• Food Science and Biotechnology
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• v.14 no.1
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• pp.49-57
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• 2005

Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.

• Journal of Microbiology and Biotechnology
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• v.24 no.7
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• pp.936-942
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• 2014

Using racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide, an intermediate for the chiral pyrethroid insecticide Esfenvalerate, as a sole nitrogen source in a minimal medium, several strains with high enatioselectivity (${\geq}98%$) were isolated by enrichment techniques. One of the strains, LG 31-3, was identified as Burkholderia multivorans, based on physiological and morphological tests by a standardized Biolog station for carbon source utilization. A novel amidase was purified from B. mutivorans LG 31-3 and characterized. The enzyme exhibited (S)-selective amidase activity on racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide. Addition of the racemic amide induced the production of the enantioselective amidase. The molecular mass of the amidase on SDS-PAGE analysis was shown to be 50 kDa. The purified amidase was subjected to proteolytic digestion with a modified trypsin. The N-terminal and internal amino acid sequences of the purified amidase showed a high sequence homology with those deduced from a gene named YP_366732.1 encoding indole acetimide hydrolase from Burkholderia sp. 383.

• Food Science of Animal Resources
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• v.43 no.5
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• pp.877-888
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• 2023

We studied the proteolysis and conducted a sensory evaluation of fermented sausages using strains derived from Kimchi [Pediococcus pentosaceus-SMFM2021-GK1 (GK1); P. pentosaceus-SMFM2021-NK3 (NK3)], Doenjang [Debaryomyces hansenii-SMFM2021-D1 (D1)], and spontaneous fermented sausage [Penicillium nalgiovense-SMFM2021-S6 (S6)]. Fermented sausages were classified as commercial starter culture (CST), mixed with GK1, D1, and S6 (GKDS), and mixed with NK3, D1, and S6 (NKDS). The protein content and pH of GKDS and NKDS were significantly higher than those of CST on days 3 and 31, respectively (p<0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the NKDS had higher molecular weight proteins than the GKDS and CST. The myofibrillar protein solubility of the GKDS and NKDS was significantly higher than that of the CST on day 31 (p<0.05). The GKDS displayed significantly higher pepsin and trypsin digestion than the NKDS on day 31 (p<0.05). The hardness, chewiness, gumminess, and cohesiveness of the GKDS were not significantly different from those of the CST. The GKDS exhibited the highest values for flavor, tenderness, texture, and overall acceptability. According to this study, sausages fermented using lactic acid bacteria (GK1), yeast (D1), and mold (S6) derived from Korean fermented foods displayed high proteolysis and excellent sensory evaluation results.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.17 no.2
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• pp.101-108
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• 1984

In order to study the optimal conditions of processing and storage for boiled and dried anchovy (Engraulis japonica) with high protein digestibility, the contents of trypsin indigestible substrate (TI) and in vitro apparent protein digestibility were determined. Peroxide value (PoV), TBA number and nonenzymatic brown pigments, that accounted for important antinutritional factors, were also measured and confirmed the relationship between those factors and formation of TI or in vitro protein apparent digestibility. The results were as follows; Samples boiled for 5 minutes showed the lower content of TI than the other samples boiled for 0.5 min. or 1 min. Hot air dried products had a lower TI content in comparison with the other dried ones such as sun dried or freeze dried products. It was revealed that the lower temperature ($8{\pm}1^{\circ}C$) did not affect to a great degree of forming TI and falling in vitro digestibility comparing to high temperature ($26{\pm}1^{\circ}C$) during storage. The lowest TI content (0.173 mg/g solid) was noted in the samples for 5 minutes and then sun drying after 56 days storage at $9{\pm}1^{\circ}C$. A rapid decrease of in vitro protein digestibility occurred within 0.5 min. of boiling and showed the value $85.3\%$. Freeze dried samples possessed the highest in vitro protein digestibility ($85.9\%$), when compared to sun dried or hot air dried products. Fat oxidation and nonenzymatic browning were proceeded with the various boiling times, drying methods and storing temperatures. It was noted that boiling for 5 minutes and freeze drying accelerate the fat oxidation significantly. More nonenzymatic brown pigments was developed in samples boiled for shorter time (0.5 min.) and that stored at high temperature ($26{\pm}1^{\circ}C$) than the other products. Therefore, fat oxidation and nonenzymatic browning assumed to be a major inhibitory reaction in enzyme digestion and those might be an important role in forming TI in boiled and dried anchovy products during processing and storage.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.151-157
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• 2003

Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

• Korean journal of applied entomology
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• v.47 no.4
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• pp.457-465
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• 2008

Eight Bacillus thuringiensis strains activated against mosquito larva were compared their characterization. Spherical-shaped parasporal inclusion of B. thuringiensis subsp. tohokuensis CAB167 was observed by phase-contrast microscopy and scanning electron microscopy. $LC_{50}$ values of B. thuringiensis subsp. tohokuensis CAB167 against Culex pipiens molestus, Culex pipiens pallens, and Aedes aegyti were 173, 190 and 580 ng/ml, respectively. B. thuringiensis subsp. tohokuensis CAB167 had a parasporal inclusion containing 4 major protein components, for example, 135, 80, 49 and 28-kDa by SDS-PAGE. Otherwise, after trypsin digestion of parasporal inclusion, SDS-PAGE was showed new protease-resistant peptides at 72 and 63-kDa. Activated toxins of isolated CAB167 were different from other reference strains on a serological by immuno-diffusion test.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.4
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• pp.430-436
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• 2010

The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

• The Korean Journal of Zoology
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• v.34 no.2
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• pp.148-158
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• 1991

The purpose of this research was to establish the culture condition for dissociated submandibu -lar gland (SG) cells. After trypsin digestion of SG from 3-4 weeks old mice, dissociated cells were cultured in 1OO/o fetal bovine serum-Dulbecco's modified Eagle's medium (FBS-DME) or 0.5-2% low protein serum replacement-DME (LPSR-DME) on plastic surface to form monolayer. The effects of FBS, LPSR and hormones on the growth and function of cultured SG cells were examined. SG cells dissociated by enzyme were successfully cultured and were characterized as epithelial-like cells by light and electron microscope. The maximal DNA synthesis of cultured SG cells was achieved by DME containing 5-10% FBS. The same results were obtained when the effects of LPSR on cell proliferation were examined up to a LPSR concentration of 2%. SG cells cultured in 20/o LPSR-DME expressed a population doubling time of 42.5 hrs and a saturation density of 1.2 $\times$10 5cell/cm$^2$. Dihydrotestosterone (DHT) in medium did not influence on the DNA synthesis of the cultured SG cells, but stimulated protein synthesis of the SG cells. Thyroxine (T4) stimulated protein synthesis of the SCI cells markedly in a dose-dependent fashion. EGF secretion by the cultured SG cells increased significandy by DHT and or T4 trearment. This finding indicated that secretion of EGF by the SG cells was under the control of the hormones such as androgen and thyroid hormones. It seems to be that the culture condition described here can be used as a useful tool for further research on the SCI cells.

• Journal of Dairy Science and Biotechnology
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• v.38 no.3
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• pp.161-167
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• 2020

Cheese pizzas fortified with casein phosphopeptide (CPP) and calcium were subjected to an in vitro digestion to assess whether CPP could prevent the precipitation of calcium. The total calcium content of the cheese pizzas was adjusted to 1,000 mg per pizza (~370 g) with the addition of calcium originating from eggshells. Two levels of trypsin-digested caseins (367 and 459 mg), with a CPP content of ~20%, were added to each pizza. The in vitro digested pizzas were then centrifuged and the supernatant was mixed with Na₂HPO₃ at 37℃ to estimate the possible soluble effect of CPP on calcium. After 24 h of reaction, the solution was centrifuged and the calcium content in the resultant supernatant was analyzed by inductively coupled plasma-atomic emission spectroscopy. One-way statistical analyses showed that CPP had a positive effect on the solubilization of calcium against phosphate (p<0.05). Cheese pizza supplemented with 459 mg of CPP powder was able to prevent precipitation of calcium by 98.8%, whereas no CPP-added cheese pizza solubilized 86.4% of the calcium. A sensory test was also carried out, revealing that panelists could not discern the bitter taste of the CPP added to the pizzas.

• The Korean Journal of Community Living Science
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• v.27 no.4
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• pp.863-873
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• 2016

The current study was undertaken to determine the effects of the ethanol extracts of loquat (Eriobotrya japonica Lindl.) seeds, flesh or leaves on the differentiation of 3T3-L1 cells and male pig preadipocytes. The cell number was measured with the MTT assay after trypsin digestion. The cell differentiation was determined by measuring the glycerol-3-phosphate dehydrogenase (GPDH) activity and triglyceride(TG) content. No cytotoxicity was observed from the loquat flesh and leaf ethanol extracts at concentrations of 5, 10, 25, 50, 100 or $200{\mu}g/mL$ in 3T3-L1 cells and pig preadipocytes. However, the cell viability of neither cell line were affected by up $50{\mu}g/mL$ of loquat seed ethanol extract. Treatment with the loquat seed and leaf ethanol extracts significantly suppressed the terminal differentiation of both cell lines in a dose-dependent manner, as confirmed by the decrease in the glycerol-3-phosphate dehydrogenase(GPDH) activity and TG content. Treatment with the loquat seed and leaf ethanol extracts inhibited the GPDH activity and reduced the TG content of both cell types more effectively than that with the loquat flesh ethanol extract. The most potent anti-adipogenic effect was obtained in the case of the ethanol extract of loquat seeds.