This study was conducted to investigate anti-inflammatory and wrinkle improvement effects of Sophora flavescens Aiton water extracts (SWE) treated with proteolytic enzyme. The antioxidant activity of proteolyzed Sophora flavescens Aiton water extracts (SWE-E) showed increased total polyphenol content, total flavonoid content, electron-donating ability and ABTS+ free radical scavenging activity compared with SWE. To investigate the anti-inflammatory effects, the inhibition of NO production was assessed in RAW 264.7 cells induced by LPS. The SWE-E showed an increased anti-inflammatory effect compared with SWE at the same concentration. The anti-wrinkle effect was evaluated by the rate of collagenase and elastase inhibitory activity, which was determined by the MMP-1 mRNA measured in human dermal fibroblast (HDF) by quantitative polymerase chain reaction (qPCR). Collagenase and elastase are important enzymes that play roles in wrinkle formation, and SWE-E showed a significantly higher collagenase inhibition rate than non-treated extracts. The MMP-1 mRNA in HDF cells decreased in a dose-dependent manner. Moreover, SWE was shown to be a non-irritant in the BCOP assay, which is an alternative method to in vivo eye irritation test. Taken together, these results suggest that proteolytic enzyme could enhance the antioxidant activity, as well as the anti-inflammatory and anti-wrinkle effects of SWE, and that SWE-E could be used as a cosmeceutical ingredient.

본 연구는 동결 건조한 krill (Euphausia superba) meal을 함량별 섭취시켰을 때 7주령된 Sprague Dawley계 수컷 흰쥐의 혈청 중 alkaline phosphatase (ALP), aminotransferase (AST, ALT) 및 lactate dehydrogenase (LDH) 등 혈청 간 기능 효소활성과 장기 조직의 불소 함량에 미치는 영향을 확인하였다. 기본식이를 급여한 대조군인 CG군을 비롯하여 10%, 20%, 30%의 krill meal을 첨가한 급여군을 각각 KM10군, KM20군, KM30군으로 구분하여 4군의 급여군으로 나누어 5주간 실험 사육한 결과는 다음과 같다. 혈청 중 alkaline phosphatase (ALP), aminotransferase (AST, ALT) 및 lactate dehydrogenase(LDH)의 효소 활성은 대조군인 CG군 보다 krill meal을 함량별 첨가 급여군에서 감소되는 것으로 나타났다. 흰쥐의 혈청 및 장기조직(간, 뇌, 심장, 신장)의 불소 함량은 krill meal의 함량에 따라 불소 함량도 증가되는 것으로 나타났다.

The purpose of this study was to develop an indirect enzyme-linked immunosorbent assay (indirect ELISA) based on a monoclonal antibody (MAb) that is specific to mackerel thermal stable-soluble protein (TSSP), that can be used for the rapid detection of mackerel in processed marine foods. Among the four MAbs (3A5-1, 2, 9, and 12) developed in previous studies, the 3A5-2 MAb that showed high specificity and sensitivity were selected and used to develop the indirect ELISA method. The detection range of the indirect ELISA was 0.02%-0.001% and the detection limit of 0.001% was shown. No cross-reaction to other marine products and food ingredients was observed by the indirect ELISA. Processed marine foods containing mackerel with ≥ 0.3 O.D. value at 405 nm were estimated as positive samples by the indirect ELISA. Therefore, the indirect ELISA can be used as a rapid and sensitive method to identify mackerel authenticity and adulteration in processed marine foods.

The objectives of this study were to characterize the physicochemical properties of ginger (Zingiber officinale Rosc.) and the optimum extract processing condition to increase the solubilization efficiency of 6-gingerol and polysaccharides disintegration. The physicochemical properties were investigated under high-pressure enzyme (HP) and enzyme (WB) treatment conditions such as reaction time (1, 2, 3 h), pressure (50, 70, 100 MPa) and sample types, and cell wall degradation enzyme (hemicellulase, cellulase, pectinase, glucosidase, etc.) The effect of high-pressure enzyme treatment depending on sample types was significantly highest in the freeze-dried ginger powder. The optimum condition for high-pressure enzyme treatment was with Pectinex Ultra SP-L (Pec) enzymes for 2 h at 50oC and 100 MPa. Water soluble indexes increased 4.6 and 3.8 times more compared to CON (15.35%) while total polyphenol contents increased by 1.8 and 1.7 times compared to CON (1.43%). The total contents of indicator components such as 6, 8, 10-gingerol and shogaol was 1.53%, increasing 2.7 times more compared to CON (0.57%) with a significant difference (p<0.001). The high-pressure and enzymatic approach described in this study would be beneficial to food industries for developing ginger functional product and materials.

The outbreaks of foodborne diseases associated with bacterial contamination are still critical issues all over the world. To ensure food safety, the diagnosis of pathogenic bacteria on site at early state of contamination are required. Escherichia coli O157:H7 (E. Coli O157:H7) is one of the major factor causing foodborne diseases. We introduce a sandwich type colorimetric detection method integrated with chitosan-coated starch magnetic polymer beads(CS@SMBs) that can separate and concentrate bacteria in aqueous environment. For signal amplification, horseradish peroxidase-conjugated antibody (HRP-Antibody) and 3,3',5,5'- tetramethylbenzidine (TMB) were employed as enzyme label and chromogenic substrate, respectively. We demonstrate that CS@SMBs not only show a good magnetic sensitivity, but also can capture a variety of bacteria regardless of Gram-negative and Gram-positive, which offer possibility for separation of the broad range of bacteria from food matrix. Our approach successfully captures E. coli O157:H7 with detection limit of 101 CFU/mL through naked eye, making promise of fast, on-site, and sensitive detection of pathogenic bacteria.

This manuscript explains the effective determination of urea by redox cyclic voltammetric analysis, for which a modified polypyrrole-graphene oxide (PPY-GO, GO 20% w/w of PPY) nanocomposite electrode was developed. Cyclic voltammetry measurements revealed an effective electron transfer in 0.1 M KOH electrolytic solution in the potential window range of 0 to 0.6 V. This PPY-GO modified electrode exhibited a moderate electrocatalytic effect towards urea oxidation, thereby allowing its determination in an electrolytic solution. The linear dependence of the current vs. urea concentration was reached using square-wave voltammetry in the concentration range of urea between 0.5 to 3.0 μM with a relatively low limit of detection of 0.27 μM. The scanning electron microscopy was used to characterize the morphologies and properties of the nanocomposite layer, along with Fourier transform infrared spectroscopy. The results indicated that the nanocomposite film modified electrode exhibited a synergistic effect, including high conductivity, a fast electron-transfer rate, and an inherent catalytic ability.

The effect of the ionization energy on the common house mosquito, Culex pipiens, pupa was investigated as the hatching rate, longevity and detoxification enzyme activity. The longevity of adult emerging in the pupa decreased depending on the exposure dose of Gray (Gy) in Cx. pipiens. The hatching rate of the F1 generation egg derived from Cx. pipiens pupa treated up to 70 Gy did not show any difference with the control, but the eggs did not hatch 100% at 120 Gy exposure. These results showed that Cx. pipiens was sterilized at 120 Gy. Several detoxifying enzyme (GST, non-specific esterase and MFO) activity of Cx. pipiens adult emerged from pupa irradiated with 120 Gy was investigated. The difference in activity was observed in female than in male irradiated with electron beam. As a result, electron beam irradiation seems to affect enzyme activity as well as induce infertility of Cx. pipiens.

The common bed bug, Cimex lectularius, possesses a cholinesterase expressed exclusively in the salivary gland (ClSChE). In this paper, we investigated the molecular structure, tissue distribution patterns, and biochemical properties of ClSChE and showed that ClSChE exists as a soluble monomeric form or a soluble dimeric form connected by a disulfide bridge. Immunohistochemical analysis confirmed that ClSChE was expressed in the epithelial cells of both the salivary gland and the duct. In addition, the secretion of monomeric ClSChE through the proboscis during feeding was detected by western blotting using a ClSChE-specific antibody. To predict the role of ClSChE injected into the tissue of an animal host, we analyzed the extent of sequestration and hydrolysis of acetylcholine (ACh)/choline (Ch) by ClSChE by ultra-performance liquid chromatography-tandem mass spectrometry. Kinetic analysis revealed that ClSChE possesses extremely low Km (high affinity to ACh) and Vmax values. These findings suggest that ClSChE functions as a sequestering enzyme specific to ACh (not to Ch) by having a very strong affinity to ACh but an extremely long turnover time.

The aim of this study was to select various fungal strains indigenous to Korea that have the potential to produce cellulases, including filter paper activity (FPase), endo-β-1,4-glucanase (EG), and β-glucosidase (BGL). Among the 25 species of Ascomycetes and the 32 species of Basidiomycetes tested in this study, the Bjerkandera adusta KUC10565, Heterobasidion orientale KUC10556, Hyphoderma praetermissum KUC10609, and Trichoderma harzianum KUC1716 all exhibited remarkably high FPase activity. In addition, the T. harzianum KUC1716 showed high levels of EG and BGL activity. This strain has been selected for further study because of their enzymatic potential.

To evaluate effects of ligninolytic enzyme type on the mycelial response and ligninolytic enzyme production during interspecific interactions among wood-rotting fungi, 4 fungal strains, Trichophyton rubrum LKY-7, Trichophyton rubrum LSK-27, Pycnoporus cinnabarinus, and Trichoderma viride, were selected. Regarding ligninolytic enzyme production, LKY-7 secreted laccase and manganese peroxidase (MnP), P. cinnabarinus secreted only laccase, and LSK-27 secreted only MnP in glucosepeptone medium, while T. viride did not produce any ligninolytic enzymes. In the co-culture of LKY-7 with P. cinnabarinus, the formation of aerial mycelium was observed and the enhancement of laccase activity owing to interspecific interaction appeared to be very low. In the co-culture of LKY-7 and P. cinnabarinus with LSK-27, a hypha-free clear zone was observed, which resulted in deadlock, and increased laccase or MnP activity was detected at the interaction zone. The interaction responses of LKY-7, P. cinnabarinus, and LSK-27 with T. viride were characterized by the formation of mycelial barrages along the interface. As mycelial barrages were observed at the T. viride territory and no brownish pigment was observed in the mycelial barrages, it is suggested that laccase and MnP are released as part of an offensive response, not as a defensive response. The co-culture of P. cinnabarinus with T. viride lead to the highest enhancement in laccase activity, yielding more than 14-fold increase in laccase activity with respect to the mono-culture of P. cinnabarinus. MnP activities secreted by LKY-7 or LSK-27 was generally low in interspecific interactions.

Physiological responses and activities of antioxidative enzymes were investigated in pepper(Capsicum annuum) seedlings subjected to low temperature stress. The seedlings were exposed to 7℃, 11℃, or 15℃ for 4 h during the early seedling stage of pepper plants. The results showed that plant height and chlorophyll content were unaffected by the treatments. Polyphenol content in seedlings exposed to 7℃ was higher than that at 11℃ and 15℃ for 2 h. After 3 h of treatment, the flavonoid content was higher in pepper seedlings exposed to 7℃ than 11℃ and 15℃, which was slightly higher than that detected in the control. The H2O2 content increased remarkably with increasing exposure time to low temperature(i.e., 7℃) as compared to that at 11℃ and 15℃. The levels of antioxidant enzymes superoxide dismutase(SOD), catalase(CAT), ascorbate peroxidase(APX), and peroxidase(POD) fluctuated. These results provide basic information that can be utilized to maintain efficient temperature in greenhouses for sustainable growth of pepper under severe low temperature conditions.

The purpose of this study was that the optimal hydrolysis conditions of endo- and exo-type enzymes were selected to utilize organic cheese byproducts. Optimal substrate concentration and optimum enzyme ratio were measured by using 4 kinds of endo-type enzymes (alcalase, neutrase, protamex, and foodpro alkaline protease) and two exo-type enzymes (flavourzyme and prozyme 2000P) for whey protein hydrolysis were analyzed using liquid chromatography. As a result, the optimal endo-type enzyme through the first enzyme reaction was selected as alcalse, and as a result of the secondary enzyme reaction, flavourzme was selected as the Exo type enzyme. The concentration of whey protein substrate for optimal primary and secondary enzyme reactions was 10%. In addition, the optimum ratio of enzyme was 0.5% of alcalase and 0.2% of flavourzyme, which showed low molecular weight chromatography pattern compared to 2% of alcalase and 1% of flavourzyme hydrolyzate. Therefore, hydrolyzing the endo-type enzyme alcalase at a concentration of 0.5% for 10 hours and then hydrolyzing the exo-type enzyme flavouryme at a concentration of 0.2% for 4 hours was considered to be the optimum condition.

암, 당뇨, 심장질환 등의 성인병 예방에 좋은 효과를 제공하기 때문에 선진국에서는 통곡의 섭취를 권장하고 있으나, 곡물이 겨층을 포함하고 있어 식미가 거친 단점이 있다. 초록통쌀은 호숙기에 수확하여 조직이 경화되지 않고 각종 미네랄, 비타민, 식이섬유 등 영양성분이 풍부한 것으로 알려져 있다. 본 연구는 고령자의 영양보충 및 식감 개선을 위한 초록통쌀 현미의 영양성분 및 효소처리에 따른 이화학적 특성변화를 분석하였다. 국내 다 소비품종인 삼광, 조평, 추청 3종의 초록통쌀의 영양성분을 비교 분석하였으며, 4종의 효소 처리(0.1 ~ 3%)에 따른 신속호화점도계, 전자현미경, 아밀로펙틴 함량 등의 이화학적 특성 변화를 측정하였다. 삼광, 조평, 추청의 주요 영양성분의 함량은 100g 당 비타민 B1이 144, 100, 105ug, 식이섬유가 21, 13, 12g, 폴리페놀이 63, 52, 56mg으로 삼광이 가장 높은 것으로 확인되었으며, 비타민 B2, 나이아신, 칼륨, 인, 마그네슘, 총 엽록소도 높은 것으로 나타났다. 효소농도가 증가함에 따라 경도가 감소하였으나, 0.3% 이상에서는 유의적 차이가 없는 것으로 나타났다. 대조군 및 각 효소처리에 따른 식미적 합성을 나타내는 신속호화점도계 peak 값은 760, 724, 708, 754, 761 cP이었고, 시간변화에 따른 품질특성을 나타내는 Breakdown 값은 157, 166, 151, 100, 99 cP로 두 항목 모두 셀룰로오스계열 효소 D 처리 시 식미가 가장 우수한 것으로 확인되었다. 전자현미경 확인결과 효소처리에 따른 물성변화로 입자간 간극이 커짐을 확인하였다. 전분의 분자량 분포특성인 아밀로펙틴 측쇄사슬 길이는 모든 처리군 평균값이 15.71~15.99 DP로 효소처리에 따른 아밀로펙틴 함량 변화는 없는 것으로 나타났다. 이러한 결과는 셀룰로오스 계열 효소처리를 통하여 물성조절이 가능하고 식미도 우수한 것으로 나타나, 초록통쌀은 저작능력이 약한 고령자가 쉽게 섭취할 수 있고, 영양개선을 위해 활용할 수 있음을 제시하고 있다.

Carbon-based magnetic nanostructures in several instances have resulted in improved physicochemical and catalytic properties when compared to multi-wall carbon nanotubes (MWCNTs) and magnetic nanoparticles. In this study, magnetic MWCNTs with a structure of NixZnxFe2O4/MWCNT as peroxidase mimics were fabricated by the one-pot hydrothermal method. The structure, composition and morphology of the nanocomposites were characterized with X-ray diffraction (XRD), Fourier transform infrared spectroscopy and transmission electron microscopy. The magnetic properties were investigated with a vibrating sample magnetometer. The peroxidase-like catalytic activity of the nanocomposites was investigated by colorimetric and electrochemical tests with 3,3´,5,5´-tetramethylbenzidine (TMB) and H2O2 as the substrates. The results show that the synthesis of the nanocomposites was successfully performed. XRD analysis confirmed the crystalline structures of the NixZnxFe2O4/ MWCNT nanohybrids and MWCNTs. The main peaks of the NixZnxFe2O4/MWCNTs crystals were presented. The Ni0.25Zn0.25Fe2O4/MWCNT and Ni0.5Zn0.5Fe2O4/MWCNT nanocatalysts showed nearly similar physicochemical properties, but the Ni0.5Zn0.5Fe2O4/MWCNT nanocatalyst was more appropriate than the Ni0.25Zn0.25Fe2O4/MWCNT nanocatalyst in terms of the magnetic properties and catalytic activity. The optimum peroxidase-like activity of the nanocatalysts was obtained at pH 3.0. The Ni0.5Zn0.5Fe2O4/MWCNT nanocatalyst exhibited a good peroxidase-like activity. These magnetic nanocatalysts can be suitable candidates for future enzyme-based applications such as the detection of glucose and H2O2.

The entomopathogenic fungus Metarhizium anisopliae is one of potent biological control agents against a variety ofinsect pests. In this study, we investigated enzyme production of M. anisopliae strains A and B. They produced extracellularenzymes for degrading the epidermis of Monochamus alternatus, a crucial mediator of the pinewood nematode Bursaphelenchusxylophilus. With Q-TOF MS/MS analysis, 29 kDa protein, a major band on a SDS-PAGE gel, was identified as subtilisin-likeserine protease PR1A. M. anisopliae A produced an extracellular enzyme more efficiently than M. anisopliae B: however,enzyme activities targeted for the cuticle were comparable. Our results suggest that the two strains of M. anisopliae havethe biological potential against M. alternatus with insecticidal protease production.

This study was conducted to certify the effect of aqueous extracts from fifty medicinal plants on the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in vitro. Each aqueous extract was prepared by combining one-part medicinal plants with twenty-parts distilled water at 80°C for 8 h. Among the fifty medicinal plants, Allium sativum L. and Cinnamomum cassia Presl were regarded as an effective anti-hangover substance. Allium sativum L. extract increased ALDH activity more than 2 times compared with ADH activity, enhancing the acetaldehyde degradation. Cinnamomum cassia Presl extract dramatically inhibited ADH activity compared with ALDH activity, thus potently decreasing the acetaldehyde formation. ADH and ALDH activities were proportionally inhibited according to the increased concentration of Cinnamomum cassia Presl extract. The aqueous extract of Cinnamomum cassia Presl at a concentration of 45.33 μg/mL inhibited ADH activity by 52.8% and ALDH activity by 11.0%.

Phospholipase A2 (PLA2) hydrolyzes phospholipids at sn-2 position to release free fatty acids. PLA2 consists of a superfamily mainly categorized as secretory PLA2 (sPLA2), cellular Ca 2+ -dependent PLA2 (cPLA2), and cellular Ca 2+ -independent PLA2 (iPLA2). We are the first to report iPLA2 in insect. Here an objective of our study is to purify a recombinant iPLA2 protein (SeiPLA2) of Spodoptera exigua using bacterial expression system. An open reading frame of SeiPLA2 was cloned into an expression vector and then transformed into Escherichia coli BL21. Over-expression with IPTG yielded recombinant SeiPLA2 (rSeiPLA2), which was then purified by an affinity chromatography using Ni-NTA column. The purified rSeiPLA2 gave significant PLA2 activity using a pyrene substrate. Its activity was inhibited by an iPLA2 specific inhibitor (BEL), but not by sPLA2 inhibitor (BPB) or cPLA2 inhibitor (MAFP