Cadaverine Involved in Urate Degrading Activity (Uricase Activity) in Soybean Radicles

Cadaverine, a 5-carbon diamine, was identified as the cofactorof uricase activity previously found in soybean seedlings. Thesubstance purified from freeze dried hypocotyls was subjectedto liquid chromatography, mass spectrometry, ¹H- and ¹³C-nuclearmagnetic resonance spectrometry for identification. The concentrationsof cadaverine in 3-day-old radicles and hypocotyls were 2.37mM and 5.09 mM, respectively. Other polyamine concentrationswere low. Biogenic polyamines (cadaverine, putrescine, spermidineand spermine) functioned as cofactors, whereas conjugated polyamines(tyramine and histamine) and amino acids had no effect. Theaddition of catalase to the assay system counteracted the effectof cadaverine. Peroxide at appropriate concentrations actedlike cadaverine with an identical K_m value, suggesting thaturate degrading activity can be ascribed to the diamine oxidase-peroxidasesystem. (Received October 19, 1982; Accepted December 23, 1982)     

Oxidase-Peroxidase Method of Ethanol Assay in Fermented Musts and Wine Products

A new alcohol oxidase-peroxidase method of ethanol assay in fermented musts and wine products is described and compared to conventional methods routinely used in winemaking. The sensitivity, accuracy, and reliability of this method were determined. The results of ethanol determination in fermented musts and wines correlated well with the data obtained by refractometry (correlation coefficient R = 0.9595, p < 0.0001) and densitometry (correlation coefficient R = 0.9384, p < 0.0001). The proposed method is less time- and labor-consuming and allows simultaneous analysis of a series of wine samples.     

Hydrolysis of Soybean Proteins with Kamchatka Crab Hepatopancreas Enzyme Complex

To perform hydrolysis with the enzyme complex from the hepatopancreas of the Kamchatka crab, a protein mixture was isolated from soybean meal by extraction at alkaline pH values. Extractable low-molecular impurities were removed by ultrafiltration and precipitation of proteins with alcohol. The amino acid composition of the obtained protein extract turned out to be similar to the composition of the fish meal traditionally used in the production of fish feeds. Analysis of the products of fermentolysis by DDS-electrophoresis, HPLC, and mass spectrometry showed a high degree of hydrolysis of soybean proteins. Depending on the time of fermentolysis, the hydrolysates contained up to 60% (18 h of hydrolysis) of free amino acids (the fraction of the weight of the hydrolyzed protein mixture) and short peptides (2–20 amino acid residues).     

Extracellular Enzyme Activity in a Willow Sewage Treatment System

This paper presents the results of studies on the activity of extra-cellular enzymes in soil-willow vegetation filter soil which is used in the post-treatment of household sewage in an onsite wastewater treatment system located in central Poland. Wastewater is discharged from the detached house by gravity into the onsite wastewater treatment system. It flows through a connecting pipe into a single-chamber septic tank and is directed by the connecting pipe to a control well to be further channelled in the soil-willow filter by means of a subsurface leaching system. Soil samples for the studies were collected from two depths of 5?cm and 1?m from three plots: close to the wastewater inflow, at mid-length of the plot and close to its terminal part. Soil samples were collected from May to October 2009. The activity of the extra-cellular enzymes was assayed by the fluorometric method using 4-methylumbelliferyl and 7-amido-4-methylcoumarin substrate. The ranking of potential activity of the assayed enzymes was the same at 5?cm and 1?m soil depths, i.e. esterase?>?phosphmomoesterase?>?leucine-aminopeptidase?>?β-glucosidase?>?α-glucosidase. The highest values of enzymatic activity were recorded in the surface layer of the soil at the wastewater inflow and decreased with increasing distance from that point.     

Food Processing Characteristics of Soybean Proteins

Relation between sulfhydryl groups in soybean proteins and the physical properties of tofu was studied. Changes in the amount of sulfhydryl groups by heating or treatment with urea were more rapid in 11S protein as compared with 7S protein. Moreover, by changing the amount of sulfhydryl groups in proteins by N-ethylmaleimide, 2-mercapto-ethanol and dithiothreitol, the physical properties of tofu from 11S protein were more significantly effected than that from 7S protein. Namely, tofu-gel from 11S. protein got harder and stronger as the amount of sulfhydryl groups increased.

The results may suggest that tofu prepared from IIS protein has more disulfide bonds in its gel than that from 7S protein.     

Cadaverine, an Essential Diamine for the Normal Root Development of Germinating Soybean (Glycine max) Seeds

When the polyamine content of soybean (Glycine max) seeds was examined during the early stages of germination, the major polyamine in the cotyledons was found to be spermidine, followed by spermine; while very low concentrations of cadaverine were found. In the embryonic axes, however, cadaverine was the main polyamine and its content markedly increased 24 hours after the start of germination. When the germination of the seeds was performed in the presence of 1 millimolar α-difluoromethylornithine (DFMO), a marked decrease in the cadaverine content was found, while the other polyamines were not affected. This decrease of the cadaverine content was already noticeable after the first hours of germination. In the presence of DFMO, a pronounced elongation in the roots of the seedlings and a marked decrease in the appearance of secondary roots as compared with controls, was observed. This abnormal rooting of the seedlings caused by DFMO was almost completely reverted by the addition of 1 millimolar cadaverine. The latter also increased the appearance of secondary roots in the seedlings. The decrease in the cadaverine content produced by DFMO could be traced to a strong inhibition of lysine decarboxylase. A temporal correlation between the increase in cadaverine content and the increase in lysine decarboxylase activity was found. Both reached a maximum at the second day of germination. The activity of diamine oxidase, the cadaverine degrading enzyme, started to increase at the third day and reached a maximum between the fourth and fifth day of germination. DFMO increased the activity of diamine oxidase by about 25%. Hence, the large decrease in cadaverine content produced by DFMO has to be attributed to the in vivo suppression of lysine decarboxylase activity. Ornithine decarboxylase activity was also suppressed by DFMO, but putrescine and spermidine contents were not affected, except in the meristematic tissues. The obtained results suggest an important role for cadaverine in the normal rooting process of soybean seedlings.     

大豆植株不同冠层种子活力及其萌发中抗氧化酶活性

以吉农15号和吉农24号大豆(Glycine max)品种为材料,根据植株高度平均分为下部、中下部、中部、中上部和上部5个冠层,分析不同冠层种子百粒重、种子活力及萌发7天子叶中抗氧化酶活性和丙二醛含量的变化。结果表明,随着植株冠层的升高,大豆百粒重呈先略上升后逐渐下降的趋势,上部冠层种子的百粒重最小。大豆植株下部与中下部冠层种子的发芽率和发芽势较高;并且该两层的种子活力指数显著高于中上部与上部冠层,种子萌发后幼苗的茎粗也显著大于中上部及上部冠层幼苗。下部与中下部冠层种子萌发时子叶中的抗氧化酶活性高于上部,而丙二醛含量低于上部,说明该两层的大豆种子活力高,萌发后幼苗健壮且子叶新陈代谢旺盛,是优选良种的最佳冠层。     

Wind Flow Characteristics on a Soybean Leaf Compared with a Leaf Model

The purpose of this paper is to describe momentum boundary layer flow parameters on a soybean leaf [Glycine max (L.) Merrill] at various velocities of the bulk air stream and these data are compared with similar measurements on an artificial leaf. The wind structure is measured at three different bulk air velocitìes (u_∞= 39, 148 and 271 centimeters per second) on an individual soybean leaf and is compared to structural effects on an artificial leaf (flat metal plate) in a small closed-circuit wind tunnel. The boundary layers were homogeneous for the metal plate, but only at the lower velocity for the soybean leaf. The boundary layer thicknesses decrease with increasing bulk air velocity for laminar flow regimes, whereas in the turbulent flow regime the boundary layer thickness greatly increases. The effect of turbulence on the soybean leaf boundary layer made the eddy diffusivities at least three times greater than in the laminar flow regime at the calculated roughness height above the leaf surface. The structure of the leaf boundary layer flow is comparable to that of the metal plate only at the lower bulk air velocity.     

Production and Properties of a Soymilk-clotting Enzyme System from a Microorganism

Some microorganisms, including some bacteria isolated from soil, were found to secrete an extracellular soymilk-clotting enzyme. Among them, strain No. K-295G-7 showed the highest soymilk-clotting activity and stability of the production of the soymilk-clotting enzyme. The enzyme system (culture filtrate) coagulated protein in soymilk, a curd being formed at pH 5.8~6.7 and at 55~75°C. The optimum temperature for the soymilk-clotting activity was 75°C and the enzyme system was stable at temperatures below 50°C down to 35°C. About 80~100% of the original activity remained after 1 hr at pH 5~7 and 35°C.     

Root Interactions in a Maize/Soybean Intercropping System Control Soybean Soil-Borne Disease,Red Crown Rot

Background

Within-field multiple crop species intercropping is well documented and used for disease control, but the underlying mechanisms are still unclear. As roots are the primary organ for perceiving signals in the soil from neighboring plants, root behavior may play an important role in soil-borne disease control.

Principal Findings

In two years of field experiments, maize/soybean intercropping suppressed the occurrence of soybean red crown rot, a severe soil-borne disease caused by Cylindrocladium parasiticum (C. parasiticum). The suppressive effects decreased with increasing distance between intercropped plants under both low P and high P supply, suggesting that root interactions play a significant role independent of nutrient status. Further detailed quantitative studies revealed that the diversity and intensity of root interactions altered the expression of important soybean PR genes, as well as, the activity of corresponding enzymes in both P treatments. Furthermore, 5 phenolic acids were detected in root exudates of maize/soybean intercropped plants. Among these phenolic acids, cinnamic acid was released in significantly greater concentrations when intercropped maize with soybean compared to either crop grown in monoculture, and this spike in cinnamic acid was found dramatically constrain C. parasiticum growth in vitro.

Conclusions

To the best of our knowledge, this study is the first report to demonstrate that intercropping with maize can promote resistance in soybean to red crown rot in a root-dependent manner. This supports the point that intercropping may be an efficient ecological strategy to control soil-borne plant disease and should be incorporated in sustainable agricultural management practices.     

Enzyme Distribution Between the Cortex and the Infected Region of Soybean Nodules

Analysis of the distribution of enzymes of carbohydrate, carboxylicacid and nitrogen metabolism in nodule tissues prepared by protoplastisolation techniques has led to some confusion about their whereabouts.In this study nodule cortical and central (infected) tissuewas separated manually using a scalpel blade. Protein contents of whole nodules, central region and cortexwere found to be 19, 25 and 5-0 mg g^–1 fresh weight, respectively.Specific activities of glutamine synthetase (GS), phosphoenolpyruvatecarboxylase (PEPC), invertase (INV) and sucrose synthase (SS)were higher in the central region than in the cortex. The reversewas true for UDP glucose pyrophosphorylase (UDPGPP), phosphoglucomutase(PGM) and phosphoglucose isomerase (PGI). The central (infected) region represented a minimum of 65% ofthe nodule fresh weight. Calculations based on this estimateand on the activities per gram fresh weight indicate that thebulk of the potential enzyme activity is located in the centralregion. This was particularly so for SS, INV and GS where greaterthan 96% of the relevant activity was located in the centralregion. Even the enzymes which had greater specific activitiesin the cortex (UDPGPP, PGM and PGI) were, in fact, predominantlyfound in the central region (86, 81, and 85%, respectively).The conclusion from this data must be that most metabolism inthe nodule is likely to occur in the central region, althoughthis does not negate the importance of the cortex in carbonand nitrogen flux. Key words: Glycine max, soybean, nodules, cortex, infected region, enzymes     

一组纤维素分解菌复合系NSC-7的酶活表达特性

为了揭示一组具有降解纤维素和林丹双重功能的细菌复合系NSC-7的降解活性, 本文对该菌系的分解能力、纤维素酶活性和半纤维素酶活性进行测定。结果表明, NSC-7在14 d内, 可降解稻秆干重的 73.6％, 其中降解纤维素82.1％, 半纤维素58.2％, 木质素5.4％。用广泛采用的酶活测定方法测定了4种纤维素酶和半纤维素酶活性, 在培养的第8天, 内切酶、总纤维素酶、外切酶和b-糖苷酶活性都达到最大值, 分别为4.48 U/mL、7.51 U/mL、15.83 U/mL和25.78 U/mL。在培养的第5天, 半纤维素酶活性达到最高值为280.9 U/mL, 其平均值比纤维素酶活性高43.71倍。     

Characteristics of a Lytic Enzyme Induced by Bacteriophage Infection of Micrococcus lysodeikticus

A lytic enzyme induced in Micrococcus lysodeikticus strain 1 by infection with N1 bacteriophage was purified 45- to 50-fold by ammonium sulfate precipitation, acid precipitation, and selective adsorption of contaminating proteins with calcium phosphate gel. The optimal pH for activity of the enzyme was 6.5 to 7.0. Maximal activity occurred at 45 to 50 C and at an ionic strength of 0.06. The enzyme had a limited specificity and lysed cell walls of M. lysodeikticus with the release of dinitrofluorobenzene reactive groups. Living cells were lysed in the absence of phage; however, the rate of lysis increased when phage was present in excess of 10 particles per bacterial cell. Young cells were most sensitive, and the sensitivity decreased to a minimum with stationary-phase cells. Acting synergistically, lysozyme and the N1-induced lysin caused lysis of cells which were resistant to either enzyme acting independently. The N1 lysin did not exhibit proteolytic activity.     

一组纤维素分解菌复合系NSC-7的酶活表达特性

为了揭示一组具有降解纤维素和林丹双重功能的细菌复合系NSC-7的降解活性, 本文对该菌系的分解能力、纤维素酶活性和半纤维素酶活性进行测定.结果表明,NSC-7在14d内,可降解稻秆干重的73.6%,其中降解纤维素82.1%,半纤维素58.2%,木质素5.4%.用广泛采用的酶活测定方法测定了4种纤维素酶和半纤维素酶活性,在培养的第8天,内切酶、总纤维素酶、外切酶和B.糖苷酶活性都达到最大值,分别为4.48U/mL、7.51U/mL、15.83U/mL和25.78U/mL.在培养的第5天,半纤维素酶活性达到最高值为280.9U/mL,其平均值比纤维素酶活性高43.71倍.     

The Effect of Changing Temperatures During Imbibition on Ultrastructure in Germinating Pea Embryonic Radicles

Hodson, M. J., Di Nola, L. and Mayer, A. M. 1987. The effectof changing temperatures during imbibition on ultrastructurein germinating pea embryonic radicles.—J. exp. Bot. 38:525–534. Pea seeds were imbibed at 5 °C or at 25 °C for 5–6h and then germinated at the same temperature or transferredto the other temperature for 15 h. After imbibition or germinationthe embryonic axis was removed, fixed and the ultrastructureof the radicle tip examined. Exposure of the seeds to 5 °Cduring imbibition resulted in the cessation of almost all ultrastructuralchanges during subsequent germination at 5 °C. When theseeds were transferred to 25 °C development of ultrastructurewas far slower than expected. In particular, mitochondrial structurefailed to develop, there were few Golgi and little ER, and lipidbodies located near the plasma membrane disappeared or fusedmuch more slowly with the membrane. An attempt is made to relatethe observations on ultrastructure with data previously reportedon the metabolism of phospholipids in the plasma membrane andER of the embryonic axis. A tentative hypothesis to accountfor the effect of temperature of imbibition on subsequent seedgermination is proposed. Key words: Pisum sativum L. cv. Alaska, germination, imbibition, temperature, ultrastructure, lipid bodies, protein bodies, vacuolation, mitochondria     

Serum Diamine Oxidase as a Hemorrhagic Shock Biomarker in a Rabbit Model

Background

In prolonged hemorrhagic shock, reductions in intestinal mucosal blood perfusion lead to mucosal barrier damage and systemic inflammation. Gastrointestinal failure in critically ill patients has a poor prognosis, so early assessment of mucosal barrier injury in shock patients is clinically relevant. Unfortunately, there is no serum marker that can accurately assess intestinal ischemia-reperfusion injury.

Objective

The aim of this study was to assess if serum diamine oxidase levels can reflect intestinal mucosal injury subsequent to prolonged hemorrhagic shock.

Methods

Thirty New Zealand white rabbits were divided into three groups: a control group, a medium blood pressure (BP) group (exsanguinated to a shock BP of 50 to 41 mm Hg), and a low BP group (exsanguinated to a shock blood pressure of 40 to 31 mm Hg), in which the shock BP was sustained for 180 min prior to fluid resuscitation.

Results

The severity of hemorrhagic shock in the low BP group was significantly greater than that of the medium BP group according to the post-resuscitation BP, serum tumor necrosis factor (TNF)-α, and arterial lactate. Intestinal damage was significantly more severe in the low BP group according to Chiu’s scoring, claudin-1, intercellular adhesion molecule (ICAM)-1, and myeloperoxidase expression. Serum diamine oxidase was significantly increased in the low BP group compared to the medium BP and control groups and was negatively correlated with shock BP.

Conclusion

Serum diamine oxidase can be used as a serological marker in evaluating intestinal injury and shows promise as an indicator of hemorrhagic shock severity.     

Unusual Behavior of Membrane Somatic Angiotensin-Converting Enzyme in a Reversed Micelle System

Properties of the membrane and soluble forms of somatic angiotensin-converting enzyme (ACE) were studied in the system of hydrated reversed micelles of aerosol OT (AOT) in octane. The membrane enzyme with a hydrophobic peptide anchor was more sensitive to anions and to changes in pH and composition of the medium than the soluble enzyme without anchor. The activity of both forms of the enzyme in the reversed micelles significantly depended on the molarity of the buffer added to the medium (Mes-Tris-buffer, 50 mM NaCl). The maximum activity of the soluble ACE was recorded at buffer concentration of 20-50 mM, whereas the membrane enzyme was most active at 2-10 mM buffer. At buffer concentrations above 20 mM, the rate of hydrolysis of the substrate furylacryloyl-L-phenylalanyl-glycylglycine by both ACE forms was maximal at pH 7.5 both in the reversed micelles and in aqueous solutions. However, at lower concentrations of the buffer (2-10 mM), the membrane enzyme had activity optimum at pH 5.5. Therefore, it is suggested that two conformers of the membrane ACE with differing pH optima for activity and limiting values of catalytic constants should exist in the reversed micelle system with various medium compositions. The data suggest that the activity of the membrane-bound somatic ACE can be regulated by changes in the microenvironment.     

根霉12#发酵产生纤溶酶的酶学性质

溶栓疗法是血栓性疾病安全有效的治疗手段,开发新型纤溶酶具有实际应用意义.分离自南方小酒药的根霉12豆粕和麸皮为原料可产生纤溶酶.已采用盐析,疏水层析、离子交换层析和凝胶层析方法对纤溶酶分离提纯.提纯的纤溶酶比活力2143u/mg(尿激酶单位),有直接溶解血栓和激活纤溶酶原的双重溶栓作用,降解纤维蛋白α、β和γ肽链速度快;最适作用温度45℃,适宜作用pH范围6.8～8.8;等电聚焦方法测定该酶等电点8.5±0.1;只分解生色底物N-Succinvl-Ala-Ala-Pro-Phe-pNA,其米氏常数Km为O.23mmol/L,酶转换数Kcat为16.36 s-1;Molish实验和甲苯胺蓝实验均证明该酶为糖蛋白,地衣酚-硫酸法测得该酶含糖量4.70%;EDTA、PMSF、PCMB对该纤溶酶有抑制作用,说明活性中心含有巯基、金属和丝氨酸;N端12个氨基酸序列为NH2-Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly,与其它生物来源的纤溶酶相比较没有同源性.根霉12#产生的纤溶酶为新型纤溶酶,有希望开发成溶栓药物.