Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.

본 연구는 곡류별 버섯 균사체의 성장률과 효소활성을 측정하였다. 현맥 배양물이 다른 곡류와 비교했을때 균사체 성장 속도가 빨랐다. 효소 활성은 기간별로 측정하였으며 그 결과, α-amylase 효소 활성은 느타리와 상황버섯균이 우수하였으며, β-amylase활성은 느타리와 장수버섯균이 우수하였다. Protease활성은 상황버섯균을 제외하고는 곡물에 관계없이 비교적 고른 생산을 보였다. 담자균 배양에 의해서 명도는 감소하고 적색도와 황색도가 증가하였다.

본 연구는 백설기의 노화를 억제 및 저장 중 품질 향상을 위한 Novamyl의 최적 첨가량을 0.1%로 선정하고 트레할로스를 5%, 10%, 15% 첨가하여 백설기를 제조한 후 상온(25oC)에서 0, 1, 2, 3일간 저장하면서 기계적 및 관능적 특성과 노화 속도 변화를 살펴보았으며 그 결과는 다음과 같다. 수분활성도는 트레할로스의 첨가량이 증가할수록 유의적으로 감소하였으며 저장 일수에 따른 차이는 없었다. 색도는 트레할로스를 첨가한 백설기의 명도(L값)가 유의적으로 낮게 나타났으며 적색도를 나타내는 a값은 첨가군이 대조구에 비해 낮게 나타났고 황색도(b값)는 첨가군이 다소 낮은 값을 보였으며 저장 일수가 증가함에 따라 황색도가 감소하는 경향을 보였다. 기계적 조직감에 대해서는 트레할로스의 첨가량이 증가할수록 경도가 감소하였고 저장 중 씹힘성의 감소폭이 감소하였다. Avrami 방정식으로 노화속도를 측정하였으며, Avrami exponent(n) 값과 시간상수(1/k) 값을 계산한 결과 트레할로스 첨가군이 대조구에 비해 노화속도가 감소하였음을 알 수 있었다. 관능검사 결과에서 15% 첨가구가 대부분의 항목에서 우수한 결과를 보였고 특히 전반적인 기호도 및 씹힘성 항목에서 높은 선호도 값을 보였으나 10% 첨가구와의 유의적인 차이는 보이지 않았다. 이와 같은 실험결과를 통해 백설기 제조 시 Novamyl 0.1%를 처리하고 트레할로스를 첨가할 경우 노화 억제와 품질 향상의 효과가 있었으며, 특히 기계적 측정 결과와 관능검사 결과를 종합했을 때, 트레할로스를 10% 첨가가 15%의 첨가에 비해 유의적인 차이가 적어 가장 적절한 것으로 나타났다. 효소의 첨가는 백설기의 노화를 억제하는 효과가 있지만 물성적인 측면에서 조직이 과도하게 물러지는 등의 문제를 보이는데 이에 트레할로스를 적당량 첨가함으로써 노화 억제를 향상시키고 물성적인 측면과 기호도를 향상시켜 저장성이 우수하고 소비자의 입맛에 적합한 백설기의 제조가 가능하다고 사료된다.

우리나라 토양으로부터 분리된 (주와 김 2009) poly (butylene succinate-co-butylene adipate; PBSA) 분해균 Burkholderia cepacia PBSA-7, Bacillus licheniformis PBSA-8 및 Burkholderia sp. PBSA-9에서 PBSA 분해효소를 암호화하는 유전자를 분석하였다. PCR을 수행하여 B.cepacia PBSA-7과 Burkholderia sp. PBSA-9는 약

Substantial efforts have been made to manipulate ruminal environment in a hope to enhance ruminal fermentation efficiency for better ruminant productivity. Some of examples are methane inhibitors, antibiotics, microbial enzymes, fatty acids and/or lipid feeding, buffering agents, ionophores and probiotics. Of these efforts, the non-ionic surfactant (NIS) has been known for its stimulation to release enzymes from a range of anaerobic microbes. A series of studies were conducted 1) to evaluate NIS diluted with water and ethanol on in vitro ruminal fermentation and 2) to determine the influence of diluted NIS on digestibility of feedstuffs. In first experiment (Exp. 1), NIS was diluted with water or ethanol to measure its effects on in vitro microbial growth, ruminal enzyme activities and gas production by mixed ruminal microbial culture. The NIS was diluted with water or ethanol separately in a 1:5 ratio (w/v). Water and ethanol-diluted NIS with wheat flour were added with rice straw-based mixed ruminal microbial cultures at the rate of 2 ㎎ NIS/16 ㎖ McDougall buffer plus 4 ml ruminal fluid solution. The mixed ruminal microbial culture was run without any NIS addition as control. Addition of NIS either diluted with water or ethanol has significantly reduced the gas production in mixed ruminal microbial culture at 12 and 24 h of incubation. At 48 h post incubation, gas production was the highest with the addition of ethanol diluted NIS followed by water-diluted NIS and control. Carboxy methyl cellulase activity in rice straw-based mixed ruminal bacterial culture was significantly higher with the addition of ethanol-diluted NIS compared with the addition of water-diluted NIS and control at 24 and 72 h post incubation. In second in vitro experiment (Exp. 2), effects of addition of ethanol diluted NIS on dry matter (DM) digestibility of alfalfa hay, gas production, pH and cellular growth in mixed ruminal microbial culture were examined. Alfalfa hay based mixed ruminal microbial culture without any NIS addition was run as a control. The pH of mixed ruminal microbial culture was significantly lower than control at all post incubation sampling hours. In vitro DM digestibility of alfalfa hay was significantly higher with the addition of NIS compared with control. Gas production was significantly less with NIS addition compared with control at all post incubation sampling hours. Microbial growth in mixed ruminal microbial culture was significantly increased with the addition of NIS compared to control.

호열성 미생물을 검토하기 위하여 전국 각지로부터 토양과 두엄을 채취하여 그로부터 호열성 미생물을 분리하였다. 토양과 두엄으로부터 분리된 호열성 미생물 1250여 균주를 선별하였고, 이들을 대상으로 미생물이 생산하는 효소활성을 검토하여 호열성 전분 분해 효소를 생산하는 1주의 미생물을 확인하였다. 확인된 1주의 미생물을 strain 2719라 명명하였다. Strain 2719 균주는 형태학적으로 gram 양성 간균의 특징을 나타냈고, 균주의 표면은 매끄럽지 않았으며, 비교적 다양한 길이를 가지고 있었다. 또한 다른 gram 양성 간균들에 비해서 많은 수의 균사들이 각 균주들 사이에 복잡하게 얽혀있었다. 생화학적 특성을 확인한 결과 catalase 양성, glucose 발효, arabinose 발효, mannitol 발효, casein·gelatin·starch 가수분해의 특징을 가지고 있었으며, 이는 Bacillus sp.로 추정되었다. 생육 pH의 범위는 pH 6-pH 8 범위에서 생육이 가능했으며, 생육 온도의 범위는 50-70oC였다. 16S rDNA sequence 분석결과 Bacillus thermoglucosidasius의 16S rDNA와 99.52%가 일치하였으나, sequence의 일부분이 다른 부분이 있고, 생육 특성에서 약간의 차이를 보였다. 또한 gene bank에 등록되어 있는 균주들의 16S rDNA sequence들과 비교하여도 일치하는 균주는 확인되지 않았다. 이와 같은 실험결과에 따라 2719 균주는 기존에 발표되지는 않았으나, Bacillus thermoglucosidasius와 매우 유사한 균주로 판단되어 Bacillus thermoglucosidasius 2719로 명명하였다.

활성글루텐을 첨가한 쌀가루에 검의 첨가가 쌀가루의 신속점도측정기(RVA) 호화양상과 farinograph 반죽특성에 미치는 영향을 조사하였다. 검의 첨가에 의해 쌀가루의 RVA 최고점도는 증가한 반면 setback은 약간 감소함을 나타내었다. 활성글루텐을 첨가한 쌀가루에 검의 첨가는 farinograph 수분흡수율을 증가시키고 반죽의 안정도를 증가시켜 반죽을 강하게 하는 효과를 부여하였다. 쌀가루에 활성글루텐을 14% 첨가하고 검, 유화제 및 효소제를 복합처리하여 쌀빵을 제조한 후 쌀빵의 부피와 저장 중 경도변화를 조사하였다. 쌀가루에 첨가하는 활성글루텐의 양이 감소함에 따라 쌀빵의 부피가 감소하였지만 검, 유화제 및 효소제를 복합첨가 함으로써 쌀빵의 부피를 현저히 개선시킬 수 있었다. 활성글루텐만을 첨가한 쌀빵은 저장 중에 경도가 급격히 상승하였으나 검, 유화제 및 효소제를 복합첨가하여 제조한 쌀빵은 저장 중에 경도의 증가 폭이 현저하게 감소함을 나타내었다. 따라서 적절한 검, 유화제, 효소제의 복합처리는 쌀빵의 부피와 경도 변화 등 쌀빵의 품질에 매우 긍정적인 영향을 주는 것으로 평가되었다.

표고균사가 액체배지나 한천배지에서 생장하여 숙성되는 동안 갈변되는 현상을 나타낸다. 표고균사는 접종 25일부터 갈변이 시작되어 30일부터 균총 전반에 걸쳐 이루어지기 시작해 접종 40일까지 갈변이 완전히 이루어진다. 이때 균사내의 효소의 활력을 조사한 결과 phenloxidase계통의 효소들은 laccase는 접종 15일에 가장 높았으며 갈변이 되면서 점점 감소되었으나 tyrosinase는 갈변이 이루어지는 30일부터 급격히 증가하였고 peroxidase는 등전점 전기영동에 의하여 조사한 바 갈변이 이루어지는 30일부터 서서히 증가하였다. 등전점전기영동에 의해 조사된 phosphatase효소는 esterase, acid phosphatase, alkaline phosphatase를 조사하였으며 균사의 갈변이 일어나기 시작하는 접종 30일까지는 증가되었으나 그 이후 갈변이 이루어지는 과정에서는 급격히 감소되었다.

This study investigated the influences of the addition of lactic acid bacteria inoculants and an enzyme during the crush processing of com silage on the nutritive value, nutrient intake and aerobic deterioration. The fermentation quality of the silages showed that although all of the silages were of good quality, the V-score of U was 91.0 points and significantly lower than those of other silages (P＜0.01). On the chemical composition of silages, the crude protein concentration of U was 7.1%DM and significantly lower than those of other silages (P＜0.05). On the digestibility, the nutritive value and nutrient intake did not significantly differ among the silages examined. After silo opening, a rapid rise in heat was seen from the 6th after opening for 11C38 and SLA, and from the 9th in U and SL, respectively.

유채(Brassica napus L.)에서 황 결핍이 흡수와 동화에 대한 영향을 알아보고자 농도를 세가지 수준(1 mM , 대조구; 0.1mM , 결핍; 0 mM , 무공급)으로 25시간 처리한 후 흡수량, 식물조직내의 nitrate reductase (NR) 및 glutamine syntheetase (GS) 활성을 분석하였다. 25시간 처리과정에서 황결핍 조건하에서의 의 흡수는 대조구와 큰 차이를 나타내지 않은 반면, 황 무공급구에서는 의 흡수는 유

In this paper, the electrochemical non-enzyme immunosensor has been developed for the determination of salmonella antigen, using inverse voltammetry. For the estimation of salmonella antigen concentration, the nanoparticles synthesized by microemulsion method were conjugated with salmonella antigen. Then, the immunocomplex between antibody immobilized on the transducer surface and antigen containing a magnetic nanoparticles was formed. From the linear relationship between the reduction peak current of Fe(III) and salmonella antigen concentration, it is suggested that the electrochemical non-enzyme biosensor is applicable to detect salmonella antigen in the concentration range of

The Black Soldier Fly (BSF, Hermetia illucens) was widely distributed throughout Korea. This insect was mainly found in the vicinity of and in cattle sheds, manure sheds, living waste dump grounds, and food waste dump grounds. This fly is a kind of a beneficial fly because BSF adults do not go into houses, they do not regurgitate on human food, they do not bite, bother or pester humans in any way and they are not associated in any way with the transmission of disease. But their greatest attribute lies their ability to eat and digest raw waste. They can devour, for example, a large, raw, Irish potato and others in just a few hours. Unlike many other flies, since the BSF larvae have very powerful mouth parts and digestive enzymes, they can ingest raw waste far more efficiently than any other known species of fly. On this study, to investigate whether feeding strategy of the BSF larvae involves extra-oral digestion or not, and to better understand this process, the salivary glands and a few tissue from the BSF were dissected and subjected to morphological and preliminary enzyme characterization.

Protease from various sources have been studied biotecnologically. For biotechnological applications, one highly preferred enzyme is protease. There have been no reports of cloned genes encoding digestive proteases in the Laccotrephes japonenis, Ranatra unicolor, Muljarus japonicus. These insects are considered to be a predator of aquatic insects. RT-PCR was used to amplify cDNA fragments for digestive proteases from total RNA the hole body of the insects. The flanking sequences of the 5'- and 3'- end of the these genes were characterized by RACE-PCR. Sequence analysis showed that these genes contained complete ORF. The deduced amino acid sequences of these protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin-like serine proteinase, 54% to Triatonatoma infestans salivary trypsin. To generate Laccotrephes japonensis serine protease, the DNA fragment coding for serine protease is cloning into suttle vector pBACⅠ, named pBAC1-JG and infected to Spodoptera frugiperda (sf9) insect cell. The cDNA encoding JG was expressed as a 32-kDa polypeptide in baculovirus infected insect cells and the recombinant protein showed activity in the protease enzyme assay using gelatin as a substrate.

A mathematical model was written for simulating the removal of phenol from wastewater in enzyme-loaded membrane reactor (EMR). The numerical simulation program was developed so as to predict the degradation of phenol through an EMR. Numerical model proves to be effective in searching for optimal operating conditions and creating an optimal microenvironment for the biocatalyst in order to optimize productivity. In this study, several dimensionless parameters such as Thiele Modulus (Φ2, dimensionless Michaelis-Menten constant (ξ), Peclet number (Pe) were introduced to simplify their effects on system efficiency. In particular, the study of phenol conversion at different feed compositions shows that low phenol concentrations and high Thiele Modulus values lead to higher reactant degradation.

Conjugated nanocrystals using CdSe/ZnS core/shell nanocrystal quantum dots modified by organic linkers and glucose oxidase (GOx) were prepared for use as biosensors. The trioctylphophine oxide (TOPO)-capped QDs were first modified to give them water-solubility by terminal carboxyl groups that were bonded to the amino groups of GOx through an EDC/NHS coupling reaction. As the glucose concentration increased, the photoluminescence intensity was enhanced linearly due to the electron transfer during the enzymatic reaction. The UV-visible spectra of the as-prepared QDs are identical to that of QDs-MAA. This shows that these QDs do not become agglomerated during ligand exchanges. A photoluminescence (PL) spectroscopic study showed that the PL intensity of the QDs-GOx bioconjugates was increased in the presence of glucose. These glucose sensors showed linearity up to approximately 15 mM and became gradually saturated above 15 mM because the excess glucose did not affect the enzymatic oxidation reaction past that amount. These biosensors show highly sensitive variation in terms of their photoluminescence depending on the glucose concentration.

The morecular cloning, gene structure, expression and enzyme activity of a serine-like proteas frome Laccotrephes Japonensis were examined. In this study, RT-PCR was used to amplify cDNA fragments for serine-like proteases from total RNA the hole body of Laccotrephes japonensis. The flanking sequences of the 5'- and 3'- end of the this gene were characterized by RACE-PCR. Sequence analysis showed that this gene contained an 963bp ORF encoding 321 amino acids. The deduced amino acid sequence of this protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin precuror LlsgP4, 54% to Triatonatoma infestans salivary trypsin. To generate Laccotrephes japonensis serine-like protease, the DNA fragment coding for serine protease is cloning into suttle vector pBACⅠ, named pBAC1-JG and infected to Spodoptera frugiperda (sf9) insect cell. The cDNA encoding JG was expressed as a 32-kDa polypeptide in baculovirus infected insect cells and the recombinant JG showed activity in the protease enzyme assay using gelatin as a substrate.