• Archives of Pharmacal Research
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• 제18권5호
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• pp.325-331
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• 1995

The objective of this study was to find a rational dosage form for vaginal mucosal delivery of LHRH. Vaginal absorption of LHRH was estimated by measuring its ovulation inducting effect in rat and in vitro vaginal membrane permeation study in rabbit. THe effects of different hydrogel bases, such as Polycarbophil and Pemulen compared with solutions on vaginal membrane permeation of LHRH were investigated. Sodium laurate, disodium ethylenediamine brane permeaiton of LHRH were investigated. Sodium ethylenediamine tetraacetate (EDTA) and sodium tauro-24, 25-dihydrofusidate (STDHF), which are effective peptidase inhibitors were chosen as additives to a LHRH hydrogel delivery system and LHRH solutions. A Polycarbophil compared with a solution formulation 3.4 times increase in LHRH vaginal membrane permeability compared with a solution formulation. Vaginal membrane permeability from the Polycarbophil was greater than that from Pemulen hydrogels. This may be due to the larger bioadhesive values. LHRH solution with EDTA(2%), STDHF(1%) and sodlaurate(0.5%) showed 4.1 times, 4.8 times and 6.0 times of ovulation inducing activity compared with control. These results suggest that enzyme inhibition effect of EDTA, STDHF and sod, laurate may be result in substantial enhancement of vaginal absorption. By administraiton of Polycarbophil hydrogels containing LHRH the ovulation inducing activity was 3.3 times greater than the solutions. This result indicates the bioadhesive hydrogels as well as peptidase in hibition significantly improved absorption of LHRH. By coadministration with these inhibitors the ovulation inducing activity of Polycarbophi hydrogel containing LHRH was comparable with subcutaneous administration in ovulation inducing activity.

• Bulletin of the Korean Chemical Society
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• 제6권1호
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• pp.23-29
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• 1985

The charge-transfer complexing properties of 1-methyl nicotinamide (MNA), an acceptor, and adenine, a donor, were investigated in water and SDS micellar solutions in relation to the intramolecular interaction in nicotinamide adenine dinucleotide ($NAD^+$). The spectral and thermodynamic parameters of MNA-indole and methyl viologen-adenine complex formations were determined, and the data were utilized to evaluate the charge-transfer abilities of MNA and adenine. The electron affinity of nicotinamide was estimated to be 0.28 eV from charge-transfer energy $of{\sim}300$ nm for MNA-indole. The large enhancement of MNA-indole complexation in SDS solutions by entropy effect was attributed to hydrophobic nature of indole. The complex between adenine and methyl viologen showed an absorption band peaked near 360 nm. The ionization potential of adenine was evaluated to be 8.28 eV from this. The much smaller enhancement of charge-transfer interaction involving adenine than that of indole in SDS solutions was attributed to weaker hydrophobic nature of the donor. The charge-transfer energy of 4.41 eV (280 nm) was estimated for nicotinamide-adenine complex. The spectral behaviors of $NAD^+$ were accounted to the presence of intramolecular interaction in $NAD^+$, which is only slightly enhanced in SDS solutions. The replacement of nicotinamide-adenine interaction in $NAD^+$ by intermolecular nicotinamide-indole interaction in enzyme bound $NAD^+$, and guiding role of adenine moiety in $NAD^+$ were discussed.

• Journal of Applied Biological Chemistry
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• 제57권3호
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• pp.197-203
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• 2014

Various parameters such as solvent selection, concentration, solid/liquid ratio, soaking time, temperature, stationary, shaking conditions, and repeated extractions were investigated in order to determine the optimum extraction conditions of ${\beta}$-glucosidase from bagasse fermented by mixed culture of Aspergillus niger NRC 7A and Aspergillus oryzae NRRL 447. Among various solvents tested, non ionic detergents gave the best results than the inorganic or organic salt solutions and distilled water. The optimum conditions for extraction of ${\beta}$-glucosidase were 30 min soaking time at $40^{\circ}C$ under shaking condition at 150 rpm, with solid/liquid ratio 1:15 (w/v), which yielded $2882.74{\pm}95.52U/g$ fermented culture (g fc) of enzyme activity. With repeated washes under the above optimum conditions, the results showed that enzyme extracted in the $1^{st}$ and $2^{nd}$ washes represents about 90% of the total activity.

• 동아시아식생활학회지
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• 제16권6호
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• pp.702-706
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• 2006

The aim of this study was to identify the onuses of abnormal precipitation in buckwheat extracts and to suggest the preventive solutions. Abnormal precipitation was formed by the coagulations of small round droplets, and increased when poor quality or old buckwheat used. It was found that, unlike poor quality buckwheat, extracts made from fresh buckwheat showed almost no saccharifying enzyme activity and a lower number of microorganisms. The addition of branched starch to the extracts restricted the occurrence of abnormal precipitation and microorganisms and imparted stability to the extracts.

• Fisheries and Aquatic Sciences
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• 제18권2호
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• pp.229-233
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• 2015

We developed a new softening technology applicable to the main body of the jumbo squid Dosidicus gigas; this will aid in squid consumption by elderly individuals and those who have masticatory and dysphagia problems. Protease solutions were injected into jumbo squid and hardness was measured using a texture analyzer. Seven enzymes were tested. Jumbo squid became progressively softer during bromelain and collupulin treatment; the hardness attained $5.6{\times}10^3N/m^2$ at bromelain concentrations of 1.00% (w/v) and $6.7{\times}10^3N/m^2$ at collupullin concentrations of 1.00% (w/v). The extents of tissue softening after bromelain and collupulin injection to 0.1%, 0.25%, 0.50%, and 1.00% (all w/v) were evaluated; the squid retained its shape after steaming for 10 min at $100^{\circ}C$ to inactivate the enzymes. Thus, the results of this research indicate that enzyme injection softens the texture of jumbo squid.

• Geomechanics and Engineering
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• 제20권2호
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• pp.165-174
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• 2020

The enzyme-induced carbonate precipitation (EICP) method has been investigated to improve the hydro-mechanical properties of natural soil deposits. This study was conducted to explore the stiffness evolution during various stress scenarios. First, the optimal concentration of urea, CaCl₂, and urease for the maximum efficiency of calcite precipitation was identified. The results show that the optimal recipe is 0.5 g/L and 0.9 g/L of urease for 0.5 M CaCl₂ and 1 M CaCl₂ solutions with a urea-CaCl₂ molar ratio of 1.5. The shear stiffness of EICP-treated sands remains constant up to debonding stresses, and further loading induces the reduction of S-wave velocity. It was also found that the debonding stress at which stiffness loss occurs depends on the void ratio, not on cementation solution. Repeated loading-unloading deteriorates the bonding quality, thereby reducing the debonding stress. Scanning electron microscopy and X-ray images reveal that higher concentrations of CaCl₂ solution facilitate heterogeneous nucleation to form larger CaCO₃ nodules and 11-12 % of CaCO₃ forms at the interparticle contact as the main contributor to the evolution of shear stiffness.

• 한국미생물·생명공학회지
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• 제9권4호
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• pp.213-218
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• 1981

순수분리정제된 $\beta$-galactosidase의 분자량은 Sephadex G-200 gel filtration법에서 130,000이며 SDS-polyacrylamide gel electrophoresis에서 130,000외에 70,000이 확인되어 이 효소는 70,000인 2개의 subunit로 구성되어 있다고 판단되었다. 효소의 안정pH는 4.5~7.0이며 효소활성의 최적 pH는 4.7 최적온도는 5$0^{\circ}C$였다. 효소활성 및 안정제로 1가, 2가 금속ion을 필요로 하지않았으며 C $u^{++}$ 1mM에서 59%, galactose 100mM에서 48% 저해되었다. 5% lactose용액, 시유 및 10% skim milk 용액에 이 정제된 $\beta$-galactosidase 10units/$m\ell$를 사용하여 5$0^{\circ}C$에서 4시간 동안 반응시켰을 때 lactose가 각각 69.5%. 88.7% 및 72.6%가 glucose와galactose로 전환된 결과를 얻었다.

• Korean Membrane Journal
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• 제8권1호
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• pp.58-66
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• 2006

The ${\beta}-glucosidase$ from olive fruit is of particular interest compared to the ones from other sources because it has shown to have high specifity to convert the oleuropein into dialdehydes, which have antibacterial activity and are of high interest for their application in the food and pharmaceutical fields. The enzyme is not yet commercially available and advanced clean and safe technologies for its purification able to maintain the functional stability are foreseen. The purification of this protein from fruit extracts has been already tempted by electrophoresis but either enzyme deactivation or high background with unclear profiles occurred. In this work, fruit extracts obtained from the ripening stage that showed the highest enzyme activity have been processed by diafiltration and ultrafiltration. Asymmetric membranes made of polyamide or polysulphone having 50 and 30 kDa molecular weight cut-off, respectively, were tested for the diafiltration process. Ultrafiltration membranes made of polyethersulfone with 4 kDa molecular weight cut-off were used to concentrate the dia-filtered permeate solutions. The efficiency of the separation processes was evaluated byenzyme activity tests using the hydrolysis of p-D-nitrophenyl-${\beta}$-D-glucopyranoside (pNPGlc) as reaction model. Qualitative and quantitative electrophoresis were applied to analyze the composition of protein solution before and after the membrane separation; in addition dot blot and western blot analyses were applied to verify the presence of ${\beta}-glucosidase$ in the processed fractions. The overall results showed that the ${\beta}-glucosidase$ functional stability was preserved during the membrane operations and the removal of 20 kDa proteins allowed to increase the specific activity of the enzyme of about 52% compared to the one present in the initial fruit extract.

• Molecules and Cells
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• 제21권2호
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• pp.308-313
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• 2006

Amygdalin is a cyanogenic glycoside compound which is commonly found in the pits of many fruits and raw nuts. Although amygdalin itself is not toxic, it can release cyanide (CN) after hydrolysis when the pits and nuts are crushed, moistened and incubated, possibly within the gastrointestinal tract. CN reversibly inhibits cellular oxidizing enzymes and cyanide poisoning generates a range of clinical symptoms. As some pits and nuts may contain unusually high levels of amygdalin such that there is a sufficient amount to induce critical CN poisoning in humans, the detection of abnormal content of amygdalin in those pits and nuts can be a life-saving measure. Although there are various methods to detect amygdalin in food extracts, an enzyme immunoassay has not been developed for this purpose. In this study we immunized New Zealand White rabbits with an amygdalin-KLH (keyhole limpet hemocyanin) conjugate and succeeded in raising anti-sera reactive to amygdalin, proving that amygdalin can behave as a hapten in rabbits. Using this polyclonal antibody, we developed a competition enzyme immunoassay for determination of amygdalin concentration in aqueous solutions. This technique was able to effectively detect abnormally high amygdalin content in various seeds and nuts. In conclusion, we proved that enzyme immunoassay can be used to determine the amount of amygdalin in food extracts, which will allow automated analysis with high throughput.

• Biomolecules & Therapeutics
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• 제10권2호
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• pp.104-113
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• 2002

The effects of enzyme inhibitors and penetration enhancers on the permeation of leucine enkephalin (Leu-Enk) and its synthetic analog, [${D-ala}^2$]-leucine enkephalinamide (YAGFL) across the nasal, rectal and vaginal mucosae were evaluated. Enzyme inhibitors and penetration enhancers employed for Leu-Enk permeation study were amastatin(AM), thimerosal(TM) and ethylenediaminetetraacetic acid disodium salt(EDTA), and sodium taurodihydrofusidate (STDHF). Those for YAGFL permeation study were TM, benzalkonium chloride(BC) and EDTA, and STDHF, sodium deoxycholate(SDC), sodium glycholate(SGC), glycyrrhizic acid ammonium salt (GAA), L-$\alpha$-Iysophosphatidylcholine(LPC) and mixed micelle (MM, STDHF: linoleic acid = 15 mM : 5 mM). The addition of TM alone on the donor and receptor solutions for Leu-Enk permeation study across all the three kinds of mucosae failed to inhibit the degradation; it completely degraded in 6 hrs, and no permeation occurred. However, with addition of three kinds of inhibitors together, the fluxes across nasal, rectal and vaginal mucosae were $\20.7{pm}2.5$>/TEX>,$\0.3{pm}0.05$>/TEX> and $\1.4{pm}0.5$ $\mu$\mid$textrm{m}$/$\textrm{cm}^2$/hr, respectively. Moreover, the addition of STDHF in the presence of the above three inhibitors enhanced permeation across nasal, rectal and vaginal mucosae 1.3, 15 and 1.3 times, respectively. YhGFL also degraded in the donor and receptor solutions rapidly as time went. With mixed inhibitors of TM and EDTA, the percents of YAGFL remaining in the donor solutions facing nasal, rectal and vaginal mucosae were 69.7, 69.8 and 79.8%, respectively; the percent permeated increased to 10, 2.1 and 5.7%, respectively. The addition of STDHF in the presence of either BC/EDTA or TW/EDTA increased the permeation 2.2, 11.0 and 2.9 times, and 2.21, 14.0 and 2.7 times for nasal, rectal and vaginal mucosae, respectively. With SDC, SGC, GAA, LPC ud MM in the presence of TM/EDTA increased permeation; especially, they increased permeation across vaginal mucosae effectively, and the enhancement factors were 12.5, 7.6, 8.7, 5.7 and 5.5, respectively. The degradation extent of YAGFL was correlated with protein concentrations in the epidermal and serosal extracts. The flux of YAGFL across nasal mucosa increased dose-dependently.