31 results about "Burkholderia gladioli" patented technology

The invention discloses a recombinant carbonyl reductase derived from Burkholderia gladioli ZJB-12126 and a coding gene thereof, a recombinant vector containing the gene, a recombinant gene engineering bacterium converted from the recombinant vector and application in preparing the recombinant carbonyl reductase.2-benzoylaminomethyl-3-one butyrate, N,N-bis-methyl-3-one-3-(2-thienyl)propanamide, ethyl 4-chloroacetoacetate (COBE) and tert-butyl (R)-6-cyano-5-hydroxy-3-carbonylhexanoate used as substrates are subjected to biological catalytic reaction to prepare high-optical-purity (2S,3R)-2-benzamidomethyl-3-hydroxybutyrate, (S)-N,N-bis-methyl-3-hydroxy-3-(2-thienyl)propanamide, ethyl (S)-4-chloro-3-hydroxybutyrate and tert-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate.

The invention provides Burkholderia gladioli ZIB-12126, which is a new bacterial strain capable of catalyzing asymmetrical reduction of carbonyl, and application of the bacterial strain in preparation of (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester by racemizing 2-benzoyl aminomethyl-3-hydroxybutyric acid ester in a microbial conversion mode. The bacterial strain is preserved in the Chinese Typical Culture Collection Center with an address of Wuhan College, Wuhan, China and the zip code of 430072, the preservation serial number of CCTCC No. M 2012379 on September 255th, 2012. The bacterial strain provided by the invention is used for synthesizing the (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester through biologic conversion, with gentle reaction condition and environmental-friendliness; and more importantly, the product configuration is mainly (2S,3R) configuration, and step of turning the product configuration by using a chemical method is not needed, so that the operation process is simple and has good industrial application prospects.

The invention discloses a nucleotide sequence and kit for detecting Burkholderia gladioli and belongs to the field of plant inspection and quarantine. Primers of fluorescence quantitative PCR (polymerase chain reaction) for detecting Burkholderia gladioli has oligonucleotide sequences shown in SEQ ID No.1 and SEQ ID No.2. The invention has the advantages that: with the fluorescence quantitative PCR detection method and kit provided by the invention, the detection of Burkholderia gladioli is specific, sensitive and rapid.

The invention discloses a preparation method of a burkholderia gladioli molecular standard sample. The preparation method comprises the step of carrying out freeze drying on extracted genome nucleic acid of burkholderia gladioli, wherein the freeze drying conditions are as follows: a freeze dryer is started, and a cooling trap is started when temperature of a drying chamber is lowered to -35 DEG C; when the temperature of the cooling trap is lowered to -40 DEG C, a nucleic acid sample prefrozen for 2 hours at the temperature of -80 DEG C is put into the drying chamber and then vacuumized; when vacuum degree is reduced to 0.5Torr, freezing is finished; and the sample is dried at the temperature of 15 DEG C, the sample is taken out when the vacuum degree is reduced to 0.1Torr, a penicillin bottle is tightened to obtain a burkholderia gladioli genome nucleic acid standard sample, and the sample is kept in a dark place and stored at the temperature of 20 DEG C. The molecular standard sample disclosed by the invention has good physical and chemical properties and can be detected by adopting a specific primer through PCR process, and sensitivity reaches 10<-4>. Detection test shows that the standard sample has good DNA integrity, uniformity and stability, can be applied to import and export detection as a positive reference material, and accuracy of a detection result is improved.

The invention discloses a recombinant carbonyl reductase gene derived from Burkholderia gladioli ZJB-12126 and a codase thereof, a recombinant vector containing the gene, a recombinant gene engineering bacterium converted from the recombinant vector and application in catalyzing asymmetric reduction of prochiral carbonyl compound. 2-benzoylaminomethyl-3-one butyrate, tert-butyl (R)-6-cyano-5-hydroxy-3-carbonyl hexanoate and ethyl 4,4,4-trifluoro acetoacetate used as substrates are subjected to biological catalytic reaction to prepare high-optical-purity (2S,3R)-2-benzoylaminomethyl-3-hydroxy-butyrate, tert-butyl 6-cyano-(3R,5R)-dihydroxy hexanoate and ethyl (S)-4,4,4-trifluoro-3-hydroxy-butanoate. The recombinant Escherichia coli can be used as a biocatalyst to perform the biological catalytic reaction, thereby providing an alternative new enzyme source for biological catalytic synthesis of the drug chiral intermediate.

The invention discloses a protein fingerprint model for Burkholderia gladioli and application thereof. The model is drafted according to the mass-to-charge ratios of the protein of Burkholderia gladioli and the peak intensity coefficients of the protein, wherein the mass-to-charge ratios include 3568.4, 3864.3, 4802.0, 5233.8, 6829.7, 7133.9, 7422.1, 7701.7, 9602.7 and 10153.8. A detection method comprises the following steps: preparation of a sample protein chip; detection with matrix-assisted laser desorption ionization time-of-flight mass spectrometer; and comparison of the protein fingerprint of the sample. The protein fingerprint model can qualitatively detect whether a sample contains Burkholderia gladioli or not, can be used for detection of commercial Burkholderia gladioli and is especially applicable to departments like a port inspection and quarantine part with detection requirements for rapidness, high flux and the like.

The invention discloses a multifunctional compound biofertilizer and belongs to the field of biotechnology. The multifunctional compound biofertilizer comprises fermented liquid of burkholderia gladioli, rahnella aquatilis and azotobacter, having effective functions of phosphorus solubilizing and nitrogen fixation, and can increase effective phosphorus content and nitrogen content needed for growth of plants in soil and improve soil fertility; IAA secreted from the biofertilizer can improve growth of plants; besides, the compound biofertilizer can generate high-activity chitinase, effectivelyinhibit breeding of pathogenic fungus, reduce diseases, further increase and supplement beneficial microorganism of soil and improve status of soil, and thus, the compound biofertilizer has good development prospect in terms of reducing usage of chemical fertilizer, improving soil fertility, improving growth plants and the like.

The invention discloses screening and application of burkholderia gladioli real-time quantitative polymerase chain reaction (qRT-PCR) reference genes and primers thereof. According to obtained transcriptome analysis results and reported reference genes, 12 candidate reference genes with relatively stable expression are preliminarily screened, and gene names are shown as follows: atpD, clpP, clpX,ftsZ, gyrB, lpxC, pyrG, recA, rpoB , rpoD, thyA and 16S; for the candidate genes, amplification primers are designed; reference genes under different culture conditions (temperature, initial pH valueof medium, culture time, treatment with NaCl different in concentration) of burkholderia gladioli are screened; geNorm, NormFinder and Bestkeeper software is employed to analyze qRT-PCR results; and finally, the reference genes with most stable expression under different culture conditions and NaCl treatment condition are obtained by screening. The invention is beneficial to the stability and reliability of analysis and study of gene expression of the burkholderia gladioli under different conditions.

The invention relates to a Burkholderia gladioli BSP-1 strain highly yielding alkaline lipase. The strain is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No.10533. The strain is from rotten onion, and has the advantages of stable enzyme production, wide source of a raw material, and low production cost. The invention also provides a method for preparing the alkaline lipase through using the strain. The method comprises the steps of seed culture, fermentation culture and separating purification. A fermentation medium is optimized in the method, and the lipase prepared in the invention has strong vitality reaching up to 145.67U / mL, has extremely good stability in alkaline environment, has good tolerance to short chain alcohols, and has wide application prospects in the fields of biomass energy, organic synthesis, drug chiral resolution, and tool enzymes.

The invention belongs to the technical field of biological control on plant diseases, and relates to an endophytic burkholderia gladioli PJB25 strain and an application thereof. According to the invention, the endophytic burkholderia gladioli PJB25 is separated and screened from healthy peanuts. The endophytic burkholderia gladioli PJB25 is safe and non-toxic. The endophytic burkholderia gladioli PJB25 is preserved in Guangdong Microbial Culture Collection Center on June 9, 2020, with preservation number of GDMCC No: 61053. The strain has a significant bacteriostatic effect on entry quarantine pests such as cylindrocladium parasiticum, and can significantly reduce the occurrence of black rot in peanuts. Meanwhile, the strain has an inhibiting effect on major fungal diseases such as peanut root rot, peanut sheath blight, peanut stem rot and peanut anthracnose threatening production of crops such as peanuts. The strain is wide in antibacterial spectrum, and provides a new choice for biological control on plant fungal diseases. The endophytic burkholderia gladioli PJB25 can be used for broad-spectrum microbial agents / bacterial fertilizers with good development and application prospects, and has important significance on green control on plant diseases.

The invention provides a burkholderia gladioli LAMP (loop-mediated isothermal amplification) constant-temperature rapid detection method. The method comprises the following steps: 1, preparing a DNA (deoxyribonucleic acid) sample to be detected and LAMP constant-temperature amplification specific primers; wherein the LAMP isothermal amplification specific primer is obtained by selecting a burkholderia gladioli specific gene sequence for synthesis, and the gene adopts a burkholderia gladioli (pseudomonas arenata subsp. Oryzae) hydrolase (cayF) gene; the LAMP isothermal amplification specific primers comprise an outer primer F3, an outer primer B3, an inner primer FIB, an inner primer BIP, a loop primer LF and a loop primer LB; and 2, carrying out LAMP isothermal amplification on the to-be-detected DNA sample by adopting the LAMP isothermal amplification reaction system, and detecting a fluorescence signal of an amplification product to obtain a detection result. The Burkholderia gladioli detection kit has good specificity and high sensitivity on Burkholderia gladioli, and can be used for rapid detection of Burkholderia gladioli in food.

The invention discloses a duplex PCR (polymerase chain reaction) method for synchronously detecting Burkholderia gladioli pv.alliicola and Erwinia chrysanthemi as quarantine bacteria. In order to establish the duplex PCR method for synchronously detecting Burkholderia gladioli pv.alliicola and Erwinia chrysanthemi as the quarantine bacteria, a pair of specific primers B3 / B6 is designed, the designed primers are bound with specific primers ADE1 / ADE2 for detecting Erwinia chrysanthemi, a duplex PCR system is built through condition optimization, and specificity and sensitivity verification is performed. With the application of the duplex PCR system, specific bands can be obtained from genomic DNA of Burkholderia gladioli pv.alliicola and Erwinia chrysanthemi and artificial bacteria-bearing samples by amplification. Results indicate that the duplex PCR method can detect Burkholderia gladioli pv.alliicola and Erwinia chrysanthemi simultaneously and can be applied to detection of two typesof quarantine bacteria of tulip bulbs imported from ports.

The invention provides a method, fluorescent PCR primers and probe for detecting burkholderia gladioli and a bongkrekic acid toxin-producing strain. The fluorescent PCR primers and probe for detecting the burkholderia gladioli include a PBA-F primer with a sequence as shown in SEQ ID NO:1, a PBA-R primer with a sequence as shown in SEQ ID NO:2, and a PBA fluorescent probe with a sequence as shown in SEQ ID NO:3. According to the technical scheme, the burkholderia gladioli and thebongkrekic acid toxin-producing strain are subjected to real-time fluorescent PCR detection by adopting a two-step method; the blank of molecular detection is filled; the method can be used for rapidly screening the burkholderia gladioli and the bongkrekic acid toxin-producing strain in foods; and the method is good in specificity, high in repeatability and high in sensitivity, can check hidden dangers in time and has relatively high practical value.

The invention discloses a recombinant carbonyl reductase sourced from Burkholderia gladioli ZJB-12126 as well as an encoding gene of the recombinant carbonyl reductase, a recombinant vector containing the gene, a recombinant gene engineering bacterium obtained by transforming the recombinant vector, and an application of the recombinant carbonyl reductase in asymmetrically reducing prochiral carbonyl compounds. According to the recombinant carbonyl reductase disclosed by the invention, by respectively taking 2-benzamidomethyl-3-ketone butyrate, 4-chloroacetoacetic acid ethyl ester (COBE), (R)-6-cyano-5-hydroxyl-3-carbonyl caproate tert-butyl ester and (4S)-3-[5-(4-fluorophenyl)-1,5-dioxopentyl]-4-phenyl-2-oxazolidinone as substrates, (2S, 3R)-2-benzamidomethyl-3-polyhydroxybutyrate, (S)-4-chloro-3-ethyl hydroxybutyrate, 6-cyano-(3R, 5R)-dihydroxyhexanoic acid tert-butyl ester and (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxyl valeryl]-4-phenyl-1,3-oxo-azacyclopentane-2-ketone with high optical purity are prepared by virtue of biocatalytic reaction, wherein the biocatalytic reaction can be performed by using recombinant escherichia coli as a biocatalyst, and thus optional new enzyme sources can be provided for biocatalytic synthesis of chiral intermediates of medicines.

The present invention relates to a novel protein comprising novel genes that is extracted from Burkholderia gladioli strain NGJ1. A nucleotide sequence encoding the novel protein is represented by sequence SEQ ID No. 1 and amino acid sequence of the novel protein is represented by the sequence SEQ ID No. 2 is further provided. A nucleotide sequence and the amino acid sequence are obtained from genetically engineered Bg_9562 gene. The novel protein as well as encoding gene is adapted for broad spectrum anti-fungal and mycophagous activities.

The invention provides Burkholderia gladioli ZIB-12126, which is a new bacterial strain capable of catalyzing asymmetrical reduction of carbonyl, and application of the bacterial strain in preparation of (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester by racemizing 2-benzoyl aminomethyl-3-hydroxybutyric acid ester in a microbial conversion mode. The bacterial strain is preserved in the Chinese Typical Culture Collection Center with an address of Wuhan College, Wuhan, China and the zip code of 430072, the preservation serial number of CCTCC No. M 2012379 on September 255th, 2012. The bacterial strain provided by the invention is used for synthesizing the (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester through biologic conversion, with gentle reaction condition and environmental-friendliness; and more importantly, the product configuration is mainly (2S,3R) configuration, and step of turning the product configuration by using a chemical method is not needed, so that the operation process is simple and has good industrial application prospects.

The invention discloses a recombinant carbonyl reductase gene derived from Burkholderia gladioli ZJB-12126 and a codase thereof, a recombinant vector containing the gene, a recombinant gene engineering bacterium converted from the recombinant vector and application in catalyzing asymmetric reduction of prochiral carbonyl compound. 2-benzoylaminomethyl-3-one butyrate, tert-butyl (R)-6-cyano-5-hydroxy-3-carbonyl hexanoate and ethyl 4,4,4-trifluoro acetoacetate used as substrates are subjected to biological catalytic reaction to prepare high-optical-purity (2S,3R)-2-benzoylaminomethyl-3-hydroxy-butyrate, tert-butyl 6-cyano-(3R,5R)-dihydroxy hexanoate and ethyl (S)-4,4,4-trifluoro-3-hydroxy-butanoate. The recombinant Escherichia coli can be used as a biocatalyst to perform the biological catalytic reaction, thereby providing an alternative new enzyme source for biological catalytic synthesis of the drug chiral intermediate.

The invention provides an LAMP (loop-mediated isothermal amplification) double-strand detection probe and freeze-dried microspheres of burkholderia gladioli as well as a preparation method and a detection method of the freeze-dried microspheres, and belongs to the field of gene detection. The double-chain detection probe comprises a light-emitting probe and a quenching probe, the light-emitting probe is located on the FIP, the 5'end of the FIP sequence is marked with a fluorescent light-emitting group, the quenching probe is complementary with the F1C sequence, and the 3 'end of the quenching probe is marked with a fluorescent quenching group. The LAMP detection freeze-dried microsphere comprises the double-stranded detection probe, the freeze-dried microsphere and the detection method, the burkholderia gladioli 16-23sRNA is taken as a detection target, a double-stranded probe-constant temperature color-changing system is adopted, a result can be interpreted by combining fluorescent quantitative PCR (Polymerase Chain Reaction) atlas analysis and a naked eye observation method, the probe is high in specificity, the color-changing color contrast is obvious, misjudgment is not easy to generate, and the detection result is accurate. The dual systems reflect each other, and the detection accuracy is high.

The invention discloses a nucleotide sequence and kit for detecting Burkholderia gladioli and belongs to the field of plant inspection and quarantine. Primers of fluorescence quantitative PCR (polymerase chain reaction) for detecting Burkholderia gladioli has oligonucleotide sequences shown in SEQ ID No.1 and SEQ ID No.2. The invention has the advantages that: with the fluorescence quantitative PCR detection method and kit provided by the invention, the detection of Burkholderia gladioli is specific, sensitive and rapid.

The invention relates to a high-yield alkaline lipase-producing Burkholderia strain (Burkholderia gladioli BPS‑1), which is preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.10533. The bacterial strain is derived from rotting onions, has stable enzyme production, wide sources of raw materials, and low production cost. The invention also provides a method for preparing alkaline lipase by the bacterial strain, which comprises the steps of seed cultivation, fermentation cultivation and separation and purification. The method optimizes the fermentation medium, and the prepared lipase has strong activity, up to 145.67U / mL, excellent stability in an alkaline environment, and good tolerance to short-chain alcohols. It is used in biomass energy, organic It has broad application prospects in the fields of synthesis, chiral resolution of drugs and tool enzymes.

The invention relates to the technical field of microorganisms, in particular to burkholderia gladioli MEJ03 and application thereof. The preservation number of the burkholderia gladioli MEJ03 is CCTCC (China Center for Type Culture Collection) NO: M20211229. The burkholderia gladioli MEJ03 can be used for efficiently degrading phthalic acid ester, especially di (2-ethylhexyl) phthalate, has phosphorus dissolving activity, can be used for dissolving insoluble phosphate calcium phosphate and ground phosphate rock, and can also be used for promoting plant growth.

The invention relates to burkholderia gladioli A1-07 and application of the burkholderia gladioli A1-07 in preparation of ferulic acid. The preparation method of the ferulic acid comprises the steps of preparing crude enzyme powder, hydrolyzing oryzanol and separating. The oryzanol conversion rate of the ferulic acid preparation method reaches 67%, the yield of the ferulic acid preparation method can reach more than 56%, and the purity of the obtained ferulic acid product is 22-31% by weight. Therefore, the ferulic acid preparation method disclosed by the invention is simple and convenient to operate, high in production efficiency and environment-friendly, and has a good industrial application prospect.

The invention belongs to the technical field of biological control of plant diseases, and relates to a strain of endophytic Burkholderia gladioli (Burkholderia gladioli) PJB25 and application thereof. The present invention isolates and screens an endophytic Burkholderia gladiolus PJB25 from healthy peanuts, which is safe and non-toxic. The strain was preserved in the Guangdong Microbial Culture Collection Center on June 9, 2020, and the preservation number is GDMCC No: 61053. The strain has a strong antibacterial effect on the entry quarantine harmful organism peanut black rot, and can significantly reduce the occurrence of peanut black rot; at the same time, it has a strong antibacterial effect on peanut root rot, peanut sheath blight, peanut stem rot, peanut anthracnose, etc. The important fungal diseases that threaten the production of peanuts and other crops all have inhibitory effects, and the antibacterial spectrum is wide, which provides a new option for the biological control of plant fungal diseases. The endogenous Burkholderia gladiolus PJB25 of the present application can be used as a broad-spectrum microbial agent / bacterial fertilizer with good development and application prospects, and is of great significance to the green prevention and control of plant diseases.

The invention discloses a burkholderia gladioli LAMP (loop-mediated isothermal amplification) detection primer group, a kit and a preparation method of freeze-dried microspheres, and belongs to the field of nucleic acid detection. The detection primer group is designed aiming at 16S-23S rRNA conserved sequences of burkholderia gladioli, and comprises an outer primer F3, an outer primer B3, an inner primer FIP, an inner primer BIP, a loop primer LF and a loop primer LB. The preparation method of the freeze-dried microspheres containing the detection primer group comprises the following steps: mixing the detection primer group with an LAMP reaction base solution to obtain an amplification reaction system; and mixing the amplification reaction system with a freeze-drying protective agent, dropwise adding into liquid nitrogen, quickly freezing into balls, and freeze-drying. According to the detection method, whether the burkholderia gladioli exists or not can be simply and efficiently detected by utilizing the freeze-dried microspheres, so that whether a detection sample contains the rice yeast acid toxin or not is judged, the specificity is high, and false positive does not occur easily.

The invention belongs to the technical field of biological control of plant diseases, and discloses a litchi endophytic Burkholderia gladiolus and its application in preventing and treating litchi anthracnose and litchi frost blight. The name of the strain is Burkholderia gladioli (Burkholderia gladioli) SZPT16, and it is preserved in the Guangdong Microbial Culture Collection Center, referred to as GDMCC. Institute, the deposit number is GDMCC No:61168, and the deposit date is August 25, 2020. The bacterial strain of the present invention is derived from litchi endophytic bacteria, and the bacterial strain and its biological preparations have a strong inhibitory effect on litchi anthracnose and litchi downy mildew, and have excellent biocontrol effects; And its processed products are safer and more reliable, and have good development and application prospects.

The invention discloses Burkholderia gladioli P8 and application thereof, and belongs to the technical field of microorganisms, the Burkholderia gladioli P8 is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation date is July 13, 2021, and the preservation registration number is CGMCC No.22825; the invention shows that the strain has the best inorganic phosphorus dissolving effect when the temperature is 35 DEG C, the pH is 7.5, the carbon source is glucose, the nitrogen source is ammonium sulfate, the C / N ratio is 40: 1 and the salt concentration is 1.5%, and the strain can secrete phosphatase and phytase, dissolve insoluble organic phosphorus and secrete plant hormones and siderophores to promote photosynthesis, respiration and substance and energy metabolism of plants, the invention can be used for solving the problems of slow seedling root development and slow seedling growth of the tamarind.

The invention discloses a method for reducing rice yeast acid and application. The method comprises the step of adding one or more of sodium chloride, baking soda and / or acetic acid into an aqueous solution for soaking the edible mushrooms. The method for reducing the rice yeast acid is safe, simple, low in cost and suitable for daily life, in the process of treating edible mushrooms, generation of the rice yeast acid by the Burkholderia gladioli coconut poison pathogenic variants can be remarkably inhibited, all needed materials can be taken from kitchen seasonings, and the method can be widely applied to prevention of rice yeast acid poisoning in daily life.

The invention discloses a Burkholderia gladioli P7 and application thereof, and belongs to the technical field of microorganisms. The Burkholderia gladioli P7 is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation date is July 13, 2021, and the preservation registration number is CGMCC No.22824; and the strain has the best organophosphorus decomposing effect when the temperature is 35 DEG C, the pH value is 7.5, the carbon source is glucose, the nitrogen source is ammonium sulfate, the C / N ratio is 40: 1 and the salt concentration is 2.5%, can secrete phosphatase and phytase, dissolve insoluble organophosphorus and secrete plant hormones and siderophores, promotes plant photosynthesis, respiration and substance and energy metabolism, and can be used for solving the problems of slow seedling root development and slow seedling growth of the tamarind.