Journal of Microbiology

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Exploring the biosynthetic potential of Korean Actinobacteria for antibacterial metabolite discovery: Sehong Park, Hyun-Woo Je, Yujin Cha, Boncheol Gu, Yeojeong Cho, Jin-Il Kim, Ji Won Seo, Seung Bum Kim, Jino Son, Moonsuk Hur, Changmin Sung, Min-Kyu Oh, Hahk-Soo Kang; J. Microbiol. 2025;63(9):e2504002. Published online September 30, 2025; DOI: https://doi.org/10.71150/jm.2504002

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Abstract PDF Supplementary Material: Actinobacteria, a phylum of Gram-positive bacteria, are renowned for their remarkable ability to produce antibacterial natural products. The National Institute of Biological Resources (NIBR) of Korea maintains a collection of Korean native actinobacteria. In this study, we explored the phylogenetic and biosynthetic diversity of the NIBR actinobacteria collection to assess its potential as a source of new antibacterial natural products. A 16S rDNA-based phylogenetic analysis revealed a high level of genetic diversity within the collection, with a predominance of Streptomyces, along with rare actinobacterial genera such as Kitasatospora and Micromonospora. Additionally, genetic network analysis of biosynthetic gene clusters (BGCs) from 15 sequenced NIBR actinobacterial strains demonstrated extensive BGC diversity, with many clusters identified as cryptic. Screening of culture extracts for antibacterial activity, followed by dereplication of active extracts, suggested the presence of potentially novel antibacterial natural products. Activity-guided isolation and whole-genome sequencing of the active strain KU57 led to the isolation of one new and three known svetamycin congeners along with their BGC. Overall, our findings highlight the NIBR actinobacteria collection as a valuable source for the discovery of new antibacterial natural products.

Journal Articles

[PROTOCOL]Analyzing viral epitranscriptomes using nanopore direct RNA sequencing: Ari Hong , Dongwan Kim , V. Narry Kim , Hyeshik Chang; J. Microbiol. 2022;60(9):867-876. Published online August 24, 2022; DOI: https://doi.org/10.1007/s12275-022-2324-4

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RNA modifications are a common occurrence across all domains of life. Several chemical modifications, including N6- methyladenosine, have also been found in viral transcripts and viral RNA genomes. Some of the modifications increase the viral replication efficiency while also helping the virus to evade the host immune system. Nonetheless, there are numerous examples in which the host's RNA modification enzymes function as antiviral factors. Although established methods like MeRIP-seq and miCLIP can provide a transcriptome- wide overview of how viral RNA is modified, it is difficult to distinguish between the complex overlapping viral transcript isoforms using the short read-based techniques. Nanopore direct RNA sequencing (DRS) provides both long reads and direct signal readings, which may carry information about the modifications. Here, we describe a refined protocol for analyzing the RNA modifications in viral transcriptomes using nanopore technology.

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Ines Benčik, Lara Saftić Martinović, Tea Mladenić, Saša Ostojić, Sanja Dević Pavlić
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Whole-genome sequencing analysis of Shiga toxin-producing Escherichia coli O22:H8 isolated from cattle prediction pathogenesis and colonization factors and position in STEC universe phylogeny: Wanderson Marques Da Silva , Mariano Larzabal , Flavia Figueira Aburjaile , Nahuel Riviere , Luisina Martorelli , James Bono , Ariel Amadio , Angel Cataldi; J. Microbiol. 2022;60(7):689-704. Published online June 22, 2022; DOI: https://doi.org/10.1007/s12275-022-1616-z

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Abstract PDF

Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen capable of causing illness in humans. In a previous study, our group showed that a STEC isolate belonging to O22:H8 serotype (strain 154) can interfere with STEC O157:H7 colonization both in vitro and in vivo. Using whole-genome sequencing and genomic comparative, we predicted a subset of genes acquired by O22:H8 strain 154 through horizontal gene transfer that might be responsible for the phenotype previously described by our group. Among them were identified genes related to the pathogenesis of non-LEE (locus of enterocyte effacement) STEC, specific metabolic processes, antibiotic resistance and genes encoding for the T6SS-1 that is related to inter-bacterial competition. In addition, we showed that this strain carries stx1c and stx2dact, a mucus-inducible variant. The results obtained in this study provide insights into STEC genomic plasticity and the importance of genomic islands in the adaptation and pathogenesis of this pathogen.

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Wanderson Marques Da Silva, Libia Yael Smith, Flavia Figueira Aburjaile, Mariano Larzabal, Sandy Ingrid Aguiar Alves, Pablo Farace, Carlos Willian Dantas, Mateus Matiuzzi Costa, Bertram Brenig, Rommel Thiago Jucá Ramos, Vasco Azevedo, Angel Cataldi
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Lipocalin2 as a potential antibacterial drug against Acinetobacter baumannii infection: Daejin Lim , Su-Jin Park , Ha Young Kim , Minsang Shin , Miryoung Song; J. Microbiol. 2022;60(4):444-449. Published online March 28, 2022; DOI: https://doi.org/10.1007/s12275-022-2007-1

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Abstract PDF

Available antibiotics to treat Acinetobacter baumannii infection is limited due to increasing resistance and the emergence of multiple drug-resistant strains. Hence, discovering effective agents against A. baumannii to reduce the number of infectionrelated deaths is imperative. In search of novel and alternative antibiotics, the antibacterial function of lipocalin2 (Lcn2) was investigated to treat systemic infections of A. baumannii using a mouse neutropenia model. We observed a significant increase in serum Lcn2 levels upon bacterial injection into the mouse, and the administration of recombinant Lcn2 (rmLcn2) extended their survival. Such protective effects were also observed in rmLcn2-pretreated macrophages, where rmLcn2 reduced the survival of the pathogen inside the macrophages. The underlying molecular mechanism of Lcn2 protection was also investigated. We observed that pretreatment of the Raw- 264.7 macrophages with rmLcn2 markedly altered the expression of tonB3, which encodes a component of the transporter for ferrisiderophores in A. baumannii. However, the expression of katG, the gene encoding catalase, remained unaffected. These indicate that Lcn2-mediated defense against the pathogen is related to nutritional immunity rather than reactive oxygen species (ROS) production. Furthermore, the addition of rmLcn2 in infected mice diminished bacterial burden in multiple organs and enhanced the expression of tonB3 in the liver, spleen, and lungs of the infected mice. Increased survival rate due to rmLcn2 treatment declined when the infection model was established using lcn2-defective (lcn2-/-) mice, which indicated the necessity of endogenous Lcn2. Therefore, the antibacterial function of Lcn2 can be exploited to develop an alternative therapeutic agent against A. baumannii.

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Jessica R. Sheldon, Lauren E. Himmel, Dillon E. Kunkle, Andrew J. Monteith, K. Nichole Maloney, Eric P. Skaar, David S. Weiss
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Gold nanoparticle-DNA aptamer-assisted delivery of antimicrobial peptide effectively inhibits Acinetobacter baumannii infection in mice: Jaeyeong Park , Eunkyoung Shin , Ji-Hyun Yeom , Younkyung Choi , Minju Joo , Minho Lee , Je Hyeong Kim , Jeehyeon Bae , Kangseok Lee; J. Microbiol. 2022;60(1):128-136. Published online December 29, 2021; DOI: https://doi.org/10.1007/s12275-022-1620-3

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Abstract PDF

Acinetobacter baumannii causes multidrug resistance, leading to fatal infections in humans. In this study, we showed that Lys AB2 P3-His–a hexahistidine-tagged form of an antimicrobial peptide (AMP) loaded onto DNA aptamer-functionalized gold nanoparticles (AuNP-Apt)–can effectively inhibit A. baumannii infection in mice. When A. baumannii-infected mice were intraperitoneally injected with AuNP-Apt loaded with Lys AB2 P3-His, a marked reduction in A. baumannii colonization was observed in the mouse organs, leading to prominently increased survival time and rate of the mice compared to those of the control mice treated with AuNP-Apt or Lys AB2 P3-His only. This study shows that AMPs loaded onto AuNP-Apt could be an effective therapeutic tool against infections caused by multidrug-resistant pathogenic bacteria in humans.

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Repositioning of a mucolytic drug to a selective antibacterial against Vibrio cholerae: In-Young Chung† , Bi-o Kim† , Hye-Jeong Jang† , You-Hee Cho; J. Microbiol. 2020;58(1):61-66. Published online January 2, 2020; DOI: https://doi.org/10.1007/s12275-020-9590-9

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Drug repositioning, the approach to explore existing drugs for use in new therapeutic indications, has emerged as an alternative drug development strategy. In this study, we found that a mucolytic drug, N-acetylcysteine (NAC) showed antibacterial activity against Vibrio cholerae. NAC can provide acid stress that selectively inhibited the growth of V. cholerae among other bacterial pathogens. To address the antibacterial mechanism of NAC against V. cholerae, six acr (acetylcysteine- resistant) mutants were isolated from 3,118 random transposon insertion clones. The transposon insertion sites of the six mutants were mapped at the five genes. All these mutants did not display NAC resistance under acidic conditions, despite their resistance to NAC under alkaline conditions, indicating that the NAC resistance directed by the acr mutations was independent of the unusual pH-sensitivity of V. cholerae. Furthermore, all these mutants displayed attenuated virulence and reduced biofilm formation, suggesting that the acr genes are required for pathogenesis of V. cholerae. This study validates the relevance of drug repositioning for antibacterials with new modes of action and will provide an insight into a novel antibacterial therapy for V. cholerae infections to minimize side effects and resistance emergence.

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Identification of brevinin-1EMa-derived stapled peptides as broad-spectrum virus entry blockers
Mi Il Kim, Thanh K. Pham, Dahee Kim, Minkyung Park, Bi-o Kim, You-Hee Cho, Young-Woo Kim, Choongho Lee
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Brevibacillus antibioticus sp. nov., with a broad range of antibacterial activity, isolated from soil in the Nakdong River: Ahyoung Choi , Young Ho Nam , Kiwoon Baek , Eu Jin Chung; J. Microbiol. 2019;57(11):991-996. Published online October 28, 2019; DOI: https://doi.org/10.1007/s12275-019-9325-y

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A Gram-stain-positive, aerobic, motile, and rod-shaped bacterial strain designated TGS2-1T was isolated from sediment soil in the Nakdong River, Republic of Korea. The optimal growth of strain TGS2-1T was observed at 28°C and pH 7.0 without NaCl supplementation. Strain TGS2-1T revealed antibiosis against various bacteria, including Staphylococcus aureus KCCM 4051, CCARM 3089 (methicillin resistant strains), Enterococcus faecalis KCCM 11814, Escherichia coli KCTC 2443, Candida albicans KACC 7270, and Filobasidium neoformans KCTC 7902. Phylogenetic analyses based on the 16S rRNA gene sequences indicated that strain TGS2-1T belonged to the genus Brevibacillus and shared 93.8–99.7% sequence similarity with Brevibacillus species. Whole-genome sequencing of strain TGS2-1T revealed a genome size of 6.2 Mbp and DNA G + C content of 47.0 mol%. The TGS2-1T genome shared an average nucleotide identity and digital DNA-DNA hybridization of 74.6–93.3% and 18.6–67.1%, respectively, with six related Brevibacillus genomes. The major fatty acid constituents of strain TGS2-1T were anteiso-C15:0 (62.3%) and anteiso-C17:0 (10.8%). Cells of strain TGS2-1T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, seven unidentified aminophospholipids, and five unidentified lipids. The isoprenoid quinone detected in the strain was menaquinone-7 (MK-7). Based on data obtained from this polyphasic taxonomic study, strain TGS2-1T represents a novel species belonging to genus Brevibacillus, for which the name B. antibioticus sp. nov. is proposed. The type strain is TGS2-1T (= KCCM 90326T = NBRC 113840T = FBCC-B2501).

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Traditional processing increases biological activities of Dendrobium offificinale Kimura et. Migo in Southeast Yunnan, China
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Antimicrobial effect and proposed action mechanism of cordycepin against Escherichia coli and Bacillus subtilis: Qi Jiang , Zaixiang Lou , Hongxin Wang , Chen Chen; J. Microbiol. 2019;57(4):288-297. Published online March 30, 2019; DOI: https://doi.org/10.1007/s12275-019-8113-z

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The detailed antibacterial mechanism of cordycepin efficacy against food-borne germs remains ambiguous. In this study, the antibacterial activity and action mechanism of cordycepin were assessed. The results showed that cordycepin effectively inhibited the growth of seven bacterial pathogens including both Gram-positive and Gram-negative bacterial pathogens; the minimum inhibitory concentrations (MIC) were 2.5 and 1.25 mg/ml against Escherichia coli and Bacillus subtilis, respectively. Scanning electron microscope and transmission electron microscope examination confirmed that cordycepin caused obvious damages in the cytoplasmatic membranes of both E. coli and B. subtilis. Outer membrane permeability assessment indicated the loss of barrier function and the leakage of cytoplasmic contents. Propidium iodide and carboxyfluorescein diacetate double staining approach coupled with flow cytometry analysis indicated that the integrity of cell membrane was severely damaged during a short time, while the intracellular enzyme system still remained active. This clearly suggested that membrane damage was one of the reasons for cordycepin efficacy against bacteria. Additionally, results from circular dichroism and fluorescence analysis indicated cordycepin could insert to genome DNA base and double strand, which disordered the structure of genomic DNA. Basis on these results, the mode of bactericidal action of cordycepin against E. coli and B. subtilis was found to be a dual mechanism, disrupting bacterial cell membranes and binding to bacterial genomic DNA to interfere in cellular functions, ultimately leading to cell death.

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Review

REVIEW] Antibacterial strategies inspired by the oxidative stress and response networks: So Youn Kim , Chanseop Park , Hye-Jeong Jang , Bi-o Kim , Hee-Won Bae , In-Young Chung , Eun Sook Kim , You-Hee Cho; J. Microbiol. 2019;57(3):203-212. Published online February 26, 2019; DOI: https://doi.org/10.1007/s12275-019-8711-9

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Abstract PDF

Oxidative stress arises from an imbalance between the excessive accumulation of reactive oxygen species (ROS) and a cell’s capability to readily detoxify them. Although ROS are spontaneously generated during the normal oxygen respiration and metabolism, the ROS generation is usually augmented by redox-cycling agents, membrane disrupters, and bactericidal antibiotics, which contributes their antimicrobial bioactivity. It is noted that all the bacteria deploy an arsenal of inducible antioxidant defense systems to cope with the devastating effect exerted by the oxidative stress: these systems include the antioxidant effectors such as catalases and the master regulators such as OxyR. The oxidative stress response is not essential for normal growth, but critical to survive the oxidative stress conditions that the bacterial pathogens may encounter due to the host immune response and/or the antibiotic treatment. Based on these, we here define the ROS-inspired antibacterial strategies to enhance the oxidative stress of ROS generation and/or to compromise the bacterial response of ROS detoxification, by delineating the ROSgenerating antimicrobials and the core concept of the bacterial response against the oxidative stress.

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Journal Articles

The antibacterial activity of E. coli bacteriophage lysin lysep3 is enhanced by fusing the Bacillus amyloliquefaciens bacteriophage endolysin binding domain D8 to the C-terminal region: Shuang Wang , Jingmin Gu , Meng Lv , Zhimin Guo , Guangmou Yan , Ling Yu , Chongtao Du , Xin Feng , Wenyu Han , Changjiang Sun , Liancheng Lei; J. Microbiol. 2017;55(5):403-408. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6431-6

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Abstract PDF

Bacteriophage endolysin is one of the most promising anti-biotic substitutes, but in Gram-negative bacteria, the outer membrane prevents the lysin from hydrolyzing peptidogly-cans and blocks the development of lysin applications. The prime strategy for new antibiotic substitutes is allowing lysin to access the peptidoglycan from outside of the bacteria by reformation of the lysin. In this study, the novel Escherichia coli (E. coli) phage lyase lysep3, which lacks outside-in cata-lytic ability, was fused with the N-terminal region of the Bacillus amyloliquefaciens lysin including its cell wall bind-ing domain D8 through the best manner of protein fusion based on the predicted tertiary structure of lysep3-D8 to ob-tain an engineered lysin that can lyse bacteria from the out-side. Our results showed that lysep3-D8 could lyse both Gram- negative and Gram-positive bacteria, whereas lysep3 and D8 have no impact on bacterial growth. The MIC of lysep3-D8 on E. coli CVCC1418 is 60 μg/ml; lysep3-D8 can inhibit the growth of bacteria up to 12 h at this concentration. The bac-tericidal spectrum of lysep3-D8 is broad, as it can lyse of all of 14 E. coli strains, 3 P. aeruginosa strains, 1 Acinetobacter baumannii strain, and 1 Streptococcus strain. Lysep3-D8 has sufficient bactericidal effects on the 14 E. coli strains tested at the concentration of 100 μg/ml. The cell wall binding do-main of the engineered lysin can destroy the integrity of the outer membrane of bacteria, thus allowing the catalytic do-main to reach its target, peptidoglycan, to lyse the bacteria. Lysep3-D8 can be used as a preservative in fodder to benefit the health of animals. The method we used here proved to be a successful exploration of the reformation of phage lysin.

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Antibacterial compound produced by Pseudomonas aeruginosa strain UICC B-40, an endophytic bacterium isolated from Neesia altissima: Rina Hidayati Pratiwi , Iman Hidayat , Muhammad Hanafi , Wibowo Mangunwardoyo; J. Microbiol. 2017;55(4):289-295. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6311-0

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This study’s aim was to determine the identity of antibacte-rial compounds produced by Pseudomonas aeruginosa strain UICC B-40 and describe the antibacterial compounds’ me-chanisms of action for damaging pathogenic bacteria cells. Isolation and identification of the compounds were carried out using thin layer chromatography (TLC), nuclear mag-netic resonance (NMR) spectroscopy and liquid chromato-graphy mass spectrometry (LC-MS) analyses. Antibacterial activity was assayed via minimum inhibitory concentration (MIC) and the antibacterial compound mechanism was ob-served morphologically through scanning electron micros-copy (SEM). This study successfully identified the (2E,5E)- phenyltetradeca-2,5-dienoate antibacterial compound (mole-cular weight 300 g/mol), composed of a phenolic ester, fatty acid and long chain of aliphatic group structures. MIC values for this compound were determined at 62.5 μg/ml against Staphylococcus aureus strain ATCC 25923. The mechanism of the compound involved breaking down the bacterial cell walls through the lysis process. The (2E,5E)-phenyltetradeca- 2,5-dienoate compound exhibited inhibitory activity on the growth of Gram-positive bacteria.

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Research Support, Non-U.S. Gov'ts

Antibacterial effects of N-acetylcysteine against endodontic pathogens: Ji-Hoi Moon , Young-Suk Choi , Hyeon-Woo Lee , Jung Sun Heo , Seok Woo Chang , Jin-Yong Lee; J. Microbiol. 2016;54(4):322-329. Published online April 1, 2016; DOI: https://doi.org/10.1007/s12275-016-5534-9

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The success of endodontic treatment depends on the eradication of microorganisms from the root canal system and the prevention of reinfection. The purpose of this investigation was to evaluate the antibacterial and antibiofilm efficacy of N-acetylcysteine (NAC), an antioxidant mucolytic agent, as an intracanal medicament against selected endodontic pathogens. Minimum inhibitory concentrations (MICs) of NAC for Actinomyces naeslundii, Lactobacillus salivarius, Streptococcus mutans, and Enterococcus faecalis were determined using the broth microdilution method. NAC showed antibacterial activity, with MIC values of 0.78–1.56 mg/ml. The effect of NAC on biofilm formation of each bacterium and a multispecies culture consisting of the four bacterial species was assessed by crystal violet staining. NAC significantly inhibited biofilm formation by all the monospecies and multispecies bacteria at minimum concentrations of 0.78–3.13 mg/ml. The efficacy of NAC for biofilm disruption was evaluated by scanning electron microscopy and ATP-bioluminescence quantification using mature multispecies biofilms. Preformed mature multispecies biofilms on saliva-coated hydroxyapatite disks were disrupted within 10 min by treatment with NAC at concentrations of 25 mg/ml or higher. After 24 h of treatment, the viability of mature biofilms was reduced by > 99% compared with the control. Moreover, the biofilm disrupting activity of NAC was significantly higher than that of saturated calcium hydroxide or 2% chlorhexidine solution. Within the limitations of this in vitro study, we conclude that NAC has excellent antibacterial and antibiofilm efficacy against endodontic pathogens and may be used as an alternative intracanal medicament in root canal therapies.

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Antibacterial metabolites from the Actinomycete Streptomyces sp. P294: Huining Su , Hongwei Shao , Keqin Zhang , Guohong Li; J. Microbiol. 2016;54(2):131-135. Published online February 2, 2016; DOI: https://doi.org/10.1007/s12275-016-5311-9

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The Actinomycete strain P294 was isolated from soil and identified as Streptomyces sp. based upon the results of 16S rRNA sequence analysis. Three compounds obtained from the solid fermentation products of this strain have been determined by 1D, 2D NMR and HRMS experiments. These compounds include two new compounds angumycinones C (1) and D (2), and the known compound X-14881 E (3). All compounds were assayed for antibacterial and nematicidal activity. The results showed the three compounds had different degrees of inhibitory activity against several target bacteria but no significant toxicity against the nematode Caenorhabditis elegans.

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Infirmity and death from diseases caused by unsafe food are a continual hazard to communal health safety and socio-economic growth throughout the world. Chemical preservatives are associated with health hazards and toxicity issues. In the study reported here, 200 soil isolates from Western Himalayan region in India were screened for potential antibacterial activity against food-borne pathogens. This study led to the isolation of a bacterial strain belonging to the Genus Bacillus and was designated as RPT-0001. The associated antibacterial activity was sensitive to pronase E treatment. Bioassay-guided fractionation using reverse phase high performance liquid chromatography (RP-HPLC) led to isolation of the antibacterial peptide designated as RPT-0001. The molecular weight of RPT-0001 was determined by electro- spray ionization mass spectroscopy (ESI-MS) as 276.9 Da. RPT-0001 was inhibitory to both Gram-negative and Grampositive food-borne bacteria tested. The characteristics of RPT-0001 do not match with that of any other known antibacterial peptides produced by Bacillus sp. or related genera. Purified RPT-0001 was successfully used in synthesis of silver nanoparticles effective against food-borne pathogenic bacteria. The antibacterial peptide and silver nanoparticles synthesized utilizing it as a capping and reducing agent hold promising potential in food preservation, in packaging material and as a therapeutic agent in the treatment of foodborne infections.

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Antimicrobial Effects of Herbal Extracts on Streptococcus mutans and Normal Oral Streptococci: Sung-Hoon Lee; J. Microbiol. 2013;51(4):484-489. Published online August 30, 2013; DOI: https://doi.org/10.1007/s12275-013-3312-5

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Streptococcus mutans is associated with dental caries. A cariogenic biofilm, in particular, has been studied extensively for its role in the formation of dental caries. Herbal extracts such as Cudrania tricuspidata, Sophora flavescens, Ginkgo biloba, and Betula Schmidtii have been used as a folk remedy for treating diseases. The purpose of this study was to evaluate and compare the antibacterial activity of herbal extracts against normal oral streptococci, planktonic and biofilm of S. mutans. Streptococcus gordonii, Streptococcus oralis, Streptococcus salivarius, Streptococcus sanguinis, and S. mutans were cultivated with brain heart infusion broth and susceptibility assay for the herbal extracts was performed according to the protocol of Clinical and Laboratory Standard Institute. Also, S. mutans biofilm was formed on a polystyrene 12-well plate and 8-well chamber glass slip using BHI broth containing 2% sucrose and 1% mannose after conditioning the plate and the glass slip with unstimulated saliva. The biofilm was treated with the herbal extracts in various concentrations and inoculated on Mitis-Salivarius bacitracin agar plate for enumeration of viable S. mutans by counting colony forming units. Planktonic S. mutans showed susceptibility to all of the extracts and S. mutans biofilm exhibited the highest level of sensitivity for the extracts of S. flavescens. The normal oral streptococci exhibited a weak susceptibility in comparison to S. mutans. S. oralis, however, was resistant to all of the extracts. In conclusion, the extract of S. flavescens may be a potential candidate for prevention and management of dental caries.

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Influence of Culture Conditions and Medium Composition on the Production of Antibacterial Compounds by Marine Serratia sp. WPRA3: Mahtab Jafarzade , Nur Ain Yahya , Fatemeh Shayesteh , Gires Usup , Asmat Ahmad; J. Microbiol. 2013;51(3):373-379. Published online June 28, 2013; DOI: https://doi.org/10.1007/s12275-013-2440-2

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This study was undertaken to investigate the influence of culture conditions and medium components on production of antibacterial compounds by Serratia sp. WPRA3 (JX020764) which was isolated from marine water of Port Dickson, Malaysia. Biochemical, morphological, and molecular characteristics suggested that the isolate is a new candidate of the Serratia sp. The isolate showed strong antimicrobial activity against fungi, Gram-negative and Gram-positive bacteria. This bacterium exhibited optimum antibacterial compounds production at 28°C, pH 7 and 200 rev/min aeration during 72 h of incubation period. Highest antibacterial activity was obtained when sodium chloride (2%), yeast extract (0.5%), and glucose concentration (0.75%) were used as salt, nitrogen, and carbon sources respectively. Different active fractions were obtained by Thin-Layer Chromatography (TLC) and Flash Column Chromatography (FCC) from ethyl acetate crude extracts namely OCE and RCE in different culture conditions, OCE (pH 5, 200 rev/min) and RCE (pH 7/without aeration). In conclusion, the results suggested different culture conditions have a significant impact on the types of secondary metabolites produced by the bacterium.

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Copper as an Antimicrobial Agent against Opportunistic Pathogenic and Multidrug Resistant Enterobacter Bacteria: Wen-Xiao Tian , Shi Yu , Muhammad Ibrahim , Abdul Wareth Almonaofy , Liu He , Qiu Hui , Zhu Bo , Bin Li , Guan-lin Xie; J. Microbiol. 2012;50(4):586-593. Published online July 21, 2012; DOI: https://doi.org/10.1007/s12275-012-2067-8

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Infections by Enterobacter species are common and are multidrug resistant. The use of bactericidal surface materials such as copper has lately gained attention as an effective antimicrobial agent due to its deadly effects on bacteria, yeast, and viruses. The aim of the current study was to assess the antibacterial activity of copper surfaces against Enterobacter species. The antibacterial activity of copper surfaces was tested by overlying 5×106 CFU/ml suspensions of representative Enterobacter strains and comparing bacterial survival counts on copper surfaces at room temperature. Iron, stainless steel, and polyvinylchloride (PVC) were used as controls. The mechanisms responsible for bacterial killing on copper surfaces were investigated by a mutagenicity assay of the D-cycloserin (cyclA gene), single cell gel electrophoresis, a staining technique, and inductively coupled plasma mass spectroscopy. Copper yielded a significant decrease in the viable bacterial counts at 2 h exposure and a highly significant decrease at 4 h. Loss of cell integrity and a significantly higher influx of copper into bacterial cells exposed to copper surfaces, as compared to those exposed to the controls, were documented. There was no increase in mutation rate and DNA damage indicating that copper contributes to bacterial killing by adversely affecting cellular structure without directly targeting the genomic DNA. These findings suggest that copper’s antibacterial activity against Enterobacter species could be utilized in health care facilities and in food processing plants to reduce the bioburden, which would increase protection for susceptible members of the community.

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NOTE] Antibiotic Resistance and Probiotic Properties of Dominant Lactic Microflora from Tungrymbai, an Ethnic Fermented Soybean Food of India: Sharmila Thokchom , Santa Ram Joshi; J. Microbiol. 2012;50(3):535-539. Published online June 30, 2012; DOI: https://doi.org/10.1007/s12275-012-1409-x

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The present investigation was conducted to assess lactic acid bacteria present in traditionally fermented food of ethnic tribes in India for probiotic properties, antibacterial activity, and antibiotic tolerance behavior. Enterococcus sp., Lactobacillus sp., and Lactococcus sp. showed antibacterial activity against Bacillus cereus MTCC 430, Staphylococcus aureus subsp. aureus MTCC 740, and Salmonella enterica ser. paratyphi A MTCC 735. Lactococcus sp. and Lactobacillus sp. could tolerate acidic conditions (pH 2) and high bile salt concentration (4000 ppm). The lactic microflora were found to be sensitive to most common antibiotics, except for cloxacillin (5 μg), cephalexin (30 μg), and cephalothin (30 μg).

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Effect of Lactobacillus reuteri on the Proliferation of Propionibacterium acnes and Staphylococcus epidermidis: Mi-Sun Kang , Jong-Suk Oh , Seok-Woo Lee , Hoi-Soon Lim , Nam-Ki Choi , Seon-Mi Kim; J. Microbiol. 2012;50(1):137-142. Published online February 27, 2012; DOI: https://doi.org/10.1007/s12275-012-1286-3

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While it is generally accepted that Propionibacterium acnes is involved in the development of acne, other bacteria including Staphylococcus epidermidis have also been isolated from the acne lesion. The interaction between Lactobacillus reuteri, a probiotic bacterium, and acnegenic bacteria is unclear. This study examined the effects of L. reuteri on the proliferation of P. acnes and S. epidermidis. Human-derived L. reuteri strains (KCTC 3594 and KCTC 3678) and rat-derived L. reuteri KCTC 3679 were used. All strains exhibited significant inhibitory effects on the growth of P. acnes and S. epidermidis. The proliferation of P. acnes was decreased by 2-log scales after incubation with L. reuteri for 24 h. In addition, the proliferation of S. epidermidis was decreased by 3-log scales after incubation with L. reuteri for 24 h, whereas the growth of L. reuteri was unaffected by P. acnes or S. epidermidis. Among the L. reuteri strains examined, L. reuteri KCTC 3679 had the strongest inhibitory effect on the growth of P. acnes and S. epidermidis, followed by L. reuteri KCTC 3594 and L. reuteri KCTC 3678. Interestingly, reuterin, an antimicrobial factor, was produced only by L. reuteri KCTC 3594. The most pronounced the antibacterial activities of L. reuteri were attributed to the production of organic acids. Overall, these results suggest that L. reuteri may be a useful probiotic agent to control the growth of bacteria involved in acne inflammation and prevent acne.

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Assessment of Resistomycin, as an Anticancer Compound Isolated and Characterized from Streptomyces aurantiacus AAA5: Rajendran Vijayabharathi , Per Bruheim , Trygve Andreassen , Duraisamy Senthil Raja , Palanisamy Bruntha Devi , Sathyaseelan Sathyabama , Venkatesan Brindha Priyadarisini; J. Microbiol. 2011;49(6):920-926. Published online December 28, 2011; DOI: https://doi.org/10.1007/s12275-011-1260-5

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A new actinomycete strain, isolated from humus soils in the Western Ghats, was found to be an efficient pigment producer. The strain, designated AAA5, was identified as a putative Streptomyces aurantiacus strain based on cultural properties, morphology, carbon source utilization, and analysis of the 16S rRNA gene. The strain produced a reddish-brown pigmented compound during the secondary metabolites phase. A yellow compound was derived from the extracted pigment and was identified as the quinone-related antibiotic resistomycin based on ultraviolet–visible spectrophotometry, fourier transform infrared spectroscopy, liquid chromatography and mass spectroscopy, and nuclear magnetic resonance analyses. The AAA5 strain was found to produce large quantities of resistomycin (52.5 mg/L). It showed potent cytotoxic activity against cell lines viz. HepG2 (hepatic carcinoma) and HeLa (cervical carcinoma) in vitro, with growth inhibition (GI50) of 0.006 and 0.005 μg/ml, respectively. The strain also exhibited broad antimicrobial activities against both Gram-positive and Gram-negative bacteria. Therefore, AAA5 may have great potential as an industrial resistomycin-producing strain.

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Epidemiological Features and Resistance Pattern in Uropathogens Isolated from Chronic Bacterial Prostatitis: Tommaso Cai , Sandra Mazzoli , Francesca Meacci , Vieri Boddi , Nicola Mondaini , Gianni Malossini , Riccardo Bartoletti; J. Microbiol. 2011;49(3):448-454. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-0391-z

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Abstract PDF

Chronic bacterial prostatitis (CBP) is, usually, caused by uropathogens, especially gram-negative bacilli, although infection is sometimes due to Gram-positive and atypical microorganisms. A recent increasing in prevalence of Gram-positive strains has been reported. The aim of this study was to explore the epidemiological features and resistance rates in uropathogens isolated from CBP outpatients in last 10 years. All consecutive outpatients with demonstrated CBP attending a single Sexually Transmitted Disease centre from January 1997 and December 2008, were enrolled and underwent microbiological cultures in first void early morning urine, midstream urine, expressed prostatic secretion, and post prostate massage urine. Prevalence of different bacterial strains was stratified in four different periods: 1997-1999, 2000-2002, 2003-2005, 2006-2008. Any changes observed in epidemiological features and resistance rates in uropathogens over the whole study period have been analyzed. The present study has been planned, thus, as in vitro study. From 6,221 patients, 4,601 Gram-positive and 1,620 Gram-negative bacterial strains have been isolated. Enterococcus faecalis and Escherichia coli strains are the first and second frequent pathogens found, respectively. Significant differences between E. faecalis prevalence in the 1997-1999 and 2006-2008 periods were found. E. coli showed a significant difference between prevalence in 1997-1999 and 2006-2008 periods. Gram-positive organisms showed a decreasing of susceptibility to ciprofloxacin as well as Gram-negative strains, while a good susceptibility to the levofloxacin was evidenced. E. faecalis prevalence seemed to be raised in 2006-2008 periods. Moreover, a decreasing of activity of ciprofloxacin and a good activity profile of levofloxacin have been reported.

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Evaluation of Antibacterial Activity against Salmonella Enteritidis: Gaëlle Legendre , Fabienne Faÿ , Isabelle Linossier , Karine Vallée-Réhel; J. Microbiol. 2011;49(3):349-354. Published online June 30, 2011; DOI: https://doi.org/10.1007/s12275-011-0162-x

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Abstract PDF: Salmonella enterica serovar Enteritidis is a well-known pathogenic bacterium responsible for human gastrointestinal enteritis mainly due to the consumption of eggs and egg-products. The first aim of this work was to study several virulence factors of a strain isolated from egg content: SEovo. First, bacterial growth was studied at several temperatures and cell morphology was observed by scanning electronic microscopy. These experiments showed Salmonella’s ability to grow at low temperatures and to produce exoproducts. Next, Salmonella motility was observed performing swimming, twitching, and swarming tests. Results indicated a positive flagellar activity and the cell ability to differentiate and become hyperflagellated under specific conditions. Moreover, SEovo adherence and biofilm formation was carried out. All of these tests enabled us to conclude that SEovo is a potential pathogen, thus it can be used as a model to perform antibacterial experiments. The second part of the study was dedicated to the evaluation of the antibacterial activity of different molecules using several methods. The antibacterial effect of silver and copper aluminosilicates was tested by two different kinds of methods. On the one hand, the effect of these two antibacterial agents was determined using microbiological methods: viable cell count and agar-well diffusion. And on the other hand, the antibacterial activity was evaluated using CLSM and SYTO Red/SYTOX Green dyeing. CLSM allowed for the evaluation of the biocide on sessile cells, whereas the first methods did not. Results showed that adhered bacteria were more resistant than planktonic counterparts and that CLSM was a good alternative to evaluate antibacterial activity on fixed bacteria without having to carry out a removing step.

Research Support, Non-U.S. Gov'ts

Inhibitory Effect of the Essential Oil from Chamaecyparis obtusa on the Growth of Food-Borne Pathogens: Mi-Jin Park , Won-Sil Choi , Ha-Young Kang , Ki-Seob Gwak , Geun-Shik Lee , Eui-Bae Jeung , In-Gyu Choi; J. Microbiol. 2010;48(4):496-501. Published online August 20, 2010; DOI: https://doi.org/10.1007/s12275-010-9327-2

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Abstract PDF

In this study, the antibacterial activity of essential oil from Chamaecyparis obtusa (Sieb. et Zucc) leaves and twigs was investigated. The test strains were Klebsiella pneumoniae, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, Escherichia coli, Legionella pneumophila, and Methicilline-resistant Staphylococcus aureus. Antibacterial activity was estimated by measuring bacterial growth inhibition. Histopathological examination was also performed. C. obtusa oil distinctly inhibited the growth of all test strains and exhibited the strongest antibacterial activity against L. monocytogenes. It was chromatographically divided into several fractions. The fractions were further tested against antibacterial activity and their chemical compositions were analyzed. The fraction containing terpinen-4-ol (TA) showed high antibacterial activity toward all strains tested. Tests with authentic samples showed that TA played a major role in the antibacterial activity of C. obtusa oil, and in a mice test, the oil actively minimized inflammation by S. aureus.

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Note] Antibacterial Activity of Recombinant hCAP18/LL37 Protein Secreted from Pichia pastoris: Soon-ja Kim , Renshu Quan , Sung-Jin Lee , Hak-Kyo Lee , Joong-Kook Choi; J. Microbiol. 2009;47(3):358-362. Published online June 26, 2009; DOI: https://doi.org/10.1007/s12275-009-0131-9

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Abstract PDF

Human antimicrobial peptide CAP18/LL37 (hCAP18/LL37) was expressed in Pichia pastoris and its antibacterial activity was tested against pathogenic bacteria. The full length ORF of hCAP18/LL37 was cloned into the pPICZαA vector followed by integration into the genomic AOX1 gene of P. pastoris. Agar diffusion assay demonstrated that the different hCAP18/LL37 transformants showed various antibacterial activities against Staphylococcus aureus, Micrococcus luteus, and Salmonella gastroenteritis. The secreted form of hCAP18/LL37 exhibited its maximum activity after 72 h incubation with 2% methanol in MM media, not in BMM. This result suggests that the yeast secreted expression system can be used as a production tool of antimicrobial peptides for industrial or pharmaceutical application.

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Journal Article

Inhibitory Effect of Methyl Gallate and Gallic Acid on Oral Bacteria: Mi-Sun Kang , Jong-Suk Oh , In-Chol Kang , Suk-Jin Hong , Choong-Ho Choi; J. Microbiol. 2008;46(6):744-750. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0235-7

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This study examined the ability of methyl gallate (MG) and gallic acid (GA), the main compounds of gallotannins in Galla Rhois, to inhibit the proliferation of oral bacterial and the in vitro formation of Streptococcus mutans biofilms. The antimicrobial activities of these compounds were evaluated in vitro using the broth microdilution method and a beaker-wire test. Both MG and GA had inhibitory effects on the growth of cariogenic (MIC<8 mg/ml) and periodontopathic bacteria (MIC=1 mg/ml). Moreover, these compounds significantly inhibited the in vitro formation of S. mutans biofilms (MG, 1 mg/ml; GA, 4 mg/ml; P<0.05). MG was more effective in inhibiting bacterial growth and the formation of S. mutans biofilm than GA. In conclusion, MG and GA can inhibit the growth of oral pathogens and S. mutans biofilm formation, and may be used to prevent the formation of oral biofilms.

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Polyphenols Attenuate Highly-Glycosylated Haemoglobin-Induced Damage in Human Peritoneal Mesothelial Cells
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Reshaped as polyester-based nanoparticles, gallic acid inhibits platelet aggregation, reactive oxygen species production and multi-resistant Gram-positive bacteria with an efficiency never obtained
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Research Support, Non-U.S. Gov't

Protective Effect of Polygoni Cuspidati Radix and Emodin on Vibrio vulnificus Cytotoxicity and Infection: Jong Ro Kim , Dool-Ri Oh , Mi Hye Cha , Byoung Sik Pyo , Joon Haeng Rhee , Hyon E. Choy , Won Keun Oh , Young Ran Kim; J. Microbiol. 2008;46(6):737-743. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0232-x

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Abstract PDF

Vibrio vulnificus, a good model organism of bacterial septicemia, causes fatal septicemia manifesting a fulminating course and a high mortality rate within days. In order to identify new natural substances preventing V. vulnificus infection, a plant library was screened for inhibiting cytotoxicity to host cells by using Trypan blue staining and LDH assay. We found that Polygoni Cuspidati Radix potently suppressed the acute death of HeLa and RAW264.7 cells in a dose dependent manner. Further studies revealed that Polygoni Cuspidati Radix inhibited V. vulnificus growth and survival in HI broth and seawater, respectively. We confirmed that Polygoni Cuspidati Radix contained high level of emodin by thin layer chromatography (TLC). Emodin showed direct antibacterial activity against V. vulnificus. In addition, emodin prevented the morphologic damages and acute death of HeLa cells caused from V. vulnificus. The safety of Polygoni Cuspidati Radix and emodin to host cells was confirmed by MTT assay. Polygoni Cuspidati Radix and emodin protected mice from V. vulnificus infection.

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Journal Article

Synergistic Antibacterial Effect between Silybin and N,N’-Dicyclohexylcarbodiimide in Clinical Pseudomonas aeruginosa Isolates: Hyun Jun Jung , Dong Gun Lee; J. Microbiol. 2008;46(4):462-467. Published online August 31, 2008; DOI: https://doi.org/10.1007/s12275-008-0138-7

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Abstract PDF

Silybin is a composition of the silymarin group as a hepatoprotective agent, and it exhibits various biological activities, including an antibacterial activity. In this study, the effects of a combination of silybin with N,N′-dicyclohexylcarbodiimide (DCCD) against clinical isolates of Pseudomonas aeruginosa were investigated. In the results of susceptibility assay, silybin showed more potent antibacterial activity in methicilin-resistant Staphylococcus aureus (MRSA) than in P. aeruginosa, but DCCD significantly increased the antibacterial activity of silybin in P. aeruginosa. The antibacterial activity of silybin was affected by the strong action of multidrug-resistant pumps rather than by a permeable disruption of lipopolysaccharide and silybin showed a remarkable synergistic activity in combination with some antibiotic agents against drug-resistant bacteria. Therefore, silybin has a potential as a combination therapeutic agent for treatment of infectious diseases by multidrug-resistant bacteria.

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Research Support, Non-U.S. Gov't

Antibacterial Characteristics of Curcuma xanthorrhiza Extract on Streptococcus mutans Biofilm: Jung-Eun Kim , Hee-Eun Kim , Jae-Kwan Hwang , Ho-Jeong Lee , Ho-Keun Kwon , Baek-Il Kim; J. Microbiol. 2008;46(2):228-232. Published online June 11, 2008; DOI: https://doi.org/10.1007/s12275-007-0167-7

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Abstract PDF

This study evaluated the antibacterial effects of a natural Curcuma xanthorrhiza extract (Xan) on a Streptococcus mutans biofilm by examining the bactericidal activity, inhibition of acidogenesis and morphological alteration. Xan was obtained from the roots of a medicinal plant in Indonesia, which has shown selective <br><br>antibacterial effects on planktonic S. mutans. S. mutans biofilms were formed on slide glass over a 72 h period and treated with the following compounds for 5, 30, and 60 min: saline, 1% DMSO, 2 mg/ml chlorhexidine (CHX), and 0.1 mg/ml Xan. The Xan group exposed for 5 and 30 min showed significantly fewer colony forming units (CFU, 57.6 and 97.3%, respectively) than those exposed to 1% DMSO, the negative control group (P<0.05). These CFU were similar in number to those slides exposed to CHX, the positive control group. Xan showed similar bactericidal effect to that of CHX but the dose of Xan was one twentieth that of CHX. In addition, the biofilms treated with Xan and CHX maintained a neutral pH for 4 h, which indicates that Xan and CHX inhibit acid production. Scanning electron microscopy showed morphological changes in the cell wall and membrane of the Xan-treated biofilms; an uneven surface and a deformation in contour. Overall, natural Xan has strong bactericidal activity, inhibitory effects on acidogenesis, and alters the microstructure of S. mutans biofilm. In conclusion, Xan has potential in anti-S. mutans therapy for the prevention of dental caries.

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Mi-Sun Kang, Jong-Suk Oh, In-Chol Kang, Suk-Jin Hong, Choong-Ho Choi
The Journal of Microbiology.2008; 46(6): 744. CrossRef

Journal Article

Antibacterial Effect of Electrolyzed Water on Oral Bacteria: Sung-Hoon Lee , Bong-Kyu Choi; J. Microbiol. 2006;44(4):417-422.; DOI: https://doi.org/2411 [pii]

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Abstract PDF: This study investigated the antibacterial effect of electrolyzed water on oral bacteria both in vitro and in vivo. Tap water was electrolyzed in a water vessel using platinum cell technology. The electrolyzed tap water (called Puri-water) was put in contact with five major periodontopathogens or toothbrushes contaminated with these bacteria for 30 sec. In addition, Puri-water was used as a mouthwash for 30 sec in 16 subjects and the antibacterial effect on salivary bacteria was evaluated. Puri-water significantly reduced the growth of all periodontopathogens in culture and on toothbrushes, and that of aerobic and anaerobic bacteria in saliva, when compared to the effect of tap water. It also significantly reduced mutans streptococci growing on mitis salivarius-bacitracin agar. Our results demonstrate that the electrolyzed tap water is effective as a mouthwash and for toothbrush disinfection.

Research Support, Non-U.S. Gov't

Hydrogen Peroxide produced by Two Amino Acid Oxidases Mediates Antibacterial Actions: Hongmin Zhang , Qiuyue Yang , Mingxuan Sun , Maikun Teng , Liwen Niu; J. Microbiol. 2004;42(4):336-339.; DOI: https://doi.org/2102 [pii]

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Abstract PDF: The antibacterial actions of two amino acid oxidases, a D-amino acid oxidase from hog kidney and a L-amino acid oxidase from the venom of Agkistrodon halys, were investigated, demonstrating that both enzymes were able to inhibit the growth of both Gram-positive and Gram-negative bacteria, and that hydrogen peroxide, a product of their enzymatic reactions, was the antibacterial factor. However, hydrogen peroxide generated in the enzymatic reactions was not sufficient to explain the degree to which bacterial growth was inhibited. A fluorescence labeling assay showed that both of these two enzymes could bind to the surfaces of bacteria. To the best of our knowledge, this is the first report regarding the antibacterial activity of the D-amino acid oxidases.

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