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Leucine Aminopeptidase (leucine + aminopeptidase)
Selected AbstractsCommunity composition and activity of prokaryotes associated to detrital particles in two contrasting lake ecosystemsFEMS MICROBIOLOGY ECOLOGY, Issue 3 2006Charles Lemarchand Abstract The composition, distribution and extracellular enzyme activities of bacteria attached to small (2,50 ,m in size) transparent exopolymer and Coomassie-stained proteinaceous particles (TEP and CSP) were examined in two lakes of different trophic status located in the Massif Central of France. TEP concentrations (104,106 particle per L) were significantly higher in the more productive lake and were significantly related to chlorophyll a concentrations. The majority of TEP and CSP were colonized by bacteria that constituted 2.6% and 7.4% of the total 4,,6-diamidino-2-phenylindole-stained bacteria in lakes Pavin and Aydat, respectively. In both lakes, the composition of particle-associated bacteria was different from that of free-living bacteria, the Betaproteobacteria and Bacteroidetes (i.e. former Cytophaga,Flavobacteria group) being the dominant groups on particles. We also found that 2,5 ,m TEP were more colonized than 2,5 ,m CSP in the two lakes, and that TEP colonization was higher in the less productive lake. Measurements of Leucine aminopeptidase and ,-glucosidase activities in fractionated lake water (0.2,1.2, 1.2,5 and >5 ,m fractions) indicated that proteolytic activity was always higher and that particle-associated bacteria have higher enzymatic activities than free-living bacteria. The glycolytic activities in the 1.2,5 and >5 ,m fractions were related to the abundance of TEP. We conclude that small freshwater detrital organic particles constitute microhabitats with high bacterial activities in pelagic environments and, undoubtedly, present significant ecological implications for the prokaryotic community structure and function in aquatic ecosystems. [source] Nutritional, physiological, and histological responses in Atlantic salmon, Salmo salar L. fed diets with genetically modified maizeAQUACULTURE NUTRITION, Issue 3 2007G.-I. HEMRE Abstract The objective of this study was to evaluate whether standard fish meal diets prepared with increasing levels of genetically modified (GM; 150 and 300 g kg,1) maize (event MON810®) as a starch source, showed any nutritional or physiological adverse effects on Atlantic salmon, Salmo salar L. postsmolt. The diets with low or high inclusions of GM maize and its near-isogenic parental line (nongenetically modified; nGM maize), were balanced with Suprex maize (Reference) to obtain compositional equivalency of diet starch, sugars and all other nutrients. Total starch level in all diets was 160 g kg,1. After 82 days of feeding, fish growth was high in all groups, however fish fed the GM maize showed slight but significant lower feed intake, which was followed by slight but significant lower specific growth rate and final body weights, compared with fish fed nGM maize, none of the groups varied significantly from fish fed the Reference diet. There was no variation in feed conversion ratios (FCR), protein and lipid efficiency ratios (PER and LER), or protein- and lipid-productive values (PPV and LPV) in this study. No significant effect of maize type was detected on apparent digestibility coefficients (ADC) of dry matter, protein or lipid. Hematological analysis and plasma nutrients varied within normal ranges for Atlantic salmon in all diet groups, except for somewhat elevated aspartate aminotransferase (ASAT) values in all groups. Hepatosomatic index (HSI) with values ranging from 1.37 to 1.60, was significantly higher for the high GM maize group compared with the high nGM maize group but not when compared with the Reference diet group. Lowered spleen (SSI) and head-kidney somatic indices (H-KSI) were registered when fed GM compared with nGM maize, the Reference treatment was however, equal to both. Distal intestine somatic index (DISI) was significantly higher for GM maize-fed fish compared with nGM maize-fed fish, but not significantly different from the Reference diet group. Histological evaluation of the mid- and distal intestine, liver, spleen and head-kidney did not reveal any diet-related morphological changes. Maltase activities in the mid- and distal intestinal tissue homogenates were affected by diet, the fish fed high GM maize having higher activities compared with high nGM maize-fed fish. Leucine aminopeptidase (LAP) and alkaline phosphatase (AP) activities were not affected by diet. Sodium-dependent d -glucose uptake in brush border membrane vesicles (BBMV) isolated from pyloric caeca of fish fed high GM maize was significantly higher than that found in fish fed the analogous diet with high nGM maize. Based on the present findings, the conclusions made are: Atlantic salmon smolts fed GM maize (event MON810®), its near-isogenic parental line and suprex maize (Reference diet), all resulted in high growth rates, ADC and feed utilization. Health, when evaluated by means of mortality (low), normal ranges of blood and plasma parameters, except somewhat elevated ASAT values and minor variations in organ sizes, were considered good in all diet groups. The changes in the glucose transport mechanism and intestinal maltase enzyme activity in the gastrointestinal tract warrant further studies. [source] Functional microbial community response to nutrient pulses by artificial groundwater recharge practice in surface soils and subsoilsFEMS MICROBIOLOGY ECOLOGY, Issue 3 2010Kirsten Schütz Abstract Subsurface microorganisms are essential constituents of the soil purification processes associated with groundwater quality. In particular, soil enzyme activity determines the biodegradation of organic compounds passing through the soil profile. Transects from surface soil to a depth of 3.5 m were investigated for microbial and chemical soil characteristics at two groundwater recharge sites and one control site. The functional diversity of the microbial community was analyzed via the activity of eight enzymes. Acid phosphomonoesterase was dominant across sites and depths, followed by l -leucine aminopeptidase and ,-glucosidase. Structural [e.g. phospholipid fatty acid (PLFA) pattern] and functional microbial diversities were linked to each other at the nonwatered site, whereas amendment with nutrients (DOC, NO3,) by flooding uncoupled this relationship. Microbial biomass did not differ between sites, whereas microbial respiration was the highest at the watered sites. Hence, excess nutrients available due to artificial groundwater recharge could not compensate for the limitation by others (e.g. phosphorus as assigned by acid phosphomonoesterase activity). Instead, at a similar microbial biomass, waste respiration via overflow metabolism occurred. In summary, ample supply of carbon by flooding led to a separation of decomposition and microbial growth, which may play an important role in regulating purification processes during groundwater recharge. [source] Exoenzyme activities as indicators of dissolved organic matter composition in the hyporheic zone of a floodplain riverFRESHWATER BIOLOGY, Issue 8 2010SANDRA M. CLINTON Summary 1. We measured the hyporheic microbial exoenzyme activities in a floodplain river to determine whether dissolved organic matter (DOM) bioavailability varied with overlying riparian vegetation patch structure or position along flowpaths. 2. Particulate organic matter (POM), dissolved organic carbon (DOC), dissolved oxygen (DO), electrical conductivity and temperature were sampled from wells in a riparian terrace on the Queets River, Washington, U.S.A. on 25 March, 15 May, 20 July and 09 October 1999. Dissolved nitrate, ammonium and soluble reactive phosphorus were also collected on 20 July and 09 October 1999. Wells were characterised by their associated overlying vegetation: bare cobble/young alder, mid-aged alder (8,20 years) and old alder/old-growth conifer (25 to >100 years). POM was analysed for the ash-free dry mass and the activities of eight exoenzymes (,-glucosidase, ,-glucosidase, , -N-acetylglucosaminidase, xylosidase, phosphatase, leucine aminopeptidase, esterase and endopeptidase) using fluorogenic substrates. 3. Exoenzyme activities in the Queets River hyporheic zone indicated the presence of an active microbial community metabolising a diverse array of organic molecules. Individual exoenzyme activity (mean ± standard error) ranged from 0.507 ± 0.1547 to 22.8 ± 5.69 ,mol MUF (g AFDM),1 h,1, was highly variable among wells and varied seasonally, with the lowest rates occurring in March. Exoenzyme activities were weakly correlated with DO, DOC and inorganic nutrient concentrations. 4. Ratios of leucine aminopeptidase : ,-glucosidase were low in March, May and October and high in July, potentially indicating a switch from polysaccharides to proteins as the dominant component of microbial metabolism. 5. Principal components analysis indicated that there were patch effects and that these effects were strongest in the summer. 6. DOM degradation patterns did not change systematically along hyporheic flowpaths but varied with overlying forest patch type in the Queets River hyporheic zone, suggesting that additional carbon inputs exist. We hypothesise that the most likely input is the downward movement of DOM from overlying riparian soils. Understanding this movement of DOM from soils to subsurface water is essential for understanding both the hyporheic metabolism and the carbon budget of streams and rivers. [source] Characterization of the proteases in the midgut of the xylophagous larvae of Oemona hirta (Coleoptera: Cerambycidae)INSECT SCIENCE, Issue 5 2009Brian David Shaw Abstract, The protein digestive capability of the larvae of the longhorn beetle (Oemona hirta, Coleoptera: Cerambycidae, Fabricius, 1775) was investigated. This species feeds only on wood where there is a high proportion of vascular tissue. The pH of the midgut, the major digestive organ, was alkaline and protein hydrolysis was maximal at alkaline pH. Use of specific synthetic peptide substrates showed that the major protease activities were the endopeptidases, trypsin and chymotrypsin-like activity, and the exopeptidase, leucine aminopeptidase and the pH curves corresponded to that with protein substrate. Studies using a range of serine protease inhibitors as well as specific inhibitors of metalloproteases, cysteine proteases and aspartate proteases confirmed a serine protease-based digestive system similar to earlier reports of sapwood-feeding Cerambycids. Control of these insect pests using protease inhibitors is discussed. [source] Expression of brush border enzymes in response to lead exposure in rat intestineJOURNAL OF APPLIED TOXICOLOGY, Issue 5 2005Priya Kapur Abstract The effect of feeding lead (50 mg kg,1 body weight) daily for 7 days on the development of various brush border enzymes in the intestine has been studied. The activities of brush border sucrase (P < 0.001), lactase (P < 0.001), , -glutamyl transpeptidase (P < 0.05) and leucine aminopeptidase were reduced (P < 0.05), whereas the alkaline phosphatase level was augmented (P < 0.05) in lead fed rats compared with controls. Kinetic studies with sucrase revealed a low Vmax (0.224 in control and 0.160 units mg,1 protein in lead exposed) with no change in Km (12.6,13.5 mm). Western blot analysis for alkaline phosphatase yielded intense staining of enzyme protein in lead fed rats compared with controls, however, the intensity of the antigen signal was reversed for sucrase under these conditions. These findings suggest that ingestion of lead may interfere with the crypt cell differentiation process thus affecting enzyme functions in the rat intestine. Copyright © 2005 John Wiley & Sons, Ltd. [source] Changes in immune and enzyme histochemical phenotypes of cells in the intestinal mucosa of Atlantic salmon, Salmo salar L., with soybean meal-induced enteritisJOURNAL OF FISH DISEASES, Issue 2 2000A M Bakke-McKellep Extracted soybean meal (SBM) in the diet for Atlantic salmon, Salmo salar L., causes an inflammatory response in the distal intestine. The morphological changes of the epithelial cells and a characterization of the inflammatory cell infiltrate of the distal intestinal mucosa were studied using a panel of enzyme and immunohistochemical markers. The salmon (average body weight 927 g) used in the study were fed either a fishmeal-based diet (control diet) or a diet in which 30% of the fishmeal protein was replaced with SBM protein (SBM diet). In salmon fed SBM, there were markedly reduced enzyme reactivities in the distal intestinal epithelial cells, both in the brush border [5,-nucleotidase (5,N), Mg2+-ATPase, alkaline phosphatase (ALP) and leucine aminopeptidase (LAP)] and in the intracellular structures [alkaline and acid phosphatase, non-specific esterase (NSE) and alanine aminopeptidase (AAP)]. There appeared to be an increased presence of cells of monocytic lineage, including macrophages, as well as neutrophilic granulocytes and immunoglobulin (Ig) M in the lamina propria of the SBM-fed fish. The mid intestine showed little response to the diet. The results suggest that toxic/antigenic component(s) of SBM affect the differentiation of the distal intestinal epithelial cells and may help explain the reduced nutrient digestibilities previously reported in salmonids fed extracted SBM. [source] Effect of Yam (Dioscorea alata Compared to Dioscorea japonica) on Gastrointestinal Function and Antioxidant Activity in MiceJOURNAL OF FOOD SCIENCE, Issue 7 2006Cheng-Chin Hsu ABSTRACT:, Effects of Chinese yam (Dioscorea alata) and Japanese yam (Dioscorea japonica) on gastrointestinal functions including intestinal microflora and intestinal enzymes' activities, as well as antioxidant protection against lipopolysaccharide (LPS)-induced oxidative damage, in Balb/cA mice were examined. In part I, mice were fed yam-supplemented diet for 4 or 8 wk, and killed with carbon dioxide. In part II, mice were fed yam-supplemented diet for 4 wk, and followed by intraperitoneal LPS treatment (i.p. 4 mg/kg bodyweight). The intake of Chinese yam and Japanese yam significantly changed intestinal microflora, in which the colony numbers of Bifidobacterium and Lactobacillus were increased and the colony numbers of Clostridium perfringens were decreased (P < 0.05). The intake of both Chinese and Japanese yams also significantly elevated the activity of leucine aminopeptidase and lipase (P < 0.05), and the activities of sucrase and maltase were increased only in 20% yam-treated groups (P < 0.05). The preintake of yam significantly alleviated subsequent LPS-induced oxidative injury by decreasing lipid oxidation level and fibronectin production and elevating superoxide dismutase activity (P < 0.05). Both Chinese and Japanese yams contained dietary fibers, polyphenols, and flavonoids, which may contribute to the observed gastrointestinal function and antioxidant protection. These results suggest that both Chinese yam and Japanese yam were beneficial for intestinal health and oxidation prevention. [source] EXTRACELLULAR ENZYMES OF THE MICROCYSTIS AERUGINOSA PCC 7813 STRAIN ARE INHIBITED IN THE PRESENCE OF HYDROQUINONE AND PYROGALLOL, ALLELOCHEMICALS PRODUCED BY AQUATIC PLANTS,JOURNAL OF PHYCOLOGY, Issue 6 2009Dariusz Dziga Several cyanobacterial species have a high potential to dominate in marine environments and freshwater reservoirs, and the ecological and physiological reasons for this phenomenon are not understood comprehensively. In this study, the ability of a Microcystis aeruginosa Kütz. strain to produce free dissolved enzymes was documented. We have observed that this highly toxic strain releases alkaline phosphatase, leucine aminopeptidase, and ,-glucosidase into the ambient environment. Additionally, the inhibitory activity of selected phenols produced by aquatic plants on the activity of these enzymes was analyzed. The investigated compounds, pyrogallol and, to a lesser degree, hydroquinone, decreased the activity of extracellular enzymes produced by M. aeruginosa, with leucine aminopeptidase being the most sensitive to the inhibitors. The noncompetitive character of enzymatic inhibition suggests that the polyphenols produced by aquatic plants are able to influence the activity of different extracellular or membrane-bound enzymes. [source] l -leucine aminopeptidase production by filamentous Aspergillus fungiLETTERS IN APPLIED MICROBIOLOGY, Issue 6 2005K.M. Nampoothiri Abstract Aims:, To screen various filamentous fungi belonging to Aspergillus spp. producing leucine and methionine aminopeptidases. Methods and Results:, Twenty-eight Aspergillus strains representing 14 species within the genus were screened for l -leucine aminopeptidase (LAP) production in two media in shake flask fermentation. Two Aspergillus sojae (NRRL 1988 and NRRL 6271) and one Aspergillus oryzae (NRRL 6270) strains were selected as the best producers for further studies. The peak LAP activities were 2·61, 2·59 and 1·30 IU ml,1 for the three fungi on days 2, 5 and 4 respectively. In addition to LAP, l -methionine aminopeptidase (MAP) activity was also detected. Apart from submerged fermentation, the highest LAP yields by solid-state fermentation were 11·39, 17·40 and 13·02 IU g,1 dry matter for the above fungi. The temperature and pH optimum of the enzyme was found to be in the range of 65,75°C at pH 8·0,9·0 for all three fungi. Metal ions, Co2+ and Fe2+ in 2 mmol l,1 concentration apparently enhanced the relative enzyme activity and heat stability. Conclusions:, Two A. sojae (NRRL 1988 and NRRL 6271) and one A. oryzae (NRRL 6270) strains were found to be the best producers of LAP and MAP. The preliminary characterization studies revealed that the enzyme is considerably thermostable and belongs to the class metalloenzymes. Significance and Impact of the Study:, A good number of aspergilli were screened and the ability of the fungal aminopeptidase to release a particular N-terminal amino acid along with its high thermal stability, makes them interesting for controlling the degree of hydrolysis and flavour development for a wide range of substrate. [source] Effect of dietary administration of probiotics on growth and intestine functionality of juvenile Senegalese sole (Solea senegalensis, Kaup 1858)AQUACULTURE NUTRITION, Issue 2 2009M.A. SÁENZ de RODRIGÁÑEZ Abstract The effects of the dietary administration of two bacterial probiotic strains (Ppd11 and Pdp13) from the Alteromonadaceae family for 60 days, were assessed by measuring growth and feed efficiency, activities of leucine aminopeptidase and alkaline phosphatase and structural changes in the intestine of juvenile Senegalese sole. In addition, the profile of intestinal microbiota was studied by Denaturing Gradient Gel Electrophoresis. Growth and nutrient utilization were significantly higher in fish receiving probiotics than in those fed the control diet. No differences were observed in proximal composition between treatments, though higher lipid muscle content was measured in fish receiving Pdp13. Those fish also exhibited higher activities of AP when compared to Ppd11 and control groups. The profile of intestinal microbiota clearly separated those fish receiving probiotics from those of the control group. Microscopical examination revealed accumulation of lipid droplets in the enterocytes of fish receiving the control diet, but not in those fed on probiotics. Interactions between those structural changes and growth performance are discussed. [source] Effects of protein hydrolysate in weaning diets for Atlantic cod (Gadus morhua L.) and Atlantic halibut (Hippoglossus hippoglossus L.)AQUACULTURE NUTRITION, Issue 2 2009A. KVÅLE Abstract The study aims to test whether predigested dietary protein enhances the utilization of formulated diets at weaning, and also whether it stimulates intestinal maturation. In this study, Atlantic cod [Gadus morhua L.; 41 days posthatch (dph)] and Atlantic halibut [Hippoglossus hippoglossus L.; 63 days postfirst feeding (dpff)] were weaned onto diets with graded levels of protein hydrolysate. By increasing the inclusion of dietary protein as hydrolysate from 0 to 400 g kg,1, cod increased the rate of survival from 7 ± 1% to 18 ± 2% (82 dph; regression, P = 4*10,7). In halibut, the survival rate decreased from 57 ± 9% to 22 ± 7% as the inclusion of protein in the form of hydrolysate increased from 0 to 450 g kg,1 (119 dpff; regression, P = 8*10,5). Growth was not affected in any of the species. Results in specific activities of the intestinal enzymes leucine aminopeptidase (LAP) and alkaline phospatase (AP) supported the results in survival in halibut and partly also in cod, by showing increased activities in groups with increased survival (anova, P < 0.05). The lower optimal level of hydrolysed protein in halibut than in cod is suggested mainly because of a slower feeding practice in halibut, which allows more extensive nutrient leaching before ingestion. [source] Cloning, expression, crystallization and preliminary X-ray crystallographic analysis of leucine aminopeptidase (LAP) from the pepA gene of Xanthomonas oryzae pv. oryzaeACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2009Kim-Hung Huynh Xanthomonas oryzae pv. oryzae (Xoo) causes the serious disease bacterial blight in rice. The pepA (Xoo0834) gene from Xoo is one of around 100 genes that have been selected for the design of antibacterial drugs. The pepA gene encodes leucine aminopeptidase (LAP), an exopeptidase that catalyzes the hydrolysis of leucine residues from the N-terminus of a protein or peptide. This enzyme was expressed in Escherichia coli, purified and crystallized, and preliminary X-ray structural studies have been carried out. The LAP crystal diffracted to 2.6,Å resolution and belonged to the cubic space group P213. The unit-cell volume of the crystal was compatible with the presence of two monomers in the asymmetric unit. [source] | ||||||||||||||||