• 한국식품저장유통학회지
• /
• 제2권1호
• /
• pp.173-183
• /
• 1995

This paper was carried out to investigate changes in chromatograms of polysacctatides and soluble pectins on Sephadex G-50 and non-cellulosic neutral sugars of polysaccharides isolated from cell wall of persimmon fruits treated with polygalacturonase and $\beta$-galactosidase in vitro. The chromatogram pattern of soluble pectins extracted from cell wall treated with $\beta$-galactosidase on Sephacryl S-500 column were similar to those of untreatment, but contents of soluble pectins treated with $\beta$-galactosidase were different from those of untreatment. The patterns of chromatograms In soluble pectins extracted from cell wall treated with polygalacturonase were more complex and lower molecular polymer than those of other cell wall-degrading enzyme treatments. Non-cellulosic neutral sugar of polysaccharides in fraction I of soluble material treated with polygalacturonase was rhamnose, those in fraction II were similar to those in fraction III and contents of arabinose, xylose and glucose were higher than contents of other non-cellulosic neutral sugars. Non-cellulosic neutral sugars of polysaccharides in fraction I in soluble material by $\beta$-galactosidase treatment were rhamnose, arabinose, galactose and mannose. Content of glucose of polysaccharides in fraction II was higher than that in fraction I . Non-cellulosic neutral sugars treated with mixed enzyme were rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose. Compositions of non-cellulosic neutral sugars of polysaccharides in fraction I were similar to those in fraction II and III.

• Journal of Microbiology and Biotechnology
• /
• 제33권1호
• /
• pp.28-34
• /
• 2023

Endoribonuclease YbeY is specific to the single-stranded RNA of ribosomal RNAs and small RNAs. This enzyme is essential for the maturation and quality control of ribosomal RNA in a wide range of bacteria and for virulence in some pathogenic bacteria. In this study, we determined the crystal structure of YbeY from Staphylococcus aureus at a resolution of 1.9 Å in the presence of zinc chloride. The structure showed a zinc ion at the active site and two molecules of tricarboxylic acid citrate, which were also derived from the crystallization conditions. Our structure showed the zinc ionbound local environment at the molecular level for the first time. Molecular comparisons were performed between the carboxylic moieties of citrate and the phosphate moiety of the RNA backbone, and a model of YbeY in complex with a single strand of RNA was subsequently constructed. Our findings provide molecular insights into how the YbeY enzyme recognizes singlestranded RNA in bacteria.

• 한국식품영양과학회지
• /
• 제43권3호
• /
• pp.404-410
• /
• 2014

배(Pyrus pyrifolia)의 과피와 과심에 복합효소처리 의해 물리화학적 특성 및 기능성에 미치는 영향을 조사하였다. 배의 과피와 과심에 4종의 복합효소를 처리할 경우 과피의 가용성 고형분 함량이 증가하였고 과심은 유의적으로 증가하지 않았다. 과피의 가용성 고형분 함량은 Pectinex로 처리하였을 때 가장 높은 증대효과를 나타냈다. 또한 배의 과피와 과심에 Pectinex로 처리하였을 때 높은 환원당 추출 수율을 얻을 수 있었다. 총당 함량은 대부분의 효소처리군에서 증가하여 효소에 의해 탄수화물이 분해되어 당 함량이 증가하는 것을 확인할 수 있었다. 배의 과피에 Viscozyme으로 처리할 경우 총 폴리페놀 함량, 총 플라보노이드 함량, DPPH radical 소거능, ABTS radical 소거능 및 SOD 유사활성이 가장 높은 증대효과를 보였다. 또한 과심에 Celluclast로 처리할 경우 총 폴리페놀 함량이 증가하였고, 총 플라보노이드 함량은 Ultraflo 처리군을 제외한 나머지 효소처리군에서는 비슷한 양상으로 증가하였다. 또한 과심에 4종의 복합효소처리 시 DPPH radical 소거능과 ABTS radical 소거능은 유의적으로 증가하지 않았고 SOD 유사활성은 모두 증가하였다. 결과적으로 배의 과피와 과심에 탄수화물 분해 복합효소를 처리함으로써 고형분, 환원당 및 항산화 물질의 추출률을 증가시키고 항산화 활성을 향상시킬 수 있음을 확인하였다.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제3권2호
• /
• pp.103-108
• /
• 1998

A hydrophobic substrate triolein was hydrolyzed by lipase in a mono-phase reaction system containing cyclodextrin(CD) as emulsifier. The triolein was transformation to an emulsion-like state in the CD containing reaction system in contrast to the oil-droplet like state without CD due to the formation of an inclusion complex between the lipids and CDs. The hydyrolysis reaction increased substantially in the CD containing reaction system, and the optimum reaction conditions including the amount of lipase, ${\beta}$-CD concentration, and mixing ratio of triolein and ${\beta}$-CD, were determined. The performance of the enzyme reaction in a mono-phase reaction system was compared with that of a two-phase reaction system which used water immiscible hexane as the organic solvent. The role of a CD in the mono-phase reaction system was elucidated by comparing the degree of the inclusion complex formation with triolein and oleic acid, Km and Vmax values, and product inhibition by oleic aicd in aqueous and CD containing reaction systems. The resulting enhanced reaction seems to be caused by two phenomena; the increased accessibility of lipase to triolein and reduced product inhibition by oleic acid through the formation of an inclusion complex.

• Bulletin of the Korean Chemical Society
• /
• 제25권10호
• /
• pp.1477-1483
• /
• 2004

We present the tandem mass spectrometry applications carried out to elucidate the coordination structure of Zn(II) bound lysine ternary complexes, $(Zn+Lys+Lys-H)^+$, which is a good model system to represent a simple (metallo)enzyme-substrate complex (ES). In particular, experimental efforts were focused on revealing the involvement of a lysine side chain ${\varepsilon}$-amino group in the coordination of $Zn^{2+}$ divalent ions. MS/MS fragmentation pattern showed that all the oxygen species within a complex fell off in the form of $H_2O$ in contrast to those of other ternary complexes containing amino acids with simple side chains (4-coordinate geometries, Figure 1a), suggesting that the lysine complexes have different coordination structures from the others. The participation of a lysine basic side chain in the coordination of Zn(II) was experimentally evidenced in MS/MS for $N{\varepsilon}$-Acetyl-L-Lys Zn(II) complexes with acetyl protection groups as well as in MS/MS for the ternary complexes with one $NH_3$ loss, $(Zn+Lys+Lys-NH_3-H)^+$. Detailed structures were predicted using ab initio calculations on $(Zn+Lys+Lys-H)^+$ isomers with 4-, 5-, and 6-coordinate structures. A zwitterionic 4-coordinate complex (Figure 7d) and a 5-coordinate structure with distorted bipyramidal geometry (Figure 7b) are found to be most plausible in terms of energy stability and compatibility with the experimental observations, respectively.

• 약학회지
• /
• 제53권4호
• /
• pp.222-227
• /
• 2009

Human topoisomerase I (Topo I) plays a pivotal role in cell replication, transcription and repair and, therefore, is an important anti-cancer target. 20S-camptothecin (CPT) is a representative Topo I inhibitor. Compounds belonging to CPT family inhibit the religation step of Topo I-DNA by binding to the DNA cleavage site. Computational docking studies with Surflex-Dock were carried out to investigate the binding modes between Topo I-DNA binary complex structure and the ligand such as 20S-CPT and 9,10-substituted 20S-CPT analogues. The docking results demonstrated that most of the compounds with $IC_{50}$ value under $0.5{\mu}M$ intercalated exactly between the -1 and +1 DNA bases, deeply toward the cleavage site. The complex was stabilized by hydrogen-bonding and hydrophobic interactions with both the enzyme and the DNA. The compounds with $IC_{50}$ value above $0.5{\mu}M$ were poorly docked and did not intercalate. In addition, the docking results confirmed the overall correlation between the $IC_{50}$ values and docking scores, indicating the possible use of the modeling for the prediction of biological activity and design of potential inhibitors.

• 한국수산과학회지
• /
• 제33권1호
• /
• pp.55-59
• /
• 2000

무지개 송어의 뇌하수체 및 배양액에 존재하는 GTH II 농도를 측정하기 위해 Avidin- Biotin complex를 이용한 sandwich EIA 계을 개발했다. Protein A sepharose affinity chromatography을 통해서 얻어진 연어 GTH II의 rabbit IgG에 biotinylation시킨 것 (Biotin-salmon GTH II rabbit IgG)을 제2 항체로 사용하였고, Non-Biotin salmon GTH II rabbit IgG는 단지 protein A sepharose affinity chromatography에서 얻어진 IgG를 제 1 항체로 사용하였다. EIA는 sandwich법에 의해서 이루어졌으며, 효소반응 기질로는 TMB(3,3'5,5-tetramethylbenzidine)를 이용했으며, 반응후 450 nm의 흡광도에서 automatic microplate reader로 측정하였다. 그 결과, $0.12\;{\~}\;125\;ng/ml$의 범위에서 용량반응곡선을 얻었으며, 측정감도 (최소 검출량)는 거의 0.58 ng/ml 정도 였다. 그리고 뇌하수체 추출물 및 배양액 각각의 희석곡선은 GTH II 표존곡선과 일치 하였다. 또한 이러한 GTH II의 표준곡선는 뇌하수체내 다른 peptide hormone와는 교차반응을 거의 나타내지 않았다. Testosterone을 처리한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여 sGnRH에 의한 GTH II 분비량을 본 sandwich EIA계와 RIA계를 비교 조사한 결과, 거의 같은 분비량을 나타냈을 뿐만아니라 같은 분비 pattern을 나타냈다. 이러한 결과로부터 본 sandwich법 EIA계에 의해서 연어과 어류의 뇌하수체 추출물 및 뇌하수체 배양액 중의 GTH II 함량 및 분비량을 측정하는데 있어서 안정된 assay계라고 생각되어진다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권9호
• /
• pp.1339-1347
• /
• 2003

An experiment was conducted to investigate the effects of microbial phytase ($Natuphos^{(R)}$) supplementation in combination with carbohydrases (composed of enzymes targeted to soybean meal (SBM) dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{(R)}$) to corn-soybean meal based diet (CSD) and complex diet (CD) with a partial replacement of SBM with rape seed meal (RSM) and cotton seed meal (CSM) on growth performance and nutrient digestibility of growing pigs. A total of 168 growing pigs averaging $13.18{\pm}1.77kg$ of initial body weight was arranged as a $2{\times}2$ factorial design with main effects of diet types (corn-SBM based diet (CSD) and complex diets (CD; 5% of SBM was replaced with 2.5% of RSM and 2.5% of CSM in diet for phase I (0 to 3 weeks) and 6% of SBM was replaced with 3% of RSM and 3% of CSM in diet for phase II (4 to 7 weeks))) and enzyme supplementation (none and 0.1% of phytase (500 FTU/kg diet) and 0.1% of carbohydrases). The diet with enzyme application were formulated to have a 0.18% unit lower aP than diets without enzyme application. Each treatment had three replicates with 14 pigs per replicate. To determine supplementation effect of phytase and carbohydrases on ileal amino acid digestibility of SBM, RSM and CSM, a total of 18 T-cannulated pigs (initial body weight; $13.52{\pm}1.24kg$) were assigned to six dietary treatments in the present study. Dietary treatments in metabolic trial included 1) SBM diet, 2) SBM diet+with enzymes (phytase (500 FTU/kg) and carbohydrases at 0.1%, respectively), 3) CSM diet, 4) CSM diet+enzymes, 5) RSM diet and 6) RSM diet+enzymes. During whole experimental period (0 to 7 wks), there was no difference in growth performance between diets (CSD and CD). However, dietary phytase and carbohydrases supplementation significantly improved gain/feed ratio (G:F) of growing pigs. During the phase II (4-7 weeks), dietary phytase and carbohydrases supplementation significantly improved all fecal nutrient digestibilities (Dry matter (DM), gross energy (GE), crude protein (CP), crude fat (CF), calcium (Ca) and phosphorus (P)). Dietary phytase and carbohydrases supplementation improved significantly overall ileal amino acid digestibilities of SBM, RSM and CSM based diets (p<0.05). The simultaneous inclusion of phytase and carbohydrases in both of CSD and CD reduced feed cost per kg body weight gain (FCG). Also, results suggest that 2.5 to 3% of RSM and CSM, respectively, might be used as a protein source in growing pig diets without having an adverse effect on the growth performance and nutrient digestibility and simultaneous phytase and carbohydrases addition improves nutritional value of SBM, RSM and CSM by improving ileal amino acid digestibilities.

• 한국수산과학회지
• /
• 제23권2호
• /
• pp.109-124
• /
• 1990

A rapid processing method for fish sauce of high quality stability and favorable flavor was investigated using mackerel waste as starting material. The chopped waste was homogenized with water and hydrolyzed by commercial proteolytic enzymes such as Complex enzyme-2000($2.18\cdot10^4$ U/g solid, Pacific Chem. Co.) and Alcalase ($1.94\cdot10^4$ U/g solid, Novo) in a cylindrical vessel with 4 baffles and 6-bladed turbine impeller. Optimal pH and temperature for the hydrolysis with Complex enzyme-2000 were 8.0 and $50^{\circ}C$, and those with Alcalase were 9.0 and $55^{\circ}C$. In both cases, the reasonabe amount of added water and enzyme concentration based on the waste weight were $40\%,\;3\%$ and hydrolyzing time was 100 min. Thermal treatment of the hydrolysate with $6\%$ of invert sugar for 2 hours at $90^{\circ}C$ was adequated to inactivation of the enzymes and pasteurization of the hydrolysate. Flavor, taste and color of the hydrolysate were improved during the thermal treatment in which the browning reaction products might participate and result in antioxidative and bactericidal effects. Combined use of $0.005\%$ of Caryophylli flos with $6\%$ of invert sugar was also effective for the improvement of taste. Yield of the fish sauce based on the total nitrogen of the raw waste was $93.7\~94.9\%$, and $87.6\~87.9\%$ of the total nitrogen in the fish sauce was in the from of amino nitrogen. The pH, salinity and histamine content of the fish sauce prepared with $15\%$ of table salt were $6.1\~6.2$, $14.0\~14.5\%$ and less than $10mg\%$, respectively. The fish sauce was stable on bacterial growth during the storage of 60 days at $26\pm3^{\circ}C$ and the quality was also maintained.

• 한국수산과학회지
• /
• 제23권2호
• /
• pp.125-136
• /
• 1990

To develope a rapid processing method for fish sauce, processing conditions of fish sauce from sardine waste was investigated. The chopped waste was homogenized and hydrolyzed by commercial proteolytic enzymes such as Complex enzyme-2000($2.18\cdot10^4$ U/g solid) and Alcalase($1.94\cdot10^4$ U/g solid) in a cylindrical vessel with 4 baffles and 6-bladed turbine impeller. Optimal temperature for the case of hydrolysis with Complex enzyme-2000 was 50 and that with Alcalase was $55^{\circ}C$. In both cases, the reasonable pH, amount of water for homo-genization, enzyme concentration and hydrolyzing time were 8.0, $40\%$ (W/W), $3\%$ and 100 min, respectively. Heating of the filtrated hydrolysate for 2 hours at $90^{\circ}C$ with $6\%$ of invert sugar was suitable for pasteurization of the hydrolysate and inactivation of enzymes. Flavor, taste and color of the hydrolysate was improved during the thermal treatment in which the browning reaction products might participate and result in antioxidative and bactericidal effects. Combined use of $0.005\%$ of Caryophylli flos with invert sugar was also effective for the improvement of taste. Yield of the fish sauce based on the total nitrogen in the raw sardine waste was $91.2\~92.3\%$ and $87.2\~87.8\%$ of the total nitrogen in the fish sauce was in the form of amino nitrogen. The pH, salinity and histamine content of the fish sauce prepared with $15\%$ of table salt were $6.1\~6.2$, $14.2\~14.4\%$ and less than $10mg\%$, respectively. The fish sauce was stable during the storage of 60 days at $26\pm3^{\circ}C$ on bacterial growth and its quality was also maintained.