75 results about "Fungal cell walls" patented technology

The invention relates to a method for controlling plant diseases caused by the fungi Botrytis cinerea and Alternaria alternata, by applying to a growing plant or to fruit or vegetables before or after harvesting, a composition which comprises an effective amount for controlling said fungi of at least one oligosaccharide ingredient, active against Botrytis cinerea and Alternaria alternata, and selected from oligosaccharides obtainable by hydrolysis of chitin, beta -glucan and other similarly active polysaccharides, excluding chitosan, of cell walls of fungi, yeasts, marine plants and exoskeletons of arthropods, The composition also forms part of the invention, as does an analogous method and composition for a method for controlling plant diseases caused by Botrytis cinerea, and utilizing at least one oligosaccharide ingredient, active against Botrytis cinerea selected from oligosaccharides obtainable by hydrolysis of chitosan, and having a molecular weight within the range of about 500 to about 10,000 daltons, provided that in this instance the composition excludes acetic acid.

The invention relates to a fruit and vegetable compound anti-staling agent, which is well-mixed oyster white soliquoid or micro emulsion and comprises the ingredients or some of the ingredients of 5-500ppmB of A biological bactericide, 50-5000ppmC of chemical bactericide, 50-5000000 enzyme activity unit / LD of catabolic enzyme of fungal cell wall, 10-10000ppmE of chelant, 10-10000 ppmF of antioxidant, 50-5000ppmG of thickener, 20-2000ppmH of solubilizer and water. The invention is characterized in that after the compound fruit and vegetable anti-staling liquid is applied to fruits, such as garlic sprout, banana, strawberry, bramble, pineapple, apple, pear, winter date, plum and orange and other fruits of berry type, the invention can obviously restrain rot of the fruits and vegetables at appropriate preservation temperature and under other conditions, thereby prolonging preservation life of the fruits and vegetables.

The invention relates to a trichoderma pseudokoningii (SMF2) strain and application thereof, which belong to the technical field of microbiology. The invention discloses the trichoderma pseudokoningii (SMF2) strain preserved in China Center for Type Culture Collection on February 24, 2009 with the preservation number being CCTCCNO: M209031, and the application of the strain to preparation of soil microbial agents of trichoderma preparations. The trichoderma pseudokoningii (SMF2) strain disclosed by the invention can be used for producing plant disease fungus cell wall digestive enzymes (CWDEs) and antibiotics of peptaibols at high yield, the CWDEs and the peptaibols cooperate in situ, and the inhibition effect of the trichoderma pseudokoningii strain to the plant disease fungus is further enhanced.

A reporter system reflecting the transport process that transports GPI-anchored proteins to the cell wall was constructed and compounds inhibiting this process were discovered. Further, genes conferring resistance to the above compounds were identified and methods of screening for compounds that inhibit the activity of the proteins encoded by these genes were developed. Therefore, through the novel compounds, the present invention showed that antifungal agents having a novel mechanism, i.e. inhibiting the process that transports GPI-anchored proteins to the cell wall, could be achieved.

A reporter system reflecting the transport process that transports GPI-anchored proteins to the cell wall was constructed and compounds inhibiting this process were discovered. Further, fungal genes conferring resistance to the above compounds were identified and methods of screening for compounds that inhibit the activity of the proteins encoded by these genes were developed. These genes encode proteins participating in fungal cell wall synthesis. Therefore, through the novel compounds, the present invention showed that antifungal agents having a novel mechanism, i.e. inhibiting the process that transports GPI-anchored proteins to the cell wall, could be achieved.

A reporter system reflecting the transport process that transports GPI-anchored proteins to the cell wall was constructed and compounds inhibiting this process were discovered. Further, genes conferring resistance to the above compounds were identified and methods of screening for compounds that inhibit the activity of the proteins encoded by these genes were developed. Therefore, through the novel compounds, the present invention showed that antifungal agents having a novel mechanism, i.e. inhibiting the process that transports GPI-anchored proteins to the cell wall, could be achieved.

Cellobio-oligosaccharides (CBO) produced by the dextransucrase-catalyzed transglycosylation reaction of sucrose and cellobiose were discovered to be effective as antifungal agents against dental caries and against fungi who rely on glucan as an integral part of the cell wall, e.g., A. terreus. The cellobio-oligosaccharides were found to be inhibitors of β-(1,3)-glucan synthase, an important enzyme involving in fungal cell wall component synthesis. The CBO caused structural changes in the growing fungal cells. In addition, the CBO were shown to be effective as anti-cariogenic agents in preventing bacterial adherence to teeth by inhibiting the formation of the bacterial plaque (glucans), e.g., that formed by Streptococcus mutans. Cellobio-oligosaccharides produced by dextransucrase were analyzed and shown to have a degree of polymerization (DP) ranging from 3 to 6 glucosyl groups. Examples of these cellobio-oligosaccharides produced by this method include, but are not limited to, trisaccharides such as α-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-D-glucopyranose and α-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl-(1→4)-D-glucopyranose.

The invention discloses a SLS lysis solution which can quickly extract fungal DNA. The SLS lysis solution contains following components (calculated by the total volume of solution): 1 - 3 g / 100ml of N-lauroyl sodium sarcosinate, 150 - 300mmol / L of Tris-HCl, 150 - 300mmol / L of NaCl, and 50 - 100mmol / L of EDTA; and the pH value is ranged from 7.5 to 8.5. The SLS lysis solution of the invention takes N-lauroyl sodium sarcosinate as the main functional component to make up the DNA extracting solution; and the DNA extracting solution can be used for extracting fungal DNA. Due to the excellent biologic degradability, the DNA extracting solution can be used for directly breaking the cell wall of pathogenic fungi, thereby avoiding the complicated physical processing of the cell-wall breaking in traditional method in extracting fungal DNA, and greatly simplifying the extracting of DAN.

The invention discloses a method for extracting fungal oil. The method comprises the steps of carrying out enzymolysis on a fungal cell wall through an enzyme-ultrasonic wave combined method (namely carrying out enzymolysis on the fungal cell wall by assisting chitinase and helicase by utilizing an ultrasonic wave); then adding SDS (Sodium Dodecyl Sulfate) to further split a fungal cell; adding isopropanol and n-pentane for one-time or multiple-time grease extraction. The method disclosed by the invention is simple in process, complete in fungal cell wall breaking and high in oil extraction rate as high as more than 95%. According to the method, a used main extraction solvent is relatively low in n-pentane boiling point and latent heat of vaporization and low in toxicity. Compared with the traditional fungal oil extraction method, the fungal oil extraction method disclosed by the invention has the advantages of low energy consumption and high environmental safety.

The present invention discloses a fungal fluorescent staining solution which is simple in operation, good in staining effect, high in detection rate, stable in system and easy to preserve. Fluoresceinin the fungal fluorescent staining solution can bind with beta-polysaccharides on the fungal cell wall with high affinity such as chitin and cellulose so that fungal components in a sample can be labeled and fungal morphology can be clearly observed under a fluorescence microscope. With the addition of an anti-interference fluorescent auxiliary in the staining solution, the binding rate of the fluorescent staining solution with the fungal cell wall is improved; moreover, the fluorescence intensity is enhanced under the ultraviolet excitation so that the propagules such as fungal hyphae and spores emit bright blue-white fluorescence; under the microscope observation, the outline of the fungi is clearly visible while the stability of the staining solution system is improved; and the fluorescence quenching risk of the staining solution is reduced. The staining solution is capable of one-step staining; staining can be completed in a few seconds; the operation is simple; the detection timeis saved; and the detection rate of the fungi is also improved.

The invention relates to the technical field of enzyme preparation, in particular to comprehensive enzyme powder with a lipolysis function and a preparation method of the comprehensive enzyme powder with the lipolysis function. The comprehensive enzyme powder is prepared from, by weight, 1-5 parts of compound decomposition enzyme, 10-30 parts of natto, 50-70 parts of vegetables, 80-100 parts of fruits, 10-30 parts of edible fungi, 10-15 parts of medicinal and edible traditional Chinese medicine extracts, 5-10 parts of mushroom extracting liquid and 40-80 parts of pineapple powder by addition of enzyme strains. Compared with the prior art, the comprehensive enzyme powder with the lipolysis function has the advantages of diversity, balance and easiness in absorption. The preparation method has the advantages that fungal polysaccharides are extracted through combination of ultrasonic waves and an enzymolysis method, fungal cell wall crushing effects are enhanced by ultrasonic waves to increase polysaccharide dissolution rate, cell walls are subjected to further enzymolysis by the enzymolysis method to enable intercellular polysaccharides to be released sufficiently, and accordingly the preparation method is efficient, time saving, labor saving, higher in polysaccharide yield and purity and suitable for large-scale production and application.

The invention discloses a method for amplifying antibody marking signal or nucleic acid probe marking signal, and the main scheme is that: lipopolysaccharide of gram negative bacteria and an antibody are crosslinked, or fungi (1,3)-beta-D-glucan of fungi cell wall and the antibody or a nucleic acid probe are crosslinked, and then tachypleus amebocyte lysate and the gram negative bacteria lipopolysaccharide or fungi cell wall (1,3)-beta-D-glucan which is crosslinked with the antibody are specifically reacted, thereby improving the sensitivity of the antibody or nucleic acid probe detection. The system for amplifying antibody marking signal or nucleic acid probe marking signal can detect object substrates lower than pg level content.

The invention relates to the field of microorganism and molecular biology, and in particular relates to a method for detecting trace fungus by using single-cell sequencing and a fungus detection kit produced by using the method. The method comprises the steps of extraction of trace fungus cells, extraction of fungus protoplasm by fragmentation of fungus cell wall, extraction and amplification of trace fungus protoplasm gDNA, database creation of gDNA, genome sequencing, bioinformatic analysis comparison, and judgment on the type of the detected fungus. The method disclosed by the invention realizes effective detection on trace fungus, and can be directly applied to separation, detection and identification of trace difficult fungus samples or mixed samples and deep study of genetic information. The fungus detection method and the kit are applicable to the fields of industrial production, environmental monitoring, air detection, soil detection, water quality detection, food detection, drug detection, cosmetic detection, health care products detection and medical detection.

The invention provides a method for extracting fungus total protein. A simple and feasible method is adopted, the characteristics that the fungal cell wall is thick and tough and the intracellular protein is difficult to release are overcome, and high-quality fungus proteome is successfully extracted. The components of an extracting solution are simple; the extracting solution I comprises 7-8M urea, 2-2.5M thiourea, 1-2m MEDTA (ethylenediamine tetraacetic acid), 10mMTris-HCL (trihydroxy methyl-aminomethane hydrochloride with the pH of 7.4) and 10ul protease inhibitor mixture; the extracting solution II comprises 2-4 percent of CHAPS (3-[3-(cholylaminopropyl) dimethylamino]CHAPS, 0.5 percent of SDS (sodium dodecyl sulfate) and 2-4 percent of DTT (dithiothreitol). The method is simple, convenient and rapid and is low in equipment requirement; and moreover, the extraction process of total protein can be finished within three hours, lots of total protein can be obtained and is used for proteome analysis, and the method has high actual application value.

The invention provides a Trichoderma harzianum strain which is preserved in the general microbiology center of management committee of microorganisms deposit, with an accession number being CGMCC No. 5202. The Trichoderma harzianum strain can dissolve cell walls of pathogen by using self-producing fungal cell wall degrading enzymes and penetrate into the pathogen cells, so that the growth of the pathogenic fungi is inhibited and then dead, thereby realizing prevention and treatment effects of diseases, and highly-efficiently preventing and treating plant diseases caused by fungi. The Trichoderma harzianum strain provided by the invention is large in sporulation amount, easy to cultivate, suitable for industrial mass production, wide in using range and strong in practicality.

The invention discloses a trichoderma citrinoviride strain. The trichoderma citrinoviride strain is preserved in China General Microbiological Culture Collection Center (CGMCC), and the preservation number of the trichoderma citrinoviride strain is CGMCC No. 8722. The trichoderma citrinoviride strain has the advantages that the trichoderma citrinoviride strain dissolves pathogenic fungus cell walls through the self-produced fungal cell wall degrading enzyme, the trichoderma citrinoviride strain seeps into the cells of pathogenic fungi to form a large amount of branches and sexual structures, the growth of the pathogenic fungi is inhibited, and the pathogenic fungi die; the pathogenic fungi of pepper damping-off can be efficiently inhibited by the trichoderma citrinoviride strain, pepper damping-off is reduced, and the trichoderma citrinoviride strain can replace chemical pesticides to inhibit pepper damping-off while environmental protection is achieved.

The invention discloses a genetic conversing method of exogenous gene for Gibberalla fujikuroi through electroporation technique, which is characterized by the following: utilizing cell wall degraded enzyme of fungi to dispose the cell wall; obtaining the protoplast to purify; blending purified protoplast and fitful exogenous gene / DNA solution; proceeding electric shock effect; making protoplasm adsorb the exogenous DNA.

The invention discloses fusion chitinase capable of efficiently degrading alpha-chitin and a related biological material and application thereof. The fusion chitinase is a protein obtained by fusing beta-N-acetyl hexosaminase and chitinase, wherein the chitinase activity of the fusion chitinase is higher than that of the beta-N-acetyl hexosaminase and that of the chitinase; the chitinase is D1) or D2), wherein the D1) is a protein with 53rd-322nd amino acid residues of which the amino acid sequence is shown in SEQ ID No.2; and the D2) is a fusion protein obtained by fusing a protein tag at the carboxyl terminal or / and the amino terminal of the protein shown in D1). The fusion chitinase has a degradation effect on various types of chitin, and has relatively high activity on colloid chitin, alpha-chitin and fungal cell wall chitin; and the fusion chitinase has the capability of completely hydrolyzing chitin and generating N-acetyl glucosamine. The fusion chitinase can be widely applied to biological medicine, biological agriculture, cosmetics, energy industry, etc.

A novel fungal enzyme having lysozyme activity has been isolated. The invention further relates to a fungal polypeptide having lysozyme activity and belonging to the GH25 family, wherein the enzyme is selected from the group consisting of (a) a polypeptide comprising an amino acid sequence, which has at least 80% identity with amino acids 1 to 233 of SEQ ID NO:2; (b) a polypeptide comprising an amino acid sequence, which has at least 80% identity with the polypeptide encoded by the lysozyme encoding part of the nucleotide sequence inserted into a plasmid present in strain DSM 16084; (c) a polypeptide which is encoded by a nucleotide sequence which hybridizes under high stringency conditions with a polynucleotide probe consisting of the complementary strand of nucleotides 84 to 782 of SEQ ID NO:1; or (d) a fragment of (a), (b) or (c) that has lysozyme activity.

A composition suitable for inhibiting the outgrowth of fungi is provided comprising a first ingredient which inhibits the biogenesis of a normal fungal cell wall and a second ingredient which is capable of perturbing the structure of the cellular membrane of said fungi, so that either the cellular integrity is essentially lost or cell division cannot take place, or both. The first ingredient preferably inhibits the anchorage of cell wall proteins in the cell wall of the fungi and is suitably a beta-(1,6)-glucose polysaccharide, preferably beta-gentiobiose or a pustulan fragment. The second ingredient is suitably a natural microbial membrane affecting substance (MMAS), preferably MB-21. The composition is particularly suitable as a preservative for inhibiting the outgrowth of fungi in food products, such as sauces, dressings, ketchups, and soups, but is also useful for preventing or inhibiting undesired fungal growth on other products such as personal health care products, e.g. soap bars.

A compound comprising one or more polysaccharide moieties each independently represented by the formula β(1→4)-[GlcNH—R]n-2,5-anhydromannose, wherein n is a positive integer from 3 to 500, and R is H or an acyl group, is described. The compound can be manufactured by (a) reacting chitosan with an acylating agent sufficient to partially N-acylate the chitosan, yielding a modified chitin / chitosan mixed polymer; and (b) reacting the modified chitin / chitosan mixed polymer with a deaminating agent to cleave the mixed polymer at the unacylated chitosan moieties. The compound can be used to immunize against fungal infection. Antibodies specific to the compound, and the use of such antibodies to protect against fungal infection are also described.

The invention relates a method for extracting a fungal genome on the surface of aged tobacco. The method includes the two main steps of aged tobacco surface fungus enrichment and genome DNA extraction. Fungi are enriched through buffering solution soaking and gradient centrifugation, grinding and cell disruption are conducted by means of liquid nitrogen, and the DNA of the genome of fungi is extracted by means of a kit shown in the description. By means of liquid nitrogen grinding, fungal cell walls can be fully split, extraction efficiency of the DNA of the fungi is improved, the DNA isolation kit in the description includes the innovation patent inhibitor removal technology, and the yield of the genome can be effectively increased. The method can be used for efficiently and quickly extracting the DNA of the fungal genome on the surface of aged tobacco.

The invention belongs to the technical field of genetic engineering, and in particular relates to a method for improving the antibacterial activity of antifungal protein. The method comprises the following steps of: connecting a DNA (deoxyribonucleic acid) sequence of a coding chitin binding domain with a cDNA (complementary DNA) sequence of coded antifungal protein by a correct reading frame; then cloning to a protein expression vector; expressing fused protein by using engineering bacteria; carrying out protein purification by further utilizing a commercialized medium; and detecting the chitin binding capacity of the purified protein and analyzing the antibacterial activity. The invention can improve the antibacterial efficiency of the antifungal protein. By applying the invention, the activity of the traditional antifungal protein and the targeted binding capacity of the protein on fungal cell walls are improved.

The invention discloses vaginal secretion microbial cell fluorescence detection dye liquor which is easy and convenient to operate, clear in dyeing effect, high in detection rate, not prone to precipitation, stable in system and easy to store. The vaginal secretion microbial cell fluorescence detection dye liquor comprises two reagents: a solution A and a solution B, fluorescein in the solution Acan be combined with beta-polysaccharide, such as chitin and cellulose, on fungal cell walls with high affinity, and fungal components existing in a sample are marked. The solution B is composed of water, fluorescein and a fluorescent auxiliary agent, and the fluorescein can fluorescently mark cells and pathogenic microorganisms in vaginal secretion. The anti-interference fluorescent auxiliary agents are respectively added into the reagents, so that the combination rate of fluorescent dyes, cells and microorganisms can be improved, the fluorescence intensity is enhanced, the cell and microorganism contours are clear and visible, the stability of a staining solution component system is improved, and the staining solution fluorescence quenching risk is reduced. The dye liquor is simple and rapid to operate, clean in background, clear in image and easy to judge, and the accuracy of vaginitis and other gynecological leucorrhea detection items can be greatly improved through clinical use.

The invention provides a multienzyme compound capable of preventing and treating fungus infection. The multienzyme compound of the invention can be used in daily prevention of fungus infection, and can kill fungi selectively without generation of toxic or side effects, or drug resistance. The multienzyme compound of the invention can treat fungus infection individually. Because the multienzyme compound can digest fungus cell walls, when cooperated with other antifungal medicaments to treat fungus infection, the multienzyme compound of the invention can promote penetration of other medicaments, enhance treatment effects of other medicaments effectively, reduce dosage and application amount, lower toxic and side effects, and reduce generation probability of drug resistance. In addition, the multienzyme compound of the invention can be selectively added with antifungal medicaments to improve treatment effects.

The invention relates to the culture of fungi and a preparing method of separating and extracting chitosan from mycelium. Actinomucor taiwanensis fungi are used as raw materials. After plate inscription, shaking culture and culture in fermentor, bacterial liquid is obtained. Then, the centrifugation is carried out on the obtained bacterial liquid to obtain wet cells. After separating, drying, crushing, alkali treatment, acid treatment and drying, the chitosan is obtained. The cell culturing method is simple; the process for preparing the chitosan is easy, the alkali treatment with low concentration during the deacetylation process greatly reduces pollutions on the environment, the production cost is low, and the yield is stable.

The invention discloses a method, solution and kit for simultaneously detecting multiple microbial genomes. The method comprises the following steps: high-throughput sequencing is carried out on a to-be-detected sample by adopting a microbial RNA detection process; before the sequencing, microbial cell lysis treatment is performed on a sample to be detected in advance, i.e., fungal cell wall lyticenzyme is adopted to perform cell wall lysis treatment on the sample to be detected, and meanwhile, an RNA enzyme inhibitor and dithiothreitol are added into a treatment solution; after cell wall cracking treatment is completed, physical grinding is further conducted; and then nucleic acid extraction and microbial RNA detection processes are carried out. According to the method disclosed by the invention, microbial genome nucleic acid including RNA and DNA in one sample can be comprehensively detected only through one process; the microbial genome detection cost and workload are reduced, andthe detection efficiency is improved; and the sample consumption is reduced, so that the microbial genome nucleic acid information of special samples or precious samples with a relatively small sampleamount can be obtained more completely.