• Membrane Journal
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• 제31권2호
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• pp.87-100
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• 2021

With a striking increase in the level of contamination and subsequent degradations in the environment, detection and monitoring of contaminants in various sites has become a crucial mission in current society. In this review, we have summarized the current research areas in membrane-based colorimetric sensors for trace detection of various molecules. The researches covered in this summary utilize membranes composed of cellulose fibers as sensing platforms and metal nanoparticles or fluorophores as optical reagents. Displaying decent or excellent sensitivity, most of the developed sensors achieve a significant selectivity in the presence of interfering ions. The physical and chemical properties of cellulose membrane platforms can be customized by changing the synthesis method or type of optical reagent used, allowing a wide range of applications possible. Membrane-based sensors are also portable and have great mechanical properties, which enable on-site detection of contaminants. With such superior qualities, membrane-based sensors examined in the researches were used for versatile purposes including quantification of heavy metals in drinking water, trace detection of toxic antibiotics and heavy metals in environmental water samples. Some of the sensors exhibited additional features like antimicrobial ability and recyclability. Lastly, while most of the sensors aimed for a detection enabled by naked eyes through rapid colour change, many of them investigated further detection methods like fluorescence, UV-vis spectroscopy, and RGB colour intensity.

• Korean Journal of Microbiology
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• 제51권4호
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• pp.448-452
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• 2015

The bacterial strain that was isolated from chinese cabbage rhizosphere, showed inhibition of yeast growth. This strain was identified as Pseudomonas fluorescens BB2 by API 20NE test and 16S rRNA gene sequence analysis. P. fluorescens BB2 strain produced antibiotics against yeast as a secondary metabolite effectively when the culture was carried out in YM medium with 3% glucose at $20^{\circ}C$. The protein antibiotic of BB2 strain which was concentrated by ammonium sulfate precipitation and n-butanol extraction inhibited the growth of yeast with the minimal inhibitory concentration of $10{\mu}g/ml$ against Candida albicans KCTC 7965, and the growth of yeast was completely inhibited at $80{\mu}g/ml$. The hydrophilic fraction of n-butanol extraction inhibited the growth of Bacillus cereus ATCC 21366, showed orange halo on chrome azurol S plate, which means the fraction contained iron chelating siderophore. The results of crystal violet uptake through the cell membrane showed that membrane permeability was increased about 9% than control, when the concentration of hydrophobic antibiotic against yeast C. albicans was $60{\mu}g/ml$. As a result, the antibiotic produced by P. fluorescens BB2 against yeast Candida is considered antimicrobial peptide, and this is the first report in the genus Pseudomonas.

• Journal of Life Science
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• 제12권6호
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• pp.803-811
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• 2002

The present study was carried out for research and development of natural antimicrobial from extract of Stachys sieboldii MIQ against food borne bacteria. The hexane extract of Stachys sieboldii MIQ at 250$\mu\textrm{g}$/$m\ell$ per disc showed 15 ~ 20 mm inhibition zone against gram positive and gram negative barteria. Minimum inhibitory concentration (MIC) of hexane extract was 250${\mu}g$/$m\ell$against Bacillus cereus, 250~500${\mu}g$/$m\ell$against Listeria monocytogenes, 500${\mu}g$/$m\ell$ against Staphylococcus aureus, Psedomonas aeruginosa. Observation by transmission electron microscope, showed that disruption of the cell wall assumed to be due to the bactericidal activity. In addition, the membrane integrity of the sensitive cells was disrupted by exposure to Stachys sieboldii MIQ extract on the D-$\beta$-galatosidase activity as substrate of O-nitrophenol-$\beta$-D-galacto-pyranoside. The hexane extract of Stachys sieboldii MIQ was very stable on the pH and thermal stability.

• Korean Journal of Fisheries and Aquatic Sciences
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• 제39권3호
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• pp.269-277
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• 2006

This study prepared functional oyster hydrolysates using two-step enzymatic hydrolysis and investigated their functional properties. To prepare two-step enzymatic hydrolysates (TSEH), oysters were hydrolyzed using 1% Protamex (PR) at $40^{\circ}C$ and pH 6.0 for 1 hr before sequential treatment with one of the following enzymes for 1 hr: Alcalase (AL), Flavourzyme (FL), Neutrase (NE), pepsin (PE), and trypsin (TR). The PRAL, PRNE and PRTR hydrolysates had significantly greater angiotensin I converting enzyme (ACE) inhibitory activity than did PR and the other TSEHs. Only the antioxidant activity of the PRNE hydrolysate was significantly different (p<0.05), while none of the TSEHs had antimicrobial activity. The oyster hydrolysate prepared by sequential treatment with Protamex and Neutrase (PRNE) had the best ACE inhibitory activity and antioxidant activity, with $IC_{50}$ values of 0.40 and 0.94 mg/mL, respectively. The PRNE hydrolysate was processed through an ultrafiltration (UF) series with molecular weight cut-off (MWCO) membranes of 3, 5, 10, and 30 kDa, and the ACE inhibitory, antioxidant, and antimicrobial activities of the permeates were determined. The permeate through the 3-kDa MWCO membrane had greater ACE inhibitory activity and antioxidant activity than did the other PRNE permeates, with $IC_{50}$ values of 0.11 and 0.40 mg/mL, respectively.

• Korean Journal of Food Science and Technology
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• 제45권3호
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• pp.370-375
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• 2013

Propolis is a resinous mixture found in the tree buds, sap flows, and other botanical sources, which is used by honey bees in the construction of their hives. Antimicrobial effects of propolis were evaluated against Streptococcus mutans KCTC 3065, S. sobrinus KCTC 3308, S. sobrinus KCTC 5134, and Porphyromonas gingivalis KCTC 5352 by an agar diffusion assay. Sensitivity of these microorganisms to propolis was evaluated in broth containing different concentrations of propolis at $37^{\circ}C$, followed by observation using transmission electron microscopy (TEM). Propolis inhibited all oral microorganisms tested at the minimum inhibitory concentration (MIC) of $0.14mg/{\mu}L$ in the agar diffusion assay. Treatment with 0.06 and $0.22mg/{\mu}L$ of propolis had a bactericidal effect in a concentration- and treatment time-dependent manner against the tested microorganisms. TEM of propolis-treated S. mutans KCTC 3065 and P. gingivalis KCTC 5352 revealed structural damage of the cell membrane. The activity of propolis was affected by heat and pH treatment. The results indicate that propolis shows antibacterial activity against oral microorganisms and that it has potential for future applications in the food industry.

• Bulletin of the Korean Chemical Society
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• 제33권1호
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• pp.115-122
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• 2012

Apidaecin Ib had strong antimicrobial activity against several tested Gram-negative bacteria including Escherichia coli, Enterobacter cloacae, and Shigella flexneri (MECs; $0.3-1.5{\mu}g/mL$), but showed no activity against all the tested Gram-positive bacteria including Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus and one yeast, Candida albicans (MECs; > $125{\mu}g/mL$). Interestingly, this peptide showed potent antibacterial activity only against Edwardsiella species (MECs; $0.6-3.6{\mu}g/mL$) among the tested fish pathogenic bacteria through a bacteriostatic process and showed no significant hemolytic activity. Apidaecin Ib took an unordered structure in all environments and also had very weak membrane perturbation activity even at $25{\mu}M$. Anti-Edwardsiella activity of apidaecin Ib is stronger than those of other antimicrobial polypeptides or antibiotics, but its activity is salt-sensitive. These results suggest that apidaecin Ib has Edwardsiella speciesspecific antibacterial activity and could be applied as new preventive or control additives for Edwardsiella species infection in freshwater fish aquaculture.

• Proceedings of the Korean Biophysical Society Conference
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.68-68
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• 2003

HP(2-20) (AKKVFKRLEKLEKLFSKIQNDK) derived from the N-terminus of Helicobacter pylori Ribosomal Protein L1 shows potent antimicrobial activity against bacterial, fungi and cancer cells without cytotoxic effect. In order to investigate the relationships between antimicrobial activity and the structures, several analogues have been designed and synthesized. The structures of these peptides in SDS micelles have been investigated using NMR spectroscopy and they revealed that analogue 3 has the longest, well-defined alpha-helix from Val5 to Trp19. NOESY experiments performed on HP and its analogues in nondeuterated SDS micelles show that protons in the indole ring of Trp16 are in close contact with methylene protons of SDS micelles. In order to probe the position of HP and its analogues relative to the SDS micelles, spin-labeled stearate was added. Large effects are observed for the chemical shifts and the intensities of Phe5, Glu9, Phe12, and Trp16 within the helix region by 16-doxylstearate. This result implies that 16-doxylstearate is located in the center of the micelles and the hydrophobic phase of the amphiphilic ${\alpha}$-helix is located in contact with the acyl chains of the micelles. Also, Lys3 and Lys4 at N-terminus and Lys20 at C-terminus may produce an optimal arrangement for electrostatic interactions between the sulfate head groups of the SDS and the positively charged lysyl N$\sub$3/$\^$+/. Interactions between the indole ring of Trp and the membrane, as well as the amphiphilic ${\alpha}$-helical structure of HP induced by Trp at the C-terminus may allow HP to span the lipid bilayer. These structural features are crucial for their potent antibiotic activities.

• Bulletin of the Korean Chemical Society
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• 제24권10호
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• pp.1478-1484
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• 2003

CRAMP was identified from a cDNA clone derived from a mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptide. Tertiary structure of CRAMP in TFE/$H_2O$ (1 : 1, v/v) solution has been determined by NMR spectroscopy previously and consists of two amphipathic $\alpha-helices$ from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Analysis of series of fragments composed of various portion of CRAMP revealed that an 18-residue fragment with the sequence from Gly16 to Leu33 (CRAMP-18) was found to retain antibacterial activity without cytotoxicity. The effects of two Phe residues at positions 14 and 15 of CRAMP-18 on structure, antibacterial activity, and interaction with lipid membranes were investigated by $Phe^{14,15}$ ${\rightarrow}$ Ala substitution (CRAMP-18-A) in the present study. Substitution of Phe with Ala in CRAMP-18 caused a significant reduction on antibacterial and membrane-disrupting activities. Tertiary structures of CRAMP-18 in 50% TFE/$H_2O$ (1 : 1, v : v) solution shows amphipathic ${\alpha}$-helix, from $Glu^2{\;}to{\;}Leu^{18}$, while CRAMP-18-A has relatively short amphipathic ${\alpha}$-helix from $Leu^4{\;}to{\;}Ala^{15}$. These results suggest that the hydrophobic property of $Phe^{14}{\;}and{\;}Phe^15$ in CRAMP-18 is essential for its antibacterial activity, ${\alpha}$-helical structure, and interactions with phospholipid membranes.

• Molecules and Cells
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• 제21권2호
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• pp.229-236
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• 2006

Gaegurin 4 (GGN4), a novel peptide isolated from the skin of a Korean frog, Rana rugosa, has broad spectrum antimicrobial activity. A number of amphipathic peptides closely related to GGN4 undergo a coil to helix transition with concomitant oligomerization in lipid membranes or membrane-mimicking environments. Despite intensive study of their secondary structures, the oligomeric states of the peptides before and after the transition are not well understood. To clarify the structural basis of its antibiotic action, we used analytical ultracentrifugation to define the aggregation state of GGN4 in water, ethyl alcohol, and 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). The maximum size of GGN4 in 15% HFIP corresponded to a decamer, whereas it was monomeric in buffer. The oligomeric transition is accompanied by a cooperative 9 nm blue-shift of maximum fluorescence emission and a large secondary structure change from an almost random coil to an ${\alpha}$-helical structure. GGN4 induces pores in lipid membranes and, using electrophysiological methods, we estimated the diameter of the pores to be exceed $7.3{\AA}$, which suggests that the minimal oligomer structure responsible is a pentamer.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1179-1188
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• 2023

Escherichia coli, particularly multidrug-resistant (MDR) strains, is a serious cause of healthcare-associated infections. Development of novel antimicrobial agents or restoration of drug efficiency is required to treat MDR bacteria, and the use of natural products to solve this problem is promising. We investigated the antimicrobial activity of dried green coffee (DGC) beans, coffee pulp (CP), and arabica leaf (AL) crude extracts against 28 isolated MDR E. coli strains and restoration of ampicillin (AMP) efficiency with a combination test. DGC, CP, and AL extracts were effective against all 28 strains, with a minimum inhibitory concentration (MIC) of 12.5-50 mg/ml and minimum bactericidal concentration of 25-100 mg/ml. The CP-AMP combination was more effective than CP or AMP alone, with a fractional inhibitory concentration index value of 0.01. In the combination, the MIC of CP was 0.2 mg/ml (compared to 25 mg/ml of CP alone) and that of AMP was 0.1 mg/ml (compared to 50 mg/ml of AMP alone), or a 125-fold and 500-fold reduction, respectively, against 13-drug resistant MDR E. coli strains. Time-kill kinetics showed that the bactericidal effect of the CP-AMP combination occurred within 3 h through disruption of membrane permeability and biofilm eradication, as verified by scanning electron microscopy. This is the first report indicating that CP-AMP combination therapy could be employed to treat MDR E. coli by repurposing AMP.