행됨에 따라 감소하였고, 감소 정도는 pH 및 총산도의 변화와 잘 일치하였다. 총 생균수는 발효초기에는 증가하다가 서서히 감소하는 경향을 보였으며 control group과 Leu. mesenteroides에 있어서 김치가 적당히 익은 정도의 pH및 산도를 보이는 시기와 최고의 균수에 도달하는 시기가 일치하였다. 관능검사 결과 모든 온도에서 Control group이 냄새와 맛이 가장 좋았고 그 다음 4종의 젖산균을 혼합접종한 시료 Leu. mesentenoides를 접종한 시료의 순으로 나타났으며, 온도에 있어서는 $14^{\circ}C$에서 맛과 냄새가 좋았다.

• Journal of Microbiology and Biotechnology
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• 제20권8호
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• pp.1215-1218
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• 2010

The recombinant DNA pLR5cat_PSAB, in which pediocin PA-1 structural and immunity genes (pedAB) fused with the promoter and deduced signal sequence of an ${\alpha}$-amylase gene from a bifidobacterial strain were inserted in Escherichia coli-lactobacilli shuttle vector pLR5cat, was transferred to Lactobacillus reuteri KCTC 3679 and the transformant presented bacteriocin activity. The recombinant L. reuteri KCTC 3679 transformed with the shortened pLR5cat(S)_PSAB, where a nonessential region for the lactobacilli replicon was removed, also showed bacteriocin activity. The molecular mass of the secreted pediocin PA-1 from the recombinant bacteria was the same as that of native pediocin PA-1 (~4.6 kDa) from Pediococcus acidilactici K10 on a sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. In cocultures with Listeria monocytogenes, the recombinant L. reuteri KCTC 3679 effectively reduced the viable cell count of the pathogenic bacterium by a 3 log scale compared with a control where L. monocytogenes was incubated alone.