Protein 25a2 is the antifungal peptide of cotton Verticillium wilt which was isolated from Bacillus amyloliquefaciens. The amino acid sequence of antifungal peptide 25a2 was analyzed using bioinformatics tools, and th...Protein 25a2 is the antifungal peptide of cotton Verticillium wilt which was isolated from Bacillus amyloliquefaciens. The amino acid sequence of antifungal peptide 25a2 was analyzed using bioinformatics tools, and the characters of signal peptides, transmembrane topological structura, physicochemical signatures, protein domain, secondary and tertiary structure of protein were predicted. The results showed that 25a2 was a secreted protein, the sequence of which included a signal peptide in N end and a transmembrane domain in C end. The predicted secondary structure showed that the antibacterial peptide was mainly free random coils, belonging to mixed protein, three-dimensional model of 25a2 was a compact ball. These results showed that the most possible action mechanism of antifungal peptide 25a2 might be "carpet" model.展开更多
A novel Eucommia antifungal peptide, named EAFP3, was isolated from the bark of Eucommia ul- moides by NaC1 extract, gel filtration and reverse phase high performance liquid chromatography. The molecular mass of EAFP3...A novel Eucommia antifungal peptide, named EAFP3, was isolated from the bark of Eucommia ul- moides by NaC1 extract, gel filtration and reverse phase high performance liquid chromatography. The molecular mass of EAFP3 is 4157.3 Da, and its partial amino acid sequence is -. LYQQLIAGITLNK.-. EAFP3 exerts an inhibitory activity against Candida albicans in vitro and the drug concentration required for 50% growth inhibition (IC50) is 31.25μg/mL.展开更多
Antibacterial and antifungal peptides found in houseflies (Musca domestica) in large number are indispensable components of its immune defense mechanism. In this study the anterior tip of the larvae of housefly was ...Antibacterial and antifungal peptides found in houseflies (Musca domestica) in large number are indispensable components of its immune defense mechanism. In this study the anterior tip of the larvae of housefly was cut off with a pair of fine scissors and hemolymph was collected and exuded in an ice-cold test tube. From the hemolymph an antifungal substance was isolated by solid-phase extraction combined with reverse phase-high performance liquid chromotography (RP-HPLC) and named as Musca domestica antifungal peptide-1 (MAF-1). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed its molecular weight was 17 kDa. UV absorption spectra revealed that this antifungal substance possessed the characteristics of protein peptides. Analysis by fingerprint-identification and tandem mass spectrometry suggested MAF-I was an unknown protein. Edman degradation identified the sequence of 30 amino acids of its N-terminal which matched no peptide in the MASCOT search database, indicating MAF-1 was a novel insect antifungal peptide. Mass spectrometry showed the precise molecular weight of MAF-1 was 17203.384 Da. Its isoelectric point was acidic.展开更多
This study investigated the antifungal activity and possible mode of action of Bacillus pumilus HN-10 antifungal peptide P-1 against Trichothecium roseum.The results showed that the antifungal peptide P-1 at a concent...This study investigated the antifungal activity and possible mode of action of Bacillus pumilus HN-10 antifungal peptide P-1 against Trichothecium roseum.The results showed that the antifungal peptide P-1 at a concentration of 1.0μg mL^(-1)had strong antifungal activity against T.roseum.P-1 inhibited the tricarboxylic acid cycle(TCA)pathway and the transporter pathway of NADH to coenzyme Q on the electron transport chain.P-1 significantly reduced succinate dehydrogenase(SDH),malate dehydrogenase(MDH),ATPase,mitochondrial complex enzymes I,II and IV enzyme activities on the electron transport chain,and 5'-triphosphate(ATP),5'-diphosphate(ADP),5'-monophosphate(AMP)content,and energy charge(EC);significantly increased 6-phosphofructokinase(PFK)enzyme activity.The release of Ca^(2+)(OD_(680))from the inner mitochondrial membrane and the openness of the mitochondrial permeability transition pore(MPTP)were analysed,and microscopy was performed following staining of mitochondria with JC-1.The results indicated that P-1 significantly increased the release of Ca^(2+) and the openness of MPTP,decreased the mitochondrial membrane potential,and produced green fluorescence;transcriptomics data analysis showed that there were 39 differentially expressed genes(DEGs)related to energy metabolism enzymes.The results verified by qRT-PCR were basically consistent with the transcriptome sequencing results.Thus,P-1 achieved its inhibitory effect mainly by regulating genes related to energy metabolism.展开更多
文摘Protein 25a2 is the antifungal peptide of cotton Verticillium wilt which was isolated from Bacillus amyloliquefaciens. The amino acid sequence of antifungal peptide 25a2 was analyzed using bioinformatics tools, and the characters of signal peptides, transmembrane topological structura, physicochemical signatures, protein domain, secondary and tertiary structure of protein were predicted. The results showed that 25a2 was a secreted protein, the sequence of which included a signal peptide in N end and a transmembrane domain in C end. The predicted secondary structure showed that the antibacterial peptide was mainly free random coils, belonging to mixed protein, three-dimensional model of 25a2 was a compact ball. These results showed that the most possible action mechanism of antifungal peptide 25a2 might be "carpet" model.
基金the National Natural Science Foundation of China(30660146)the Ministry of Sciences of Technology Program of China(2003CCA02600)
文摘A novel Eucommia antifungal peptide, named EAFP3, was isolated from the bark of Eucommia ul- moides by NaC1 extract, gel filtration and reverse phase high performance liquid chromatography. The molecular mass of EAFP3 is 4157.3 Da, and its partial amino acid sequence is -. LYQQLIAGITLNK.-. EAFP3 exerts an inhibitory activity against Candida albicans in vitro and the drug concentration required for 50% growth inhibition (IC50) is 31.25μg/mL.
基金Acknowledgements This work was supported by grants from the National Natural Science Foundation of China (No. 39970087), the Special Fund of Governor of Guizhou Province [QKJB(2003)04], and "Western Lights" of Chinese Academy of Sciences [KJWZ(2005)404].
文摘Antibacterial and antifungal peptides found in houseflies (Musca domestica) in large number are indispensable components of its immune defense mechanism. In this study the anterior tip of the larvae of housefly was cut off with a pair of fine scissors and hemolymph was collected and exuded in an ice-cold test tube. From the hemolymph an antifungal substance was isolated by solid-phase extraction combined with reverse phase-high performance liquid chromotography (RP-HPLC) and named as Musca domestica antifungal peptide-1 (MAF-1). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed its molecular weight was 17 kDa. UV absorption spectra revealed that this antifungal substance possessed the characteristics of protein peptides. Analysis by fingerprint-identification and tandem mass spectrometry suggested MAF-I was an unknown protein. Edman degradation identified the sequence of 30 amino acids of its N-terminal which matched no peptide in the MASCOT search database, indicating MAF-1 was a novel insect antifungal peptide. Mass spectrometry showed the precise molecular weight of MAF-1 was 17203.384 Da. Its isoelectric point was acidic.
基金financially supported by National Key R&D Program of China(2018YFD0400205).
文摘This study investigated the antifungal activity and possible mode of action of Bacillus pumilus HN-10 antifungal peptide P-1 against Trichothecium roseum.The results showed that the antifungal peptide P-1 at a concentration of 1.0μg mL^(-1)had strong antifungal activity against T.roseum.P-1 inhibited the tricarboxylic acid cycle(TCA)pathway and the transporter pathway of NADH to coenzyme Q on the electron transport chain.P-1 significantly reduced succinate dehydrogenase(SDH),malate dehydrogenase(MDH),ATPase,mitochondrial complex enzymes I,II and IV enzyme activities on the electron transport chain,and 5'-triphosphate(ATP),5'-diphosphate(ADP),5'-monophosphate(AMP)content,and energy charge(EC);significantly increased 6-phosphofructokinase(PFK)enzyme activity.The release of Ca^(2+)(OD_(680))from the inner mitochondrial membrane and the openness of the mitochondrial permeability transition pore(MPTP)were analysed,and microscopy was performed following staining of mitochondria with JC-1.The results indicated that P-1 significantly increased the release of Ca^(2+) and the openness of MPTP,decreased the mitochondrial membrane potential,and produced green fluorescence;transcriptomics data analysis showed that there were 39 differentially expressed genes(DEGs)related to energy metabolism enzymes.The results verified by qRT-PCR were basically consistent with the transcriptome sequencing results.Thus,P-1 achieved its inhibitory effect mainly by regulating genes related to energy metabolism.