A study was carried out on the changes occurring in the amino acid fraction of a hybrid ryegrass during ensilage in laboratory-scale silos to help to establish the relative roles of plant and microbial proteases on protein degradation in the silo. Herbage treatments included (i) normal grass without treatment (ii) lambda-irradiated grass (sterile) without treatment (iii) sterile, inoculated with a strain of Lactobacillus plantarum and (iv) sterile, inoculated with a strain of Lactobacillus paracasei subsp. paracasei. These treatments had a significant effect on silage amino acid profiles. Concentrations of free amino acids and the extent of amino acid catabolism varied with treatment. However, levels were notably higher in control silages after 90 days (free amino acid nitrogen constituting 54% of total amino acid nitrogen compared with 37, 32 and 22% for treatments i, ii and iv, respectively). These results indicate that the extent of protein hydrolysis during ensilage is influenced by factors other than rate of pH decline and plant protease activity, and that microbial proteases play a role. 相似文献
Protegrin antimicrobial peptides (AMP) possess a high activity against a variety of microorganisms. In the present contribution, we analyse the structural requirements of protegrin analogues reported by Ostberg and Kaznessis (Peptides 2005; 26: 197) for having antimicrobial activity against several microbial species by using interpretable QSAR models. Models were carried out using multiple linear regression (MLR) combined with genetic algorithm (GA) and smoothed amino acid sequence properties were employed for characterizing the peptide dataset. The main advantage of smoothing process is the alteration of local amino acid properties by the properties of the amino acids in the closer neighbourhood. We report models encompassing different characteristics for describing the activities against different microbial species. Our results suggest the existence of specific mechanisms of action for protegrin analogues against different microbial species. 相似文献
In order to maximize milk protein production, one must present sufficient amounts of the essential amino acids to the intestinal tract in forms that can be absorbed. We do not know the specific tissue-level amino acid requirements of lactating cows, but they are likely to be similar to the amino acid content of milk protein with requirements for other metabolic functions similar to those in nonruminants. Formulating diets to meet these amino acid requirements is complicated because much of the dietary crude protein is converted to rumen microbial protein. Knowing the amount of dietary crude protein converted to ruminal microbial protein and the amino acid content of the rumen microbes; and the proportion of ruminally undegradable protein, its postruminal digestibility and amino acid content will allow one to make a reasonable estimate of the quality of protein presented for gastrointestinal digestion and absorption. Hypothetical calculations indicate potential dietary differences in quality of protein presented for absorption. Many of these differences correspond very well with production responses observed in research trials. Failure of this system to explain production results in other studies points to areas where additional information is still needed. 相似文献
Estimates were obtained of the extent to which cysteine and methionine were incorporated into the protein of the microbes of rumen digesta without prior degradation and resynthesis. By using the amino acids labeled with both (35)S and (14)C, it was observed that a large proportion of the (35)S appeared in the sulfide pool and of the (14)C appeared in volatile fatty acids. By isolating the appropriate amino acid, obtaining the (14)C to (35)S ratio, and comparing this with the ratio in the added amino acid, the degree of direct incorporation was calculated. For cysteine it was estimated that at most 1% and for methionine, at most 11% of the amino acid in the free pool was incorporated unchanged into microbial protein. As a consequence of these findings, it is considered that the method for measuring microbial protein synthesis in rumen digesta based upon incorporation of (35)S from the free sulfide pool is not seriously affected by direct utilization of sulfur amino acids arising from dietary sources. 相似文献
Amino acids have been shown to be a potentially significant N source for the alpine sedge, Kobresia myosuroides. We hypothesised that freeze-thaw and dry-rewet events allow this plant species increased access to amino acids by disrupting
microbial cells, which decreases the size of competing microbial populations, but increases soil amino acid concentrations.
To test this hypothesis, we characterized freeze-thaw and dry-rewet events in the field and simulated them in laboratory experiments
on plant-soil microcosms. In one experiment, 15N,13C-[2]-glycine was added to microcosms that had previously been subjected to a freeze-thaw or dry-rewet event, and isotopic
concentrations in the plant and microbial fractions were compared to non-stressed controls. Microbial biomass and uptake of
the labeled glycine were unaffected by the freezing and drying treatments, but microbial uptake of 15N was lower in the two warmer treatments (dry-rewet and summer control) then in the two colder treatments (freeze-thaw and
fall control). Plant uptake of glycine-15N was decreased by climatic disturbance, and uptake in plants that had been frozen appeared to be dependent on the severity
of the freeze. The fact that intact glycine was absorbed by the plants was confirmed by near equal enrichment of plant tissues
in 13C and 15N. Plants under optimal conditions recovered 3.5% of the added 15N and microbes recovered 5.0%. The majority of the 13C and 15N label remained in a non-extractable fraction in the bulk soil. To better understand the isolated influences of environmental
perturbations on soil amino acid pools and population sizes of amino-acid utilizing microbes, separate experiments were performed
in which soils, alone, were subjected to drying and rewetting or freezing and thawing. Potential respiration of glycine and
glutamate (substrate-induced respiration; SIR) by the soil microbial communities was unaffected by a single freeze-thaw event.
Glycine SIR was decreased slightly (∼10%) by the most extreme drying treatment, but glutamate SIR was not significantly affected.
Freezing lowered the concentration of water-extractable amino acids while drying increased their concentration. We interpret
the surprising former result as either a decrease in proteolytic activity in frozen soils relative to amino acid uptake, or
a stimulation in microbial uptake by physical nutrient release from the soil. We conclude that climatic disturbance does not
provide opportunities for increased amino acid uptake by K. myosuroides, but that this plant competes well for amino acid N under non-stressed conditions, especially when soils are warm. We also
note that this alpine tundra microbial community's high resistance to freeze-thaw and dry-rewet events is novel and contrasts
with studies in other ecosystems.
Received: 24 February 1997 / Accepted: 28 August 1997 相似文献
Altitudinally-defined climate conditions provide specific vegetation types and soil environments that could influence soil microbial communities, which in turn may affect microbial residues. However, the knowledge is limited in terms of the degree to which microbial communities and residues present and differ along altitude. In this study, we examined the soil microbial communities and residues along the northern slope of Changbai Mountain, China using phospholipid fatty acid (PLFA) and amino sugar analysis, respectively. Soil samples were taken from five different vegetation belts defined by climates. Principal component analysis (PCA) revealed substantial differences in soil microbial community composition among study sites, appeared to be driven primarily by soil pH and C/N ratio on the first principal component (PC1) which accounted for 50.7% of the total sample variance. The alpine tundra was separated from forest sites on the second principal component (PC2) by a signifiscantly higher amount of fungal PLFA (18:2ω6,9). Soil pH and C/N ratio were also correlated with the ratios of Gram-positive to Gram-negative bacteria (Gm+/Gm−), glucosamine to galactosamine (GluN/GalN), and glucosamine to muramic acid (GluN/MurA). Both total PLFAs and amino sugars were positively correlated with soil organic carbon, inorganic nitrogen, available phosphorus and potassium. We concluded that soil pH and C/N ratio were the most important drivers for microbial community structure and amino sugar pattern, while substrate availability was of great importance in determining the concentrations of microbial communities and residues. These findings could be used to facilitate interpretation of soil microbial community and amino sugar data derived from measurements in latitude or managed forests. 相似文献
The combination of fluorescence in situ hybridization and microautoradiography (FISH-MAR approach) was applied to brine samples of a solar saltern crystallizer pond from Mallorca (Spain) where the simultaneous occurrence of Salinibacter spp. and the conspicuous square Archaea had been detected. Radioactively labeled bicarbonate, acetate, glycerol, and an amino acid mixture were tested as substrates for the microbial populations inhabiting such brines. The results indicated that hitherto uncultured 'square Archaea' do actively incorporate amino acids and acetate. However, Salinibacter spp. only showed amino acid incorporation in pure culture, but no evidence of such activity in their natural environment could be demonstrated. No glycerol incorporation was observed for any component of the microbial community.Communicated by W.D. Grant 相似文献
Analytical procedures have been modified to determine the abundance of muramic acid in four different Holocene sediment samples. Muramic acid is specific to the peptidoglycan moiety of the cell walls of most eubacterial pro‐karyotic organisms. The following procedure seemed to be the most appropriate for the detection of muramic acid and amino acids, including diaminopimelic acid. Hydrolysis of the samples (in 6 N HCl, 4.5 h, at 100°C) was followed by separation and purification of amino sugars and amino acids using Amberlite XAD‐2 and then Bio‐Rad AG 50W‐X8 resins. The N,O‐heptafluorobutyryl‐n‐butyl ester derivatives were prepared by esterification in acidified (3 N HCl) n‐butanol for 3 h at 100°C, followed by acylation by refluxing with heptafluorobutyric anhydride in acetonitrile (2:1 v/v) for 12 min at 150°C. The derivatives were analyzed by gas chromatography (GC) and gas chromatography‐mass spectrometry. Fast atom bombardment (FAB) ionization was used for the muramic acid derivative to determine its molecular weight and structure, d‐and l‐amino acids were separated by GC and a capillary chiral column. By using this technique a stable N,O‐heptafluo‐robutyryl‐n‐butyl ester derivative of muramic acid was identified at picogram levels in Holocene sedimentary microbial communities. It has been reported previously that microorganisms in sediments rapidly degrade muramic acid from cell walls of dead prokaryotes. Kinetic experiments revealed that muramic acid was relatively stable in intact cell walls but decomposed rapidly in the free form. These investigations noted above showed that the concentration of muramic acid may be used as an indicator of the presence of the intact cell walls of cyanobacteria and most other bacteria in Holocene microbial communities, and of microbial contamination in samples older than the Holocene. 相似文献
Six ileally cannulated pigs (mean initial body weight 34.8 kg) were used to study the effect of microbial phytase on apparent ileal digestibility of P, total N and amino acids. Three P-adequate diets (digestible P concentration 2.3 g kg(- )l) containing barley (B), soyabean meal (S) or a mixture of the two (BS) with or without phytase supplement (1000 FTU x kg(-1)) were fed to pigs using a 6 x 6 Latin square design. The addition of phytase increased (p < 0.05) apparent ileal P digestibility of diets B, S and BS by 16.5, 19.2 and 19.2%, respectively. There was no effect of phytase on the ileal digestibility of total N. Apparent ileal digestibility of amino acids tended to increase in the BS diet supplemented with phytase (mean improvement of 2.2%); but no significant difference was found for any amino acid as compared with the unsupplemented diet. To asses the additivity of apparent amino acid digestibility, the determined values for the BS diet were compared to those calculated from digestibilities found in diets B and S. There were no significant differences between the determined and calculated values. It is concluded that the addition of microbial phytase to P-adequate diets does not affect ileal amino acid digestibility in growing pigs and that the apparent amino acid digestibility values determined in single ingredients may be additive when included into a complex diet. 相似文献
Optimal culture conditions of a methylotrophic Hyphomicrobium methylovorum and improved purification of serine hydroxymethyltransferase from the bacterium were established for the large-scale preparation of the enzyme. The first crystalline serine hydroxymethyltransferase from the microbial source was obtained in the apo form and found to be homogeneous. Amino acid analysis revealed that the enzyme had higher value per subunit for acidic and neutral amino acids than that from rabbit liver. The carboxy-terminal amino acid analysis suggested the sequence -Ile-Ala-Tyr. 相似文献
The substrates Z-XLeu-(Ala)2 and Z-Phe X-(Ala)2 (Z = benzyloxycarbonyl, X = various amino acid residues) were synthesized in order to investigate the primary specificity of acid proteinases from molds and yeasts. Since these peptides are mainly susceptible to cleavage by the enzymes at the peptide bonds shown by the arrows, it was possible to determine the specificity with respect to the amino acid residues on both sides of the splitting point. Pepsin was used for comparison. The results indicated that the microbial acid proteinases exhibit specificity for aromatic or hydrophobic amino acid residues on both sides of splitting point in peptide substrates, as does pepsin. However, the microbial enzymes showed somewhat broader specificity than pepsin. The former enzymes, which possess trypsinogen-activating ability, show specificity for a lysine residue, while pepsin or Mucor rennin-like enzyme does not. Although pepsin is very specific for a tyrosine residue on the imino side of the splitting point, the microbial enzymes do not show such stringency. 相似文献
The aromatic amino acid biosynthesis pathway, together with its downstream branches, represents one of the most commercially valuable biosynthetic pathways, producing a diverse range of complex molecules with many useful bioactive properties. Aromatic compounds are crucial components for major commercial segments, from polymers to foods, nutraceuticals, and pharmaceuticals, and the demand for such products has been projected to continue to increase at national and global levels. Compared to direct plant extraction and chemical synthesis, microbial production holds promise not only for much shorter cultivation periods and robustly higher yields, but also for enabling further derivatization to improve compound efficacy by tailoring new enzymatic steps. This review summarizes the biosynthetic pathways for a large repertoire of commercially valuable products that are derived from the aromatic amino acid biosynthesis pathway, and it highlights both generic strategies and specific solutions to overcome certain unique problems to enhance the productivities of microbial hosts. 相似文献
Selenocysteine and pyrrolysine are the 21st and 22nd amino acids, which are genetically encoded by stop codons. Since a number
of microbial genomes have been completely sequenced to date, it is tempting to ask whether the 23rd amino acid is left undiscovered
in these genomes. Recently, a computational study addressed this question and reported that no tRNA gene for unknown amino
acid was found in genome sequences available. However, performance of the tRNA prediction program on an unknown tRNA family,
which may have atypical sequence and structure, is unclear, thereby rendering their result inconclusive. A protein-level study
will provide independent insight into the novel amino acid. 相似文献
The
amino acid octamers (
-Glu)8 and (
Asp)8 were synthesized in order to screen for new microbial degraders of unnatural
amino acid peptides. We have successfully isolated the microorganisms from soil acclimated to a medium containing the oligopeptides; they were classified as Klebsiella ornithinolytica, Delftia acidovorans, and α-Proteobacteria.相似文献