Effect of biobed composition, moisture, and temperature on the degradation of pesticides

Biobeds retain and degrade pesticides through the presence of a biobed mixture consisting of straw, peat, and soil. The effects of biobed composition, moisture content, and temperature on pesticide degradation were investigated in laboratory studies. Straw produced the main microbial activity in the biobed mixtures as strong positive correlations were observed between straw, respiration, and phenoloxidase content. Most pesticides investigated were dissipated by cometabolic processes, and their dissipation was correlated with respiration and/ or phenoloxidase content. More pesticides were more dissipated at biobed moisture levels of 60% water holding capacity (WHC) than at 30% and 90% WHC, while 20 degrees C gave higher dissipation rates than 2 and 10 degrees C. A straw:peat:soil ratio of 50:25:25% v/v is recommended in field biobeds since this produces high microbial activity and low pH, favorable for lignin-degrading fungi and phenoloxidase activity.     

Structure-activity relationships of derivatives of fusapyrone, an antifungal metabolite of Fusarium semitectum

Fusapyrone (1) and deoxyfusapyrone (2) are two 3-substituted-4-hydroxy-6-alkyl-2-pyrones isolated from Fusarium semitectum that have considerable antifungal activity against molds. Because of their low zootoxicity and selective action they are potentially utilizable along with biocontrol yeasts for control of postharvest crop diseases. Seven derivatives of 1 (3 and 5-10) and one derivative of 2 (4) were obtained by chemical modifications of the glycosyl residue, the 2-pyrone ring, the aliphatic chain, or a combination thereof, and a structure-activity correlation study was carried out with regard to their zootoxicity and antifungal activity. Derivatives 7-10, as well as 1, were slightly zootoxic in Artemia salina (brine shrimp) bioassays, whereas pentaacetylation of 1 into 3, 5, and 6 resulted in a strong increase in toxicity. Compound 4, the tetraacetyl derivative of 2, was as toxic as 2. Because the structural changes of 1 that resulted in an increase of biological activity in A. salina bioassay were those that affected mainly the water solubility of the molecule, it appears that toxicity is related to hydrophobicity. Compounds 1 and 2 showed strong antifungal activity toward Botrytis cinerea, Aspergillus parasiticus, and Penicilliun brevi-compactum (minimum inhibitory concentration at 24 h = 0.78-6.25 microg/mL). Among derivatives 3-10, only compounds 7, 9, and 10 retained some activity, limited to B. cinerea and at high concentration (25-50 microg/mL). None of the compounds 1-10 inhibited the growth of the biocontrol yeasts Pichia guilliermondii and Rhodotorula glutinis at the highest concentration tested (50 microg/mL).     

微酸性电解水对虾仁的杀菌效果及其动力学

为探明微酸性电解水(slightly acidic electrolyzed water,SAEW)对南美白对虾(Litopenaeus vannamei)虾仁表面杀菌效果及其动力学规律,选取料液比1:4、1:10、1:20,将虾仁分别浸洗杀菌2、5、10 min,对杀菌过程中虾仁表面及SAEW残存液中总菌落数,与有效氯质量浓度(available chlorine concentration,ACC)的变化进行测定,建立ACC衰减与微生物减灭的动力学模型,并通过颜色、硬度、pH值,及水分横向弛豫行为分析,探讨SAEW杀菌处理对虾仁品质的影响。结果表明SAEW对虾仁表面大肠杆菌有较强杀菌效果,并随处理时间的延长、作用量的增大,SAEW的杀菌效力不断增强,处理5 min时,随着料液比的增加,虾仁表面菌落数从最初的6.6 l(CFU/mL),依次降到5.0、4.7、4.4 lg(CFU/mL),料液比为1:20时,静置浸洗10 min后,虾仁表面菌落数由最初的6.6 lg(CFU/mL)降至3.9 lg(CFU/mL);同时SAEW浸洗液中残存菌落数也随处理时间的延长、作用量的增大,而不断减少,在处理2、5和10 min时,SAEW中的残存菌落数分别为4.18、3.47、2.78 lg(CFU/mL),处理时间为5 min时,随料液比的增加,SAEW中的残存菌落数分别为3.47、2.78、2.65 lg(CFU/mL);同时SAEW中ACC的消耗随处理时间的延长、而不断变大,杀菌处理5、10 min时,ACC质量浓度从初始的20.53 mg/L分别降至7.79、10.97 mg/L。动力学分析表明:SAEW在杀灭虾仁表面大肠杆菌的过程中,ACC的衰减可以用一级动力学模型描述,拟合后决定系数R2均大于0.9,而微生物的减灭遵循更为复杂的动力学模型;此外经过SAEW杀菌处理的虾仁,其颜色、pH值、硬度、以及水分的横向弛豫行为,与未处理样品相比,基本没有显著性变化。相关结果能为SAEW在水产品加工过程中的应用提供技术指导,同时也有助于SAEW杀菌技术理论的完善。     

Latent polyphenol oxidases from sago log (Metroxylon sagu): partial purification, activation, and some properties

Latent polyphenol oxidase (LPPO), an enzyme responsible for the browning reaction of sago starches during processing and storage, was investigated. The enzyme was effectively extracted and partially purified from the pith using combinations of nonionic detergents. With Triton X-114 and a temperature-induced phase partitioning method, the enzyme showed a recovery of 70% and purification of 4. 1-fold. Native PAGE analysis of the partially purified LPPO revealed three activity bands when stained with catechol and two bands with pyrogallol. The molecular masses of the enzymes were estimated by SDS-PAGE to be 37, 45, and 53 kDa. The enzyme showed optimum pH values of 4.5 with 4-methylcatechol as a substrate and 7.5 with pyrogallol. The LPPO was highly reactive toward diphenols and triphenols. The activity of the enzyme was greatly enhanced in the presence of trypsin, SDS, ethanol, and linoleic acid.     

The Quality of Stemwood of Pinus sylvestris in an Alkalised Environment

The impact of long-term dust pollution emitted from a cement plant on soil chemistry, and the concentrations of plant nutrients, lignin, cellulose and hemicellulose in the stemwood of 80–85-year-old Pinus sylvestris was investigated at different distances from the emission source. It was found that alkaline cement dust (pH 12.3–12.6) emissions for over 40 years resulted in an alkalisation (pH 6.7–7.9) of the polluted soil compared to a pH value of 3.8 in unpolluted soil. There were also nutrient imbalances in the soil, as well as certain disturbances in mineral nutrition processes and accumulation of nutrients in the tree stems. The average concentrations of K, Ca and Mg in stems were higher and those of N and P lower than in the unpolluted area. The lignin (L) content in stemwood increased, hemicellulose (Hc) decreased, while cellulose (Ce) did not change. A variation in the partitioning of L, Ce, Hc and nutrients between different sections of stems and between trees from different sample plots was found. L, Ce and Hc were not related to the internal K, Ca and Mg concentrations. Correlations were established between L, Ce, or Hc content and C content, and between L and Hc content in stem tissues. The contents of wood components were not related to N or P in the alkalised areas, but seemed to be more associated with P than with N. Alterations in the arbitrarily chosen ratio L/(Ce + Hc) indicated changes in wood quality, and a negative correlation with N/P was found in stem tissue in the polluted area, while positive correlations with N/Mg and Ca/Mg were found in the control area.     

Inhibitory effect of mimosine on polyphenoloxidase from cephalothoraxes of Pacific white shrimp (Litopenaeus vannamei)

The inhibitory effect of mimosine on polyphenoloxidase (PPO) from the cephalothorax of Pacific white shrimp (Litopenaeus vannamei) was studied. Mimosine showed inhibitory activity toward PPO from white shrimp with an apparent molecular weight of 210 kDa as evidenced by the decrease in the activity staining band, as compared to the control. An inhibition kinetic study revealed that mimosine exhibited the mixed type reversible inhibition on PPO from white shrimp with a Ki value of 3.7 mM. Mimosine showed copper (Cu2+) reduction and chelating capacity in a dose dependent manner. Mimosine could react with the intermediate browning product, thereby rendering lower red-brown color formation. Therefore, mimosine could inhibit PPO by different modes of inhibition and could be used to prevent melanosis formation in Pacific white shrimp.     

Determination of total sulfite in shrimp, potatoes, dried pineapple, and white wine by flow injection analysis: collaborative study

A method for the determination of total sulfite in shrimp, potatoes, dried pineapple, and white wine by flow injection analysis (FIA) was collaboratively studied by 8 laboratories. In the method, the sample solution is reacted with sodium hydroxide to liberate aldehyde-bound sulfite. The sample stream is acidified to produce SO2 gas, which diffuses across a Teflon membrane in the gas diffusion cell into a flowing stream of malachite green. The degree of discoloration of the malachite green is proportional to the amount of sulfite in the sample solution. Red wine was included in the study but interlaboratory precision for these samples was not satisfactory and correlation with Monier-Williams results was poor. The present method is not recommended for use with these samples. For shrimp, potatoes, dried pineapple, and white wine, average reproducibility (RSDR) of results was 25% for samples at 10 ppm SO2 and 10% for samples at greater than 50 ppm. Overall average reproducibility was 14%. Recoveries of sulfite added to samples averaged 80%. Comparison of FIA with the Monier-Williams method indicated comparable results by the 2 methods. The FIA method has been adopted official first action for determination of greater than or equal to 5 ppm total sulfite in shrimp, potatoes, dried pineapple, and white wine.     

Presence and dehydration of ikaite, calcium carbonate hexahydrate, in frozen shrimp shell.

Ikaite, calcium carbonate hexahydrate, has by means of X-ray diffraction analyses of frozen samples been identified as the mineral component of the white spots formed in the shell of frozen shrimp during storage. When the shrimp thaw and the shell material is dried and kept at room temperature, ikaite rapidly transforms into a mixture of anhydrous calcium carbonate forms. X-ray diffraction analyses and Raman spectra of synthetic ikaite as well as the dehydration product confirm the assignments, and the rate constant for dehydration is approximately 7 x 10(-)(4) s(-)(1) at ambient temperature. Differential scanning calorimetry showed that dehydration of synthetic ikaite is an entropy-driven, athermal process and confirms that a single first-order reaction is rate-determining. Ikaite is found to be stable in aqueous solution at temperatures below 5 degrees C and in the shell of frozen shrimps but decomposes on thawing to form anhydrous calcium carbonates.     

Diphenol activation of the monophenolase and diphenolase activities of field bean (Dolichos lablab) polyphenol oxidase

This paper reports a study on the hydroxylation of ferulic acid and tyrosine by field bean (Dolichos lablab) polyphenol oxidase, a reaction that does not take place without the addition of catechol. A lag period similar to the characteristic lag of tyrosinase activity was observed, the length of which decreased with increasing catechol concentration and increased with increasing ferulic acid concentration. The activation constant K(a) of catechol for ferulic acid hydroxylation reaction was 5 mM. The kinetic parameters of field bean polyphenol oxidase toward ferulic acid and tyrosine were evaluated in the presence of catechol. 4-Methyl catechol, L-dihydroxyphenylalanine, pyrogallol, and 2,3,4-trihydroxybenzoic acid, substrates with high binding affinity to field bean polyphenol oxidase, could stimulate this hydroxylation reaction. In contrast, diphenols such as protocatechuic acid, gallic acid, chlorogenic acid, and caffeic acid, which were not substrates for the oxidation reaction, were unable to bring about this activation. It is most likely that only o-diphenols that are substrates for the diphenolase serve as cosubstrates by donating electrons at the active site for the monophenolase activity. The reaction mechanism for this activation is consistent with that proposed for tyrosinase (Sanchez-Ferrer, A.; Rodriguez-Lopez, J. N.; Garcia-Canovas, F.; Garcia-Carmona, F. Biochim. Biophys. Acta 1995, 1247, 1-11). The presence of o-diphenols, viz. catechol, L-dihydroxyphenylalanine, and 4-methyl catechol, is also necessary for the oxidation of the diphenols, caffeic acid, and catechin to their quinones by the field bean polyphenol oxidase. This oxidation reaction occurs immediately with no lag period and does not occur without the addition of diphenol. The kinetic parameters for caffeic acid (K(m) = 0.08 mM, V(max) = 32440 u/mg) in the presence of catechol and the activation constant K(a) of catechol (4.6 mM) for this reaction were enumerated. The absence of a lag period for this reaction indicates that the diphenol mechanism of diphenolase activation differs from the way in which the same o-diphenols activate the monophenolase activity.     

超高压前处理对虾仁冻藏期品质的影响

为研究超高压辅助脱壳所得到的中华管鞭虾虾仁在冻藏期的品质变化,以虾仁色泽、质构、肌原纤维蛋白相对含量、表面疏水性、总巯基相对含量和Ca²⁺-ATPase活性为指标,探讨超高压前处理对虾仁品质的影响。结果表明,冻藏6个月后虾仁L^*值、a^*值和b^*值均有所变化,与对照组相比,超高压前处理对虾仁L^*值的影响不显著,但能延缓冻藏后期a^*值的增加,对保持虾肉色泽的稳定有一定作用;超高压前处理对虾仁的咀嚼性和弹性影响不显著,但能增加虾仁的硬度。超高压前处理使冻藏初期虾仁肌原纤维蛋白的表面疏水性增加,而对其溶解性、巯基含量及Ca²⁺-ATPase活性的影响不显著;冻藏后期由于虾仁蛋白冷冻变性,致使肌原纤维蛋白溶解性、巯基含量及Ca²⁺-ATPase活性下降。综上所述,超高压辅助脱壳在一定程度上有利于保持冻虾仁的色泽和硬度。本研究结果为利用超高压技术辅助脱除中华管鞭虾虾壳,生产冷冻虾仁提供了新思路。     

Polyphenol Oxidase in Wheat Grain: Whole Kernel and Bran Assays for Total and Soluble Activity

Color is a key quality trait of wheat products, and polyphenol oxidase (PPO) is implicated as playing a significant role in darkening and discoloration. In this study, total and soluble PPO activities were characterized in whole kernel assays and bran extracts. In whole kernel assays similar to AACC Approved Method 22–85, four wheat cultivars were ranked the same for both total and soluble (leached) PPO activity with L‐DOPA (diphenol) as the substrate. Total kernel PPO activity was much greater than soluble PPO activity in three hexaploid wheat cultivars, indicating that insoluble PPO was the major contributor to kernel PPO measurements. Tyrosine (monophenol) was an excellent PPO substrate in kernel assays as expected but had no activity as a substrate for soluble PPO. However, soluble PPO activity with tyrosine was activated by the addition of the diphenols chlorogenic acid and caffeic acid. When PPO was assayed in homogenized bran, 89–95% of total PPO activity remained insoluble, associated with the bran particles. The kernel assay detected <2% of PPO measured in an equivalent amount of homogenized bran. However, total PPO activity was 2‐fold higher in Klasic than in ID377s, both when measured in the kernel assay and in homogenized bran, indicating that the kernel assay was an accurate predictor of relative total extracted PPO activity in these two cultivars. Adding detergents (0.1% SDS plus 0.2% NP‐40) to the bran extraction buffer increased both soluble and insoluble PPO activity. Results indicate that relative PPO activities among wheat cultivars are similar in whole kernel and kernel leachate assays, and that the predominant insoluble fraction of PPO, which is relatively uncharacterized, may be largely responsible for wheat product discoloration.     

Role of beta-adrenoceptor signaling and AMP-activated protein kinase in glycolysis of postmortem skeletal muscle

Postmortem glycolysis is directly linked to the incidences of PSE (pale, soft, and exudative) and DFD (dark, firm, and dry) meats, which cause significant economic loss to the meat industry. However, mechanisms controlling postmortem glycolysis are unclear. The objective of this study was to determine the role of beta-adrenoceptor signaling and AMP-activated protein kinase (AMPK) in postmortem glycolysis. Eighteen 2 month old C57BL/6J female mice were randomly separated into three groups. Group I received an intraperitoneal injection of saline solution only and served as the control; group II received a saline injection and then were forced to swim for 1 min; and group III received an injection of propranolol (1 mg/kg) in saline. In addition, six C57BL/6J female AMPK knockout mice were assigned to group IV, which received a saline injection and were forced to swim for 1 min. The longissimus dorsi muscle was sampled at 0, 1, and 24 h postmortem for pH and enzyme activity measurements. The objective is to elucidate the roles of beta-adrenoceptor signaling and AMPK in the glycolysis of postmortem muscle. Results showed that AMPK activity had a major role in determining the ultimate muscle pH, with an ultimate pH for control mice of 6.16 and AMPK knockout mice of 6.48. The beta-adrenoceptor signaling is essential for initial rapid glycolysis. Blocking beta-adrenoceptor signaling prevented the initial pH decline induced by stress. Activation of beta-adrenoceptor signaling due to preslaughter stress activates glycogen phosphorylase, resulting in a rapid glycolysis shortly after slaughter. On the other hand, the activation of AMPK is important for maintaining the activity of glycogen phosphorylase and pyruvate kinase, leading to a sustained glycolysis and a low ultimate pH.     

Modelling in situ activities of enzymes as a tool to explain seasonal variation of soil respiration from agro-ecosystems

Understanding in situ enzyme activities could help clarify the fate of soil organic carbon (SOC), one of the largest uncertainties in predicting future climate. Here, we explored the role of soil temperature and moisture on SOM decomposition by using, for the first time, modelled in situ enzyme activities as a proxy to explain seasonal variation in soil respiration. We measured temperature sensitivities (Q₁₀) of three enzymes (β-glucosidase, xylanase and phenoloxidase) and moisture sensitivity of β-glucosidase from agricultural soils in southwest Germany. Significant seasonal variation was found in potential activities of β-glucosidase, xylanase and phenoloxidase and in Q₁₀ for β-glucosidase and phenoloxidase activities but not for xylanase. We measured moisture sensitivity of β-glucosidase activity at four moisture levels (12%–32%), and fitted a saturation function reflecting increasing substrate limitation due to limited substrate diffusion at low water contents. The moisture response function of β-glucosidase activity remained stable throughout the year. Sensitivity of enzymes to temperature and moisture remains one of the greatest uncertainties in C models. We therefore used the response functions to model temperature-based and temperature and moisture-based in situ enzyme activities to characterize seasonal variation in SOC decomposition. We found temperature to be the main factor controlling in situ enzyme activities. To prove the relevance of our modelling approach, we compared the modelled in situ enzyme activities with soil respiration data measured weekly. Temperature-based in situ enzyme activities explained seasonal variability in soil respiration well, with model efficiencies between 0.35 and 0.78. Fitting an exponential response function to in situ soil temperature explained soil respiration to a lesser extent than our enzyme-based approach. Adding soil moisture as a co-factor improved model efficiencies only partly. Our results demonstrate the potential of this new approach to explain seasonal variation of enzyme related processes.     

利用边缘辅助分割网络提取稻虾共作养殖田

为确保稻虾共作的安全生产、社会供应以及政府政策的有效制定，须准确获取稻虾共作种植面积、空间分布及变化信息。现有稻虾养殖田提取方法以中分辨率时序影像为数据源，提取结果存在边界粗糙且噪声较多等问题。为获取精度高、边界规整的稻虾养殖田提取结果，该研究以高分辨率影像为数据源，提出一种基于边缘辅助任务的深度学习语义分割模型——edge assisted segmentation network（EASNet）。该模型首先将稻虾养殖田特有的边缘“虾沟”作为一种辅助信息在设计的边缘辅助模块单独分割，然后将该模块的输出与主任务分割模块输出进行融合，使主任务既增强了稻虾养殖田边界结构信息，又能学习到稻虾养殖田特有的空间及语义信息。试验结果表明，在边缘辅助模块的增强下，稻虾养殖田分割结果更完整，边界更清晰，其语义精度的交并比和边界精度的F1分数分别提升了1.5%、5.8%。整体语义精度的召回率、交并比、F1分数分别达到0.970、0.964、0.930，边界精度的召回率、F1分数达到0.864、0.859，松弛边界精度的召回率、F1分数达到0.876、0.913。将训练好的EASNet模型应用到盱眙县全域，得到2020年盱眙县稻虾养殖田空间分布图，在与传统的水体季相差异法、随机森林方法提取的稻虾养殖田结果的对比中，该方法取得了总体精确度为96.71%及Kappa系数为0.934的最优结果，为基于深度学习的稻虾养殖田提取方面的应用提供参考。     

Can commercial mulches be reservoirs of invasive earthworms? Promotion of ligninolytic enzyme activity and survival of Amynthas agrestis (Goto and Hatai, 1899)

Amynthas agrestis is an exotic, invasive earthworm in North America that has been associated with horticulture settings as well as damage to forest soil. An experiment was conducted to find out whether A. agrestis, an earthworm commonly found in mulches in Vermont, stimulates ligninolytic enzymes in the presence of commercial wood mulches. Mesocosms filled with a sandy loam soil were topped with either spruce, cedar or pine mulch. Half of the mesocosms received juvenile A. agrestis, the other half did not. After 7 weeks soils were analyzed for phenoloxidase and peroxidase activity. Most A. agrestis survived and developed into adults during the incubation period. Significantly greater phenoloxidase activity was detected in soils with A. agrestis than without earthworms. Mean (standard deviation) phenoloxidase activities in the presence of A. agrestis were 0.15 (±0.10), 1.14 (±0.46), 2.71 (±0.98) μmol g⁻¹ h⁻¹ for pine, spruce and cedar respectively, and 0.012 (±0.023), 0.25 (±0.25), 0.78 (±0.45) μmol g⁻¹ h⁻¹ in the absence of A. agrestis. There was significantly greater peroxidase activity for the pine and spruce treatment when earthworms were present. Mean peroxidase activities were 0.47 (±0.21), 0.94 (±0.29), 1.20 (±0.77) μmol h⁻¹ g⁻¹ soil for pine, spruce and cedar, respectively for soils with A. agrestis and 0.15 (±0.10), 0.37 (±0.10), 0.63 (±0.30) μmol h⁻¹ g⁻¹ soil in the absence of earthworms. The increased ligninolytic activity in combination with successful maturation of juveniles into adult A. agrestis suggests that mulch can be habitat for these invasive earthworms. This finding is supported by a survey of master gardeners in Vermont and New Hampshire 20% of whom reported to have seen these earthworms mainly in their gardens and mulched beds.     

Oxidative coupling activity in soil extracts

An assay was developed in which 50 mm citrate buffer was used to extract catalysts responsible for oxidative coupling reactions in soil. The assay was based on the formation of the dimerized quinone from the oxidative coupling of 2,6-dimethoxyphenol. Oxidative coupling activity was destroyed by heating the extracts for 15 min at 100°C and was inhibited by H₂O₂ (98% at 10 mm), KCN (96% at 10 mm), dithiothreitol (100% at 0.1 mm) and 2,3-dimercapto-1-propanol (100% at 0.1 mm). In the assay, a pH of 7.0 and a temperature of 55°C were determined as optimal. Freezing the soil extracts provided the best protection against activity loss, whereas storage caused a decline in oxidative coupling activity as a function of increasing temperatures (5–50°C). If soil had been supplemented with sucrose before extraction, activity increased. Soil extracts reacted with syringaldazine, benzidine, o-dianisidine, guaiacol, and p-cresol, substrates previously used to detect phenoloxidase enzymes.     

南极磷虾捕捞初期适宜挤压脱壳工艺参数

为了验证南极磷虾挤压脱壳设备的生产效果、完善并优化船上挤压脱壳生产工艺,该文开展了不同进料速度、原料放置时间和预冷时间对得肉率和虾壳残留率影响的研究。结果表明,对于刚捕捞后未经处理的磷虾,试验设备脱壳效果理想,得肉率约为25%、虾壳残留约为5%;放置时间在120 min以内的磷虾均可用作脱壳磷虾肉的生产,越新鲜的磷虾脱壳后得到的磷虾肉品质越好;对原料进行预冷来延长品质保持时间是保证产品质量的有效方法;冷冻后磷虾得肉率明显降低,实际生产中不建议采用。研究结果可为南极磷虾脱壳技术的应用和产业化提供参考。     

Postmortem changes in pork muscle protein phosphorylation in relation to the RN genotype

Postmortem changes in pork muscle protein phosphorylation in relation to the RN(-) genotype were investigated using one-dimensional gel electrophoresis and a phosphor specific staining. The phosphorylation levels of several protein bands were found to be affected by the RN(-) genotype and to change during postmortem development. Glycogen phosphorylase, phosphofructokinase, and pyruvate kinase were found in protein bands affected by the RN(-) genotype, and the phosphorylation profile indicates that part of the increased rate and extended pH decline of the RN(-) genotype could be a consequence of phosphorylation of these key enzymes during the postmortem metabolism. The results illustrate that the protein phosphorylation level of the muscle proteins could be interpreted as a global metabolic fingerprint containing information about the activity status of the enzymes in the postmortem metabolism.     

Novel fibrinolytic enzyme in fermented shrimp paste, a traditional asian fermented seasoning

A novel fibrinolytic enzyme was purified from fermented shrimp paste, a popular seasoning used in Asian countries. The enzyme is a monomer with an apparent molecular weight of 18 kDa, and it is composed primarily of beta-sheet and random coils. The N-terminal amino acid sequence was determined to be DPYEEPGPCENLQVA. It is a neutral protease with an optimal activity from pH 3 to 7. No inhibition was observed with PMSF, Pepstatin A, E64, and 1,10-phenanthroline, but the enzyme was slightly inhibited by EDTA and Cu(2+). It was relatively specific to fibrin or fibrinogen as a protein substrate, yet it hydrolyzed none of the plasma proteins in the studies. In vitro, the enzyme was resistant to pepsin and trypsin digestion. It also had an anticoagulant activity measured with activated partial thrombin time and prothrombin time tests. The novel fibrinolytic enzyme derived from traditional Asian foods is useful for thrombolytic therapy. In addition, this enzyme has a significant potential for food fortification and nutraceutical applications, such that its use could effectively prevent cardiovascular diseases.