• Title/Summary/Keyword: Protein engineering

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Characteristics of Hydrolytic Enzymes that Produced by Bacillus subtilis CK-2 Isolated from Doenjang (된장으로부터 분리한 Bacillus subtilis CK-2가 생산하는 가수분해효소의 활성 특성)

  • Lee, Sang-Hyup;Kim, Chul-Ho
    • Journal of Life Science
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    • v.27 no.7
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    • pp.805-811
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    • 2017
  • In the previous paper, we isolated a bacterium that can hydrolyze various organic materials from soybean paste, including cellulose, lipids, starch, and protein. The activity and chemical properties of the crude enzymes produced by the isolate Bacillus subtilis CK-2 were further investigated. Cellulase showed the highest activity at pH 5.0 and $55^{\circ}C$. The stability of cellulase was maintained within the ranges of pH 5.0~10.0 and $20{\sim}50^{\circ}C$. Cellulolytic enzymes were activated by a $Co^{2+}$ ion, demonstrating the highest activity at a 0.45%(w/v) concentration of $Co^{2+}$. The optimal conditions for amylase were pH 5.0 and $50^{\circ}C$. The activity of amylase was stable within the ranges of pH 4.0~5.0 and $20{\sim}50^{\circ}C$. The $Co^{2+}$ ion was also necessary for amylase activity, which was the highest at a 0.2%(w/v) concentration of $Co^{2+}$. The optimal pH and temperature conditions of protease were pH 8.0 and $50^{\circ}C$. The activity of protease was stable within the ranges of pH 7.0~8.5 and $20{\sim}50^{\circ}C$. Protease activity was catalyzed by $Mn^{2+}$, which was the highest at a 0.125%(w/v) concentration of $Mn^{2+}$. The isolate B. subtilis CK-2 demonstrated a high activity of autolysin. Based on these results, we identified and suggested the optimal pH, temperature, and metal ion concentration in the use of the hydrolytic enzymes of B. subtilis CK-2 for industrial purposes.

A Study on the Activity of Anti-Aging by Second Fermented Snail Extract with Hericium erinaceum Mycelium (노루궁뎅이버섯 균사체를 이용한 2 차 발효달팽이 추출물의 항노화 활성에 관한 연구)

  • Zhoh, Choon-Koo;Lee, Min-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.1
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    • pp.143-154
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    • 2016
  • This study is related to develop a snail extract through a snail secondary fermentation process, getting anti-aging activity with healthy and beauty skin care scientific applications. In order to obtain a primary fermentation was incubated with Hericium erinaceus mycelium. Through the secondary fermentation process using Leuconostoc mesenteroides, was deeply described a total process of obtaining second fermented extract using snail body. Mycelium is applied in this study was extracted using Hericium erinaceus mycelium and Leuconostoc mesenteroides. The final yield of the extract was 62 wt%. Experimental results of secondary fermentation snail extract were contained with 32 wt% water, 31.5 wt% total amino acid protein, 15.7 wt% polysaccharide, 12.3 wt% fatty acid and others 8.5 wt%. In addition, in order to study about skin beauty care and anti-aging activity, we evaluated antioxidant activity with DPPH, elastin enzyme (elastase) inhibitory activity, tyrosinase inhibition rate, collagen synthetic function, fibroblast synthetic activity. First; anti-oxidative activity of secondary fermentation snail extract (IC50%) was spent with 7.27 mg/mL, control samples were spent with green tea extract was 11.8 mg/mL, common snails extract was 15.7 mg/mL, DL-a-tocopherol was 9.25 mg/mL respectively. Second; elastin enzyme inhibitory activity of secondary fermentation snail extract (IC50%) was spent with 32.5 mg/mL, control samples were also spent with green tea extract was 45.9 mg/mL, general snail extract was 67.7 mg/mL. Third; tyrosinase inhibitory activity of secondary fermentation snail extract (IC50%) was spent with 140.3 mg/mL, control samples were also spent with green tea extract was 250.7 mg/mL, general snails extract was 389.5 mg/mL, niacineamide was 125.9 mg/mL. Forth; fibroblast synthetic activity of secondary fermentation snail extract was increased with 125.6%, control samples were also spent with green tea extract was 98.9%, general snails extract was 109.5%, niacineamide was 125.9 mg/mL, DL-a-tocopherol was 96.2%. Fifth; collagen synthetic activity of secondary fermentation snail extract was increased with 118%, control samples were also spent with green tea extract was 87.3%, general snails extract was 93.2%, adenosine was 127.9%. In conclusion, on the basis of this study, in the future it is expected to be applied to the skin beauty care application and development of Korean style cosmetic products.

Changes in Pood Components of Top Shell, Omphalius pfeifferi capenteri by Thermal Processing at High Temperature (고온가열처리에 의한 바다방석고둥 (Omphalius pfeifferi capenteri)의 식품성분 변화)

  • Ha Jin Hwan;Song Dae Jin;Kim Poong Ho;Heu Min Soo;Cho Moon Lae;Sim Hyo Do;Kim Hey Suk;Kim Jin Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.166-172
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    • 2002
  • The top shell, Omphalius pfeifferi capenteri meat vacuum-packed in can (diameter$\times$height, 74.1mm$\times$50.7mm) were heated at 115$^{\circ}C$ up to $F_0$ values of 5 min, 10 min, 15 min and 20 min, and the changes in food components were studied. After 14 days storage at 37$^{\circ}C$ and 55$^{\circ}C$, no growth of microorganism and panelling were recognized from the canned meats which were sterlized at 115$^{\circ}C$ with $F_0$ value of S min and over. In the case of proximate composition of the canned meats, the moisture content decreased with the increase of $F_0$ value, while crude protein increased. The increase of volatile basic nitrogen content, pH and degree of browning and the decrease of mineral, total amino acid, free amino acid, trimethylamine oxide, total creatinine contents and yields were observed during thermal processing, In sensory evaluation on color, texture and taste in the canned meats, no significant difference was observed among a boiled sample and the canned meats heated at re value of 10 min and below. But, in the canned meats heated at $F_0$ value of over 15 min, its sensory scores decreased with the increase of $F_0$ value. From these results, the reasonable $F_0$ value for preparation of the heat-treated top shell meats was in the range of 5$\~$10 min.

Manufacturing and Characteristics of Fruit Wine from Acanthapanax sessiliflorus (오가피 열매 발효주의 제조 및 특성)

  • Choi, Jae-Myoung;Kim, Kwang-Yup;Lee, Sang-Hwa;Ahn, Jun-Bae
    • Food Engineering Progress
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    • v.14 no.1
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    • pp.1-6
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    • 2010
  • Fruit wine from Acanthopanax sessiliflorus(A. sessiliflorus) including many pharmacological components was manufactured and its functional properties were investigated. The fruit part of A. sessiliflorus contained 75.74${\pm}$0.49%(w/w) moisture, 12.51${\pm}$1.23%(w/w) crude protein, 4.20${\pm}$0.51.%(w/w) crude fat and 5.21${\pm}$1.64%(w/w) crude ash. Minerals of fruit were potassium(12.94${\pm}$0.08 mg/g), calcium(1.53${\pm}$0.06 mg/g) and magnesium(1.12${\pm}$0.05 mg/g). Initial soluble solid and fermentation temperature were 24-30$^{\circ}$Brix and 20${^{\circ}C}$ for manufacturing fruit wine from A. sessiliflorus. When initial soluble solid of a must was adjusted to more than 30$^{\circ}$Brix, ethanol production was suppressed slightly. The polyphenol content of the fruit wine fermented at 20${^{\circ}C}$(125.24${\pm}$1.86 mg/mL) was higher than those at 25${^{\circ}C}$(99.69${\pm}$2.11 mg/mL) and 30${^{\circ}C}$(95.55${\pm}$1.54 mg/mL). Electron donating activities of wines fermented at 20, 25 and 30${^{\circ}C}$ were 85.9${\pm}$2.3, 55.7${\pm}$2.5 and 55.2${\pm}$3.4%, respectively. The content of eleutheroside B increased up to 146.58${\pm}$4.10 $\mu$g/mL during fermentation. There was no significant effect of fermentation temperature on eleutheroside B content. The fruit part of A. sessiliflorus can be used as a valuable resource for development of nutraceutical foods.

Chemical Components and Antimicrobial Activity of Garlics from Different Cultivated Area (산지별 마늘의 화학성분 및 항균활성)

  • Jeong, Chang-Ho;Bae, Young-Il;Lee, Jin-Hwa;Roh, Jeang-Gwan;Shin, Chang-Sik;Choi, Jine-Shang;Shim, Ki-Hwan
    • Journal of agriculture & life science
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    • v.43 no.1
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    • pp.51-59
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    • 2009
  • The chemical components and antimicrobial activities of garlic from different area were investigated and analyzed to provide basic data for functional food materialization and processing. Hunter's values of garlic from different area were L 53.41~57.15, a -3.49~-4.38 and b 11.47~17.55. The moisture, crude protein, crude fat, nitrogen free extract, crude fiber and ash were 65.24~71.96, 6.24~9.35, 0.21~0.49, 19.01~22.72, 0.58~0.95 and 1.01~2.01%, respectively. The major minerals of garlic from different area were Na(27.22~112.03), Mg(18.17~32.56), K(242.16~569.28), Ca(28.60~63.93), P(117.72~265.21 mg%) and major free sugars were sucrose, glucose and fructose. The major amino acids of garlic from different area were proline, arglmne, glutamic acid and aspartic acid and content of total amino acid was 2,709.33~4,561.04 mg%. The ascorbic acid content of garlic from different area was 2.966~8.673 mg%. Composition of fatty acids of garlic from different area were linoleic acid, oleic acid and palmitic acid, unsaturated fatty acid and saturated fatty acid contents were 72.18~74.35 and 25.65~27.82%, respectively. Antimicrobial activities of garlic extracts as different area increased depends on concentration and showed the high antimicrobial activities against Gram(+) and Gram(-).

Development of Crushing Device for Whole Crop Silage and the Characteristics of Crushed Whole Crop Silage (총체맥류 분쇄기 개발 및 분쇄 총체맥류 사일리지의 품질 특성)

  • Lee, Sunghyoun;Yu, Byeongkee;Ju, Sunyi;Park, Taeil
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.4
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    • pp.344-349
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    • 2016
  • This study was conducted to evaluate the possibility of expanding the usage of whole crop silage from beef cattle and dairy cow to hogs and chickens. For this purpose, a crushing device was developed to crush whole crop silage. The crushed silage was sealed, and analyzed for its feed value. The silage varieties used for the experiment included Saessal barley and Geumgang wheat. Whole crop barley and wheat were crushed in the crushing system as a whole without separating stems, leaves, grains, etc.. When the crushed whole crop silages (CWCS) were analyzed, full grain, grains above 10 mm in size, grains 5~10 mm in size, and grains below 5 mm in size accounted for, 20%, 4%, 27%, and 49 %, respectively. In order to facilitate the fermentation of CWCS, inoculated some fermenter into each CWCS sample (barley or wheat). As control, another set of sample was not inoculated. Crude protein (CP), ether extract (EE), crude fiber (CF), neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, cellulose content, total digestible nutrient (TDN), and relative feed value (RFV) of fermenter-inoculated Saessal barley were 2.45 %, 1.61%, 8.95%, 16.94%, 9.52%, 1.01%, 8.51%, 81.38%, and 447.5%, respectively. The CP, EE, CF, NDF, ADF, lignin, cellulose content, TDN, and RFV in the other sample of Saessal barley without inoculation of fermenter were 2.57%, 1.62%, 9.61%, 18.25%, 10.13%, 1.10%, 9.04%, 80.90%, and 412.9%, respectively. The CP, EE, CF, NDF, ADF, lignin, cellulose content, TDN, and RFV of fermenter-inoculated Geumgang wheat sample were 2.43%, 1.27%, 10.99%, 19.49%, 11.23%, 1.46%, 9.77%, 80.03%, and 382.6%, respectively. The CP, EE, CF, NDF, ADF, lignin, cellulose content, TDN, RFV of the other set sample of Geumgang wheat sample without the inoculation of fermenter were 2.28%, 1.44%, 10.08%, 18.02%, 10.44%, 1.26%, 9.18%, 80.65%, and 416.9%, respectively. The TDN and RFV content in the fermenter-inoculated Saessal barley were 81.38% and 447.5%, respectively, while the one in the fermenter-inoculated Geumgang wheat were 80.03% and 382.6% respectively. When the feed value of whole crop barley and wheat silage without crushing process was compared to the feed value of whole crop barley and wheat silage made from crushing system, the latter appeared to be higher than the former. This could be due to the process of sealing the crushed silage which might have minimized air content between samples and shortened the golden period of fermentation. In conclusion, these results indicate that a crushing process might be needed to facilitate fermentation and improve the quality of silage when making whole crop silage.

Cellular Energy Allocation of a Marine Polychaete Species (Perinereis aibuhitensis) Exposed to Dissolving Carbon Dioxide in Seawater (해수 중 용존 이산화탄소 농도 증가가 두토막눈썹참갯지렁이(Perinereis aibuhitensis)의 세포내 에너지 할당에 미치는 영향)

  • Moon, Seong-Dae;Lee, Ji-Hye;Sung, Chan-Gyoung;Choi, Tae Seob;Lee, Kyu-Tae;Lee, Jung-Suk;Kang, Seong-Gil
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.16 no.1
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    • pp.9-16
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    • 2013
  • An experiment was conducted to evaluate the biochemical adverse effect of increased carbon dioxide in seawater on marine polychaete, Perinereis aibuhitensis. We measured the available energy reserves, Ea (total carbohydrate, protein, and lipid content) and the energy consumption, Ec (electron transport activity) of Perinereis aibuhitensis exposed for 7-d to a range of $CO_2$ concentration such as 0.39 (control =390 ppmv), 3.03 (=3,030 ppmv), 10.3 (=10,300 ppmv), and 30.1 (=30,100 ppmv) $CO_2$ mM, respectively. The cellular energy allocation (CEA) methodology was used to assess the adverse effects of toxic stress on the energy budget of the test organisms. The results of a decrease in CEA effect of increased carbon dioxide in seawater from all individual in Ea and Ec. Increase of carbon dioxide reduced pH in seawater, significantly. The chemical changes in sea- water caused by increasing $pCO_2$ might cause stresses to test organisms and changes in the cellular energy allocations. Results of this study can be used to understand the possible influence of $CO_2$ concentration increased by the leakage from sub-sea bed storage sites as well as fossil fuel combustion on marine organisms.

Characterization of a Korean Domestic Cyanobacterium Limnothrix sp. KNUA012 for Biofuel Feedstock (토착 남세균 림노트릭스 속 KNUA012 균주의 바이오연료 원료로서의 특성 연구)

  • Hong, Ji Won;Jo, Seung-Woo;Kim, Oh Hong;Jeong, Mi Rang;Kim, Hyeon;Park, Kyung Mok;Lee, Kyoung In;Yoon, Ho-Sung
    • Journal of Life Science
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    • v.26 no.4
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    • pp.460-467
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    • 2016
  • A filamentous cyanobacterium, Limnothrix sp. KNUA012, was axenically isolated from a freshwater bloom sample in Lake Hapcheon, Hapcheon-gun, Gyeongsangnam-do, Korea. Its morphological and molecular characteristics led to identification of the isolate as a member of the genus Limnothrix. Maximal growth was attained when the culture was incubated at 25℃. Analysis of its lipid composition revealed that strain KNUA012 could autotrophically synthesize alkanes, such as pentadecane (C15H32) and heptadecane (C17H36), which can be directly used as fuel without requiring a transesterification step. Two genes involved in alkane biosynthesis-an acyl-acyl carrier protein reductase and an aldehyde decarbonylase-were present in this cyanobacterium. Some common algal biodiesel constituents-myristoleic acid (C14:1), palmitic acid (C16:0), and palmitoleic acid (C16:1)-were produced by strain KNUA012 as its major fatty acids. A proximate analysis showed that the volatile matter content was 86.0% and an ultimate analysis indicated that the higher heating value was 19.8 MJ kg−1. The isolate also autotrophically produced 21.4 mg g−1 phycocyanin-a high-value antioxidant compound. Therefore, Limnothrix sp. KNUA012 appears to show promise for application in cost-effective production of microalga-based biofuels and biomass feedstock over crop plants.

Investigating the Partial Substitution of Chicken Feather for Wood Fiber in the Production of Wood-based Fiberboard (목질 섬유판 제조에 있어 도계부산물인 닭털의 목섬유 부분적 대체화 탐색)

  • Yang, In;Park, Dae-Hak;Choi, Won-Sil;Oh, Sei Chang;Ahn, Dong-uk;Han, Gyu-Seong;Oh, Seung Won
    • Korean Chemical Engineering Research
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    • v.56 no.4
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    • pp.577-584
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    • 2018
  • This study was conducted to investigate the potential of chicken feather (CF), which is a by-product in poultry industry, as a partial substitute of wood fiber in the production of wood-based fiberboard. Keratin-type protein constituted the majority of CF, and its appearance did not differ from that of wood fiber. When the formaldehyde (HCHO) adsorptivities of CF compared by its pretreatment type, feather meal (FM), which was pretreated CF with high temperature and pressure and then grounded, showed the highest HCHO adsorptivity. In addition, there was no difference between the adsorbed HCHO amounts, which was measured by dinitrophenylhydrazine method, of scissors-chopped CF and CF beated with an electrical blender. Mechanical properties and HCHO emission of medium-density fiberboards (MDF), which were fabricated with wood fiber and 5 wt% CF, beated CF or FM based on the oven-dried weight of wood fiber, were not influenced by the pretreatment type of CF. However, when the values compared with those of MDF made with just wood fiber, thickness swelling and HCHO emission of the MDF were improved greatly with the addition of CF, beated CF or FM. Based on the results, it might be possible to produce MDF with improved dimensional stability and low HCHO emission if CF, beated CF or FM is added partially as a substitute of wood fiber in the manufacturing process of MDF produced with the conventional urea-formaldehyde resin of $E_1$ grade. However, the use of CF or FM in the production of MDF has a low economic feasibility at the current situation due to the securing difficulty and high cost of CF. In order to enhance the economic feasibility, it requires to use CF produced at small to medium-sized chicken meat plants. More importantly, it is considered that the technology developed from this research has a great potential to make provision for the prohibition of animal-based feed and to dispose environmentally avian influenza-infected poultry.

Seasonal Dust Concentration and Characteristics of Windowless Broiler Building (무창 육계사의 계절별 먼지 농도와 특성 연구)

  • Choi H. C.;Yeon G. Y.;Song J. I.;Kang H. S.;Kwon D. J.;Yoo Y. H.;Barroga A. J.;Yang C. B.;Chun S. S.;Kim Y. K.
    • Journal of Animal Environmental Science
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    • v.11 no.3
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    • pp.197-206
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    • 2005
  • This study was carried out to investigate the concentration and characteristics of dust originating from windowless broiler building in each season. 12.0m width and 46m tenth with side wall height of 3.0m was investigated and capacity was 12,800 birds at a stock density of 23.2 birds per square meter. Dust concentrations in terms of total suspended particles (TSP), and particulate matter of sizes $10{\mu}m(PM10),\;2.5{\mu}m (PM2.5),\;and\;1{\mu}m(PM1)$ were measured at 30-minute intervals. On the basis of broiler age, the average dust concentration in summer in TSP as follows: 1,229 904.5 558.8 and $1,053{\mu}g/m^3$ on the broilers' first to fourth week of age, respectively. But during winter, the average dust concentration showed an increasing pattern, as follows: 465.4, 1,401, 4,497, 5,097 and $6,873{\mu}g/m^3$ on the broilers' first to fifth week of age, respectively. The maximum dust concentration of $11,132{\mu}g/m^3$ was observed on the fifth week. On a daily basis, the maximum dust concentration during summer was detected in early morning, and the minimum in the afternoon. The aerial dust particle size of $0.05\~0.35{\mu}m$ was the highest in number. But on volume basis, particle size of 16~99 un had the largest percentage in the broiler house. Crude protein of the dust $(42.8\~65.2\%)$, on dry matter basis, was higher than that $(20.5\~24.5\%)$ fed to the broilers. Heavy metal concentration of the dust also had high levels compared with that of the feed.

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