Alterations in the hepatic enzymes following experimental lead poisoning

An investigation on the influence of lead toxicity on some of the hepatic enzymes was studied in rats both after a shorter interval of 15 d and after longer intervals of 60 and 90 d. Three different doses of lead as 5, 10, and 50 mg/kg body wt were administered orally on every alternate day. Whereas significant inhibition of succinic dehydrogenase was seen following lead poisoning, the activity acid and alkaline phosphatase increased with lead intoxication. The histoarchitecture of the liver was grossly intact. Liver accumulated less lead compared to kidney at 60 and 90 d.     

Molecular and immunological characterization of the major outer membrane proteins of Brucella

Abstract Saccharomyces cerevisiae exponentially growing in basic or 0.7 M NaCl medium were isotopically labelled with ³⁵S-methionine, followed by protein separation and quantification by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) combined with computerised image analysis. The electrophoretic separation resolved about 650 proteins of which 13 displayed significant and at least 2-fold changes in rate of synthesis during saline growth. By sequencing of 2D-PAGE resolved proteins, one of the 8 induced spot, p42.9/5.5, was shown to correspond to the full length (containing the N-terminal extension) product of the GPD 1 gene encoding the cytoplasmic glycerol 3-phosphate dehydrogenase. The expression of the TDH 3 gene, glyceraldehyde 3-phosphate dehydrogenase, and the ENO 2 gene, enolase, decreased during growth in NaCl medium, declines hypothesised to have an impact on the flux to glycerol.     

Reaction profile of the last step in cytochrome P-450 catalysis revealed by studies of model complexes

 A series of oxoiron(IV) porphyrin cation radical complexes was investigated as compound I analogs of cytochrome P-450. Both the spectroscopic features and the reactivities of the complexes in oxygen atom transfer to olefins were examined as a function of only one variable, the axial ligand trans to the oxoiron(IV) bond. The results disclosed two important kinetic steps – electron transfer from olefin to oxoiron(IV) and intramolecular electron transfer from metal to porphyrin radical – which are affected differently by the axial ligands. The large kinetic barrier of the latter step in the reaction of olefins with the perchlorato-bound oxoiron(IV) porphyrin cation radical complex enabled the trapping of a reaction intermediate in which the metal, but not the porphyrin radical, is reduced. The first electron transfer step is probably followed by σ-bond formation, which readily accounts for formation of isomerized organic products at low temperatures. It is finally postulated that part of the enhanced oxygenation activities of cytochrome P-450 monooxygenases and chloroperoxidases is due to a lowering of the energy barrier for the second electron transfer step via participation of their redox-active cysteinate ligand. Received: 16 January 1997 / Accepted: 24 May 1997     

SNaPe: a versatile method to generate multiplexed protein fusions using synthetic linker peptides for in vitro applications

Understanding the structure and function of protein complexes and multi‐domain proteins is highly important in biology, although the in vitro characterization of these systems is often complicated by their size or the transient nature of protein/protein interactions. To assist in the characterization of such protein complexes, we have developed a modular approach to fusion protein generation that relies upon S ortase‐mediated and Na tive chemical ligation using synthetic Pe ptide linkers (SNaPe) to link two separately expressed proteins. In this approach, we utilize two separate linking steps – sortase‐mediated and native chemical ligation – together with a library of peptide linkers to generate libraries of fusion proteins. We have demonstrated the viability of SNaPe to generate libraries from fusion protein constructs taken from the biosynthetic enzymes responsible for late stage aglycone assembly during glycopeptide antibiotic biosynthesis. Crucially, SNaPe was able to generate fusion proteins that are inaccessible via direct expression of the fusion construct itself. This highlights the advantages of SNaPe to not only access fusion proteins that have been previously unavailable for biochemical and structural characterization but also to do so in a manner that enables the linker itself to be controlled as an experimental parameter of fusion protein generation. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.     

干旱条件下钙离子对一氧化氮诱导黄瓜不定根发生的影响

以黄瓜品种‘新春4号’为材料,研究干旱胁迫下一氧化氮(NO)和钙离子(Ca²⁺)处理下黄瓜的生根指标、内源Ca²⁺荧光强度以及抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT、抗坏血酸过氧化物酶APX)活性,分析干旱条件下黄瓜不定根发生过程中NO和Ca²⁺之间的关系.结果表明: 200 μmol·L^-1 氯化钙(CaCl₂)和0.05%聚乙二醇(PEG)共处理显著提高了干旱条件下黄瓜不定根的根长和根数;添加Ca²⁺螯合剂(EGTA)和通道抑制剂(BAPTA/AM)处理显著降低了干旱条件下NO诱导的不定根根数和根长.干旱条件下,NO和CaCl₂处理提高了黄瓜下胚轴内源Ca²⁺荧光强度;而NO清除剂(cPTIO)处理的Ca²⁺荧光强度显著低于NO处理.干旱条件下,NO和CaCl₂处理显著提高了黄瓜下胚轴抗氧化酶活性;而Ca²⁺抑制剂或螯合剂处理显著降低了NO诱导的抗氧化酶活性.由此可见,干旱条件下Ca²⁺参与了NO调控黄瓜抗氧化酶活性,缓解了干旱胁迫对不定根形成产生的伤害,进而促进了不定根的发生.     

Influence of high inorganic selenium and manganese diets for fattening pigs on oxidative stability and pork quality parameters

Data are scarce regarding combined high Se and Mn supplementation in livestock diets, however, as Se and Mn are functionally related as cofactors of glutathione peroxidase (GPx) and Mn-superoxide dismutase (SOD), respectively, beneficial synergistic effects on oxidative stability of tissues may result. This experiment evaluated the effect of an oversupply of Se and Mn within European legal limits compared with recommendations on performance, oxidative stability of the organism and meat quality in a randomised complete block design. A total of 60 crossbred gilts were fed maize–barley–soya bean meal diets formulated in a 2×2 factorial approach with inorganic Se (0.2 v. 0.5 mg/kg Se dry matter (DM)) or inorganic Mn (20 v. 150 mg/kg Mn DM) from 31 to 116 kg BW. Se supplementation reduced feed intake, whereas high Mn diets impaired average daily gain (P<0.05). Qualitative carcass characteristics were impaired by Se and Mn predominantly in the semimembranosus muscle. Activity of GPx in liver was increased by high Se diets (P<0.05). Mn supplementation increased catalase (CAT) activity in liver, GPx in plasma and total antioxidative capacity (TAC) in muscle, whereas it decreased CAT activity in plasma (P<0.05). Cu/Zn-SOD in muscle showed higher activity in high-Se-low-Mn diets but lower activity when both high Se and Mn were combined (Se×Mn P<0.05). Mn supplementation increased Mn concentration in longissimus thoracis et lumborum, but simultaneously reduced Se concentration (P<0.05). Upon retail display, Mn increased lipid oxidation more pronouncedly (higher thiobarbituric acid reactive substances; P<0.05) than Se (P<0.10). Despite some positive effects (Mn increased TAC, Se increased GPx, Se and Mn increased tenderness), no synergistic effects of high Se and Mn diets or an overall beneficial impact on meat quality, especially during storage, could be observed. Including the negative effects on performance, feeding Se and Mn up to the maximum legal level cannot be recommended.     

The Importance of Amine-degrading Enzymes on the Biogenic Amine Degradation in Fermented Foods: A review

Biogenic amines (BAs) are a class of harmful compounds often be found in high protein foods, especially naturally fermented foods. BAs derive from free amino acid decarboxylation through microbial activities and can cause toxic effects (headache, heart palpitations, vomiting) on humans, depending on individual sensitivity. Indigenous amine-degrading strains or strains producing amine-degrading enzymes (ADEs) have drawn great attention since they play an important role in affecting BA accumulation, and enzymes/genes involved in the biosynthetic mechanisms. They also help maintain the sensory quality of the final products. Besides, due to ADEs’ harmless catalytic products, they can be further utilized in fermented foods and beverages to reduce BAs. This review describes in detail the mechanisms of BAs formation, as well as the diversity of ADEs able to degrade BAs in a model or real food systems. A deeper knowledge of this issue is crucial because ADEs’ activities are often associated with strains rather than species or genera. Moreover, this information can help to improve the selection and characterization of strains for further applications as starters or bioprotective cultures, to obtain high-quality foods with reduced BAs contents.     

Membrane—vanadium interaction: A toxicokinetic evaluation

Vanadium is an important trace metal widely distributed in environment. Interaction of vanadate with skeletal muscle sarcolemma and basement membrane has been focussed. Scatchard analysis indicated the presence of more than one binding site for vanadate. Vanadate inhibits sarcolemmal and intestinal brush border membrane enzymes in a non-competitive manner. Membrane-vanadium interaction may lead to several structural and functional changes. The binding of vanadium to basement membrane may have some protective role.     

The enzymatic basis for pesticide bioremediation

Enzymes are central to the biology of many pesticides, influencing their modes of action, environmental fates and mechanisms of target species resistance. Since the introduction of synthetic xenobiotic pesticides, enzymes responsible for pesticide turnover have evolved rapidly, in both the target organisms and incidentally exposed biota. Such enzymes are a source of significant biotechnological potential and form the basis of several bioremediation strategies intended to reduce the environmental impacts of pesticide residues. This review describes examples of enzymes possessing the major activities employed in the bioremediation of pesticide residues, and some of the strategies by which they are employed. In addition, several examples of specific achievements in enzyme engineering are considered, highlighting the growing trend in tailoring enzymatic activity to a specific biotechnologically relevant function.