• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.5
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• pp.442-448
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• 2006

Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

• Journal of Microbiology
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• v.45 no.5
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• pp.402-408
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• 2007

A strain producing a potent protease was isolated from turban shell. The strain was identified as Bacillus sp. S17110 based on phylogenetic analysis. The enzyme was purified from culture supernatant of Bacillus sp. S17110 to homogeneity by ammonium sulfate precipitation, SP-Sepharose, and DEAE-Sepharose anion exchange chromatography. Protease activity of the purified protein against casein was found to be stable at pH 7 to pH 10 and around $50^{\circ}C$. Approximately 70% of proteolytic activity of the enzyme was detected either in the presence of 100 mM SDS or Tween 20. The enzyme activity was enhanced in the presence of $Ca^{2+},\;Zn^{2+},\;Mg^{2+}$, but was inhibited by EDTA, indicating that it requires metal for its activity. The purified enzyme was found to be a monomeric protein with a molecular mass of 75 kDa, as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography. The purified enzyme was analyzed through peptide fingerprint mass spectra generated from matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and a BLAST search, and identified as immune inhibitor A (inhA) deduced from nucleotide sequence of B. cereus G9241. Since InhA was identified as protease that cleave antibacterial proteins found in insect, inhA-like protease purified from Bacillus sp. S17110 might be pathogenic to sea invertebrates.

• Journal of Marine Bioscience and Biotechnology
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• v.14 no.1
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• pp.9-24
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• 2022

Low-molecular weight peptides derived from fish collagen exhibit several bioactivities, including antioxidant, antiwrinkle, antimicrobial, antidiabetic, and antihypertension effects. These peptides are also involved in triglyceride suppression and memory improvement. This study aimed to investigate the optimal processing condition for preparing low-molecular weight peptides from flounder skin, and the properties of the hydrolysate. The optimal processing conditions for peptic hydrolysis were as follows: a ratio of pepsin to dried skin powder of 2% (w/w), pH of 2.0, and a temperature of 50℃. Peptic hydrolysate contains several low-molecular weight peptides below 300 Da. Gly-Pro-Hyp(GPHyp) peptide, a process control index, was detected only in peptic hydrolysate on matrix-assisted laser desorption/ionization-time-of-flight(MALDI-TOF) spectrum. 2,2'-azinobis-(3-3-ethylbenzothiazolline-6- sulfonic acid(ABTS) radical scavenging activity of the peptic hydrolysate was comparable to that of 1 mM ascorbic acid, which was used as a positive control at pH 5.5, whereas collagenase inhibition was five times higher with the peptic hydrolysate than with 1 mM ascorbic acid at pH 7.5. However, the tyrosinase inhibition ability of the peptic hydrolysate was lower than that of arbutin, which was used as a positive control. The antibacterial effect of the peptic hydrolysate against Propionibacterium acne was not observed. These results suggest that the peptic hydrolysate derived from a flounder skin is a promising antiwrinkle agent that can be used in various food and cosmetic products to prevent wrinkles caused by ultraviolet radiations.

• Journal of the Korean Applied Science and Technology
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• v.36 no.3
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• pp.685-699
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• 2019

Antioxidant, antibacterial, and skin penetration tests were conducted to investigate the skin absorption of Pyrus serotina var leaf extracts using polymer micelles and their applicability to cosmetic ingredients. Total polyphenol content was found to be $118.83{\pm}9.39mg/g$ in Pyrus serotina var leaf ethanol extract and $106.89{\pm}4.45mg/g$ in Pyrus serotina var leaf hydrothermal extract. The DPPH radical scavenging activity was found to be the highest radical scavenging activity of $74.39{\pm}7.48%$ of the Pyrus serotina var leaf ethanol extract at the concentration of 500 mg/L. The SOD-like activity was $91.62{\pm}0.43%$, the highest value at the concentration of 1,000 mg/L in the hydrothermal extract. After the experiment, antioxidation, wrinkle improvement and whitening activity were confirmed, and the Pyrus serotina var leaf extract was highly likely to be realized as antioxidant and antibacterial material. In the skin penetration experiment with the Pyrus serotina var leaf ethanol extract, the permeation amount of total accumulated tannic acid was found to be Formulation 2 ($55.45{\mu}g/cm^2$), Formulation 1 ($46.43{\mu}g/cm^2$), Formulation 0 ($34.36{\mu}g/cm^2$). In the liposome's skin penetration experiment containing pear leaf hydrothemal extract, the total amount of accumulated tannic acid permeation was found to be Formulation 5 ($75.01{\mu}g/cm^2$), Formulation 4 ($64.01{\mu}g/cm^2$) and Formulation 3 ($36.60{\mu}g/cm^2$). Through this study, we confirmed the possibility of antioxidant and wrinkle effects of Pyrus serotina var leaf extract. In addition, as a result of skin penetration through the production of polymer micelles and liposome containing Pyrus serotina var leaf extract, It will be more usable in cosmetic industry.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.52-67
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• 1998

The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.

• Journal of Mushroom
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• v.6 no.1
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• pp.32-37
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• 2008

This study was carried out to investigate the growth inhibition of Propionibacterium acnes by mycelial culture broth of Paecilomyces japonica in the Mulberry leaf extract. The growth inhibition effect of P. japonica mycelial culture broth against P. acnes in various concentration of Mulberry leaf extract was the most effective in 3% Mulberry leaf extract. The inhibition effect of P. japonica mycelial culture broth against P. acnes according to the culture time was the moust effective after 25 days mycelial cultivation. As the treating amount of the mycelial culture broth was increased, the growth inhibition effect against P. acnes was increased gradually. The growth inhibition effect of mycelial culture broth against P. acnes according to the time of heat treatment was active by 45min at $100^{\circ}C$, while it was inactive more than 60min at $100^{\circ}C$. This result suggested that the antibacterial materials are possible to be glycoside or peptides. Taken together P. japonica mycelial culture in the Mulbarry leaf extract has a possibility to be an element of skin-care cosmetics regulating the acnes.

• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.104-110
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• 2011

Actinomycetes, Gram positive soil bacteria, are valuable microorganisms which produce useful secondary metabolites including antibiotics, antiparasitic substances, anti-cancer drugs, and immunosuppressants. Although a major family of actinomycetes, known as streptomycetes, has been intensively investigated at the molecular level for several decades, a potentially valuable and only recently isolated non-streptomycetes rare actinomycetes (NSRA) family has been poorly characterized due to lack of proper genetic manipulation systems. Here we report that a PCR-based genome screening strategy was performed with approximately 180 independently isolated actinomycetes strains to isolate potentially valuable NSRA strains. Thanks to this simple PCR-based genome screening strategy we were able to identify only seven NSRA strains, followed by 16S rRNA sequencing for confirmation. Through further bioassays, one potentially valuable NSRA strain (tentatively named Nocardiopsis species MMBL010) was identified which possessed both antifungal and antibacterial activities, along with the presence of polyketide synthase and non-ribosomal peptide synthase genes. Moreover, Nocardiopsis species MMBL010, which was intrinsically recalcitrant to genetic manipulation, was successfully transformed via E. coli-driven conjugation. These results suggest that PCR-based genome screening, followed by the establishment of an E. coli-driven conjugation system, is an efficient strategy to maximize potentially valuable compounds and their biosynthetic genes from NSRA strains isolated from various environments.

• Microbiology and Biotechnology Letters
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• v.38 no.3
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• pp.255-262
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• 2010

Strain BCNU 2001 was isolated from soil samples collected from Tea-baek Mountain area. The biochemical characteristics and 16S ribosomal RNA gene sequences of the isolate revealed that the strain belonged to the Pseudomonas aeruginosa. The supernatants had an antimicrobial effect on various kind of bacteria and fungi. Especially BCNU 2001 was able to greatly inhibit the growth of Micrococcus luteus, Proteus mirabilis, Proteus vulgaris, and Aspergillus niger, and its inhibition zone was measured as 18.5 mm against Micrococcus luteus, 19.0mm against Proteus mirabilis, 17.0mm against Proteus vulgaris, and 13.5 mm against Aspergillus niger, respectively. Hexane and dichloromethane extracts of BCNU 2001 exhibited significant activity against bacteria, and dichloromethane and ethylacetate extracts showed significant activity against fungi. Pseudomonas strain BCNU 2001 was also determined to have antimicrobial peptide against various microorganisms including Gram positive bacteria, Gram negative bacteria and fungi. The obtained results may provide preliminary support for the usefulness of Pseudomonas strain BCNU 2001.