Burkholderia sp. OS17의 항균활성 증진을 위한 배양최적화

남영호,최아영,황병수,정유진,Nam, Young Ho,Choi, Ahyoung,Hwang, Buyng Su,Chung, Eu Jin 한국미생물학회 2018 미생물학회지 Vol.54 No.4

본 연구는 담수환경에서 항균활성을 보유한 미생물을 발굴하고, 활성 증진을 위해 배양조건을 최적화하는 것이다. 상주시 중동면 오상저수지에서 시료를 채취하여 38종의 미생물을 순수분리하였다. 16S rRNA 염기서열 분석에 근거하여 Proteobacteria강(22종), Actinobacteria강(7종), Bacteroidetes강(6종), Firmicutes강(3종)으로 구성되어있는 것을 확인하였다. 메티실린내성 황색포도상구군 등 10종의 유해미생물에 대한 항균활성을 보유한 Burkholderia sp. OS17 균주를 선발하였다. 항균활성 증진을 위한 상용배지, 온도, 초기 pH별 생육 및 항균활성 비교실험을 수행하였다. OS17 균주는 YPD 배지, $35^{\circ}C$, pH 6.5로 배양했을 때 가장 활성이 높았다. LB, NB, TSB, R2A 배지와 $20^{\circ}C$, $25^{\circ}C$ 배양했을 때는 생장은 가능하나 항균활성이 전혀 없었다. 이전결과를 바탕으로 YPD 배지, $35^{\circ}C$에서 배양하면서 5 L fermenter를 이용하여 생육, 항균활성, pH 확인을 통해 배양 48시간을 최적 배양시간으로 선정하였다. 항균활성을 보유한 미생물의 배양 최적화는 항균물질 생산에 영향을 미치고, 이는 상업적 응용에 이점으로 작용할 수 있다. In this study, we isolated and identified bacteria from freshwater and soil collected from Osang reservoir, to screen antimicrobial bacteria against various pathogenic bacteria. 38 strains were isolated and assigned to the class Proteobacteria (22 strains), Actinobacteria (7 strains), Bacteroidets (6 strains), and Firmicutes (3 strains) based on 16S rRNA gene sequence analysis. Among them, strain OS17 showed a good growth inhibition against 5 methicillin-resistant Staphylococcus aureus subsp. aureus strains and Bacillus cereus, Bacillus subtilis, Filobasidium neoformans. As a result of the 16S rRNA gene sequence analysis, strain OS17 show the high similarity with Burkholderia ambifaria $AMMD^T$, B. diffusa $AM747629^T$, B. tettitorii $LK023503^T$ 99.8%, 99.7%, 99.6%, respectively. We investigated cell growth and antimicrobial activity according to commercial culture medium, temperature, pH for culture optimization of strain OS17. Optimal conditions for growth and antimicrobial activity in strain OS17 were found to be: YPD medium, $35^{\circ}C$ and pH 6.5. When the strain was cultured in LB, NB, TSB, R2A media at $20^{\circ}C$ and $25^{\circ}C$, the antimicrobial activity did not show. Culture filtrate of strain OS17 showed antimicrobial activity against 5 MRSA strains, Bacillus cereus, Bacillus subtilis, and Filobasidium neoformans with inhibition zones from 2 to 8 mm. Optimal reaction time was 48 h in YPD medium, 100 rpm and 0.3 vvm in 2 L-scale fed-batch fermentation process for antimicrobial activity. Culture optimization of strain OS17 can be improved on antimicrobial activity. Therefore, the antimicrobial activity of Burkholderia sp. OS17 had potential as antibiotics for pathogens including MRSA.

Preparation, characterization, and food application of rosemary extract-loaded antimicrobial nanoparticle dispersions

Lee, Kyung Hyun,Lee, Ji-Soo,Kim, Eun Suh,Lee, Hyeon Gyu Elsevier 2019 LWT- Food science and technology Vol.101 No.-

<P><B>Abstract</B></P> <P>Rosemary extract (RE) was nanoencapsulated with the natural antimicrobial polysaccharide chitosan and γ-poly glutamic acid (γ-PGA) to improve the antimicrobial activity of RE. Three types of RE nanoparticles (RENPs) were generated with different chitosan concentrations (0.1–1.8 mg/mL) and particle sizes (200–600 nm). Nanoencapsulation significantly increased the antimicrobial activity of RE against <I>Bacillus subtilis</I>, and the highest activity was observed for the largest particles, which were prepared with the highest chitosan concentration. Interestingly, the antimicrobial activity of the RE nanoparticle dispersions (RENPDs), the mixture of RENPs and free RE was significantly higher than those of free RE and RENPs, despite the same RE concentration. When the RENPDs were applied to barley tea, all types of RENPDs exhibited significantly higher antimicrobial activity than free RE, reducing the population of <I>Bacillus subtilis</I> by more than 0.5–3.6 log CFU/mL. Therefore, this study suggests the potential of not only RE-loaded chitosan/γ-PGA NPs but also NPDs as a natural preservative in food.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Rosemary extract (RE) was nanoencapsulated with chitosan (CS) to improve antimicrobial activity. </LI> <LI> Blank CS nanoparticles (NPs) without any core material had antimicrobial activity. </LI> <LI> CS synergistically enhanced the antimicrobial activities of the encapsulated RE. </LI> <LI> RE NP dispersions (NPDs) had significantly higher antimicrobial activity than free RE and RE NPs. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Honeybee (<i>Apis cerana</i>) major royal jelly protein 4 exhibits antimicrobial activity

Kim, Bo Yeon,Lee, Kwang Sik,Jung, Boknam,Choi, Yong Soo,Kim, Hye Kyung,Yoon, Hyung Joo,Gui, Zhong-Zheng,Lee, Jungkwan,Jin, Byung Rae 한국응용곤충학회 2019 Journal of Asia-Pacific Entomology Vol. No.

<P><B>Abstract</B></P> <P>Major royal jelly proteins (MRJPs) are important protein components of bee royal jelly (RJ) and exhibit various biological and pharmacological activities. The antimicrobial activities of the royalisin and the jelleines contained within MRJP 1 and MRJP 2 in RJ have been elucidated. However, the antimicrobial effects of other MRJPs remain largely unknown. In this study, we demonstrated the antimicrobial activity of the Asiatic honeybee (<I>Apis cerana</I>) MRJP 4 (AcMRJP4). Recombinant AcMRJP4 was expressed as a 63-kDa protein in baculovirus-infected insect cells. We examined the antimicrobial activity of recombinant AcMRJP4 against bacteria, fungi, and yeast. The mechanisms underlying the antimicrobial activity of AcMRJP4 were assessed using western blot analysis, immunofluorescence staining, and scanning electron microscopy. Recombinant AcMRJP4 bound to the cell walls of bacteria, fungi, and yeast and induced structural damage in the microbial cell walls. AcMRJP4 has an antimicrobial role and exhibits a broad spectrum of antimicrobial activities against bacteria, fungi, and yeast. We demonstrated that AcMRJP4 functions as an antimicrobial agent with activity against bacteria, fungi, and yeast. Together, our data identified a novel function of MRJP 4 as an antimicrobial agent.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The cDNA of the honeybee (<I>Apis cerana</I>) major royal jelly protein 4 (AcMRJP4) was cloned. </LI> <LI> AcMRJP4 exhibits antimicrobial activities against bacteria, fungi, and yeast. </LI> <LI> AcMRJP4 functions as an antimicrobial agent. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Antimicrobial Activity of Antimicrobial Peptide LPcin-YK3 Derived from Bovine Lactophoricin

( Ji-sun Kim ),( Ji-ho Jeong ),( Jang-hee Cho ),( Dong-hee Lee ),( Yongae Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.8

We previously reported on lactophoricin (LPcin), a cationic α-helical antimicrobial peptide derived from bovine milk, which has antimicrobial effects on Candida albicans as well as Grampositive and Gram-negative bacteria. In this study, we designed the LPcin-YK3 peptide, a shorter analog of LPcin, and investigated its antimicrobial activity. This peptide, consisting of 15 amino acids with + 3 net charges, was an effective antimicrobial agent against the on the Gram-positive strain, Staphylococcus aureus (MIC: 0.62 μg/ml). In addition, the hemolytic activity assay revealed that the peptide was not toxic to mouse and human erythrocytes up to 40 μg/ml. We also used circular dichroism spectroscopy to confirm that peptide in the presence of lipid has α-helical structures and later provide an overview of the relationship between each structure and antimicrobial activity. This peptide is a member of a new class of antimicrobial agents that could potentially overcome the problem of bacterial resistance caused by overuse of conventional antibiotics. Therefore, it could be used as a therapeutic or natural additive, particularly in the cosmetics industry.

편백정유의 항산화와 항균활성에 관한 연구

정지영 ( Ji Young Jung ),김한석 ( Han Suk Kim ),김경란 ( Kyeong Ran Kim ) 대한미용학회 2010 대한미용학회지 Vol.6 No.4

Human body and the scalp of internal, external or when symptoms occur, which often make acne and cause inflammation, Demodex in the skin, such as parasitic skin flora Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes on the wall directly by stimulating hair follicles go into the dermis, causing skin irritation leads to inflammation. In this study, using Hinoki Cypress Oil Antioxidant and antimicrobial activity of skin flora and to learn, skin toner manufacturer in the clinical trials for skin flora, and antimicrobial activity by measuring the absorbance, Salmonella typhimurium Ames test carried out by using the functions were verified. Results of this study Hinoki Cypress Oil on DPPH free radical scavenging activity of 5 μL/mL in the 7.93±0.07%, 10 μL/mL and 15.49±0.18%, 20 μL/mL and 17.57±0.23%, 40 μL/mL and 20.98±0.54%, 80 μL/mL in the 28.06±0.69% appeared to have antioxidant benefits were concentration-dependent manner. However, as a result of low levels of Hinoki Cypress Oil scavenging of SC50 did not represent a positive daejomuljilin the BHT 0.1 mg/mL of the scavenging rate of 74.89±0.64% respectively. Hinoki Cypress Oil Minimum inhibitory concentration (MIC) in S. aureus appeared as a 0.02%, S. epidermidis and P. acnes is determined by the 0.01% S. aureus was higher than the Sensibility. Minimum bactericidal concentration (MBC) of S. epidermidis were determined from the 0.03%, 0.04% did not show a survival Strains. P. acnes in the 0.04% was 99.9% almost completely inactivated S. aureus of the Minimum bactericidal concentration (MBC) that was not identified. Antimicrobial activity in verification Hinoki Cypress Oil 10% from 20% show restraint in the fungus P. acnes on the two antimicrobial S. aureus and S. epidermidis to the antimicrobial activity than the three strains appear to be a very good chance that the antimicrobial functions Hinoki Cypress Oil concentration increases, respectively. Antimicrobial for skin flora and growth inhibitory concentration of skin flora skin toner P. acnes, S. epidermidis, S. aureus appeared in order, the higher the concentration, application time is longer, showed higher antimicrobial. Using the genetic toxicological safety of Salmonella typhimurium in comparison with the results of control samples by the colony counts could not see the difference between the speech was considered so. In this study, Hinoki Cypress Oil on the skin flora Antioxidant and antimicrobial activity with low concentrations found to be very high in the scalp and skin which are used in functional cosmetics have been suggested as a possibility.

Antioxidant and Antimicrobial Activities of Extracts from a Medicinal Plant, Sea Buckthorn

Jeong, Jong-Hyun,Lee, Ji-Won,Kim, Kyoung-Su,Kim, Ju-Sung,Han, Sang-No,Yu, Chang-Yeon,Lee, Ju-Kyong,Kwon, Yong-Soo,Kim, Myong-Jo The Korean Society for Applied Biological Chemistr 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.1

This study was performed to evaluate the antioxidant and antimicrobial activities from the methanol extracts of the root and stem of Hippophae rhamnoides and their further partitioned fractions including hexane, ethyl acetate, butanol, and water. Antioxidant activities of the plant parts were measured by 1,1-dephenyl-2-picryl-hydrazyl (DPPH), superoxide dismutase (SOD)-like activity, and ferric thiocyanate (FTC) assays, and compared to standard antioxidants of butylated hydroxyl anisole, butylated hydroxyl toluene, $\alpha$-tocopherol, and ascorbic acid. Antimicrobial activity of the plant extracts and fractions were evaluated by determining the minimum inhibitory concentration values. DPPH assay showed that the overall strong antioxidant activities from the methanol extracts and fractions. The values of SOD-like activities in hexane fractions of the root and stem were close to the measurement of a reference, ascorbic acid at 1,000 ppm. The methanol extract of the root in FTC assay showed a remarkable antioxidant and free radical scavenging activity. The extracts and fractions of the root and stem showed better antimicrobial activity than compared antimicrobial agents, (+)-catechin, ketoconazol, and mycostantin. This study indicates that the plant root and stem contain a variety of compounds contributing to antioxidant and antimicrobial activity, which could be used for food additives and the development of useful natural compounds.

소목 추출물의 항균력 및 살균소독력

이진영,민경진,Lee, Jin-Young,Min, Kyung-Jin 한국환경보건학회 2011 한국환경보건학회지 Vol.37 No.2

Antimicrobial activity and bactericidal activity of Caesalpinia sappan L. extracts were investigated against five food-borne pathogens, E. coli, S. aureus, S. typhimurium, B. cereus and L. monocytogenes. Methanol extract of Caesalpinia sappan L. revealed antimicrobial activities against five pathogens. In particular, by paper disc diffusion the highest activity was shown against L. monocytogenes. Antimicrobial activities of methanol extracts showed the most potent activities, but hexane fraction had no activity. Fractions of ethyl acetate and butanol turned out to have higher antimicrobial activities against Gram(+) bacteria than Gram(-) bacteria. The minimum inhibitory concentration against five food-borne pathogens was 1.563 mg/ml on Gram(+) bacteria and 3.125 mg/ml on Gram(-) bacteria. The result of antimicrobial activity in a shaking flask method showed that bacterial growth rate fell by more than 99.999% at 3.125 mg/ml of methanol extract. The highest rate of viable reduction (99.998%) was shown at 0.781 mg/ml of methanol extract against L. monocytogenes. After five minutes of reaction between test strains and methanol extracts, the growth rates of five kinds of bacteria were reduced by more than 99.999% at a concentration of 100 mg/ml. Therefore, it is suggested that methanol extracts of Caesalpinia sappan L. can be developed as a natural sanitizer or disinfectant.

왕겨초액 처리지의 항균 특성

민춘기(Choonki Min),조중연(Joongyeon Jo),신준섭(Junsub Shin),이세은(Seeun Lee) 한국펄프·종이공학회 2010 한국펄프·종이공학회 학술발표논문집 Vol.2010 No.10

Antimicrobial activities of Acidic Liquid with Carbonized Rice Hull(ALCRH) and paper treated with ALCRH were investigated to apply ALCRH for paper as a natural antimicrobial agent. ALCRH showed antimicrobial activity for both bacteria and yeast, with higher performance for bacteria than for yeast. Antimicrobial activity was not developed on paper coated with ALCRH by bar coater probably due to the evaporation of antimicrobial compounds of ALCRH from the paper surface with time. It was necessary for ALCRH to penetrate into paper to develop antimicrobial activity. Saturation of paper by dipping it in ALCRH was recommended as one of the ways to make antimicrobial paper.

감초 성분의 항균활성, 정량 및 방부력에 관한 연구

김혜진 ( Hye Jin Kim ),장하나 ( Ha Na Jang ),배정윤 ( Jeong Yun Bae ),하지훈 ( Ji Hoon Ha ),박수남 ( Soo Nam Park ) 한국미생물생명공학회(구 한국산업미생물학회) 2014 한국미생물·생명공학회지 Vol.42 No.4

본 연구에서는 감초성분인 glycyrrhizin, liquiritin, liquiritigenin, isoliquiritigenin의 항균활성을 측정하고, 원산지별 감초 추출물에서 항균활성 성분인 isoliquiritigenin과 liquiritigenin을 정량하여 비교평가하였다. 또한 challenge test를 통하여 가장 뛰어난 항균활성을 나타낸 isoliquiritigenin과 합성 방부제인 메틸 파라벤과의 방부력을 비교하였다. 감초 성분의 항균활성을 disc diffusion assay를 통해 측정한 결과, Liquiritigenin과 isoliquiritigenin이 B. subtilis, P. acnes, E. coli, P. aeruginosa에 대해 저해활성을 보였다. 특히 감초성분 중 큰 항균활성을 나타낸 isoliquiritigenin과 합성 방부제로 사용되는 메틸 파라벤과 프로필 파라벤의 최소저해농도를 비교한 결과 isoliquiritigenin이 강력한 항균활성을 나타내는 것을 확인하였다. HPLC를 통해 원산지별 감초 추출물 isoliquiritigenin과 liquiritigenin을 정량한 결과, 이들의 함량은 한국산 감초가 중국산 감초와 우즈베키스탄산 감초보다 높은 것을 확인하였다. 이를 통해 한국산 감초가 우수한 항균력을 나타내는데 isoliquiritigenin, liquiritigenin 함량이 영향을 미치는 것으로 판단된다. Isoliquiritigenin의 B. subtilis, P. acnes, E. coli, P. aeruginosa에 대한 challenge test를 통해 메틸 파라벤 보다 isoliquiritigenin이 뛰어난 방부력을 나타냄을 확인하였다. 이상의 본 연구 결과로부터 감초 성분의 항균활성을 확인하였고, 원산지별 감초(한국, 중국, 우즈베키스탄) 중 한국 감초가 우수한 항균력을 나타내는데 isoliquiritigenin과 liquiritigenin 함량이 영향을 미치는 것으로 사료된다. 또한 감초 성분 중 가장 우수한 항균활성을 나타낸 isoliquiritigenin은 파라벤류를 대체 할 수 있는 천연 보존제로써 이용 가능성이 있다고 판단된다. The present study was aimed at investigating the antimicrobial activities of licorice’s active ingredients. Four samples of licorice ingredients (glycyrrhizin, liquiritin, liquiritigenin, and isoliquiritigenin) were evaluated for their antimicrobial activities against six skin microorganisms. The bioassay applied for determining the antimicrobial effects employed a disc diffusion assay, the minimum inhibitory concentration, and the challenge test. The ingredients showed antibacterial activities. Especially, isoliquiritigenin has significant antimicrobial activities against two Gram-positive (Bacillus subtilis, Propionobacterium acnes) and two Gramnegative (Escherichia coli, Pseudomonas aeruginosa) bacteria. These samples had much higher antimicrobial activities than synthetic preservatives. Our results reveal that liquiritigenin and isoliquiritigenin could be useful compounds for the development of antibacterial agents for the preservation of cosmetics and foods. The two flavonoids, liquiritigenin and isoliquiritigenin, sourced from Korea, China, Uzbekistan, were quantified using HPLC. The results demonstrated that Korean licorice has two flavonoids (liquiritigenin, isoliquiritigenin) in much higher quantities than was observed in the licorice obtained from the two other countries. Thus, isoliquiritigenin and Korean licorice extract represent new candidates for antimicrobial agents.

Antioxidant and Antimicrobial Activities of Extracts from a Medicinal Plant, Sea Buckthorn

( Jong Hyun Jeong ),( Ji Won Lee ),( Kyoung Su Kim ),( Ju Sung Kim ),( Sang No Han ),( Chang Yeon Yu ),( Ju Kyong Lee ),( Yong Soo Kwon ),( Myong Jo Kim ) 한국응용생명화학회 2010 Applied Biological Chemistry (Appl Biol Chem) Vol.53 No.1

This study was performed to evaluate the antioxidant and antimicrobial activities from the methanol extracts of the root and stem of Hippophae rhamnoides and their further partitioned fractions including hexane, ethyl acetate, butanol, and water. Antioxidant activities of the plant parts were measured by 1,1-dephenyl-2-picryl-hydrazyl (DPPH), superoxide dismutase (SOD)-like activity, and ferric thiocyanate (FTC) assays, and compared to standard antioxidants of butylated hydroxyl anisole, butylated hydroxyl toluene, α-tocopherol, and ascorbic acid. Antimicrobial activity of the plant extracts and fractions were evaluated by determining the minimum inhibitory concentration values. DPPH assay showed that the overall strong antioxidant activities from the methanol extracts and fractions. The values of SOD-like activities in hexane fractions of the root and stem were close to the measurement of a reference, ascorbic acid at 1,000 ppm. The methanol extract of the root in FTC assay showed a remarkable antioxidant and free radical scavenging activity. The extracts and fractions of the root and stem showed better antimicrobial activity than compared antimicrobial agents, (+)-catechin, ketoconazol, and mycostantin. This study indicates that the plant root and stem contain a variety of compounds contributing to antioxidant and antimicrobial activity, which could be used for food additives and the development of useful natural compounds.