• BMB Reports
• /
• v.38 no.5
• /
• pp.526-532
• /
• 2005

A lectin with in-vitro anticancer activity against established human cancer cell lines has been purified by affinity chromatography on asialofetuin-linked amino activated silica beads from the tubers of Arisaema tortuosum, popularly known as Himalayan Cobra lily, a monocot plant from the family Araceae. The bound Arisaema tortuosum lectin (ATL) was eluted with glycine-HCl buffer, pH 2.5. ATL was effectively inhibited by asialofetuin, a complex desialylated serum glycoprotein as well as by N-acetyl-D-lactosamine, a disaccharide. It gave a single band corresponding to a subunit molecular weight of 13.5 kDa in SDS-PAGE, pH 8.8 both under reducing and non reducing conditions. When subjected to gel-filtration on Biogel P-200, it was found to have a molecular weight of 54 kDa, suggesting a homotetramer structure, in which individual polypeptides are not bound to each other with disulfide bonds. ATL is a glycoprotein with 0.9% carbohydrate content, stable up to $55^{\circ}C$ and at pH 2 to 10. The lectin had no requirement for divalent metal ions i.e. $Ca^{2+}$ and $Mn^{2+}$ for its activity. However, as reported for other monocot lectins, ATL gave multiple bands in isoelectric focusing and Native PAGE, pH 8.3. The lectin was found to inhibit in vitro proliferation of human cancer cell lines HT29, SiHa and OVCAR-5.

• The Journal of the Korea Contents Association
• /
• v.10 no.2
• /
• pp.25-34
• /
• 2010

Consumer's buying loyalty for goods can be changed by exposure to new stimulus, so different with others and esthetical packaging design is necessary for drawing the eyes of consumer at this time. In this study, we made up a questionnaire and analyzed thirties who is judged as a substantial consumer for recognition of newly launched seasoning packaging design. They answered there is no specific character comparatively in existing seasoning packaging design and gave a high appraisal on symbolism of calligraphy logotype which is produced by attributes but got a low appraisal on readability. And they appraised Cheong-jeong-won Maat-seon-saeng higher than Da-si-da San-deul-e on safety, quality preservation, and exhibitivity and chose safety and preservation among those as essential element inducing to buy.

• Journal of Plant Biotechnology
• /
• v.6 no.4
• /
• pp.235-239
• /
• 2004

We studied the effect of genotype, explant, age of explant, medium (plant growth regulators and gelling agents), and standardized an efficient regeneration protocol for inbred lines of Chinese cabbage (Brassica rapa ssp. pekinensis). Of the different concentrations of NAA and BA tested, the combination of $5\;\cal{mg/L}\;BA\;and\;0.5\;\cal{mg/L}$ NAA gave the highest frequency of shoot regeneration. The cotyledonary explants had more shoot regeneration frequency ($\ge40\%$) than the hypocotyl ex-plants. Besides, the cotyledonary explants, excised from the 4-day old seedlings, showed higher shoot regene-ration ($56.7\%$). Of the three gelling agents and their concentrations used, 16 g/L agar was found to be the best for shoot regeneration. Shoot regeneration frequency in-creased significantly by supplementing the medium with $4\;\cal{mg/L}\;of\;AgNO_3$ MS medium devoid of NAA showed higher frequency of rooting in the regenerated shoots than the ones supplemented with NAA. Our improved regeneration protocol will be especially useful for the genetic transformation of Chinese cabbage inbred lines to develop transgenic hybrids.

• BMB Reports
• /
• v.35 no.3
• /
• pp.273-282
• /
• 2002

Sp3 is a bifunctional transcription factor that has been reported to stimulate or repress the transcription of numerous genes. Although the size of Sp3 mRNA is 4.0kb, the size of the known Sp3 cDNA sequence is 3.6kb. Thus, Sp3 functional studies have been performed with an artificially introduced start codon, and thus an amino-terminus that differs from the wild-type. Ideally, full-length cDNA expression vectors with the appropriate start codon should be utilized for these studies. Using 5'rapid amplification of cDNA ends, a full-length Sp3 cDNA clone was generated and the sequence verified in nine cell lines. No AUG initiation codon was present. However, stop codons were present in all three frames 5' to the known coding sequence. In vitro translation of this full-length cDNA clone produced the expected three isoforms-one at 100 kDa and two in the mid 60 kDa range. Electrophoretic mobility shift assays showed that the protein products had the ability to bind to the Sp1/3 consensus sequence. In vitro studies, using our Sp3 clone and site directed mutagenesis, identified the translation initiation site for the larger isoform as AUA. AUA has not been previously described as an endogenous initiation codon in eukaryotes.

• Journal of Korean Neurosurgical Society
• /
• v.63 no.6
• /
• pp.723-729
• /
• 2020

Objective : The use of oblique lateral interbody fusion at the L5-S1 level (OLIF51) is increasing, but no study has directly compared OLIF51 and transforaminal lumbar interbody fusion (TLIF) at the L5-S1 level. We evaluated the usefulness of OLIF51 by comparing clinical and radiologic outcomes with those of TLIF at the same L5-S1 level. Methods : We retrospectively reviewed and compared 74 patients who underwent OLIF51 (OLIF51 group) and 74 who underwent TLIF at the L5-S1 level (TLIF51 group). Clinical outcomes were assessed with the visual analogue scale for back pain and leg pain and the Oswestry Disability Index. Mean disc height (MDH), foraminal height (FH), disc angle (DA), fusion rate, and subsidence rate were measured for radiologic outcomes. Results : The OLIF51 group used significantly higher, wider, and larger-angled cages than the TLIF51 group (p<0.001). The postoperative MDH and FH were significantly greater in the OLIF51 group than in the TLIF51 group (p<0.001). The postoperative DA was significantly larger in the OLIF51 group than in the TLIF51 group by more than 10º (p<0.001). The fusion rate was 81.1% and 87.8% at postoperative 6 months in the OLIF51 and TLIF51 groups, respectively, and the TLIF51 group showed a higher fusion rate (p<0.05). The subsidence rate was 16.2% and 25.3% in the OLIF51 and TLIF51 groups, respectively, and the OLIF51 group showed a lower subsidence rate (p<0.05). Conclusion : OLIF51 was more effective for the indirect decompression of foraminal stenosis, providing strong mechanical support with a larger cage, and making a greater lordotic angle with a high-angle cage than with TLIF.

• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.5
• /
• pp.598-606
• /
• 2010

This study investigated the nutritional quality of soybean meal (SBM) fermented by Aspergillus ($FSBM_A$) and/or followed by Lactobacillus fermentation ($FSBM_{A+L}$). Both fermented products significantly improved protein utilization of SBM with higher trichloroacetic acid (TCA) soluble true protein content, in vitro protein digestibility and available lysine content, especially in $FSBM_{A+L}$. Moreover, $FSBM_{A+L}$ produced a huge amount of lactic acid resulting in lower pH as compared to the unfermented SBM or soybean protein concentrate (SPC) (p<0.05). $FSBM_A$ and $FSBM_{A+L}$ raised 4.14% and 9.04% of essential amino acids and 5.38% and 9.37% of non-essential amino acids content, respectively. The ${\alpha}$-galactoside linkage oligosaccharides such as raffinose and stachyose content in $FSBM_A$ and $FSBM_{A+L}$ decreased significantly. The results of soluble protein fractions and distribution showed that the ratio of small protein fractions (<16 kDa) were 42.6% and 63.5% for $FSBM_A$ and $FSBM_{A+L}$, respectively, as compared to 7.2% for SBM, where the ratio of large size fractions (>55 kDa, mainly ${\beta}$-conglycinin) decreased to 9.4%, 5.4% and increased to 38.8%, respectively. There were no significant differences in ileal protein digestibility regardless of treatment groups. SPC inclusion in the diet showed a better protein digestibility than the SBM diet. In summary, soybean meal fermented by Aspergillus, especially through the consequent Lactobacillus fermentation, could increase the nutritional value as compared with unfermented SBM and is compatible with SPC.

• Biomedical Science Letters
• /
• v.6 no.4
• /
• pp.261-268
• /
• 2000

Fibrinolytic enzyme (FE-2) was purified from the fruiting bodies of Tricholoma saponaceum using DEAE-Cellulose chromatography and Mono-S column chromatography, The enzyme has a molecular weight of 18.23 kDa and include Zn$^{2+}$ ion as found by ICP/MS. The N-terminal amino acid sequence of the enzyme was A-L-Y-V-G-X-S-P-X-Q-Q-S-L-L-V It has a pH optimum at pH 7.5, suggested that FE-2 was a neutral pretense. The activity of FE-2 was highly inhibited by EDTA and 1,10-phenanthroline, indicating that the enzyme is a metalloprotease. The activity of FE-2 was increased by $Mg^{2+}$, Zn$^{2+}$, Fe$^{2+}$, and Co$^{2+}$, but the enzyme activity was totally inhibited by Hg$^{2+}$. No inhibition was found with PMSF, E-64, pepstatin and 2-mercaptoethanol. The enzyme hydrolyzed both $A\alpha$ and B$\beta$ chains of human fibrinogen. The $\gamma$ chain was resistant to hydrolysis by FE-2.

• Mycobiology
• /
• v.40 no.3
• /
• pp.173-180
• /
• 2012

A ${\beta}$-glucosidase from Penicillium italicum was purified with a specific activity of 61.8 U/mg, using a chromatography system. The native form of the enzyme was an 88.5-kDa tetramer with a molecular mass of 354 kDa. Optimum activity was observed at pH 4.5 and $60^{\circ}C$, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and $65^{\circ}C$, respectively. Its activity was inhibited by 47% by 5 mM $Ni^{2+}$. The enzyme exhibited hydrolytic activity for p-nitrophenyl-${\beta}$-D-glucopyranoside (pNP-Glu), p-nitrophenyl-${\beta}$-D-cellobioside, p-nitrophenyl-${\beta}$-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-${\beta}$-D-lactopyranoside, p-nitrophenyl-${\beta}$-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a ${\beta}$-glucosidase. The $k_{cat}/K_m\;(s^{-1}mM^{-1})$ values for pNP-Glu and cellobiose were 15,770.4 mM and 6,361.4 mM, respectively. These values were the highest reported for ${\beta}$-glucosidases. Non-competitive inhibition of the enzyme by both glucose ($K_i=8.9mM$) and glucono-${\delta}$-lactone ($K_i=11.3mM$) was observed when pNP-Glu was used as the substrate. This is the first report of non-competitive inhibition of ${\beta}$-glucosidase by glucose and glucono-${\delta}$-lactone.

• The Korean Journal of Mycology
• /
• v.36 no.2
• /
• pp.183-188
• /
• 2008

A fibrinolytic serine protease was purified from the fruiting bodies of an edible mushroom, Albatrellus confluens. The enzyme had a molecular mass of 30086.41 Da, as measured by MALDI-TOF mass spectrometry. The N-terminal amino acid sequence of the enzyme was Glu-Thr-Val-Thr-Glu-Thr-Thr-Ala -Pro-Trp-Gly-Leu-Ser-Arg-Ile. It displayed optimal activity at $50^{\circ}C$ and within a pH range of $8.0{\sim}10.0$, suggesting that the enzyme is an alkaline protease. The enzyme was stable up to $30^{\circ}C$. The enzyme displayed a strong substrate specificity for the synthetic peptide, N-Suc-Ala-Ala-Pro-Phe pNA. The enzyme activity was completely inhibited by addition of PMSF, indicating that the enzyme is a serine protease. No inhibition was observed following addition of E-64, pepstatin, or EDTA. The activity of the purified enzyme was decreased in the presence $Fe^{2+}$ or $Co^{2+}$, and the enzyme was completely inhibited by addition of $Hg^{2+}$. From these results, we propose that Albatrellus confluens could be used for biofunctional foods development and has potential therapeutic value for the treatment of vascular diseases.

• Korean Journal Plant Pathology
• /
• v.13 no.4
• /
• pp.225-231
• /
• 1997

Activities of polygalacturonase, laccase, and intra- and extra-cellular $\beta$-glucosidase produced by 20 Botrytis cinerea isolates in liquid culture media containing cucumber cell was as a carbon source were measured and their relationships to the pathogenicity were analyzed. No significant correlations between these enzyme activities and the pathogenicity of B. cinerea were found. Mycelial growth rate on Bayendamm media, however, was higthly correlated with the pathogenicity (r=0.522) anong these isolates. Immuno-blot analysis of the culture filtrate using antibody against against exo-polygalacturonase revealed that only one band with molecular weight of 66 kDa was detected amone 34 tested isolates. It appears that these enzymes may not be primary factors in dermining the pathogenicity of B. cinerea.