• Journal of Microbiology and Biotechnology
• /
• 제22권10호
• /
• pp.1381-1387
• /
• 2012

We applied enzymatic hydrolysis and tangential flow filtration (TFF) to purify a novel anticancer peptide from Mytilus coruscus (M. coruscus) and investigated its anticancer properties. To prepare the peptide, eight proteases were employed for enzymatic hydrolysis. Pepsin hydrolysates, which showed clearly superior cytotoxic activity on prostate cancer cells, were further purified using a flow filtration system using a TFF and consecutive chromatographic methods. Finally, a novel anticancer peptide was obtained, and the sequence was identified as Ala-Phe-Asn-Ile-His-Asn-Arg-Asn-Leu-Leu. The peptide from M. coruscus effectively induced cell death on prostate, breast and lung cancer cells but not on normal liver cells. This is the first report of an anticancer peptide derived from the hydrolysates of M. coruscus.

• 한국축산식품학회지
• /
• 제43권4호
• /
• pp.625-638
• /
• 2023

Among various biological agents, bacteriocins are important candidates to control Listeria monocytogenes which is a foodborne pathogen. In this study, a novel bacteriocin, named agilicin C7, was isolated from Ligilactobacillus agilis C7 showing inhibitory activity against L. monocytogenes. Agilicin C7 biosynthesis gene was characterized by bioinformatics analyses and heterologously expressed in Escherichia coli for further study. The anti-listeria activity of recombinant agilicin C7 (r-agilicin C7) was lost by proteases and α-amylase, suggesting that agilicin C7 is a glycoprotein. r-Agilicin C7 has wide pH and thermal stability and is also stable in various organic solvents. It destroyed L. monocytogenes by damaging the integrity of the cell envelope. These properties of r-agilicin C7 indicate that agilicin C7 is a novel amylase-sensitive anti-listerial Class IId bacteriocin. Physicochemical stability and inhibitory activity against L. monocytogenes of r-agilicin C7 suggest that it can be applied to control L. monocytogenes in the food industry, including dairy and meat products.

• Food Science and Biotechnology
• /
• 제16권6호
• /
• pp.1055-1059
• /
• 2007

Two monoterpene glucosides, paeoniflorin and albiflorin, in peony (Paeonia lactiflora) were purified from 70% ethanol extract of Paeoniae Radix by diethyl ether washing and n-butanol partition, acetone dissolution, and gradient preparative HPLC. After the whole course of purification, yield of paeoniflorin, albiflorin, and the sum of them were 75.0, 38.8, and 68.7%, respectively, together with the corresponding purity of 96.2, 93.8, and 96.0%.

• Journal of Microbiology and Biotechnology
• /
• 제28권11호
• /
• pp.1946-1954
• /
• 2018

The aim of this study was to isolate and characterize a lytic Yersinia enterocolitica-specific phage (KFS-YE) as a biocontrol agent. KFS-YE was isolated and purified with the final concentration of ($11.72{\pm}0.03$) log PFU/ml from poultry. As observed by transmission electron microscopy, KFS-YE consisted of an icosahedral head and a contractile tail, and was classified in the Myoviridae family. KFS-YE showed excellent narrow specificity against Y. enterocolitica only. Its lytic activity was stable at wide ranges of pH (4-11) and temperature ($4-50^{\circ}C$). The latent period and burst size of KFS-YE were determined to be 45 min and 38 PFU/cell, respectively. KFS-YE showed relatively robust storage stability at -20, 4, and $22^{\circ}C$ for 40 weeks. KFS-YE demonstrated a bactericidal effect in vitro against Y. enterocolitica and provided excellent efficiency with a multiplicity of infection as low as 0.01. This study demonstrated the excellent specificity, stability, and efficacy of KFS-YE as a novel biocontrol agent. KFS-YE may be employed as a practical and promising biocontrol agent against Y. enterocolitica in food.

• 식품과학과 산업
• /
• 제50권1호
• /
• pp.67-73
• /
• 2017

Control of food pathogens is critical in food safety field. Norovirus is one of the major causes of gastroenteritis and food poisoning worldwide, however, currently, there is not a vaccine or a specific drug available for its treatment. There are several methods to inactivate norovirus during food processing by chemical and physical treatments, however, the use of natural substance has been suggested as an optional strategy due to their safety and consumer preference. In this study supported by Ministry of Food and Drug Safety in Korea, we identified novel plant-derived substances with significant anti-norovirus activities. The aim of this project was to determine the antiviral activity of a wide range of natural substances, including plant-derived extracts and essential oils, using a norovirus surrogate system, human norovirus replicon-bearing cells, and mouse in vivo experiments. During the activity screening test, we identified novel anti-norovirus substances or oils using plaque assay with MNV-1. Six selected substances were formulated into an optimum mixture and used as an ingredient for salad sauce of which anti-novovirus activity was confirmed(pending for patent and paper submission). The potential application of selected natural substances as a metal surface sanitizer was also tested. Interestingly, the mixture of selected natural compounds showed a significant inhibitory effect against norovirus. These results suggest that these substances may be used as food ingredient with anti-norovirus antivity or components for surface sanitizers to prevent norovirus contamination.

• Molecules and Cells
• /
• 제41권4호
• /
• pp.301-310
• /
• 2018

LysR-type transcriptional regulators (LTTRs) contain an N-terminal DNA binding domain (DBD) and a C-terminal regulatory domain (RD). Typically, LTTRs function as homotetramers. VV2_1132 was identified in Vibrio vulnificus as an LTTR that is a homologue of HypT (also known as YjiE or QseD) in Escherichia coli. In this study, we determined the crystal structure of full-length VV2_1132 at a resolution of $2.2{\AA}$, thereby revealing a novel combination of the domains in the tetrameric assembly. Only one DBD dimer in the tetramer can bind to DNA, because the DNA binding motifs of the other DBD dimer are completely buried in the tetrameric assembly. Structural and functional analyses of VV2_1132 suggest that it might not perform the same role as E. coli HypT, indicating that further study is required to elucidate the function of this gene in V. vulnificus. The unique structure of VV2_1132 extends our knowledge of LTTR function and mechanisms of action.

• 한국환경농학회:학술대회논문집
• /
• 한국환경농학회 2009년도 정기총회 및 국제심포지엄
• /
• pp.119-126
• /
• 2009

In recent years, research at the Environmental Microbial and Food Safety Laboratory (EMFSL), Agricultural Research Service (ARS) has focused on the development of novel image-based sensing technologies to address agro-food safety concerns, and transformation of these novel technologies into practical instrumentation for industrial implementations. The line-scan-based hyperspectral imaging techniques have often served as a research tool to develop rapid multispectral methods based on only a few spectral bands for rapid online applications. We developed a newer line-scan hyperspectral imaging platform for high-speed inspection on high-throughput processing lines, capable of simultaneous multiple inspection algorithms for different agro-food safety problems such as poultry carcass inspection for wholesomeness and apple inspection for fecal contamination and defect detection. In addition, portable imaging devices were developed for in situ identification of contamination sites and for use by agrofood producer and processor operations for cleaning and sanitation inspection of food processing surfaces. The aim of this presentation is to illustrate recent advances in the above agro.food safety sensing technologies.

• 한국생명과학회:학술대회논문집
• /
• 한국생명과학회 2003년도 제39회 학술심포지움
• /
• pp.23-37
• /
• 2003

A novel inhibitor against maltose-producing a-amylase was prepared via stepwise degradation of a high molecular weight acarbose (HMWA) using Thermus maltogenic amylase (ThMA). The structure of the purified inhibitor was determined to be ${\alpha}$-D-glucopyranosyl-${\alpha}$-acarviosinyl-D-glucopyranose (GlcAcvGlc). Progress curves of p-nitrophenyl-${\alpha}$-D-maltoside (PNPG2) hydrolysis by various amylolytic enzymes, including maltogenase (MGase), ThMA, and cyclodextrinase(CDase) I-5, in the presence of acarbose or GlcAcvGlc indicated a slow-binding mode of inhibition. The inhibition potency of GlcAcvGlc for MGase, ThMA, and CDase I-5 was 3 orders of magnitude higher than that of acarbose.

• Preventive Nutrition and Food Science
• /
• 제17권4호
• /
• pp.274-279
• /
• 2012

For the quantitative analysis of genetically modified (GM) maize in processed foods, primer sets and probes based on the 35S promoter (p35S), nopaline synthase terminator (tNOS), p35S-hsp70 intron, and zSSIIb gene encoding starch synthase II for intrinsic control were designed. Polymerase chain reaction (PCR) products (80~101 bp) were specifically amplified and the primer sets targeting the smaller regions (80 or 81 bp) were more sensitive than those targeting the larger regions (94 or 101 bp). Particularly, the primer set 35F1-R1 for p35S targeting 81 bp of sequence was even more sensitive than that targeting 101 bp of sequence by a 3-log scale. The target DNA fragments were also specifically amplified from all GM labeled food samples except for one item we tested when 35F1-R1 primer set was applied. A reference plasmid pGMmaize (3 kb) including the smaller PCR products for p35S, tNOS, p35S-hsp70 intron, and the zSSIIb gene was constructed for real-time PCR (RT-PCR). The linearity of standard curves was confirmed by using diluents ranging from $2{\times}10^1{\sim}10^5$ copies of pGMmaize and the $R^2$ values ranged from 0.999~1.000. In the RT-PCR, the detection limit using the novel primer/probe sets was 5 pg of genomic DNA from MON810 line indicating that the primer sets targeting the smaller regions (80 or 81 bp) could be used for highly sensitive detection of foreign DNA fragments from GM maize in processed foods.

• Journal of Microbiology and Biotechnology
• /
• 제25권12호
• /
• pp.2090-2099
• /
• 2015

Recently, the cardiovascular disease has been widely problematic in humans probably due to fibrin formation via the unbalanced Western style diet. Although direct (human plasmin) and indirect methods (plasminogen activators) have been available, bacterial enzyme methods have been studied because of their cheap and mass production. To detect a novel bacterial fibrinolytic enzyme, 111 bacterial strains with fibrinolytic activity were selected from kimchi. Among them, 14 strains were selected because of their stronger activity than 0.02 U of plasmin. Their 16S rRNA sequence analysis revealed that they belong to Bacillus, Leuconostoc, Propionibacterium, Weissella, Staphylococcus, and Bifidobacterium. The strain B. subtilis ZA400, with the highest fibrinolytic activity, was selected and the gene encoding fibrinolytic enzyme (bsfA) was cloned and expressed in the E. coli overexpression system. The purified enzyme was analyzed with SDS-PAGE, western blot, and MALDI-TOF analyses, showing to be 28.4 kDa. Subsequently, the BsfA was characterized to be stable under various stress conditions such as temperature (4-40oC), metal ions (Mn²⁺, Ca²⁺, K²⁺, and Mg²⁺), and inhibitors (EDTA and SDS), suggesting that BsfA could be a good candidate for development of a novel fibrinolytic enzyme for thrombosis treatment and may even be useful as a new bacterial starter for manufacturing functional fermented foods.