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

Streptozotocin(STZ)을 45mg/kg.b.w의 용량으로 흰쥐의 미정맥에 투여 한 후 유발된 당뇨 흰쥐에게 1일 1회 7일간 1,000mg/kg의 용량으로 도라지 뿌리 에탄올 추출물을 투여 후 glucose 함량과 당대사에 관여하는 효소인 glucose-6-phosphatase(G-6-Pase) glucose-6-phosphate dehydrogenase (G-6-PDH), glucokinase(GK)활성과 glycogen 함량, triglyceride(TG), total cholesterol등의 지질대사에 관여하는 물질들을 측정하였다 그 결과 도라지 뿌리 에탄올 추출물 투여군이 glucose, TG, total cholesterol등의 함량과 G-6-Pase 활성의 유의적인 감소(p<0.05)를 나타내었으며 glycogen 함량과 HDL-cholesterol, G-6-PDH, GK의 활성이 유의적인 증가(p<0.05)를 나타내었다. 이와 같이 도라지 뿌 리 에탄올 추출물이 항당뇨 개선효과를 갖는 유효성분을 함유하고 있음을 알 수 있었다.

Lipoxygenase(LOX) 결핍 콩(05C4 및 LS, LOX2,3-free)과 일반콩(태광, LOX1,2,3-present)으로 제조된 된장의 급이가 고지방-콜레스테롤 식이성 흰쥐의 체내 지질성분 및 항산화 효소계에 미치는 영 향을 알아보고자 하였다. SD계 흰쥐는 정상군, 고지방-콜레스테롤 급이군(HFC), 고지방-콜레스테롤 식이+5% 된장 분말 급이군[HFC+D1, 태광된장; HFC+D2, 05C4된장; HFC+D3, LS된장)으로 구분하여 5주간 사육되었다. In vitro에서 된장의 콜레스테롤 흡착활성은 4mg/mL 농도에서 LS된장이 태광된장 에 비해 1.5배 높았다. 식이효율은 정상군에 비해 대조군에서 유의적으로 높았으며, LS된장 급이군 (HFC+D3)은 대조군에 비해 유의적으로 감소되는 경향이었다. Rö hrer 및 T.M.지수와 체지방 함량은 대조군에 비해 된장 급이군에서 모두 유의적으로 감소되었으며, 특히 Lee지수 및 BMI는 LOX 결핍콩 된장 급이군에서 유의적으로 감소되었다. 혈청 총 지질 및 중성지방 함량은 대조군에 비해 된장 급이군 에서 유의적으로 감소되었으나, 총 콜레스테롤, HDL-, LDL-콜레스테롤 및 심혈관질환 위험지수는 LS 된장 급이군만 유의적인 감소를 보였다. 혈청 AST 및 ALT 활성은 HFC+D1 및 HFC+D2간에는 유의차 가 없었고, HFC+D3은 HFC+D1 및 HFC+D2에 비해 유의적으로 낮은 수준이었다. 간 조직에서 총 지 질 함량은 HFC+D3이 대조군에 비해 약 25.2%의 감소되었으며(p<0.05), 중성지방 및 총 콜레스테롤 함량은 HFC+D2 및 HFC+D3이 대조군에 비해 유의적인 감소를 보였다. 분변 중의 중성지방 함량은 된 장 급이군 모두 대조군에 비해 26.2~40.3%의 증가를 보였으며(p<0.05), 총 콜레스테롤 함량은 LS된장 급이군만 유의적으로 증가하였다. 간 조직 중 SOD 활성은 대조군에 비해 05C4 및 LS된장 급이군에서 유의적으로 증가하였으며, catalase 및 GSH-px 활성은 된장 급이군 모두 대조군에 비해 유의적으로 증가하였다. UDPGT 활성은 된장의 급이 시 정상군과 유사한 수준으로 증가되었으며, 특히 LS된장 급 이군은 정상군에 비해 유의적으로 증가하였다. 따라서 LOX 결핍 콩으로 제조된 된장은 체내 지질 개선 및 항비만 활성에 긍정적인 도움이 될 것으로 판단된다.

To increase the collagen recovery rate, bromelain (PB) and a microbial enzyme (PM) were used to treat to pork skin with single agent or combinations. The quality of collagen from the pork skin was evaluated by enzymatic treatments. The highest results for the solid contents and pork skin recovery rate obtained with the microbial-enzyme-bromelain mixtue (PMB) were 13.60% and 18.05% respectively. The result also showed that the color was affected by different types of enzyme treatments. Although PM treatment showed the highest result in the protein content of 251.30 mg/100 g, PMB treatment was the highest in the test of collagen content of 37.73 g/100 g among the treatments. However bands of the pork skin were detected widely at 130 kDa and 170 kDa ranges in SDS-PAGE. The band of PB treatment showed at the range of below 17 kDa, changed into a smaller molecular weight. The collagen content test of the pork skin by the treatments, collagen contents with combination treatment of pork skin with PMB (0.5%) resulted the highest in 43.76 g/100 g. Also the fat content at the above treatment was reduced to 11.12% compared to the other treatments. With these results of this experiment, we conclude that the enzymatic treatments were effective for the processing property of pork skin like enhancing the yield of collagen

This study was aimed to develop a gruel using rice extrudates and to evaluate physicochemical properties of Tarakjuk (milk porridge; MP) prepared with milk (M) and rice powder (RP, control), rice extrudate (RE), or enzyme treated rice extrudates (ETR). Pasting property of ETR was significantly different from those of RP or RE with significantly low peak viscosity, breakdown and setback values in the ETR sample. Viscosity of MP prepared with ETR was also significantly low, as compared to that of control (> 900 cP). The total solids and spreadability of MP with ETR was higher than those with RP or RE. Hunter color values varied significantly depending on enzyme treatment levels in ETR samples with enzyme dose-dependent increase in b-value. Average starch digestibility of ETR sample was higher by 10.2% than that of control sample. DPPH radical scavenging activities of ETR samples were greater than those of RE or RP. These results indicated that ETR could be beneficial for preparing easy-drink and diet food with higher starch digestibility and fluidity, especially for gastric tube-fed patients.

Phospholipase A2 (PLA2) catalyzes the committed step for eicosanoid biosynthesis. Two cellular PLA2 genes (iPLA2 and cPLA2) have been identified in Spodoptera exigua. PLA2 activity varied among developmental stages and different tissues. There was a positive correlation between PLA2 activities and their expression levels. Interestingly, there was significant inhibition by sPLA2 inhibitor, suggesting a presence of sPLA2 in S. exigua. An entomopathogenic bacterium, Xenorhabdus hominickii, infection suppressed expression of PLA2 genes and its extract inhibited enzyme activities. Oxindole of a bacterial metabolite of X. hominikii inhibited PLA2 activity and expression.

The present study investigated the effect of enzyme inclusion on silage quality using meta-analysis tool. A total of 16 research papers reporting the effect of enzyme application on silage quality were employed in the meta-analysis of this study. Mixed model for integrating quantitative results from multiple studies was used first to calculate the predicted error of each study. Individual error from the estimated model was the applied into standard deviation of each study to calculate the mean difference. Finally, summary effect was determined using standard mean difference (SMD) and inversed variance weighting. Mixed model analysis and SMD analysis showed the same effect patterns in all analysis items. Enzyme inclusion in silage significantly (p < 0.05) altered all silage quality characteristics investigated compared to control when enzyme was not included. Our results showed that enzyme treatment increased dry matter content, preserved crude protein effectively, and elevated water soluble carbohydrate content. However, the pH value, acetic acid, propionic acid, neutral detergent fiber, and acid detergent fiber contents in silage with enzyme inclusion were lower than those of the control.

α-Amylase 및 protease 활성을 동시에 가지는 1-B-12 균주를 된장으로부터 분리하였다. 16S rRNA 유전자 염기서열 분석결과, Bacillus licheniformis로 동정되어 B. licheniformis 1-B-12 균주로 명명하였다. 효소활성 측면에서 현미(50%), 미강(30%) 및 대두(20%)의 조합이 최적의 기질 조합 조건으로 확인되었다. 기능성이 부가된 효소식품을 제조하기 위하여 Lactobacillus casei GW140 균주를 도입하였는데, B. licheniformis 1-B-12 균주와의 혼합배양 조건은 2단발효(B. licheniformis 1-B-12 접종 > 24 h 발효 > L. casei GW140 접종 > 12 h 추가 발효)가 적합한 것으로 나타났다. 이러한 최적 조건으로 생산된 효소식품을 동결건조하여 25℃에서 보관하였을 때 시험 마지막 날인 45일까지 효소의 활성이 유지되었다.

Grains are nutritious foods that richly contain fiber, vitamins and minerals but it would be difficult to chew for some elderly people with weakened chewing ability. This study was aimed to find the appropriate conditions for reducing hardness of grains by enzyme treatment. To select appropriate enzyme, each grain (white rice, brown rice, barely, sorghum, and job’s tears) was soaked for one hour in enzyme solution at optimum activation temperature. Grains were soaked for one hour in selected enzyme solution at different concentration (0.1, 0.3, 0.5 and 0.7%) under warming condition of cooker at 70℃, steamed in electric rice pressure cooker, rapidly cooled at -18℃ for one hour, placed at 20℃ for another hour, and then hardness was measured by texture analyzer. The initial hardness of white rice, brown rice, barely, sorghum, and job’s tears were 6.5 × 104, 7.1 × 104, 7.0 × 104, 7.8 × 104 and 7.7 × 104 N/m 2 , respectively. The lowest hardness of grains, 4.0 × 104, 5.0 × 104, 4.6 × 104, 5.7 × 104 and 4.7 × 104 N/m 2 were obtained with 0.3% of enzyme D solution while the hardness was not changed significantly with higher concentration. These results suggest that some hard grains treated with enzyme would become softer than white rice, and thus these grains may be used to prepare a healthier meal for elderly people having mastication problem.

Drosophila suzukii is an economically important pest of fruit in America and Europe as well as in Asia. Sparassol (1) and methyl orsellinate (2) are produced by Sparassis cripta and S. latifolia during culture. Fumigant and contact toxicities of synthetic sparassol (1) methyl orsellinate (2) and methyl 2,4-dimethoxy-6-methylbenzoate (DMB; 3) were investigated. The possible mode of action of the compounds was pursued by assessing their inhibition activities against acetylcholinesterase (AChE) and glutathione S-transferase (GST). Little or no fumigant activity was observed from tested compounds. In contact toxicity activity, LD50 (μg/fly) values are 5.29, 1.18, 0.02 for male, and 11.14, 2.27, 0.05 for female, respectively. In enzyme inhibition assay, DMB (3) which easily synthesized from sparassol could be an alternative agent for controlling insect pest. In addition, Sparassis species could be an industrial resource of sparassol.

The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed

땅콩을 발아시켜 새싹으로 키우게 되면 땅콩에 함유되어 있는 resveratrol과 asparagine 성분이 크게 증가된다는 보고가 있다. 이러한 사실을 바탕으로 땅콩새싹 추출물을 이용한 숙취해소음료를 개발하였 다. 본 연구에서는 땅콩새싹 추출물을 이용한 숙취해소 음료가 항산화 및 알코올분해효소 활성에 미치는 영향을 조사하기 위해서, 총 페놀 함량과 DPPH radical 소거활성 및 ADH와 ALDH 효소활성을 in vitro 및 in vivo 실험을 통하여 확인하였다. in vitro 실험결과 총 페놀 함량과 DPPH radical 소거활성에 있 어서는 두 가지 모두 땅콩새싹 숙취해소음료가 제품A보다는 약 6배, 제품B보다는 약 2배 정도로 높은 활성을 보였으며, ADH와 ALDH 효소활성은 제품A 보다 각각 약 40배와 약 7배 정도로 높게 나타났으 며, 제품B 보다는 약 20배와 약 2배 정도로 높게 나타났다. in vivo 실험을 통한 혈중 알코올농도는 시 간이 경과함에 따라 빠르게 감소하는 경향을 보였고, 아세트알데히드는 시간이 경과함에 따라 대조군에 비하여 빠르게 분해되는 것을 확인할 수 있었다. 따라서 땅콩새싹을 이용한 숙취해소 음료는 시중에 판 매되고 있는 숙취해소음료와 비교해 볼 때 항산화 및 알코올분해효소 활성에 효과적이라 판단된다.

본 연구는 효소식품과 효소표방식품 (기타가공식품, 음료베이스, 기타발효음료, 액상차) 98건에 대한 α-아밀라아제, β-아밀라아제 활성과 당 함량을 조사하였다. 효소식품과 기타가공품의 α-아밀라아제 활성은 각각 4.9~53,854.6 U/g, 2.9~1,182.7 U/g으로 같은 유형간에 큰 차이가 있었다. 발효식품의 α-아밀라아제 활성은 각각 0.1~1.7 U/g이었다. 효소식품, 기타가공품 그리고 발효식품의 β-아밀라아제 평균 활성은 각각 126.0 U/g, 5.6 U/g, 10.5 U/g으로 효소표방식품은 효소식품보다 훨씬 낮은 활성을 나타냈다. 평균 당 함량은 효소식품 22.4 g/100 g, 기타가공품 14.8 g/100 g, 음료베이스 46.9 g/100 g, 기타발효음료류 41.1 g/100 g, 액상차 39.5 g/100 g으로 발효식품에서 높은 당 함량을 나타냈다. α-아밀라아제 활성과 유당 함량은 효소식품에서 통계적으로 강한 상관관계(r = 0.644)를 나타냈고 기타가공식품에서는 매우 강한 상관관계(r = 0.903)를 나타냈다. β-아밀라아제 활성과 유당 함량은 효소식품에서 통계적으로 강한상관관계(r = 0.648)를 나타냈고 기타가공식품에서는 강한상관관계(r = 0.757)를 나타냈다. 효소식품과 기타가공품에서 α-아밀라아제 활성과 β-아밀라아제 활성 사이에는 매우 강한 상관관계(r = 0.869, r = 0.760)를 나타냈다. 즉, α-아밀라아제 활성과 β-아밀라아제 활성 사이에 비례관계가 성립함을 알 수 있었다.

The objective of this study was to evaluate the efficiency of sperm cryosurvival in boar sperm separated by Percoll containing antioxidant enzymes. The boar semen was collected into a pre-warmed (37℃) thermos bottle by gloved-hand method and was separated by 65% Percoll with superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) before freezing. The frozen sperm was thawed at 38.5℃ for 45 sec in water-bath for sperm characteristic analysis. The sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction, Rhodamine123/PI double staining for mitochondrial integrity and were analyzed using flow cytometry. In results, sperm viability, acrosome reaction and mitochondrial integrity were improved in separated sperm groups compared with unseparated sperm by Percoll (UP) group. Especially, viability was significantly higher in sperm separated by Percoll containing 400 IU CAT group compared with other groups (P<0.05). And acrosome reaction was decreased in sperm separated by Percoll with 300 IU SOD, 400 IU CAT and 0.5 mM GSH groups compared with other groups, however, there were no significantly difference mitochondrial integrity among sperm separated by Percoll with antioxidant enzymes. In conclusion, we suggest that use of Percoll containing antioxidant enzymes for sperm separation will be beneficial for sperm cryopreservation in pigs.

고추 추출액의 수율 및 생리적 기능성을 향상시키기 위하 여 고추에 cellulase(C 처리군), pectinase(P 처리군), amylase(A 처리군)를 각각 또는 이들 효소들을 혼합(CP, CA, PA 및 CPA 처리군) 첨가하여 2~8시간동안 가수분해시킨 후 가열처리하 고, 여과시킨 추출액에 대하여 이화학적 및 관능적 평가를 측 정하였다. 효소 처리 고추 추출액의 수율은 효소 처리를 하지 않았을 때 38.84% 정도로 매우 낮게 나타났지만, 효소처리군 이 높은 수율을 보였고, 효소 단독처리군보다는 병용처리 시 수율이 증가하였으며, 효소 처리 시간이 길어질수록 유의적으 로 수율이 증가하는 것으로 나타났다. 특히 cellulase + pectinase + amylase(CPA) 복합처리군은 추출수율이 74.37%까지 증가 하였다. 가용성 고형분의 함량변화는 대조군의 경우에는 8.51% 를 나타내었으나, 효소처리군은 대조군보다 높은 함량을 나 타내었으며, CA 혼합처리군과 CPA 혼합처리군이 가장 높은 가용성 고형분 함량을 보였다. 환원당의 함량 또한 효소 처리에 의하여 증가하는 것으로 나타났다. 효소 처리에 의한 색도 의 변화에서는 대조군과 효소처리군 간에 색도의 변화를 보 이지는 않았다. 효소 처리한 고추 추출액에 대한 관능검사 결 과에서는 전반적으로 효소 처리를 하지 않은 대조군에 비하 여 효소 처리 시 기호도 면에서 우수한 것으로 나타나, 고추 추출물 제조를 위한 효소 처리는 수율 및 기호도 증진을 위한 좋은 방법으로 사료된다.

Insect immunity is innate and consists of cellular and humoral immune responses. Cellular immune response usually requires hemocyte-spreading behavior, which is accompanied by cytoskeletal rearrangement. A glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), catalyzes an oxidation reaction of glyceraldehyde-3-phosphate to 1,3-biphosphoglycerate in the cytosol. Another function of GAPDH in mammalian cell is to bind C-terminal α-tubulin to facilitate cytoskeletal arrangement. An immunoprecipitation (IP) of viral protein, CpBV-CrV1, against hemocyte protein lysate revealed that CpBV-CrV1 binds to GAPDH, identified by MALDI-TOF analysis. RNA interference (RNAi) of GAPDH significantly suppressed cellular immune response, but neither RNAi of hexokinase nor aldolase suppressed the cellular immune response. A common molecular motif of CpBV-CrV1 and a-tubulin at C-terminal region supported the IP analysis. To test the role of α-tubulin motif in CpBV-CrV1, point mutations of CpBV-CrV1 were applied and resulted in loss of the biological activity of CpBV-CrV1. Furthermore, an immunofluorescence assay indicates CpBV-CrV1 colocalized with a-tubulin in hemocytes collected from Plutella xylostella parasitized by Cotesia plutellae possessing C. plutellae bracovirus (CpBV). This result suggests that GAPDH plays a critical role in hemocyte-spreading behavior during immune challenge, and it is a molecular target of the pathogenic virus.