• Journal of the Korean Chemical Society
• /
• v.5 no.1
• /
• pp.33-37
• /
• 1961

We have shown that carboxy-peptidase destroys the biological activity of angiotensin octa-and deca-peptides. Since Proline occurs as the seventh amino acid from the amino end of the chain and since carboxypeptidase does not cleave proline from a peptid chain, it is evident that the heptapeptid H.asp-arg-val-tyr-ileu-his-pro.OH is formed by this hydrolysis. This peptide must then be biologically inactive. In order to determine whether the phenyl group of the C-terminal amino acid was the necessary requirement for biological activity of the octapeptide, $ala^8$ angiotensin octapeptide(amino acids of peptides numbered from amino end) was synthesized. For this synthesis the four dipeptides were prepared: carbobenzoxy-L-prolyl-L-alanine-P-nitrobenzyl-ester, m.p. $134-135^{\circ}C,$ carbobenzoxy-L-isoleucyl-imidazole benzyl-L-histidine methyl ester, m.p. $114-116^{\circ}C,$ carbobenzoxy-L-valyl-L-tyrosine hydrazide and carbobenzoxy B-benzyl-L-aspartyl-nitro-L-arginine. The first three dipeptides were obtained as crystalline compounds. Imidazole-benzyl-L-histidine was used in the hope that it would block the histidine imidazole against side reactions in steps subsequent to the formation of the C-terminal tetrapeptide. Also, it was through that the imidazole benzylated peptides would be easier to crystallize. This, however, was not the case. The tetrapeptide, carbobenzoxy-L-isoleucyl-L-im, benzyl-histidyl, L-prolyl-L-alanine-nitrobenzyl ester was not obtained in a crystalline form. Neither could the mono-or dihydrobromide of the tetrapeptide free base be induced to crystallize. Carbobenzoxy-L-valyl-L-tyrosine azide was condensed with the tetrapeptide free base to yield the protected hexapeptide; carbobenzoxy-L-valyl-L-tyrosyl-L-isoleucyl-L-im, benzyl, histidyl-L-Prolyl-L-alanine-nitrobenzyl ester. Upon removal of the carbobenzoxy group with hydrogen bromide in acetic acid an amorphous free base hexapeptide ester was obtained. This compound gave the correct C, H, N analysis and contained the six amino acids in the correct ratio. The octapeptide was obtained by condensing this hexapeptide with carbobenzoxy-B-benzyl-L-aspartyl-nitro, L-arginine using the mixed anhydride method of condensation. This amorphous product was proven to be homogenous by chromatography in two solvent systems and upon hydrolysis yielded the eight amino acids in correct ratio. The five protecting groups were removed from the octapeptide by hydrogenolysis over palladium black catalyst. Biological assay of the free peptide indicated that it possessed less than 0.1 per cent of both pressor and oxytocic activity of the phenylalanine8 angiotensin. This suggests that the phenyl group is a point of attachment between angiotensin and its biological receptor site.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.705-708
• /
• 2001

Enzymatic synthesis and purification of sugar esters using supercritical $CO_2$ were investigated. The observed yield of suagr ester produced by transesterification of methyl glucoside and oleic acid using a lipase(Novozym 435) was 67% at 24 hours of reaction in the supercritical $CO_2$. The solubility of the fatty acids in the supercritical $CO_2$ was measured to find the conditions of supercritical separation of the fatty acids from the reaction mixture. The solubility of capric and oleic acid was 5 ${\times}$ $10^{-4}$, 1.7 ${\times}$ $10^{-4}$ mol/mol $CO_2$, respectively, varying at $35{\sim}50^{\circ}C$ and 80${\sim}$120 atm.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.9
• /
• pp.1229-1243
• /
• 2013

In this paper, an air isolate (NITT6L) has been screened based on hemolytic activity, emulsification activity, drop collapsing test, and oil displacement test, as well as lipase activity. It was found that strain NITT6L was able to reduce the surface tension of the medium from 61.5 to 39.83 mN/m and could form stable emulsions with tested vegetable oils. Morphological, biochemical, 16S rRNA sequencing analyses, and fatty acid methyl ester analysis using gas chromatography confirmed that the air isolate under study was Pseudomonas aeruginosa. Characterization of the biosurfactant using agar double diffusion assay revealed that the biosurfactant was anionic in nature, and CTAB-methylene blue assay and Molisch test revealed its glycolipid nature. The FT-IR spectrum confirmed that the crude biosurfactant was a rhamnolipid. Using unoptimized medium containing sucrose as the carbon source, the isolate was found to produce 0.3 mg/ml of rhamnolipid in batch cultivation (shake flask) at $37^{\circ}C$ and pH 7. Optimization of the medium components was carried out using design of experiments and the yield of rhamnolipid has been enhanced to 4.6 mg/ml in 72 h of fermentation.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.41 no.6
• /
• pp.409-413
• /
• 2008

A simple method for the purification of porphyran from laver Porphyra yezoensis was developed to obtain information for the development of food materials with biological functionality. Crude porphyran (CP) was extracted from dried laver in boiling water for 3 h, and then fractionated using cetylpyridinium chloride into an acidic fraction (CP-F1) and a neutral fraction (CP-F2). CP-F1 was fractionated further by fractional ethanol precipitation. Fraction CP-F1-70, precipitated at an ethanol concentration of 61-70% was the major fraction containing 68.1% of the yield from the initial fraction CP-F1. The CP-F1-70 fraction displayed a single band on Sepharose CL-4B with a molecular mass of 550 kDa, indicating a homogeneous polysaccharide. The molar ratio of galactose, 3,6-anhydro-L-galactose, 6-0-methyl-D-galactose and ester sulfate of CP-F1-70 was 1:0.32:0.07:0.53. This method is very useful for rapid and large-scale preparation of purified porphyran because it is compatible with mass production.

• Archives of Pharmacal Research
• /
• v.29 no.6
• /
• pp.459-463
• /
• 2006

Previous studies on the anticonvulsant activity of $N-Cbz-{\alpha}-aminoglutarmides$ have shown that the derivatives of $N-Cbz-{\alpha}-amino-N-alkoxy$ glutarimide have significant anticonvulsant activity. In addition, their anticonvulsant activities are dependent on the presence of N-alkoxy groups. Based on these results, a series of $N-Cbz-{\alpha}-amino-glutarimidooxy$ carboxylates derivatives (3a-e) were synthesized in moderate yield using a known synthetic procedure. Their anticonvulsant activities were evaluated using the maximal electroshock seizure (MES) test, the pentylene tetrazole induced seizure (PTZ) test, and the strychinine (Str) threshold test with the ultimate aim of developing more active anticonvulsants. None of the compounds (3a-e) tested showed anticonvulsant activity in the MES and PTZ test. However, all the compounds tested exhibited significant anticonvulsant activity in the Str. test. The most active compound in the Str. test was the methyl ester of $N-Cbz-{\alpha}-amino-glutarimidooxy$ acetic acid 3a $(ED_{50}\;=\;42.9\;mg/kg)$.

• The Journal of Internal Korean Medicine
• /
• v.26 no.3
• /
• pp.605-614
• /
• 2005

Objectives : Ojawhan was formulated to contain various natural products known to cure erectile dysfunction. This study was aimed to investigate the effects of Ojawhan on the nitric oxide synthase(NOS) activity, nitrite level, antioxidation and erectile responses in rat's corpus cavernosum penis. Methods : Ojawhan was washed, dried in the shade and crushed. The crushed Ojawhan ,was extracted 3 times, each time with 3 volumes of methyl alcohol at $60^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 62g. Ojawhan extract oral-administered 100 mg per 1 kg of body weight for 30 days. First, samples were treated with Ojawhan, then ethanol-treated rats and L-N-Nitroarginine methyl ester(L-NAME) treated rats were put with the samples. Result : The level of urethral lipid peroxide in the ethanol-Ojawhan double administered rats was decreased as low as in the normal group, while the one in the ethanol-treated group was increased. The urethral NOS activity, the level of urethral nitrite, the level of testosterone and the erectile response to cavernous nerve stimulation in the ethanol-Ojawhan double administered rats were increased as high as in the normal group while the one in the ethanol-treated group was decreased. The electile response to cavernous nerve stimulation and the level of nitrite in L-NAME ($10^{-4}$)-treated rats was restored by the administration of Ojawhan as high as in the normal group. Conclusions : Ojawhan was effective in restoring the ethanol-induced or L-NAME-induced erectile dysfunction in rats.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.1
• /
• pp.176-182
• /
• 2008

Scolopendra was known to cure erectile dysfunction. This study was aimed to investigate the effects of Scolopendra on the nitric oxide synthase activity, nitrite level, cyclic-GMP and erectile responses in rat's corpus cavernosum penis. The crushed Scolopendra was extracted 3 times, each time with 3 volumes of methyl alcohol at $60^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 14.2 g. Scolopendra extract was oral-administered 50 mg per 1 kg of body weight for 30 days. First, samples were treated with Scolopendra, and then ethanol-treated rats and L-N-Nitroarginine methyl ester (L-NAME) treated rats were fed with the samples. The level of urethral nitrite and nitric oxide synthase activity in the ethanol-Scolopendra double administered rats were as high as in the normal group, while the one in the ethanol-treated group was decreased. The level of urethral cyclic-GMP and guanylate cyclase actiyity in the ethanol-Scolopendra double administered rats were as high as in the normal group, while the one in the ethanol-treated group was decreased. The level of urethral phosphodiesterase activity in the ethanol-Scolopendra double administered rats was as low as in the normal group, while the one in the ethanol-treated group was increased. The erectile response to a cavernous nerve stimulation in L-NAME $(10^{-4})-treated$ rats was restored by the Scolopendra to the similar level seen in the normal group. The electile response to cavernous nerve stimulation in the ethanol-Scolopendra double administered rats were increased as high as in the normal group while the one in the ethanol-treated group was decreased. Scolopendra was effective in restoring the ethanol-induced or L-NAME-induced erectile dysfunction in rats.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.25 no.2
• /
• pp.234-241
• /
• 2011

Mantidis Vagina Ovorum was formulated to contain various natural products known to cure erectile dysfunction. This study was aimed to investigate the effects of Mantidis Vagina Ovorum on the nitric oxide synthase (NOS) activity, nitrite level, antioxidation and erectile responses in rat's corpus cavernosum penis. The crushed Mantidis Vagina Ovorum was extracted 3 times, each time with 3 volumes of methyl alcohol at $60^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 16.6 g. Mantidis Vagina Ovorum extract oral-administered 75 mg per 1 kg of body weight for 30 days. First, samples were treated with Mantidis Vagina Ovorum, and then cimetidine-treated rats and L-N-Nitroarginine methyl ester (L-NAME) treated rats were put with the samples. The level of urethral lipid peroxide in the cimetidine-Mantidis Vagina Ovorum double administered rats was decreased as low as in the normal group, while the one in the cimetidine-treated group was increased. The urethral NOS activity, the level of urethral nitrite, the level of testosterone and the electile response to cavernous nerve stimulation in the cimetidine-Mantidis Vagina Ovorum double administered rats were increased as high as in the normal group while the one in the cimetidine-treated group was decreased. The electile response to cavernous nerve stimulation and the level of nitrite in L-NAME ($10^{-4}$)-treated rats was restored by the administration of Mantidis Vagina Ovorum as high as in the normal group. Mantidis Vagina Ovorum was effective in restoring the cimetidine-induced or L-NAME-induced erectile dysfunction in rats.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.36 no.2
• /
• pp.149-154
• /
• 2007

An antibacterial substance to the Streptococcus mutans, a causative bacterium for decayed teeth, was isolated from the dried sea mustard, Undaria pinnatifida, and identified by GC and GC/MS. Acetone extract from the sea mustard (10.4 kg), was evaporated and partitioned to 4 fractions such as hexane, chloroform, butanol and water. The most active chloroform fraction were further purified through basic alumina, silicic acid and ODS column, successively, and finally, 3 antibacterial substances were isolated on the HPLC attached ODS column by using 95% MeOH and guided with UV detector (254 nm). Antibacterial substances (total 160mg, yield $1.5\times10^{-3}$%) had the same Rf value (0.42) on the TLC developed hexane diethyl ether acetic acid (80:30:1) and those methyl esters moved to 0.95. They were identified as the same unsaturated fatty acid, $C_{18:4,\;n-3}$ (3,6,9,12-octadecatetraenoic acid, stearidonic acid) compared relative retention times (15.5 min) with authentic fatty acid on the GC chromatogram. It was further confirmed unambiguously on the GC/MS giving molecular ion peak at m/z 290 which coincided with its methyl ester.

• Korean Chemical Engineering Research
• /
• v.53 no.2
• /
• pp.205-210
• /
• 2015

In this paper, microalgae lipid extractions were performed using conventional organic solvent and supercritical carbon dioxide (SC-$CO_2$) for biodiesel-convertible lipid fractions. The highest levels (58.31%) of fatty acid methyl ester (FAME) content in the lipid extracted by SC-$CO_2$ was obtained, and 18.0 wt.% crude lipid yield was achieved for Bligh-Dyer method. In the SC-$CO_2$ extraction, methanol as a co-solvent was applied to increase the polarity of extract. The experimental results indicated that crude lipid yield, FAME content and yield extracted by combination of SC-$CO_2$ with methanol were 12.5 wt.%, 56.32% and 7.04 wt.%, respectively, and this method could reduce the extraction time from 2 hour to 30 min when compared to SC-$CO_2$ extraction. Therefore, SC-$CO_2$ extraction is proven to be an environmentally-friendly and an effective method for lipid extraction from microalgae.