Microbial protein from leguminous and non-leguminous substrates

The biodegradation of leguminous and non-leguminous organic materials by Pleurotus sajor-caju and P. ostreatus was studied. Comparisons were made between mushroom yield on both types of substrates. The conversion percentage from dry substrate to fresh mushroom (biological efficiency) was determined. Mushrooms were analyzed for their protein content, carbohydrates, percentages of ash, fat content and crude fibre. The energy value of fruit bodies was computed. The yield obtained from leguminous substrates was significantly high as compared to that yield which was obtained from non-leguminous substrates. The lowest yield of P. sajor caju (10 g ± 0.3 g) and P. ostreatus (6.5 g ± 1.2 g) was obtained using the Bougainuillea substrate and the highest one (519 g ± 16 g, P. sajor-caju and 488 g ± 18 g, P. ostreatus) using nitrogen-fixing leguminous Leucaena leucocepholea. The nitrogen content in fruit bodies was found to be higher with leguminous substrate than with non-leguminous ones.     

Nitrogen-fixing bacteria associated with leguminous and non-leguminous plants

Nitrogen is generally considered one of the major limiting nutrients in plant growth. The biological process responsible for reduction of molecular nitrogen into ammonia is referred to as nitrogen fixation. A wide diversity of nitrogen-fixing bacterial species belonging to most phyla of the Bacteria domain have the capacity to colonize the rhizosphere and to interact with plants. Leguminous and actinorhizal plants can obtain their nitrogen by association with rhizobia or Frankia via differentiation on their respective host plants of a specialized organ, the root nodule. Other symbiotic associations involve heterocystous cyanobacteria, while increasing numbers of nitrogen-fixing species have been identified as colonizing the root surface and, in some cases, the root interior of a variety of cereal crops and pasture grasses. Basic and advanced aspects of these associations are covered in this review.     

Effects of nitrogen addition on litter decomposition, soil microbial biomass, and enzyme activities between leguminous and non-leguminous forests

Anthropogenic nitrogen (N) deposition is an expanding problem that affects the functioning and composition of forest ecosystems, particularly the decomposition of forest litters. Legumes play an important role in the nitrogen cycle of forest ecosystems. Two litter types were chosen from Zijin Mountain in China: Robinia pseudoacacia leaves from a leguminous forest (LF) and Liquidambar formosana leaves from a non-leguminous forest (NF). The litter samples were mixed into original forest soils and incubated in microcosms. Then, they were treated by five forms of N addition: NH₄ ⁺, NO₃ ^?, urea, glycine, and a mixture of all four. During a 6-month incubation period, litter mass losses, soil microbial biomass, soil pH, and enzyme activities were investigated. Results showed that mixed N and NO₃ ^?-N addition significantly accelerated the litter decomposition rates of LF leaves, while mixed N, glycine-N, and urea-N addition significantly accelerated the litter decomposition rates of NF leaves. Litter decomposition rates and soil enzyme activities under mixed N addition were higher than those under single form of N additions in the two forest types. Nitrogen addition had no significant effects on soil pH and soil microbial biomass. The results indicate that nitrogen addition may alter microbial allocation to extracellular enzyme production without affecting soil microbial biomass, and then affected litter decomposition process. The results further reveal that mixed N is a more important factor in controlling litter decomposition process than single form of N, and may seriously affect soil N cycle and the release of carbon stored belowground.     

Antifungal peptides: Origin, activity, and therapeutic potential

Antifungal peptides have been identified in a wide range of life forms which include plants, mammals, and microorganisms. Their structures are as varied as their antifungal properties. Semisynthetic and fully synthetic analogs have been developed from a few of these natural peptides that are superior to the parent compound. A few of these peptides hold promise in combating fungal infections and have entered clinical trials.     

Plant-derived antifungal proteins and peptides

Plants produce potent constitutive and induced antifungal compounds to complement the structural barriers to microbial infection. Approximately 250,000-500,000 plant species exist, but only a few of these have been investigated for antimicrobial activity. Nevertheless, a wide spectrum of compound classes have been purified and found to have antifungal properties. The commercial potential of effective plant-produced antifungal compounds remains largely unexplored. This review article presents examples of these compounds and discusses their properties.     

Apoptosis-inducing antifungal peptides and proteins

Despite the availability of various classes of antimycotics, the treatment of patients with systemic fungal infections is challenging. Therefore the development of new antifungals is urgently required. Promising new antifungal candidates are antimicrobial peptides. In the present review, we provide an overview of antifungal peptides isolated from plants, insects, amphibians and mammals that induce apoptosis. Their antifungal spectrum, mode of action and toxicity are discussed in more detail.     

Antifungal properties of Canavalia ensiformis urease and derived peptides

Ureases (EC 3.5.1.5) are metalloenzymes that hydrolyze urea into ammonia and CO₂. These proteins have insecticidal and fungicidal effects not related to their enzymatic activity. The insecticidal activity of urease is mostly dependent on the release of internal peptides after hydrolysis by insect digestive cathepsins. Jaburetox is a recombinant version of one of these peptides, expressed in Escherichia coli. The antifungal activity of ureases in filamentous fungi occurs at submicromolar doses, with damage to the cell membranes. Here we evaluated the toxic effect of Canavalia ensiformis urease (JBU) on different yeast species and carried out studies aiming to identify antifungal domain(s) of JBU. Data showed that toxicity of JBU varied according to the genus and species of yeasts, causing inhibition of proliferation, induction of morphological alterations with formation of pseudohyphae, changes in the transport of H⁺ and carbohydrate metabolism, and permeabilization of membranes, which eventually lead to cell death. Hydrolysis of JBU with papain resulted in fungitoxic peptides (∼10 kDa), which analyzed by mass spectrometry, revealed the presence of a fragment containing the N-terminal sequence of the entomotoxic peptide Jaburetox. Tests with Jaburetox on yeasts and filamentous fungi indicated a fungitoxic activity similar to ureases. Plant ureases, such as JBU, and its derived peptides, may represent a new alternative to control medically important mycoses as well as phytopathogenic fungi, especially considering their potent activity in the range of 10⁻⁶–10⁻⁷ M.     

Antifungal activity of synthetic peptides derived from Impatiens balsamina antimicrobial peptides Ib-AMP1 and Ib-AMP4

Seeds of Impatiens balsamina contain a set of related antimicrobial peptides (Ib-AMPs). We have produced a synthetic variant of Ib-AMP1, oxidized to the bicyclic native conformation, which was fully active on yeast and fungal strains; and four linear 20-mer Ib-AMP variants, including two all-D forms. We show that the all-D variants are as active on yeast and fungal strains as native peptides. In addition, fungal growth inhibition nor salt-dependency of Ib-AMP4 could be improved by more than two-fold via replacement of amino acid residues by arginine or tryptophan. Native Ib-AMPs showed no hemolytic nor toxic activity up to a concentration of 100 microM. All these data demonstrate the potential of the native Ib-AMPs to combat fungal infections.     

Mass spectrometry of peptides and proteins

This tutorial article introduces mass spectrometry (MS) for peptide fragmentation and protein identification. The current approaches being used for protein identification include top-down and bottom-up sequencing. Top-down sequencing, a relatively new approach that involves fragmenting intact proteins directly, is briefly introduced. Bottom-up sequencing, a traditional approach that fragments peptides in the gas phase after protein digestion, is discussed in more detail. The most widely used ion activation and dissociation process, gas-phase collision-activated dissociation (CAD), is discussed from a practical point of view. Infrared multiphoton dissociation (IRMPD) and electron capture dissociation (ECD) are introduced as two alternative dissociation methods. For spectral interpretation, the common fragment ion types in peptide fragmentation and their structures are introduced; the influence of instrumental methods on the fragmentation pathways and final spectra are discussed. A discussion is also provided on the complications in sample preparation for MS analysis. The final section of this article provides a brief review of recent research efforts on different algorithmic approaches being developed to improve protein identification searches.     

NMR spectroscopy of peptides and proteins

High resolution nuclear magnetic resonance (NMR) spectroscopy is the only method available for determining the three-dimensional structures of peptides and proteins in solution at atomic resolution. This article deals with a range of practical considerations associated with such studies, including sample preparation, instrumental setup, one- and two-dimensional NMR methods, interpretation of spectral data, and structure calculations.     

Gas-phase hydrolysis of proteins and peptides

A simplified gas-phase hydrolysis procedure for proteins and peptides is described. The apparatus consists of a glass vacuum desiccator, a ceramic plate, and a Teflon ring. The method was shown to give reproducible compositions for hydrolysis of human serum albumin and microanalysis of α-melanocyte stimulating hormone including the quantitation of as little as one residue of tryptophan. It minimizes sample handling and allows for the simultaneous hydrolysis of a large number of samples.     

Electron transfer in peptides and proteins

Proteins and peptides use their amino acids as medium for electron-transfer reactions that occur either in single-step superexchange or in multistep hopping processes. Whereas the rate of the single-step electron transfer dramatically decreases with the distance, a hopping process is less distance dependent. Electron hopping is possible if amino acids carry oxidizable side chains, like the phenol group in tyrosine. These side chains become intermediate charge carriers. Because of the weak distance dependency of hopping processes, fast electron transfer over very long distances occurs in multistep reactions, as in the enzyme ribonucleotide reductase.