Anaerobic digestion(AD)is widely employed for sludge stabilization and waste reduction.However,the slow hydrolysis process hinders methane production and leads to prolonged sludge issues.In this study,an efficient and...Anaerobic digestion(AD)is widely employed for sludge stabilization and waste reduction.However,the slow hydrolysis process hinders methane production and leads to prolonged sludge issues.In this study,an efficient and eco-friendly lysozyme pre-treatment method was utilized to address these challenges.By optimizing lysozyme dosage,hydrolysis and cell lysis were maximized.Furthermore,lysozyme combined with hydrothermal pretreatment enhanced overall efficiency.Results indicate that:(1)When lysozyme dosage reached 90 mg/g TS after 240 min of pretreatment,SCOD,soluble polysaccharides,and protein content reached their maxima at 855.00,44.09,and 204.86 mg/L,respectively.This represented an increase of 85.87%,365.58%,and 259.21%compared to the untreated sludge.Threedimensional fluorescence spectroscopy revealed the highest fluorescence intensity in the IV region(soluble microbial product),promoting microbial metabolic activity.(2)Lysozyme combined with hydrothermal pretreatment significantly increased SCOD,soluble proteins,and polysaccharide release from sludge,reducing SCOD release time.Orthogonal experiments identified Group 3 as the most effective for SCOD and soluble polysaccharide release,while Group 9 released the most soluble proteins.The significance order of factors influencing SCOD,soluble proteins,and polysaccharide release is hydrothermal temperature>hydrothermal time>enzymatic digestion time.(3)The lysozyme-assisted hydrothermal pretreatment group exhibited the fastest release and the highest SCOD concentration of 8,135.00 mg/L during anaerobic digestion.Maximum SCOD consumption and cumulative gas production increased by 95.89%and 130.58%,respectively,compared to the control group,allowing gas production to conclude 3 days earlier.展开更多
Eco-friendly and antimicrobial globular protein lysozyme is widely produced for several commercial applications.Interestingly,it can also be able to convert mechanical and thermal energy into electricity due to its pi...Eco-friendly and antimicrobial globular protein lysozyme is widely produced for several commercial applications.Interestingly,it can also be able to convert mechanical and thermal energy into electricity due to its piezo-and pyroelectric nature.Here,we demonstrate engineering of lysozyme into piezoelectric devices that can exploit the potential of lysozyme as environmentally friendly,biocompatible material for mechanical energy harvesting and sensorics,especially in micropowered electronic applications.Noteworthy that this flexible,shape adaptive devices made of crystalline lysozyme obtained from hen egg white exhibited a longitudinal piezoelectric charge coefficient(d-2.7 pC N^(-1))and piezoelectric voltage coefficient(g-76.24 mVmN^(-1))which are comparable to those of quartz(-2.3 pC N^(-1) and 50 mVmN^(-1)).Simple finger tapping on bio-organic energy harvester(BEH)made of lysozyme produced up to 350 mV peak-to-peak voltage,and a maximum instantaneous power output of 2.2 nW cm^(-2).We also demonstrated that the BEH could be used for self-powered motion sensing for real-time monitoring of different body functions.These results pave the way toward self-powered,autonomous,environmental-friendly bio-organic devices for flexible energy harvesting,storage,and in wearable healthcare monitoring.展开更多
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 genom...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.展开更多
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 transitio...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.展开更多
Aim To induce and express the T4 lysozyme in Pichia pastoris and test the antibacterial activity of the protein. Methods T4 lysozyme gene was inserted into expression vector pPIC9K of Pichia pastoris with the fusion a...Aim To induce and express the T4 lysozyme in Pichia pastoris and test the antibacterial activity of the protein. Methods T4 lysozyme gene was inserted into expression vector pPIC9K of Pichia pastoris with the fusion at N terminal. The recombinant plasmid was digested by Sal I and then introduced into prepared GS115 competent cells by electroporation. Positive clone and multiple inserts were screened. The secreted proteins in the supernatants were tested. In the agar holes diffusion assay, our expressed protein showed significant antibacterial circles. Results T4 lysozyme protein inhibited the growth of staphylococcus aureus and streptococcus Pneumoniae. There was no difference in the bactericidal activity and the amount of protein expression between the single and multiple copies. The antibacterial activity of expressed protein remained the same during the heat stability test. Conclusion T4 lysozyme was successfully induced and expressed in Pichia pastoris. There is no relationship between copy number and expression. T4 lysozyme protein is heat stable.展开更多
The interactions of chlorogenic acid （CA）, neochlorogenic acid （NCA） and cryptochlorogenic acid （CCA） with lysozyme （LYSO） were investigated in physiological buffer by fluorescence spectroscopy. The mechanism ...The interactions of chlorogenic acid （CA）, neochlorogenic acid （NCA） and cryptochlorogenic acid （CCA） with lysozyme （LYSO） were investigated in physiological buffer by fluorescence spectroscopy. The mechanism study indicated that CA, NCA and CCA could strongly quench the intrinsic fluorescence of LYSO through static quenching procedures with one binding site. Thermodynamic data show that the major force in the binding processes of CA to LYSO was hydrophobic interactions; for NCA, it was the hydrogen bonds and van der Waals forces, as for the CCA system, the mainly force is electrostatic force.展开更多
Lysozymes can hydrolyze bacteria and play an important role in animal digestion and innate immunity. The cDNA of a chicken-type lysozyme gene （Mdlys） was cloned from housefly （Musca domestica）. The 484 bp full-len...Lysozymes can hydrolyze bacteria and play an important role in animal digestion and innate immunity. The cDNA of a chicken-type lysozyme gene （Mdlys） was cloned from housefly （Musca domestica）. The 484 bp full-length cDNA contains a 426 bp open reading frame （ORF） that encodes MdLys of 141 amino acids. Phylogenetic analysis indicated that the MdLys was similar to chicken-type lysozymes. Spatio-temporal expression of Mdlys was analyzed by RT-PCR. The Mdlys transcript can be detected in both midgut and fat body and was expressed at a relatively lower level at the embryo stage. Mdlys mRNA was upregulated 2 h post bacterial challenge, maintained for 2 to 6 h, and slightly declined from 12 to 24 h post-injection. Western blot analysis showed that MdLys was highly expressed in midgut and was also detected in the hemolymph and fat body. MdLys expression was slightly increased in midgut after challenging with Escherichia coli or Staphylococcus aureus. Its expression was also slightly increased in the fat body after challenging with S. aureus, but no obvious change occurred after E. coli challenge. MdLys expression in the hemolymph was not affected by bacterial challenge. In the developmental stages, MdLys expression levels had no obvious change from the first instar to the pupae stage. There was also no variation under 24 h starvation stress. Recombinant MdLys displayed inhibitory activity against Gram-negative and Gram-positive bacteria. Together, these results suggest that MdLys may play an important role in the innate immunity of houseflies.展开更多
To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme (hLYZ) cDNA and kanamycin resistance gene was constructed for intramammary expression and clinic...To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme (hLYZ) cDNA and kanamycin resistance gene was constructed for intramammary expression and clinical studies. After one time acupuncture or intracisternal infusion of healthy cows with 400 μg of the p215C3LYZ vector, over 2.0 μg/ml of rhLYZ could be detected by enzymatic assay for about 3 weeks in the milk samples. Western blotting showed that rhLYZ secreted into milk samples from the vector-injected cows had molecular weight similar to that of the natural hLYZ in human colostrums. Twenty days after the primary injection, the quarters were re-injected with the same vector by quarter acupuncture and even higher concentrations of rhLYZ could be detected. Indirect competitive ELISA of milk samples showed that the vector injection did not induce detectable humoral immune response against hLYZ. Clinical studies showed that twice acupuncture of quarters with the p215C3LYZ vector had overt therapeutic effect on clinical and subclinical mastitis previously treated with antibiotics, including disappearance of clinical symptoms and relatively high microbiological cure rates. These data provide a solid rationale for using the vector to develop gene therapy for treating bovine mastitis.展开更多
This study of renaturation by dilution and size exclusion chromatogra phy (SEC) addition of urea to improve yield as well as the initial and final pro tein concentrations showed that although urea decreased the rate o...This study of renaturation by dilution and size exclusion chromatogra phy (SEC) addition of urea to improve yield as well as the initial and final pro tein concentrations showed that although urea decreased the rate of lysozyme ref o lding, it could suppress protein aggregation to sustain the pathway of correct r efolding at high protein concentration; and that there existed an optimum urea c oncentration in renaturation buffer. Under the above conditions, lysozyme was su ccessfully refolded from initial concentration of up to 40 mg/mL by dilution and 100 mg/mL by SEC, with the yield of the former being more than 40% and that of the latter being 34.8%. Especially, under the condition of 30 min interval time, i.e. τ>2(t_R2 -t_R1 ), the efficiency was increased by 25% and the renaturation buffe r could be recycled for SEC refolding in continuous operation of downstream proc ess.展开更多
Temperature-sensitive hydrogel—poly(N-isopropyl acrylamide) (PNIPA) was prepared and applied to protein refolding. PNIPA gel disks and gel particles were synthesized by the solution polymerization and inverse suspens...Temperature-sensitive hydrogel—poly(N-isopropyl acrylamide) (PNIPA) was prepared and applied to protein refolding. PNIPA gel disks and gel particles were synthesized by the solution polymerization and inverse suspension polymerization respectively. The swelling kinetics of the gels was also studied. With these prepared PNIPA gels, the model protein lysozyme was renatured. Within 24h, PNIPA gel disks improved the yield of lysozyme activity by 49.3% from 3375.2U·mg^-1 to 5038.8U·mg^-1. With the addition of faster response PNIPA gel beads, the total lysozyme activity recovery was about 68.98% in 3h, as compared with 42.03% by simple batch dilution. The novel refolding system with PNIPA enables efficient refolding especially at high protein concentrations. Discussion about the mechanism revealed that when PNIPA gels were added into the refolding buffer, the hydrophobic interactions between denatured proteins and polymer gels could prevent the aggregation of refolding intermediates, thus enhanced the protein renaturation.展开更多
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 i...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.展开更多
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 ...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.展开更多
Long-terrn injectable microspheres have some inherent disadvantages such as migration of microspheres from the originalsite an.d the burst effect. In order to avoid these problems, microsphere-loaded thermosensitive, ...Long-terrn injectable microspheres have some inherent disadvantages such as migration of microspheres from the originalsite an.d the burst effect. In order to avoid these problems, microsphere-loaded thermosensitive, hydrogel system was designed and expected to achieve a zero-order release Of biomolecular drugs in relativehigh initial drug loadings. Lysozyme, an antibacterial protein usually used to reduce prosthetic valve endocarditis,was selected as the model drug. Poly （DL-lactide-co-glycolide） （PLGA） microspheres, prepared by solvent evaporation method, were employee to encapsulate lysozyme and dispersed into thermosensitive pre-gel solution containing methylcellulose （MC）, polyethylene glycol （PEG）, sodium citrate （SC）, and sodium alginate （SA）. The mixture could act asadrug reservoir by.performing sol-gel transition rapidly if the temperature was raised from roomtemperature to 37℃. The in vitro release results showed that the burst effect was avoided due to strengthening ofdiffusion resistance in the gel. The formulation was able.to deliver lysozy.me for over.30 daysin a nearly zero-order release profile with a rate of 32.8μg.d^-1 which exhibits its remarkable potential for effective aoolication in long-term drug delivery.展开更多
The interaction of lysozyme（Lys） and gold nanoparticles was investigated via UV-vis absorption and resonance light-scattering method.There are some changes of the plasmon absorption and resonance light-scattering of...The interaction of lysozyme（Lys） and gold nanoparticles was investigated via UV-vis absorption and resonance light-scattering method.There are some changes of the plasmon absorption and resonance light-scattering of gold nanoparticles that were observed via the addition of Lys.The normalized plasmon absorption and resonance light-scattering intensity with gold nanoparticles were both linear wilh 1-20 nmol/L Lys.A simple model about the component of the gold nanoparticles and Lys complex was established and the calculated result was fitted well in their concentration ratio.Furthermore,the activity analysis of Lys showed that the interaction was weak and nondestructive.展开更多
Lysozyme reaction was developed as a novel technique for minimizing the amount of excess sludge in the sequential batch reactor(SBR).In the present work,excess sludge taken from a SBR system was treated by lysozyme re...Lysozyme reaction was developed as a novel technique for minimizing the amount of excess sludge in the sequential batch reactor(SBR).In the present work,excess sludge taken from a SBR system was treated by lysozyme reaction and then returned to the reactor.The quality of the effluent water and characteristics of the activated sludge in the SBR were analyzed to determine the effectiveness of the reduction process.The results show that excess sludge production could be reduced to almost 100%in the first30 d of operation and could be reduced to further by 40%in the succeeding 20 d or so.In these time periods,the average removal efficiencies of the chemical oxygen demand and total nitrogen are 87.38%and 52.78%,respectively,whereas the average total phosphorous in the effluent is nearly 17.18%greater than that of the effluent of the reference system.After 50 d of operation,the sludge floc size is in the range of 20 to 80μm,which was smaller than the size prior to the start of the hydrolysis and the ratio of mixed liquor volatile suspended solids/mixed liquor suspended solids increases from 86%to 90%.展开更多
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 wer...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%.展开更多
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 sys...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.展开更多
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 ...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.展开更多
The influence of water on protein conformation was investigated by simulating the molecular dynamics of a model protein lysozyme in different water systems.The lysozyme-water system with TIP3P water model and lysozyme...The influence of water on protein conformation was investigated by simulating the molecular dynamics of a model protein lysozyme in different water systems.The lysozyme-water system with TIP3P water model and lysozyme-water cluster system with six-ring water model were evaluated.In addition,the radial distribution function of solvent around lysozyme was calculated.It is found that the distribution of water molecules around lysozyme is similar to that of water clusters.The analyses of dihedral angles and disulfide bonds of lysozyme show that the conformation of lysozyme is severely damaged in the lysozyme-water cluster system compared with that in the lysozyme-water system.This difference can be attributed to the formation of larger number of intermolecular hydrogen bonds between lysozyme and water cluster.It is in agreement with the analysis that water clusters can change the degree of denaturation in the process of heat denaturation of lysozyme.展开更多
文摘Anaerobic digestion(AD)is widely employed for sludge stabilization and waste reduction.However,the slow hydrolysis process hinders methane production and leads to prolonged sludge issues.In this study,an efficient and eco-friendly lysozyme pre-treatment method was utilized to address these challenges.By optimizing lysozyme dosage,hydrolysis and cell lysis were maximized.Furthermore,lysozyme combined with hydrothermal pretreatment enhanced overall efficiency.Results indicate that:(1)When lysozyme dosage reached 90 mg/g TS after 240 min of pretreatment,SCOD,soluble polysaccharides,and protein content reached their maxima at 855.00,44.09,and 204.86 mg/L,respectively.This represented an increase of 85.87%,365.58%,and 259.21%compared to the untreated sludge.Threedimensional fluorescence spectroscopy revealed the highest fluorescence intensity in the IV region(soluble microbial product),promoting microbial metabolic activity.(2)Lysozyme combined with hydrothermal pretreatment significantly increased SCOD,soluble proteins,and polysaccharide release from sludge,reducing SCOD release time.Orthogonal experiments identified Group 3 as the most effective for SCOD and soluble polysaccharide release,while Group 9 released the most soluble proteins.The significance order of factors influencing SCOD,soluble proteins,and polysaccharide release is hydrothermal temperature>hydrothermal time>enzymatic digestion time.(3)The lysozyme-assisted hydrothermal pretreatment group exhibited the fastest release and the highest SCOD concentration of 8,135.00 mg/L during anaerobic digestion.Maximum SCOD consumption and cumulative gas production increased by 95.89%and 130.58%,respectively,compared to the control group,allowing gas production to conclude 3 days earlier.
基金supported by CURAM-Science Foundation Ireland(SFI)center for medical devices(Grant Number 13/RC/2073_P2)Irish Research Council Postdoctoral Fellowship(GOIPD/2021/928):Disposable,biodegradable,endoscopic ultrasonic imaging probe(DISPOSON)+1 种基金SFI Opportunistic Fund(no.12/RI/2345/SOF)is acknowledged for the NTEGRA Hybrid Nanoscope used in Piezoresponse Force MicroscopyOpen access funding provided by IReL.
文摘Eco-friendly and antimicrobial globular protein lysozyme is widely produced for several commercial applications.Interestingly,it can also be able to convert mechanical and thermal energy into electricity due to its piezo-and pyroelectric nature.Here,we demonstrate engineering of lysozyme into piezoelectric devices that can exploit the potential of lysozyme as environmentally friendly,biocompatible material for mechanical energy harvesting and sensorics,especially in micropowered electronic applications.Noteworthy that this flexible,shape adaptive devices made of crystalline lysozyme obtained from hen egg white exhibited a longitudinal piezoelectric charge coefficient(d-2.7 pC N^(-1))and piezoelectric voltage coefficient(g-76.24 mVmN^(-1))which are comparable to those of quartz(-2.3 pC N^(-1) and 50 mVmN^(-1)).Simple finger tapping on bio-organic energy harvester(BEH)made of lysozyme produced up to 350 mV peak-to-peak voltage,and a maximum instantaneous power output of 2.2 nW cm^(-2).We also demonstrated that the BEH could be used for self-powered motion sensing for real-time monitoring of different body functions.These results pave the way toward self-powered,autonomous,environmental-friendly bio-organic devices for flexible energy harvesting,storage,and in wearable healthcare monitoring.
基金supported by the National Natural Science Foundation of China(No.32072969)the National Key R&D Program of China(No.2022YFD2401002)+1 种基金the Natural Science Foundation of Fujian Province(No.2022 J01325)the Open Research Fund Program of Fujian Provincial Key Laboratory of Marine Fishery Resources and Eco-Environment(No.Z822280).
文摘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.
基金the China National Nature Science Foundation(Grant No.12102404)。
文摘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.
文摘Aim To induce and express the T4 lysozyme in Pichia pastoris and test the antibacterial activity of the protein. Methods T4 lysozyme gene was inserted into expression vector pPIC9K of Pichia pastoris with the fusion at N terminal. The recombinant plasmid was digested by Sal I and then introduced into prepared GS115 competent cells by electroporation. Positive clone and multiple inserts were screened. The secreted proteins in the supernatants were tested. In the agar holes diffusion assay, our expressed protein showed significant antibacterial circles. Results T4 lysozyme protein inhibited the growth of staphylococcus aureus and streptococcus Pneumoniae. There was no difference in the bactericidal activity and the amount of protein expression between the single and multiple copies. The antibacterial activity of expressed protein remained the same during the heat stability test. Conclusion T4 lysozyme was successfully induced and expressed in Pichia pastoris. There is no relationship between copy number and expression. T4 lysozyme protein is heat stable.
文摘The interactions of chlorogenic acid （CA）, neochlorogenic acid （NCA） and cryptochlorogenic acid （CCA） with lysozyme （LYSO） were investigated in physiological buffer by fluorescence spectroscopy. The mechanism study indicated that CA, NCA and CCA could strongly quench the intrinsic fluorescence of LYSO through static quenching procedures with one binding site. Thermodynamic data show that the major force in the binding processes of CA to LYSO was hydrophobic interactions; for NCA, it was the hydrogen bonds and van der Waals forces, as for the CCA system, the mainly force is electrostatic force.
基金supported by the National High Technology Research and Development Program of China (863 Program) (No. 2007AA09Z425 and 2006AA100311)the Ph.D. Program Foundation of the Ministry of Education of China (No. 20060422034).
文摘Lysozymes can hydrolyze bacteria and play an important role in animal digestion and innate immunity. The cDNA of a chicken-type lysozyme gene （Mdlys） was cloned from housefly （Musca domestica）. The 484 bp full-length cDNA contains a 426 bp open reading frame （ORF） that encodes MdLys of 141 amino acids. Phylogenetic analysis indicated that the MdLys was similar to chicken-type lysozymes. Spatio-temporal expression of Mdlys was analyzed by RT-PCR. The Mdlys transcript can be detected in both midgut and fat body and was expressed at a relatively lower level at the embryo stage. Mdlys mRNA was upregulated 2 h post bacterial challenge, maintained for 2 to 6 h, and slightly declined from 12 to 24 h post-injection. Western blot analysis showed that MdLys was highly expressed in midgut and was also detected in the hemolymph and fat body. MdLys expression was slightly increased in midgut after challenging with Escherichia coli or Staphylococcus aureus. Its expression was also slightly increased in the fat body after challenging with S. aureus, but no obvious change occurred after E. coli challenge. MdLys expression in the hemolymph was not affected by bacterial challenge. In the developmental stages, MdLys expression levels had no obvious change from the first instar to the pupae stage. There was also no variation under 24 h starvation stress. Recombinant MdLys displayed inhibitory activity against Gram-negative and Gram-positive bacteria. Together, these results suggest that MdLys may play an important role in the innate immunity of houseflies.
基金Project (Nos. BJ2001315 and BE2004611) supported by the De-partment of Science and Technology of Jiangsu Province, China
文摘To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme (hLYZ) cDNA and kanamycin resistance gene was constructed for intramammary expression and clinical studies. After one time acupuncture or intracisternal infusion of healthy cows with 400 μg of the p215C3LYZ vector, over 2.0 μg/ml of rhLYZ could be detected by enzymatic assay for about 3 weeks in the milk samples. Western blotting showed that rhLYZ secreted into milk samples from the vector-injected cows had molecular weight similar to that of the natural hLYZ in human colostrums. Twenty days after the primary injection, the quarters were re-injected with the same vector by quarter acupuncture and even higher concentrations of rhLYZ could be detected. Indirect competitive ELISA of milk samples showed that the vector injection did not induce detectable humoral immune response against hLYZ. Clinical studies showed that twice acupuncture of quarters with the p215C3LYZ vector had overt therapeutic effect on clinical and subclinical mastitis previously treated with antibiotics, including disappearance of clinical symptoms and relatively high microbiological cure rates. These data provide a solid rationale for using the vector to develop gene therapy for treating bovine mastitis.
文摘This study of renaturation by dilution and size exclusion chromatogra phy (SEC) addition of urea to improve yield as well as the initial and final pro tein concentrations showed that although urea decreased the rate of lysozyme ref o lding, it could suppress protein aggregation to sustain the pathway of correct r efolding at high protein concentration; and that there existed an optimum urea c oncentration in renaturation buffer. Under the above conditions, lysozyme was su ccessfully refolded from initial concentration of up to 40 mg/mL by dilution and 100 mg/mL by SEC, with the yield of the former being more than 40% and that of the latter being 34.8%. Especially, under the condition of 30 min interval time, i.e. τ>2(t_R2 -t_R1 ), the efficiency was increased by 25% and the renaturation buffe r could be recycled for SEC refolding in continuous operation of downstream proc ess.
基金the National Natural Science Foundation of China (No. 20276065).
文摘Temperature-sensitive hydrogel—poly(N-isopropyl acrylamide) (PNIPA) was prepared and applied to protein refolding. PNIPA gel disks and gel particles were synthesized by the solution polymerization and inverse suspension polymerization respectively. The swelling kinetics of the gels was also studied. With these prepared PNIPA gels, the model protein lysozyme was renatured. Within 24h, PNIPA gel disks improved the yield of lysozyme activity by 49.3% from 3375.2U·mg^-1 to 5038.8U·mg^-1. With the addition of faster response PNIPA gel beads, the total lysozyme activity recovery was about 68.98% in 3h, as compared with 42.03% by simple batch dilution. The novel refolding system with PNIPA enables efficient refolding especially at high protein concentrations. Discussion about the mechanism revealed that when PNIPA gels were added into the refolding buffer, the hydrophobic interactions between denatured proteins and polymer gels could prevent the aggregation of refolding intermediates, thus enhanced the protein renaturation.
基金supported in the context of the Italian Malaria Network by grants from Compagnia di San Paolo-IMIthe University of Torino Intramural FundsRegione Piemonte,Ricerca Sanitaria Finalizzata 2007 to PA
文摘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.
文摘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.
基金Supported by the National Natural Science Foundation of China （No.20576057） and Fundamental Research Foundation of Tsinghua University （JCqn2005033）.
文摘Long-terrn injectable microspheres have some inherent disadvantages such as migration of microspheres from the originalsite an.d the burst effect. In order to avoid these problems, microsphere-loaded thermosensitive, hydrogel system was designed and expected to achieve a zero-order release Of biomolecular drugs in relativehigh initial drug loadings. Lysozyme, an antibacterial protein usually used to reduce prosthetic valve endocarditis,was selected as the model drug. Poly （DL-lactide-co-glycolide） （PLGA） microspheres, prepared by solvent evaporation method, were employee to encapsulate lysozyme and dispersed into thermosensitive pre-gel solution containing methylcellulose （MC）, polyethylene glycol （PEG）, sodium citrate （SC）, and sodium alginate （SA）. The mixture could act asadrug reservoir by.performing sol-gel transition rapidly if the temperature was raised from roomtemperature to 37℃. The in vitro release results showed that the burst effect was avoided due to strengthening ofdiffusion resistance in the gel. The formulation was able.to deliver lysozy.me for over.30 daysin a nearly zero-order release profile with a rate of 32.8μg.d^-1 which exhibits its remarkable potential for effective aoolication in long-term drug delivery.
基金supported by the National Nature Sciences Foundation of China(Nos.20475004,90713013 &20775004)Shanghai Leading Academic Discipline Project(no.S30109)
文摘The interaction of lysozyme（Lys） and gold nanoparticles was investigated via UV-vis absorption and resonance light-scattering method.There are some changes of the plasmon absorption and resonance light-scattering of gold nanoparticles that were observed via the addition of Lys.The normalized plasmon absorption and resonance light-scattering intensity with gold nanoparticles were both linear wilh 1-20 nmol/L Lys.A simple model about the component of the gold nanoparticles and Lys complex was established and the calculated result was fitted well in their concentration ratio.Furthermore,the activity analysis of Lys showed that the interaction was weak and nondestructive.
基金Project(51078130)supported by the National Natural Science Foundation of ChinaProject(10C0419)supported by the the Education Department of Hunan Province,China
文摘Lysozyme reaction was developed as a novel technique for minimizing the amount of excess sludge in the sequential batch reactor(SBR).In the present work,excess sludge taken from a SBR system was treated by lysozyme reaction and then returned to the reactor.The quality of the effluent water and characteristics of the activated sludge in the SBR were analyzed to determine the effectiveness of the reduction process.The results show that excess sludge production could be reduced to almost 100%in the first30 d of operation and could be reduced to further by 40%in the succeeding 20 d or so.In these time periods,the average removal efficiencies of the chemical oxygen demand and total nitrogen are 87.38%and 52.78%,respectively,whereas the average total phosphorous in the effluent is nearly 17.18%greater than that of the effluent of the reference system.After 50 d of operation,the sludge floc size is in the range of 20 to 80μm,which was smaller than the size prior to the start of the hydrolysis and the ratio of mixed liquor volatile suspended solids/mixed liquor suspended solids increases from 86%to 90%.
基金financially supported by the National Natural Science Foundation of China (project 21077133)the Natural Foundation of Shandong Province and the Top Talent Project of China University of Petroleum (16RC17040003)
文摘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%.
基金supported the Central Public-Interest Scientific Institution Basal Research Fund, China (Y2019GH03)the Special Fund for Agro-Scientific Research in the Public Interest of China (210503114)SINOGRAIN Ⅱ (CHN-17/0019): Technological Innovation to Support Environmentally-Friendly Food Production and Food Safety Under a Changing Climate-Opportunities and Challenges for Norway-China Cooperation
文摘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.
基金surported by the National Natural Science Foundation of China(39960039)the Natural Science Foundation of Yunnan Provinee(2002C0079M),China
文摘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.
基金Supported by National Natural Science Foundation of China (No. 20676094)
文摘The influence of water on protein conformation was investigated by simulating the molecular dynamics of a model protein lysozyme in different water systems.The lysozyme-water system with TIP3P water model and lysozyme-water cluster system with six-ring water model were evaluated.In addition,the radial distribution function of solvent around lysozyme was calculated.It is found that the distribution of water molecules around lysozyme is similar to that of water clusters.The analyses of dihedral angles and disulfide bonds of lysozyme show that the conformation of lysozyme is severely damaged in the lysozyme-water cluster system compared with that in the lysozyme-water system.This difference can be attributed to the formation of larger number of intermolecular hydrogen bonds between lysozyme and water cluster.It is in agreement with the analysis that water clusters can change the degree of denaturation in the process of heat denaturation of lysozyme.
