• BMB Reports
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• v.53 no.3
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• pp.154-159
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• 2020

We investigated the effects of physalin A, B, D, and F on osteoclastogenesis induced by receptor activator of nuclear factor κB ligand (RANKL). The biological functions of different physalins were first predicted using an in silico bioinformatic tool (BATMAN-TCM). Afterwards, we tested cell viability and cell apoptosis rate to analyze the cytotoxicity of different physalins. We analyzed the inhibitory effects of physalins on RANKL-induced osteoclastogenesis from mouse bone-marrow macrophages (BMMs) using a tartrate-resistant acid phosphatase (TRAP) stain. We found that physalin D has the best selectivity index (SI) among all analyzed physalins. We then confirmed the inhibitory effects of physalin D on osteoclast maturation and function by immunostaining of F-actin and a pit-formation assay. On the molecular level, physalin D attenuated RANKL-evoked intracellular calcium ([Ca(2+)](i)) oscillation by inhibiting phosphorylation of phospholipase Cγ2 (PLCγ2) and thus blocked the downstream activation of Ca2+/calmodulin-dependent protein kinases (CaMK)IV and cAMP-responsive element-binding protein (CREB). An animal study showed that physalin D treatment rescues bone microarchitecture, prevents bone loss, and restores bone strength in a model of rapid bone loss induced by soluble RANKL. Taken together, these results suggest that physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via suppressing the PLCγ2-CaMK-CREB pathway.

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.111-113
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• 2018

In this study, purification of cold-adapted protease from Janthinobacterium sp. was investigated. First, using gradient precipitation, protease was confirmed to be deposited in the 30-80% range of ammonium sulfate. Next, DEAE-Sepharose column was used for the binding of the protease under various conditions. The optimal binding condition was found to be pH 8.5 and flow rate of 30 ml/h. Under the optimal condition, the protease was purified with 29% recovery yield. This result can be useful for the purification of other cold-adapted protein.

• The Korean Journal of Mycology
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• v.12 no.1
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• pp.35-43
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• 1984

To produce and characterize antineoplastic constituents in the submerged cultured­mycelia of Laccaria laccata, the mycelia were extracted with distilled water. Purification of the extract was carried out by acetone precipitation, by ion exchange chromatography using DEAE­Sephadex A-50, CM-Sephadex C-25 resins, and by gel filtration chromatography on Sephadex G-200. Each fraction obtained during the purification was examined for antineoplastic activity against sarcoma 180 in ICR mice. As the purification proceeded, the antineoplastic activity was markedly increased. The highly purified Fraction E showed 75% tumor inhibition ratio at a dose of 10mg/kg/day and contained 81% polysaccharide and 4% protein. The antitumor component of Fraction E stimulated an accumulation of peritoneal exudate cells including peritoneal macrophages, and is named laccaran.

• Microbiology and Biotechnology Letters
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• v.28 no.5
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• pp.251-257
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• 2000

Serratia marcescens purine nucleoside phosphorylase (PNP) was purfied to homogeneity by streptomycin sulfate treatment, Sephacry HR S-200 gel filtration chromatography and AMP-agarose affinity chromatography. The specific activity of the enzyme was increased 49-fold during purification with an overall yield of 7.0%. The molecular weight was 168kD as estimated by Sephadex G-150 gel filtration chromatography. The S. marcescens enzyme was composed of six identical subunits with subunit molecular weight of 28kD, as estimated by SDS-PAGE. The Km values of S. marcescens enzyme for inosine and deoxyinsoine were 0.38 and 1.20 mM, respectively. The ph optimum was near 8.0, and the enzyme was relatively heat-stable protein. The enzyme was inactivated com-pletely by 0.5 mM of $Cu^{ 2+}$.

• Food Science of Animal Resources
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• v.20 no.2
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• pp.125-132
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• 2000

Lactoferrin was isolated from the colostrum of Korean native cow by using several purification steps such as batch extraction, ion exchange chromatography, gel filtration chromatography and affinity chromatography. Other whey protein components that having similar molecular weight and affinity to lactoferrin were gradually removed from crude Korean native cow's lactoferrin during the purification steps. The amount of lactoferrin collected from a liter of Korean native cow's colostrum was 65mg and the recovery rate was 29.4%. The molecular weight of the purified Korean native cow's lactoferrin was estimated approximately 81,000dalton.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.446-452
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• 1993

The cholesterol oxidase produced from Bacillus sphaericus was purified and characterized. Through a series of purification procedures including DEAE-Toyoperal 650C, Sephadex G-200 and DEAE-Sephadex A-50 column chromatography, the purified enzyme was shown to have a specific activity of 0.179 units/mg protein having 31.8 fold purification and final yield of 12%. The molecular weight of the enzyme was estimated to be 47kDa and 47.tkDa by Sephadex G-200 chromatography and SDS-PAGE. The optimum temperature and pH for the enzyme were 30C and 6.0, respectively. The activity of the purified cholesterol oxidase was inhibited by Fe2+ and Hg+.

• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.559-562
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• 1998

Odontoglossum ringspot virus (ORSV) was finally purified from ORSV-infected orchid plants by diethylaminoethyl (DEAE) cellulose anion exchange column chromatography. The virus was reliably eluted by potassium chloride at the concentration from 0.1 M to 0.13 M. Partial purification was done by solubilization with Triton X-100 (allkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG; MW 8,000). The finally purified ORSV represented one distinct homogeneous band and the molecular weight of its capsid protein was about 17,500 Dalton in electrophoretic analysis. Electron microscopy showed not only intact particles ranged from 280 nm to 340 nm in length, but also segmented particles that final 140 nm to 220 nm and even disks. Enzyme-linked immunosorbent assay (ELISA) showed that final yield was 12 mg/100 g of the infected leaves. Bioassay demonstrated that the purified ORSV had the normal infectivity to orchid plants and Nicotiana glutionsa. Based on these data, anion exchange column chromatography could be efficiently applied to the purification of ORSV and other viruses similar to ORSV.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2001.04a
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• pp.38-38
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• 2001

No abstract. See Full-text

• The Microorganisms and Industry
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• v.19 no.4
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• pp.14-26
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• 1993

Optimizing protein production in recombinant E. coli strains involves manipulation of genetic and environmental factors. In designing a production system, attention must be paid to gene expression efficiency, culture conditions and bioreactor configuration. Although not much emphasis was given to the physiology of host strains in this review, an understanding of the relationship between the physiology of host cell growth and the overproduction of a cloned gene protein is of primary importance to the improvement of the recombinant fermentation processes. Sometimes it is desirable to make use of gene fusion systems, e.g. protein A, polypeptide, gutathione-S-transferase, or pneumococcal murein hydrolase fusion, to facilitate protein purification.

• Korean Journal of Environmental Biology
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• v.37 no.4
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• pp.585-591
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• 2019

Most insect-resistant LMOs have been produced by applying Cry and Vip3Aa proteins. Vip3Aa protein is activated during the vegetative stage of Bacillus thuringensis (Bt) and the inhibitory activity of the Vip3Aa protein against pathogenic attacks from lepidopteran insect species is well known. However, a risk assessment of the Vip3Aa protein compared to the Cry protein has not been conducted in South Korea. This study demonstrates a possible risk assessment method for Vip3Aa protein against honeybees (Apis mellifera). For the risk assessment of the protein, we purified the recombinant Vip3Aa protein in Escherichia coli. The survival rate and symptoms of general intoxication of 4 months honeybees were measured after Vip3Aa exposure. These results indicated that there was no significant difference in the survival rate and the symptom between Vip3Aa and the control buffer. In this study, we established standard methods of Vip3Aa protein purification and oral adult toxicity test using A. mellifera as an LMO risk assessment technique for preserving the natural ecosystem of South Korea.