• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.203-208
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• 1998

We isolated and characterized an antifungal peptide from the seeds of Phytolacca americana. Growth inhibition assay with Botrytis cinerea was used to screen inhibitory proteins from 60 different plant species. A 4 kDa antifungal peptide (Pa-AFP) inhibitory to hyphal growth of B. cinerea was found in the seeds of P. americana. The peptide Pa-AFP was purified to homogeneity by chromatographies of Sephadex G-50, DEAE-Sepharose, Sephacryl S-300, and C18 reverse-phase HPLC. Western blot analysis showed that a polyclonal antibody raised against the purified peptide cross-reacted with a 4 kDa protein in seeds but not in root and leaf tissues of P. americana. Pa-AFP inhibited the hyphal growth of Botrytis cinerea, Rihzoctonia solani, Fusarium oxysporum, and Magnaporthe grisea. Pa-AFP exhibited growth inhibition of Saccharomyces cerevisiae strain BWG7a, which was sensitive to osmotin.

• Bulletin of the Korean Chemical Society
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• v.28 no.1
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• pp.136-138
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• 2007

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.2
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• pp.164-172
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• 2001

The angiotensin-I converting enzyme (ACE) inhibitors from laver hydrolysate was isolated. Among the 13 kinds of proteases, Maxazyme NNP was most effective for preparing the high ACE inhibitory compound. In extraction conditions of ACE inhibitory peptide from laver hydrolysate, ACE inhibitory activity of hydrolysate treated with diethylether for decolorization and that of $70\%$ ethanol soluble fraction among the different ethanol concentrations were higher than other preparations. Low molecular fraction less than 3,000 dalton of layer hydrolysate separated by ultrafiltration had the highest ACE inhibitory activity, for further separation of ACE inhibitory peptide from laver hydrolysate, gel filtration chromatography (Sephadex G-25), reverse-phase HPLC (ODS & Vydac C-18) and gel permeation chromatography (Superdex Peptide HR) were performed. The molecular mass of the ACE inhibitory peptide fractions of gel permeation chromatography determined by electrospray-mass spectrometer were 413.48 (S1O2V2V1P),346.86 (S1O2V2V2P) and 320.32 (S2O6V3V1P) dalton and their amino acid sequence were Val-Gln-Gly-Asn, Thr-Glu-Thr and Phe-Arg, respectively.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.212-217
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• 2004

The gene coding for mannanase from Bacillus subtilis WL-7, a number of glycosyl hydrolase family 26, was hyperexpressed in Escherichia coli. Two recombinant plasmids, pE7MAN and pENS7, were constructed by introducing the complete mannanase gene and the mature mannanase gene lacking N-terminal signal peptide region into a expression vector pET24a(＋), respectively. The level of mannanase produced by E. coli BL21 (DE3) carrying pENS7, which included the mature mannanase gene, was considerably higher than that by E. coli BL21 (DE3)/pE7MAN. Almost mannanase produced by the recombinant E. coli carrying pENS7 at growth temperature of $37^{\circ}C$ existed as inactive enzyme of insoluble form. Growth at temperature below $31^{\circ}C$ increased the soluble fraction of mannanase having catalytic activity in the recombinant E. coli cells. The highest productivity of active mannanase was observed in cell-free extract of the recombinant E. coli grown at growth temperature ranging from $25^{\circ}C$ to $28^{\circ}C$, while mannanase activity per soluble protein of the cell-free extract was highest in the cells grown at $^31{\circ}C$.

• The Korean Journal of Nuclear Medicine
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• v.17 no.1
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• pp.41-53
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• 1983

It is already well known that many factors are involved in maintaining normal blood glucose level. The amount and components of meal are also thought to be some of the factors which affect the blood glucose and insulin levels. It is reported that as for Koreans sugar takes up over 75% out of 2,098 kcal, the average daily calorie intake per adult. It implies that Koreans take a high-sugar diet compared with Westerners who take $40\sim50%$ of sugar out of their total average daily calorie. For the purpose of studying diurnal variations in serum glucose, insulin and C-peptide of normal Korean adults based on ordinary Korean diet, we selceted 13 normal Korean male adults and divided them into two groups, Group I (7 persons) and Group II (6 persons). We put Group I on 3,100kcal and 75% sugar diet, and Group II on 2,100 kcal and 69% sugar diet per day for over 4 days. Serum glucose, insulin and C-peptide were checked every 30 minutes or every hour throughout 24 hours. Results are as follows: 1. As for serum glucose level, in the prep ran dial fasting state in the morning, $mean{\pm}S.D.$ of Group I was $91.1{\pm}8.2mg%$, while that of Group II is $82.5{\pm}4.4mg%$. Both groups showed peaks of increased glucose level at postprandial 1 hour after each meal. The peak returned to the level shown during the fasting state at postprandial 1 hour after breakfast while the relatively high glucose levels were maintained respectively even for 2 or 3 hours.after lunch and dinner. 2. As for serum insulin level, Group showed $mean{\pm}S.D.$ of $14.7{\pm}3.0{\mu}U/ml$ while Group II shows that of $7.0{\pm}2.6{\mu}U/ml$ in the fasting state. Group I particularly showed the largest peak from preprandial a half or one and half an hour to postprandial one hour of lunch, and made relatively small peaks $(47.7{\pm}10.8{\mu}U/ml)$ at postorandial 1 hour after breakfast and dinner. No such large peak was marked in Group II, though it showed relatively similar patterns of peak after each meal. 3. As for C-peptide, in the fasting state, Group I and Group II showed $3.50{\pm}1.85$ and $1.66{\pm}0.53ng/ml$ of $mean{\pm}S.D.$, respectively. Group II showed peaks parallel to those for insulin level. None out of seven in Group I showed expected increase in C-peptide based insulin secretion at a half or one and half an hour before lunch. On the contrary, C-peptide increased in 5 subjects out of seven in Group I at 11:00 p.m. when insulin did not increase. 4. According to the integrated concentration method for a measurement of 24-hour total. insulin secretion rate, the $mean{\pm}S.D.$ of Group I was $76.4{\pm}15.2$ U and that of Group II was $58.6{\pm}21.1$ U. The above results confirm that Koreans, when given ordinary diet of 2,100 kcal and 69%, sugar, show insulin secretion pattern essentially similar to that of Westerners. On the contrary, when they are put on a high-calorie diet of 3,100 kcal a day, 75% of which is sugar, insulin secretion can be increased before lunch without increase in blood glucose. These results implies that insulin secretion can be affected by some other factors. The observation that an increase in C-peptide after 11 : 00 p.m. independent of insulin level supports an assertion that insulin secretion and C-peptide secretion can be thought as being physiologically dissociable, and these changes of diurnal patterns in the levels of serum insulin and C-peptide are thought to be resulted from the large meal and high-carbohydrate diet.

• Applied Biological Chemistry
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• v.15 no.1
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• pp.19-25
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• 1972

A kind of peptide which posseses an yeaststatic activity was isolated from Astragalus membranaceus Bunge and following characteristics was obtained. 1. The isoelectric pH of this peptide was 8.2 and histidine, an alkaline amino acid, was identified from this peptide. 2. This substance showes conspicuous heat stability and does not indicate any remarkable reduction of yeaststatic activity even for 5 hours treatment at $100^{\circ}C$. or for 30 minutes at $121^{\circ}C$. 3. The inhibitory activity of the yeast growth is not originated from the yeastcidal action but yeaststatic effect of this sample. 4. The sample shows strong stability ranging from pH 2 to 10. 5. The saccharide; glucose, sucrose, maltose, gives no effect on the yeaststatic activity of the sample even high concentration, 15 percent, and also no effect gives by magnesium, calcium and phosphate salts. 6. The available concentration of this sample on the inhibition of yeast growth was located at the ppm extent, for example, the concentration of fifty percent growth inhibition to Saccharomyces cerevisiae or S. carsbergensis was 4 ppm and 3 ppm to Candida pulcherrima, 13 ppm to S. coreanus, 18 ppm to S. sake and 38 ppm to C. tropicalis. 7. On the alcohol fermentation of S. coreanus, the peptide, an yeast growth inhibitor, gives no effect at all. 8. This substance is named as Astradix-P (Astragalus membranaceus, Radix, Peptide).

• Journal of Sericultural and Entomological Science
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• v.52 no.1
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• pp.45-51
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• 2014

A cecropin-A gene was isolated from the immunized larvae of the Japanese oak silkworm, Antheraea yamamai and designed Ay-CecA. The complete Ay-CecA cDNA consists of 419 nucleotides with 195 bp open reading frame encoding a 64 amino acid precursor that contains a putative 22-residue signal peptide, a 4-residue propetide and a 37-residue mature peptide with a theoretical mass of 4046.81. The deduced amino acid sequence of the peptide evidenced a significant degree of identity (62 ~ 78% identity) with other lepidopteran cecropins. Like many insect cecropin, Ay-CecA also harbored a glycine residue for C-terminal amidation at the C-end, which suggests potential amidation. To understand this peptide better, we successfully expressed bioactive recombinant Ay-CecA in Escherichia coli that are highly sensitive to the mature peptide. For this, we fused mature Ay-CecA gene with insoluble protein ketosteroid isomerase (KSI) gene to avoid the cell death during induction. The fusion KSI-CecA protein was expressed as inclusion body. The expressed fusion protein was purified by Ni-NTA immobilized metal affinity chromatography (IMAC), and cleaved by cyanogen bromide (CNBr) to release recombinant Ay-CecA. The purified recombinant Ay-CecA showed considerably antibacterial activity against Gram-negative bacteria, E. cori ML 35, Klebsiella pneumonia and Pseudomonas aeruginosa. Our results proved that this peptide with a potent antibacterial activity may play a role in the immune response of Japanese oak silkworm.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.1
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• pp.377-381
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• 2012

The aim of this study was to screen for polypeptides binding specifically to LoVo human colorectal cancer cells using a phage-displayed peptide library as a targeting vector for colorectal cancer therapy. Human normal colorectal mucous epithelial cells were applied as absorber cells for subtraction biopanning with a c7c phage display peptide library. Positive phage clones were identified by enzyme-linked immunosorbent assay and immunofluorescence detection; amino acid sequences were deduced by DNA sequencing. After 3 rounds of screening, 5 of 20 phage clones screened positive, showing specific binding to LoVo cells and a conserved RPM motif. Specific peptides against colorectal cancer cells could be obtained from a phage display peptide library and may be used as potential vectors for targeting therapy for colorectal cancer.

• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1588-1590
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• 2012

We compared the mRNA expression profile of the Harmonia axyridis larvae that were either untreated or treated with LPS. The extracted mRNAs were subjected to ACP RT-PCR analysis using a combination of arbitrary primers and oligo (dT) primer. Among the 47 DEGs differentially expressed, we identified a cDNA showing homology with defensin-like antibacterial peptide. The cDNA showed a putative 32-residue signal sequence and a 50-residue mature peptide named harmoniasin. We also investigated the antibacterial activity of the harmoniasin analog, which exhibited potent antibacterial activities against Gramnegative and -positive bacteria strains and it also evidenced no hemolytic activity.

• Korean journal of applied entomology
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• v.35 no.4
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• pp.321-325
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• 1996

To isolate a novel gene for antibacterial peptide, an inducible clone(BmInc8) was selected by differential screening strategy from Bombyx mori cDNA library prepared from lavae injected with Escherichia coli. This clone contained a cDNA insert of 564 nucleotides and encoded 59 amino acids with an apparent molecular mass of 6.3 kDa. The cDNA sequence of BmInc8 had 61.2% identity compared to that of the bactericidin from Manduca sexta and also the deduced amino acids sequences from this insert had 65% identity compared to that of the cecropin D peptide Hyalophora cecropia. The transient expression assay of this insert using prokaryotic expression vector system revealed that the expressed peptide displayed the antibacterial activity. The cDNA sequence was deposited in GenBank under the accession number U30289.