57 results about "Fungal protein" patented technology

The invention belongs to a method for genetic transformation and regeneration of a eucalyptus, and particularly relates to the method for genetic transformation and regeneration by taking hypocotyledonary axis of an aseptic seedling of a eucalyptus urophylla as an explant. The hypocotyledonary axis of the seedling of the eucalyptus urophylla is taken as the explant, calluses are pre-cultured and induced, co-culture transformation is performed with agrobacterium tumefactions carrying exogenous genes, kanamycin is used for selection, and adventitious bud induction, bud proliferation, adventitious bud growth, rootage and transplantation are performed for getting a plant to be transformed. PCR (polymerase chain reaction) detection and RT-PCR (reverse transcription-polymerase chain reaction) detection prove that, as for the obtained plant of the eucalyptus urophylla to be transformed, the exogenous genes are introduced into a genome of the eucalyptus urophylla and expressed; the transformed plant having better resistance to phytophthora nicotianae is found in the anti-disease detection of in vitro blades of the eucalyptus urophylla of a reverse radish-anti-fungal protein (RS-AFP2); and the relevant enzyme activity determination result of the resistance shows that, after inoculation of the transformed plant, the activities of L-phenylalanin ammo-nialyase (PAL), peroxidase (POD) and polyphenol oxidase (PPO) are enhanced.

An object of the present invention is to search and identify novel antifungal proteins capable of inhibiting the growth of plant pathogenic microorganisms including Magnaporthe grisea and Rhizoctonia solani causing two major rice diseases at relatively low concentrations, and further to clone a gene for said protein. The present invention provides an antifungal protein which can be obtained from fraction(s) precipitated by ammonium sulfate precipitation using an aqueous extract from Pleurotus cornucopiae, wherein said protein has an antifungal activity against at least rice blast, and exhibits existence of a component having a molecular weight of about 15 kDa as determined by SDS-PAGE method; a gene encoding said protein and uses thereof.

Provided herein are compositions and methods for producing milk proteins in plants, which allow for safe, sustainable and humane production of milk proteins for commercial use, such as use in food compositions. The disclosure provides recombinant fusion proteins comprising a milk protein, or fragment thereof and a structured mammalian, avian, plant, or fungal protein, or fragment thereof. The disclosure also provides methods for producing the recombinant fusions proteins, and food compositions comprising the same.

The present invention relates to an antifungal protein, AlfAFP1, which is a modified form of an antifungal protein isolated from Medicago plants, the modified form exhibiting enhanced anti-fungal activity for controlling fungal pathogenesis in plants. A method for inhibiting fungal colonization of plants is described which includes preparation of nucleotide sequences encoding the modified antifungal protein, preparation of vectors containing the nucleotide coding sequence, and methods for transforming plants with the nucleotide sequences. The polypeptide can be formulated into compositions useful in controlling plant pathogenic fungi.

Disclosed herein are fungal nucleic acid sequences that encode novel polypeptides. Also disclosed are polypeptides encoded by these nucleic acid sequences, as well as derivatives, variants, mutants, or fragments of the aforementioned polypeptide, polynucleotide, or antibody. The novel leucine aminopeptidase (LAP) and other amino- and carboxypeptidases polypeptides, referred to herein as EXOX nucleic acids and proteins of the invention are useful in a variety of medical, research, and commercial applications.

The invention belongs to the technical field of organic agricultural product production, and particularly provides a method for planting organic strawberry by a micro-ecological technology and a preparation of a used micro-ecological bacteria liquid. In particular, the micro-ecological bacteria liquid is obtained by preparing a plurality of stock solutions. The components of the stock solutions comprise a photosynthetic bacterium stock solution, a beauveria bassiana stock solution, trichoderma culture, pathogenic fungus protein and a biological organic bacterial fertilizer (nutritive water). The bacteria liquid can be used as a medicament for sterilizing and killing worms and also can be used as an organic fertilizer for using. The high-quality organic strawberry which meets the requirement of OFDC (Organic Food Development Center) standard can be produced by only spraying the bacterial liquid regularly. By adjusting and controlling a micro-ecological environment of microorganism, an effective microbial community in a microbial community in a plant body after spraying the micro-ecological bacteria liquid can be changed into a dominant bacterial community, so that effects such as nutrition competition of the dominant bacterial community and the like can be fully played, the number of harmful bacterial communities is controlled below an economic harm level, and a maladjusted microecosystem can be restored into balance, so that the best economic, ecological and social benefits are obtained.

The invention discloses a group of natural antimicrobial peptides derived from fungi. Amino acid sequences of the natural antimicrobial peptides are separately shown in SEQ ID NO: 3-4. Mature peptidesequences of the antimicrobial peptides are obtained through retrieving a fungal protein database and carrying out screening analysis; proven by comparative analysis, the amino acid sequences of the natural antimicrobial peptides are obviously different from those of all currently-known antimicrobial peptides, and the natural antimicrobial peptides belong to novel antimicrobial peptides. The antimicrobial peptides are optimized through codon, and then, the recombinant expression of the antimicrobial peptides can be achieved in pichia pastoris. Proven by antimicrobial experiments, the antimicrobial peptides disclosed by the invention have remarkable bacteriostasis activity to gram-positive bacteria, particularly Staphylococcus aureus ATCC43300, Streptococcus suis CVCC 3928 and CVCC 606, canbe applied to the fields of antibacterials, food additives, cosmetics, feed additives, preservatives and the like and have broad application values and market prospects.

A diagnostic test and method is provided comprising mixing blood or another biological fluid sample with a test compound and spotting the blood on filter paper for subsequent analysis of the effect of the test compound on the blood. The biological fluid can be a cerebrospinal fluid, a peritoneal fluid, a cyst fluid, an amniotic fluid, a lavage fluid, a saliva, a cell extract or a tissue extract. The compound is chosen among an amino acid, a peptide, a protein, a carbohydrate, an oligosaccharide, a polysaccharide, a glycoprotein, a lipid, a lipoprotein, a glycosaminoglycan, a hormone, a steroid, a vitamin, a low molecular weight synthetic or natural compound which influences the blood to cause an alteration of its composition, e.g., a toxin, allergen, autoantigen, bacterial protein or polysaccharide, viral protein, fungal protein or polysaccharide, parasitic protein or polysaccharide, bacterial lipopolysaccharide or any other compound relevant to diseases.

The invention relates to the field of molecular biology, in particular to a method for improving the resistance of wheat to sharp eyespot by adopting a genetic engineering method to introduce a rhizoma gastrodiae antifungal protein gene and a rabbit defensin gene into conventional wheat and expressing. The method for improving resistance to wheat sharp eyespot comprises the following steps: constructing an expression vector pAHC-NP-GAFP of the rhizoma gastrodiae antifungal protein gene and the rabbit defensin gene; introducing the pAHC-NP-GAFP into receptor-wheat immature embryo cells through particle bombardment, performing callus induction and plant regeneration on the wheat immature embryo cells to obtain transgenic candidate plants, obtaining positive transgenic plants through molecular detection and obtaining T5-generation transgenic homozygous lines from the positive transgenic plants through inbreeding and PCR screening; performing sharp eyespot resistance screening on progenies of the T5-generation transgenic homozygous lines; and screening out the transgenic plant with improved resistance to sharp eyespot. The method has the advantages that the method has the characteristics of strong pertinence, short cultivation period and little influence on important agronomic characters compared with conventional planting-cultivating methods.

The invention discloses a promoter for a filamentous fungal protein secretion pressure feedback regulation element and feedback inhibition resistance, a plasmid, a preparation method and a transformed cell. The regulation element is a base sequence of the area from (-378bp)-(-291bp) of a filamentous fungal promoter. The regulation element is related to the protein secretion pressure feedback. The promoter against feedback inhibition is a base sequence obtained by at least losing or replacing the base of the area from (-378bp)-(-291bp) on the filamentous fungal promoter, and has promoter activity. The promoter disclosed by the invention can be efficiently expressed in filamentous fungi and particularly in the protein of aspergillus oryzae.

The invention belongs to the field of plant genetic engineering, and discloses clone and application of a fungus PcGDH (Pleurotuscystidiosus Glutamate Dehydrogenase) gene for improving efficient utilization of nitrogen. The NADP (H) (Nicotinamide Adenine Dinucleotide Phosphate) enzymatic activity of a PcGDH in-vitro positive reaction is larger than that of a reverse reaction, that is the PcGDH protein is inclined to utilize NH4+ to translate alpha-oxoglutarate into glutamic acid. In addition, the affinity of the PcGDH to NH4+ is larger than glutamic acid. By virtue of a genetic engineering technology, the PcGDH gene is over-expressed in rice in a heterogeneous manner to improve utilization of nitrogen by transgenic rice, improve growth of the transgenic rice and increase absolute plant and thousand seed weight of the transgenic rice. Therefore, the PcGDH gene for improving the rice nitrogen can be utilized to culture novel rice products with good agronomic characters.

The invention discloses a group of pyronema omphalodes natural antibacterial peptides, wherein the amino acid sequences of the pyronema omphalodes natural antibacterial peptides are shown by SEQ ID NO: 5 to 8. A fungus protein database is retrieved so as to obtain the mature peptide sequences of the antibacterial peptides are obtained by retrieving a fungus protein database and screening and analyzing; through a comparative analysis, the group of natural antibacterial peptides has an obvious difference from the amino acid sequences of all antibacterial peptides known at present, and belongs toa novel type of antibacterial peptides. The group of antibacterial peptides is optimized by a codon to realize recombination expression in pichia pastoris. The result of an antibacterial experiment indicates that the antibacterial peptide has a remarkable bacteriostatic activity against gram-positive bacteria, particularly against staphylococcus aureus ATCC43300 and streptococcus suis CVCC 3928 and CVCC 606, and can be applied to the fields such as antibacterial agents, food additives, cosmetics, feed additives and preservatives, and has a great application value and a broad market prospect.

Provided herein are compositions and methods for producing milk proteins in plants, which allow for safe, sustainable and humane production of milk proteins for commercial use, such as use in food compositions. The disclosure provides recombinant fusion proteins comprising a milk protein, or fragment thereof and a structured mammalian, avian, plant, or fungal protein, or fragment thereof. The disclosure also provides methods for producing the recombinant fusions proteins, and food compositions comprising the same.

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 discloses a method for preparing an edible fungus protein extract with antitumor efficacy. The preparation method comprises the following steps of: 1) extracting by water after crushing edible fungus sporocarp, collecting a water-soluble matter to obtain an edible fungus water-soluble extract; 2) carrying out ammonium sulfate precipitation of which the saturation level is 80% on the edible fungus water-soluble extract, collecting sediments, and obtaining the edible fungus protein extract with the antitumor efficacy after dialyzing the sediments by deionized water, wherein the edible fungus is at least one of the following funguses: auricularia polytricha, pleurotus citrinopileatus, abalone mushroom, pycnoporus cin, cordyceps militaris, pleurotus djamor, black fungus, clitocybe maxima, flammulina velutipes, oudemansiella radicata and pholiota adipose. The edible fungus protein extract disclosed by the invention has a significant proliferation inhibition role on an HepG2 cell, an MCF-7 cell and an A-549 cell, and can be used for preparing a product for resisting tumors or tumor cells.

The invention discloses a Penicillium oxalicum host strain for enhancing expression of filamentous fungi protein, which is named Penicillium oxalicum M3QDelta15A and collected to China General Microbiological Culture Collection Center, wherein the collection number is CGMCC No.9092. The invention also discloses application of the strain in producing filamentous fungi protein. The experiment proves that the engineering bacterium disclosed by the invention has low secretion background in the aspect of expressing Penicillium oxalicum exoglucanase CBHI. Compared with the expression of host with Penicillium oxalicum strain Delta15A, the Penicillium oxalicum M3QDelta15A host has high protein expression level. When using cheap starch as the carbon source to produce the protein, the Penicillium oxalicum host strain has the advantages of low production cost and short period, and has favorable industrial development and application prospects.

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 an application of a domestic silkworm cocoon antifungal protease inhibitor in fungus prevention and a separation method of the domestic silkworm cocoon antifungal protease inhibitor. The separation method comprises the following steps: layering and cutting a silkworm cocoon into pieces, then adding 100mm of Tris-HCL buffer solution with pH of 7.5, and extracting for 30 minutes under the condition that the temperature is 37 DEG C and the rotation speed is 220 r / pm, centrifuging for 15 minutes under the rotation speed of 10000rpm, filtering supernatant by utilizing a 0.22-micrometer filter membrane, to obtain the domestic silkworm cocoon antifungal protease inhibitor. The prepared protease inhibitor can inhibit the activity of fungal protease secreted by Tritirachium album limber, can inhibit the spore germination of silkworm pathogenic fungal beauveria bassiana and still can maintain the activity after being incubated for 10 minutes in boiled water. The prepared domestic silkworm cocoon antifungal protease inhibitor belongs to the natural, high-activity and high-stability antifungal protease and is non-toxic and harmless; and meanwhile, the silk used in the method is rich in resource, and a novel way is provided for diversified development of the silk.

The method for processing lily utilizing biological fermentation technology mainly includes the following steps: making dried lily into lily starch, adding corn cob powder and uniformly mixing them, enzyme activating, inoculating to make fermentation, distilling to obtain lily liquor, tail water and residue; adding cooked rice in residue, adding jujube powder, uniformly mixing them, enzyme activiting, inoculating the make fermentation, distilling, using distilled liquor, tail water and melitose to make them into lily honey liquor, adding the partial residue produced in production of lily starch into tail water produced in first distillation, boiling, inoculating to make fermentation, flavouring to obtain lily vinegar, besides utilizing residual materials and biological fermentation process to make them into lily beverage and lily fungal protein feed.

The invention discloses an engineering strain producing anti-fungal protein genes and a construction method and application thereof. The strain is pET32a(+)-Ap1-BL21, and the preservation serial number is CGMCC No.11542. According to the construction method, double enzyme digestion is carried out on anti-fungal protein AP1 genes and plasmid pET32a(+) through BamH I and XholI, and the anti-fungal protein AP1 genes and the plasmid pET32a(+) are connected through T4DNA ligase and then converted into E.coli BL21 competent cells to obtain the engineering strain pET32a(+)-Ap1-BL21 with the anti-fungal protein genes. The gene engineering strain contains the anti-fungal protein genes, induction time at the temperature of 30 DEG C is short, the expression rate (43.2%) is high, the purification protein yield (124.82 mg / L) is high, the recovery rate (92%) is high, protein has a good inhibiting effect on phytopathogen such as soybean root rot bacteria and fusarium fujikuroi after ITPG inducting, HIS column purifying and enterokinase restriction enzyme digesting are carried out, and the strain provides a novel strain resource for research and development of recombinant protein pesticide.

The invention provides an antifungal protein, which is obtained by purifying a water soluble extract of hulless oat seed, wherein the apparent molecular weight of the antifungal protein is about 35kDa. The preparation method comprises the following steps: crushing and degreasing the hulless oat seed; extracting the hulless oat seed by buffer solution to obtain a coarse protein; and obtaining the antifungal protein by the two-step chromatographic separation which is the Resource S cation exchange and Superdex 75 gel exclusion chromatographic purification. The antifungal protein has obvious effect of inhibiting the growth of white rot fungus and trichoderma viride, and the minimum inhibition for the white rot fungus is 0.42mu g, and 0.21mu g for the thetrichoderma viride. In addition, the antifungal protein also has certain effect of inhibiting the growth of fusarium. The antifungal protein can be used as a food anticorrosion additive, a crop antifungal damage spraying agent and the like, and has high practical values in the aspects of food anticorrosion, crop prophylaxis and crop disease resistance.

The invention discloses a preparation technology of edible fungus mycelium and / or an edible fungus mycelium powder, and products thereof. The preparation technology of the edible fungus mycelium and / or the edible fungus mycelium powder comprises the following steps of taking sheet food materials and other materials as raw materials, and carrying out mixing according to proportion so as to obtain an edible fungus culture material; and then, carrying out the processes of humidity controlling, microwave puffing, sterilizing, cooling, edible fungus inoculating, fungus germinating, mycelium microwave-drying and so on so as to obtain the edible fungus mycelium and / or edible fungus mycelium powder. The preparation technology of the edible fungus mycelium and / or the edible fungus mycelium powder is capable of providing more types of edible fungus mycelium and / or edible fungus mycelium powder which are convenient for eating; moreover, nutrients of edible fungus culture material and medicinal ingredients of the edible fungus mycelium are transformed and improved into microbial fungal nutrient protein powder and so on which can be eaten and absorbed by the human body on basis of biotechnology. And thus, the edible fungus mycelium and / or the edible fungus mycelium powder are low in calories, and high in contents of fungal proteins, mycelium polysaccharides, a plurality of amino acids and unsaturated fatty acids, various essential trace elements for the human body and so on so that the edible fungus mycelium and / or the edible fungus mycelium powder can be used for health-caring and preventive treatment of diseases; moreover, the preparation technology of the edible fungus mycelium and / or the edible fungus mycelium powder has the advantages that the edible fungus mycelium and / or theedible fungus mycelium powder is low in production cost, high in absolute biological conversion rate, free of medicament fumigation as well as pesticide and insecticide addition during production processes, energy-saving, and environmentally friendly.

The invention relates to a shaped vegetarian meat product comprising:a) 30-80 wt. % water;b) 5-35 wt. % oil, said oil having a solid fat content at 20 degrees Celsius (N20) of at least 1.5%;c) 2-25 wt. % protein selected from algal protein, bacterial protein, dairy protein, egg protein, fungal protein, plant protein, and combinations thereof;d) 0-40 wt. % of one or more particulate ingredients selected from herbs, spices, vegetables and combinations thereof;wherein the vegetarian meat product contains at least 4 vol. % of oil droplets having an equivalent spherical diameter in the range of 100 micrometer to 1,000 micrometer as determined by means of micro computed tomography.This shaped vegetarian meat product has a very attractive juicy appearance and texture.

Provided are transgenic plants expressing MtDef5 antifungal proteins and peptides exhibiting high levels of resistance to susceptible fungi. Such transgenic plants contain a recombinant DNA construct comprising a natural or heterologous signal peptide sequence operably linked to a nucleic acid sequence encoding these molecules. Also provided are methods of producing such plants, methods of protecting plants against susceptible fungal infection and damage, as well as compositions that can be applied to the locus of plants, comprising microorganisms expressing these molecules, or these molecules themselves, as well as pharmaceutical compositions containing these molecules. Human and veterinary therapeutic use of MtDef5 antifungal proteins and peptides to treat susceptible fungal infections are also encompassed by the invention.

The invention discloses a divalent disease-resistant gene plant expression carrier, containing starter, rabbit alexin gene, Gastrodiae anti-fungal protein gene, terminator and screening label gene, all expressed in composition form in plant cell. And its characteristic: it recombines rabbit alexin gene and Gastrodiae anti-fungal protein gene with plant cell expression carrier. Its transgenic plant cell will contain rabbit alexin protein and Gastrodiae anti-fungal protein. It contains two wider antibacterial spectrums of disease-resistant gene, but their antibacterial mechanisms are different.

The invention discloses a process for preparing pharmaceutical raw material from cultivated edible mushrooms which comprises, shredding animal sarcosoma 500 portions into meat mincing, or soaking plant fruit, root, stem or leaf 500 portions, placing into incubator, adjusting pH to be 5-6, sanitizing at high temperature, cooling down, charging 10-15 portions of edible mushroom bacterial, carrying out bacterium culturing for 20-30 days at the temperature of 20-25 deg. C, digging out the total culture material, drying to obtain the pharmaceutical raw materials.

The invention relates to a method of preparing a meat analogue, said method comprising the steps of:providing proteinaceous fibres comprising, by weight of the proteinaceous fibres, 40-80 wt. % of water and 5-50 wt. % protein selected from dairy protein, egg protein, plant protein, fungal protein and combinations thereof;providing an aqueous gelling composition, comprising by weight of the gelling composition:i. 0.01-2 wt. % xanthan gum;ii. 0.01-2 wt. % galactomannan;iii. at least 80 wt. % water;combining 100 parts by weight of the proteinaceous fibres with 2-50 parts by weight of the aqueous gelling composition.The invention further relates to a meat analogue comprising an aqueous gelling composition and to the use of the aqueous gelling composition for enhancing juiciness in meat analogues.

A diagnostic test and method is provided comprising mixing blood or another biological fluid sample with a test compound and spotting the blood on filter paper for subsequent analysis of the effect of the test compound on the blood. The biological fluid can be a cerebrospinal fluid, a peritoneal fluid, a cyst fluid, an amniotic fluid, a lavage fluid, a saliva, a cell extract or a tissue extract. The compound is chosen among an amino acid, a peptide, a protein, a carbohydrate, an oligosaccharide, a polysaccharide, a glycoprotein, a lipid, a lipoprotein, a glycosaminoglycan, a hormone, a steroid, a vitamin, a low molecular weight synthetic or natural compound which influences the blood to cause an alteration of its composition, e.g., a toxin, allergen, autoantigen, bacterial protein or polysaccharide, viral protein, fungal protein or polysaccharide, parasitic protein or polysaccharide, bacterial lipopolysaccharide or any other compound relevant to diseases.