Significantly enhanced thermal stability of HMX by phase-transition lysozyme coating

A new robust bio-inspired route by using lysozyme aqueous solution for surface modification on 1,3,5,7-tetranitro-1,3,5,7-tetrazocane(HMX)was described in this paper.HMX crystals were coated by in situ phase transition of lysozyme(PTL)molecules.The HMX decorated by PTL was characterized by SEM,XRD,FTIR and XPS,demonstrating a dense core-shell coating layer.The coverage of lysozyme on HMX crystal was calculated by the ratio of sulfur content.The surface coverage increased from 60.5% to 93.5% when the content of PTL was changed from 0.5 wt% to 2.0 wt%,indicating efficient coating.The thermal stability of HMX was investigated by in situ XRD and DSC.The thermal phase transition temperature of HMX(β to δ phase)was delayed by 42℃ with 2.0 wt% PTL coating,which prevented HMX from thermal damage and sensitivity by the effect of PTL coating.After heating at 215℃,large cracks appeared in the naked HMX crystal,while the PTL coated HMX still maintained intact,with the impact energy of HMX dropped dramatically from 5 J to 2 J.However,the impact energy of HMX with 1.0 wt% and 2.0 wt% coating content(HMX@PTL-1.0 and HMX@PTL-2.0)was unchanged(5 J).Present results potentially enable large-scale fabrication of polymorphic energetic materials with outstanding thermal stability by novel lysozyme coating.

Molecular Characterization and Antibacterial Activity of a G-Type Lysozyme in Yellow Drum(Nibea albiflora)

Lysozyme(EC3.2.1.17)plays an important role in the immune response;as a nonspecific immune factor,it can resist causative agents.Lysozyme can be divided into c-type and g-type in fish.In a previous study,through genome-wide association analysis,the g-type lysozyme gene,which is named NaLyg in yellow drum(Nibea albiflora),was found to be a key candidate gene for disease resistance in response to Vibrio harveyi infection.The cDNA of NaLyg was 1025 bp,including four exons and three introns,and its open reading frame(ORF)had a full-length of 582 bp,encoding 193 amino acids.NaLyg was found to be conserved during evolution through bioinformatic analyses.The NaLyg protein possessed a sugar binding domain and three catalytic sites,including Glu71,Asp84 and Asp101.Quantitative qRT-PCR results confirmed that NaLyg gene mRNA was visibly increased after V.harveyi infection.The NaLyg protein purified by prokaryotic expression killed some gram-negative bacterial pathogens by inducing cell wall destruction,including V.harveyi,Aeromonas hydrophila and Edwardsiella tarda.Moreover,the NaLyg protein killed two gram-positive bacteria,Bacillus subtilis and Staphylococcus aureus.Taken together,the experimental results suggested that the NaLyg protein of N.albiflora played an important role in fighting bacterial infections.

Concentration-Dependent Effect of Nickel Ions on Amyloid Fibril Formation Kinetics of Hen Egg White Lysozyme:a Raman Spectroscopy Study

Nickel,an important transi-tion metal element,is one of the trace elements for hu-man body and has a crucial impact on life and health.Some evidences show the excess exposure to metal ions might be associated with neurological diseases.Herein,we applied Raman spectroscopy to study the Ni(II)ion effect on kinetics of amyloid fibrillation of hen egg white lysozyme(HEWL)in thermal and acidic conditions.Using the well-known Raman indicators for protein tertiary and secondary structures,we monitored and analyzed the concentration effect of Ni(II)ions on the unfolding of tertiary structures and the transformation of sec-ondary structures.The experimental evidence validates the accelerator role of the metal ion in the kinetics.Notably,the additional analysis of the amide I band profile,combined with thioflavin-T fluorescence assays,clearly indicates the inhibitory effect of Ni(II)ions on the formation of amyloid fibrils with organizedβ-sheets structures.Instead,a more significant promotion influence is affirmed on the assembly into other aggregates with disordered struc-tures.The present results provide rich information about the specific metal-mediated protein fibrillation.

Coordination and Photoisomerization of Azobenzene-Amino Acid Schiff Base Copper(II) Complexes to Lysozyme

In this study, we exhibited an amino acid (arginine and threonine) derivative Schiff base copper(II) complexes incorporating an azobenzene moiety as a photoresponsive site and conjugated it to egg white lysozyme, a well-known protein, to change ligand conformation under binding to lysozyme. Among several spectroscopic investigations, ESR clearly showed that the nitrogen atom of the amino acid residue of lysozyme was bound to the paramagnetic copper(II) ion of the complex, and UV light irradiation confirmed photoisomerization of the azobenzene moiety of the ligand to cis-form. The binding mode was considered by means of spectroscopic as well as computational methods, whereas complete crystallographic verification was still a preliminary stage.

家蚕抗菌蛋白Cecropin D和Lysozyme的分离纯化及性质鉴定

经大肠杆菌E．coli K12D31诱导后的家蚕幼虫，其免疫血淋巴经CM-Sepharose CL-6B离子交换层析、Sephadex G-100凝胶过滤层析及HPLC分离后，得到2种抗菌蛋白，经液相色谱-电喷雾质谱（ESI-MS）分析鉴定，纯化的2种抗菌蛋白分别是家蚕抗菌肽eeeropin D和溶菌酶lysozyme酸性电泳（A-PAGE）测活免疫血淋巴得到抗菌蛋白活性谱：cecropin D在8h无显著表达，12h有较强抗菌活性，30h达到最高，以后逐渐降低；lysozyme在8h后检测到表达，12h到达高峰，之后逐渐下降，48h的血淋巴中未检测到明显的表达．研究认为抗菌蛋白lysozyme和cecropin D是家蚕幼虫感染细菌8h后大量表达的抗菌蛋白，主要参与细菌感染12～30h的血淋巴对细菌的清除，是参与家蚕幼虫抗菌免疫的重要蛋白．

Hemozoin triggers tumor necrosis factor alpha-mediated release of lysozyme by human adherent monocytes:new evidences on leukocyte degranulation in P.falciparum malaria

Objective:Avidly phagocytosed hemozoin(malarial pigment) alters several functions of human monocytes and stimulates generation of several cytokines.Recently,we showed that phagocytosis of hemozoin by human monocytes increases expression and activity of matrix metalloproteinase-9,a proteolytic enzyme available in specific gelatinase granules,which contain several enzymes including lysozyme.Present work investigated active lysozyme release after phagocytosis of hemozoin and its dependence on production of tumor necrosis factor alpha. Methods:After phagocytosis of hemozoin,hemozoin-containing trophozoites or control meals(opsonized nonparasitized red blood cells and latex particles),monocyte supematants were monitored for 2 hours,in presence of blocking anti-human tumor necrosis factor alpha antibodies or recombinant human tumor necrosis factor alpha cytokine in selected experiments.Lysozyme release was evaluated by a specific spectrometric assay measuring lysozyme activity after coincubation of cell supematants with suspensions of Mycrococcus Lysodeikticus,while levels of soluble tumor necrosis factor alpha were analyzed by specific enzyme-linked immunodsorbent assay. Results:Levels of lysozyme activity and soluble tumor necrosis factor alpha protein were increased in hemozoin in-or trophozoites-laden monocytes supematants.Phagocytosis per se(control meals) also increased lysozyme release,but levels were significantly lower than those obtained after phagocytosis of hemozoin or trophozoites. Interestingly,all effects on lysozyme release observed after phagocytosis were abrogated by blocking anti-human tumor necrosis factor alpha antibodies,while they were mimicked by recombinant human tumor necrosis factor alpha cytokine.Conclusions:Present work shows that phagocytosis of hemozoin promotes monocyte degranulation and enhances active lysozyme release.The effect requires tumor necrosis factor alpha mediation.

Lysozyme as an alternative to growth promoting antibiotics in swine production

Lysozyme is a naturally occurring enzyme found in bodily secretions such as tears, saliva, and milk. It functions as an antimicrobial agent by cleaving the peptidoglycan component of bacterial cell walls, which leads to cell death. Antibiotics are also antimicrobials and have been fed at subtherapeutic levels to swine as growth promoters. These compounds benefit swine producers by minimizing production losses by increasing feed efficiency and decreasing susceptibility to bacterial infection and disease. This manuscript reviews the knowledge of the effects of lysozyme, as compared to traditional subtherapeutic antibiotics in swine feed, on pig performance and health. It is clear from decades of studies that antibiotic use in feeds increases pig performance, particularly in the nursery. Similarly, lysozyme, as a feed additive, increases growth and feed efficiency. While the mechanism by which antibiotics and lysozyme improve performance is not clearly understood, both of these feed additives improve gastrointestinal health, improve the metabolic profile, and alter the gastrointestinal bacteria ecology of swine. Therefore, lysozyme is a suitable alternative to growth-promoting subtherapeutic antibiotic use in swine feed.

Corrosion Inhibition by Co-Immobilized Lysozyme and Lipase in Circulating Cooling Water System

The corrosion inhibition performance of co-immobilized lysozyme and lipase was investigated in a recirculating cooling water system. Four methods were carried out in co-immobilization, and the operating parameters were optimized by using the respond surface methodology(RSM). The corrosion inhibition performance of co-immobilized lipase and lysozyme was evaluated by weight loss measurements and electrochemical measurements. The results revealed that the optimal co-immobilization method should be the sequential immobilization of lysozyme and then lipase. The inhibition efficiency was 86.10% under the optimal co-immobilized conditions. Electrochemical data showed that co-immobilized lysozyme and lipase was a mixed-type inhibitor and the corrosion inhibition efficiency was 81%.

Analysis of Resistant Spectrum to Rice Blast in Transgenic Rice Lines Introduced Lysozyme Gene from T4 Phage

The receptor cultivar Nan29 and thirty-six T5 rice lines derived from ten T0 generation transgen-ic plants harboring lysozyme gene were challenged in the greenhouse by inoculating 63 isolates belonging to 48 races of Magnaporthe grisea from Yunnan Province. The transgenic rice lines exhibited resistance to more than 72% of isolates inoculated in this experiment, and 38.1% (24 isolates) of them could infect the receptor cultivar Nan29. The results indicated that the transgenic rice lines possessed wide-spectrum resistance against various rice blast races and the resistant spectrum of rice lines were different although some lines derived from same T0 plant. The transgenic rice lines exhibited also high resistance to leaf and neck blast in the disease field evaluation, but not all of resistant lines against leaf blast were resistant to neck blast.

Exploring the Performance of Magnetic Immobilized Lysozyme on Sludge Hydrolysis and Mechanism of Improving Dewaterability of Excess Sludge

The performance of the immobilized lysozyme and the native lysozyme on enhancing the excess sludge dewaterability was investigated.The results indicated that the specific resistance to filtration(SRF)decreased by 62.8%for native lysozyme and 53.6%for immobilized lysozyme at the enzyme dosage of 9 mg/g(dry sludge).Correlation analysis was carried out to explore the role of different extracellular polymeric substance(EPS)fractions on excess sludge dewaterability.The results illustrated that the SRF negatively correlated with protein,polysaccharide from soluble EPS(S-EPS)and loosely bound EPS(LB-EPS)and positively correlated with that from tightly bound EPS(TB-EPS).Three-dimensional excitation emission matrix(3D-EEM)fluorescence analysis combined with the scanning electron microscope(SEM)images,revealed that sludge floc structure and microbial cells were destroyed by enzymatic treatment,and that the enzymatic hydrolysis could help to improve the transformation of hydrophilic groups from TB-EPS and the performance of the excess sludge dewatering process.The assessment of hydrolysis using the immobilized enzyme provided a new insight for the safe disposal of the sludge.

Refolding of Denatured/Reduced Lysozyme Using Weak-Cation Exchange Chromatography

Oxidative refolding of the denatured/reduced lysozyme was investigated by using weak-cation exchange chromatography (WCX). The stationary phase of WCX binds to the reduced lysozyme and prevented it from forming intermolecular aggregates. At the same time urea and ammonium sulfate were added to the mobile phase to increase the elution strength for lysozyme. Ammonium sulfate can more stabilize the native protein than a common eluting agent, sodium chloride. Refolding of lysozyme by using this WCX is successfully. It was simply carried out to obtain a completely and correctly refolding of the denatured lysozyme at high concentration of 20.0 mg/mL.

Molecular characterization and functional analysis of two new lysozyme genes from soybean cyst nematode(Heterodera glycines)

Soybean cyst nematode(SCN, Heterodera glycines(I.)) is one of the most important soil-borne pathogens for soybeans. In plant parasitic nematodes, including SCN, lysozyme plays important roles in the innate defense system. In this study, two new lysozyme genes(Hg-lys1 and Hg-lys2) from SCN were cloned and characterized. The in situ hybridization analyses indicated that the transcripts of both Hg-lys1 and Hg-lys2 accumulated in the intestine of SCN. The q RT-PCR analyses showed that both Hg-lys1 and Hg-lys2 were upregulated after SCN second stage juveniles(J2 s) were exposed to the Grampositive bacteria Bacillus thuringiensis, Bacillus subtilis or Staphylococcus aureus. Knockdown of the identified lysozyme genes by in vitro RNA interference caused a significant decrease in the survival rate of SCN. All of the obtained results indicate that lysozyme is very important in the defense system and survival of SCN.

Genomic organization and evolution of ruminant lysozyme c genes

Ruminant stomach lysozyme is a long established model of adaptive gene evolution. Evolution of stomach lysozyme function required changes in the site of expression of the lysozyme c gene and changes in the enzymatic properties of the enzyme. In ruminant mammals, these changes were associated with a change in the size of the lysozyme c gene family. The recent release of near complete genome sequences from several ruminant species allows a more complete examination of the evolution and diversification of the lysozyme c gene family. Here we characterize the size of the lysozyme c gene family in extant ruminants and demonstrate that their pecoran ruminant ancestor had a family of at least 10 lysozyme c genes, which included at least two pseudogenes. Evolutionary analysis of the ruminant lysozyme c gene sequences demonstrate that each of the four exons of the lysozyme c gene has a unique evolutionary history, indicating that they participated independently in concerted evolution. These analyses also show that episodic changes in the evolutionary constraints on the protein sequences occurred, with lysozyme c genes expressed in the abomasum of the stomach of extant ruminant species showing the greatest levels of selective constraints.

Nano-silica particles enhanced adsorption and recognition of lysozyme on imprinted polymers gels

Molecular imprinting technology has a great potential to be used in protein separation and purification. In this work, lysozyme imprinted polyacrylamide gel was prepared with silica particles as a sacrificial template to generate macro-porosity for fast adsorption. The adsorption equilibrium time and adsorption capacity were 9 h and 56 mg/g respectively, which was 2 h less and 2.3-fold more than polymers without the sacrificial template. In order to test molecular imprinting polymers （MIPs） ＇ selectivity, bovine serum albumin （BSA） was chosen as interferent for binary adsorption tests. In addition, the adsorption selectivity was further investigated using different molar ratios of lysozyme to BSA with fixed total concentration of proteins, as well as using various total concentra- tions of proteins with an equimolar ratio of lysozyme to BSA. It has been proven that the total con- centration of proteins should be larger than 1.5 × 10^-7 mol/mL, when the molar ratio of BSA to Lyz is 1： 1, in order to effectively separate Lyz from the binary protein mixture. The macro-porous lyso- zyme molecularly imprinted polymers have less adsorption time, larger adsorption capacity, and bet- ter imprinting effect.

Combined Use of Co-immobilized Lysozyme and Lipase and Chemical Inhibitors in Circulating Cooling Water System

In order to improve the stability and corrosion inhibition performance of bioenzyme, lipase and lsozyme were co-immobilized on the mesoporous molecular sieve MCM-41 by the adsorption method. Then the immobilized enzymes were combined with amino trimethylene phosphonic acid and polyaspartic acid to prevent corrosion caused by circulating cooling water. The weight-loss method and electrochemical techniques were used to evaluate the performance of composite inhibitors. The co-immobilized lysozyme and lipase achieved good inhibition effects. After they were combined with amino trimethylene phosphonic acid and polyaspartic acid, the corrosion inhibition properties were further improved. The inhibition efficiency was promoted to 94.4%. During the corrosion inhibition process, the immobilized enzymes played an important role. The addition of corrosion inhibitor could inhibit the anodic dissolution and cathodic hydrogen evolution process of carbon steel at the same time. The adsorption of co-immobilized lysozyme and lipase composite inhibitor on the steel surface was a joint action involving physical adsorption and chemical adsorption.

Pr(Ⅲ) and Nd(Ⅲ) Absorption Spectroscopic Probe to Investigate Interaction with Lysozyme (HEW)

Pr（ Ⅲ ） and Nd（ Ⅲ ） can be utilized as absorption spectroscopic probes to investigate the interaction of biomolecules like Lysozyme （HEW） with Ca（ Ⅱ ） in-vitro ; the most abundant metal ion in the human body system. The spectroscopic techniques involving comparative absorption, absorption difference, and quantitative intensity analysis using 4f-4f transitions are utilized for changes in the inner sphere coordination pattern of Pr（ Ⅲ ） and Nd（Ⅲ ） in solution as well as in solid state. The present study deals with an important biomolecule in human metabolism, that is, Lysozyme （HEW）. The absorption er-Condon the probab spectral parameters such as the oscillator strength （P）, the Judd-Ofelt （Tλ） intensity parameters, and the Slatinter electronic parameters are calculated using chi square methods. The obtained results are used to determine le geometry of the complex in the solution, the nature of the bond between Pr（Ⅲ）/Nd（Ⅲ） with lysozyme, and the inner sphere coordination environment of f-f transitions. The results obtained from various experimental conditions are utilized to investigate the coordination changes in the Pr（Ⅲ）/Nd（Ⅲ） complexes caused by different coordinating sites of lysozyme, normalized bite, denticity, the solvent nature, the coordination number, the nature of bond and other parameters to mimic the interaction of the Ca（ Ⅱ ） ion with such biomolecule.

Study on Preventive Effect of Cecropin and Lysozyme on Skin Fester Disease in Trionyx sinensis

[ Objective ] This study was to investigate the preventive effect of cecropin and lysozyme on skin fester disease in Trionyx sinensis. [ Method ] By employing intramuscular injection method, the preventive effect of mixtures of cecropin （10 mg/kg） and lysozyme（ 10 mg/kg） and of cecropin（6 mg/kg） and lysozyme（ 5 mg/kg） were challenged on Aeromonas encheleia-caused skin fester disease in T. sinensis. The mortality and relative percent survival were used evaluate the preventive effect of different drugs. The serum agglutination titer, cytophagocytosis and bactericidal activity were detected to compare the effect of drugs on the immunity and disease resistance of T. sinensis. [ Result] The residue concentrations of gentamicin and lysozyme in serum were compared to evaluate the side effect of drugs used challenged. The results showed that combined use of cecropin and lysozyme gave the best preventive effect and immunity in T. sinensis, with an extremely significant difference from other treatments （ P 〈 0.01 ）. Analysis of drug residue in serum indicated that lysozyme was completely metabolized 6 days after medication, while gentamicin residue was still observed 12 days later. [ Conclusion] The combined application of cecropin and lysozyme showed better preventive effect than antibiotics in fester disease of T. sinensis. Moreover, no drug residue and side effect were observed in the combined use treatment. This method avoids the side effect of antibiotics, worthy of extension at commercial scale.

Research Advances on Nutritional Physiological Functions and Application of Lysozyme

Lysozyme is an important immune factor of animals and a hydrolase specifically acting on microbial cell wall, which exhibits various functions, such as anti-bacteria, anti-inflammation, antivirus, enhancing immunity of animals and improving gastrointestinal mieroflora. Lysozyme is non-toxic to livestock and poultry without drug residue or side effects, which is a novel green health-care feed additive. In this paper, the types, function mechanisms and physiological functions of lysozyme and its application in livestock industry were summarized, aiming at providing a scientific basis for the application of lysozyme in animal feeds.

Therapeutical effect of marine lysozyme suppository on bacterial vaginitis caused by S. aureus and E. coli

Aim To research therapeutical effect of marine lysozyme suppository on bacterial vaginitis caused by S. aureus and E. coll. Methods Lysozyme obtained from concha ostreae which were used to preparate marine lyso- zyme suppository. The identification and test of suppository was in line with the standards stated in Chinese pharma- coperia（2010 edition）. After determined its quality control, we studied its therapeutical effect on bacterial vaginitis which caused by S. aureus and E. coli, by vaginitis model in rats infection of S. aureus bacteria and E. coli. Results The preparation technology of marine lysozyme suppository was simple, convenient and clinically effective. The marine lysozyme suppository was delivered by dose 0.5, 0.25, 0. 125 g ~ kg-1, the cure rates of S. aureus infec- tion were 80% , 50% and 30% , respectively, the cure rates of E. coli infection were 90% , 60% and 30% re- spectively, the cure rates of mixed infection were 92.9% , 82. 1% and 92.9% respectively. Marine lysozyme sup- pository had an outstanding therapeutical effect on bacterial vaginitis which caused by S. aureus and E. coll. Con- clusion The preparative process of marine lysozyme suppository was practicable, and it had a good therapeutic effect on bacterial vaginitis caused by S. aureus and E. coll.

Dramatic Changes of Matrix Metalloproteinases-7 and Lysozyme in the Ulcerative Colitis of Mice Induced by Dextran Sulfate Sodium

Ulcerative colitis （UC） is a lifelong illness with profound emotional and social impacts, and could cause serious damage to large intestine, especially in colon. However, the pathogenesis of UC remained unclear. The present study attempts to find out the role of matrix metalloproteinases-7 （MMP-7） and lysozyme in the pathogenesis of UC through a mice model induced by dextran sulfate sodium （DSS）. The UC model was evaluated both by disease activity index （DAI） and the intestinal histopathology. The results show that there is a high correlation between the DAI score and the pathological changes of colon. Interleukin-6 （IL-6） serum levels and large intestinal fluids levels in UC mice are always higher than that of the control groups, which might be associated with the degree of the inflammation damage in the colon. The change tendency of the MMP-7 mRNA and protein expressions are both up-regulated firstly and then down-regulated from 1 to 5 d in the colon, but only the MMP-7 protein is up-regulated at 7 d again. The up-regulated MMP-7 levels in the early stage of UC may play a protective role through the activated defensins, while the down-regulated levels in the mid-later stage of UC may be connected with the severe lesions in the colon. However, the up-regulated MMP-7 levels in the later stage of UC in the colon may also contribute to the tissue repair or be served as a marker to CRC （colorectal cancer）. The distribution of lysozyme protein indicates that there may be Paneth-like cells in the colon. Both the changes of MMP-7 and lysozyme in the small intestine may play a protective role for the safe environment of the whole gut, especially to the colon of UC.