212 results about "Surfactin" patented technology

An isolated Bacillus amyloliquefaciens Ba-BPD1 having an Accession No. of DSM 21836 is provided. This novel strain has unique 16S ribosomal RNA sequenced as SEQ ID NO:1 and produces amylase, protease, cellulase and lipase, fibrinolytic enzyme to show their biodegradation capacities. Further, B. amyloliquefaciens Ba-BPD1 produces the antibiotic substances, such as iturin, fengycin and surfactin, and has antimicrobial capacity for inhibiting the fungal or bacterial growth. In conclusion, the novel strain of Bacillus amyloliquefaciens Ba-BPD1 and its products can be applied in agriculture, wastewater treatment, food industry and chemical industry.

The invention relates to a method for preparing a bacillus subtilis lipopeptid biosurfactant, which is characterized in that the adopted bacterial strain is efficient lipopeptid producing strain bacillus subtilis BIT09S1,BIT09S2 and BIT09A2, and the lipopeptid biosurfactant is prepared by taking cheap molasses, pulse flour, konjac gum finemeal, guar gum and gum breaking liquid of modified guar gum as carbon sources for the culture medium; adding other nitrogen sources, phosphorus sources, inorganic salt components and nutrient elements into the culture medium; and carrying out fermentation, separation and purification. The raw materials used for producing lipopeptid by the method are easily obtained and can be enormously purchased. The crude extract of the obtained lipopeptid biosurfactant Surfactin can be used in various drilling and production processes such as oil-gas field fracturing, acidizing, de-plugging, controlling and water shut-off, oily water treatment, environment renovation and the like, and the biosurfactant has wide prospect.

The invention discloses a preparing method of new type lipopeptide biosurfactant surfactin and appliance, which comprises the following steps: utilizing bacillus subtilis strain E8 (conserved in Chinese microbe conservative council common microbe center, the keeping number at 1107) to proceed microbe liquid ferment prepare surfactant surfactin; proceeding carbon source, nitrogen source and inorganic salt optimize for the needed liquid culture medium; getting fermentation liquor; centrifuging to remove thallus; acidifying supernatant; extracting rough extract; further-extracting through dissolvent; acidifying; collecting deposition; centrifuging; vacuum-drying; getting pre-refined product; freeze-drying through drainage pillar and FPLC extraction; getting product with high purity; making the output of acidifying extraction at 24-56 h reach 5-13g / L. The dominant fermentate molecular weight of high-yield strain E8 can reach 1022Da and CMC can reach 1 mu M, which is the lowest of reported surfactin CMC value.

An isolated Bacillus amyloliquefaciens Ba-BPD1 having an Accession No. of DSM 21836 is provided. This novel strain has unique 16S ribosomal RNA sequenced as SEQ ID NO:1 and produces amylase, protease, cellulase and lipase, fibrinolytic enzyme to show their biodegradation capacities. Further, B. amyloliquefaciens Ba-BPD1 produces the antibiotic substances, such as iturin, fengycin and surfactin, and has antimicrobial capacity for inhibiting the fungal or bacterial growth. In conclusion, the novel strain of Bacillus amyloliquefaciens Ba-BPD1 and its products can be applied in agriculture, wastewater treatment, food industry and chemical industry.

An oil-in-water emulsified composition suitable for external preparation for skin and cosmetic product, which comprised 0.1 to 5% by mass of (A) lipopeptide compound derived from a microorganism such as surfactin and its analogous compound, 0.05 to 1.5% by mass of (B) xanthan gum, 25 to 70% by mass of (C) oil component and (D) water, free from non-ionic surfactants and acrylic water-soluble polymers, and is excellent in feeling upon use, moisture retention, emollient property and stability as well as environmental suitability and safety for living organisms, and external preparation for skin and cosmetic product using the composition.

The invention discloses bacillus methylotrophicus F7 and an application thereof. The invention aims at providing a strain which has a plurality of beneficial properties, namely producing siderophore, cellulase and protease, producing auxin, producing ammonia and the like; the strain not only can be used for effectively inhibiting various pathogenic bacteria such as ralstonia solanacearum, fusarium oxysporum f.sp.cubense, colletotrichum gloeosporioides, fusarium oxysporum f.sp cucumerinum but also can promote plant growth. In accordance with the technical scheme, the bacillus methylotrophicus F7 is preserved in China General Microbiological Culture Collection Center (CGMCC) on February 1, 2016, with preservation number of CGMCC No.12124; in addition, five surfactin homologs, six fengycin homologs and three iturin homologs, which have an antibacterial effect, are extracted from fermentation liquid of the bacillus methylotrophicus F7; and the bacillus methylotrophicus F7 belongs to the field of agricultural biotechnology.

A process for producing Surfactin, comprising culturing a microorganism of the genus Bacillus in a liquid culture medium containing flour of beans such as soybean or an extract thereof as a nitrogen source and accumulating Surfactin in the culture broth, and a microorganism of the genus Bacillus which have an activity to produce a crude Surfactin in a concentration of from 8 to 50 g / L on culturing for 20 to 90 hours.

The invention provides bacillus amyloliquefaciens HAB-2, which has the collection number of CCTCC M 2015070. Although the key lipopeptide synthetase gene sfp gene is deleted, the surfactin substances can still be generated, indicating that the strain has a specific and distinctive biosynthetic pathway and has a remarkably high research value. The bacillus amyloliquefaciens HAB-2 has very strong ability of inhibiting pathogenic fungi, has a wide antibacterial spectrum and can inhibit multiple kinds of pathogenic fungi while the antibacterial effect is good and the antibacterial rate is high. Further research finds that the active ingredient can be directly obtained through simple treatment on the fermentation broth of the strain, and the antibacterial active ingredient is stable and has very high activity in strong acid and strong alkaline environments; the antibacterial activity is not changed after 24 hours of ultraviolet irradiation, and high temperature does not influence the antibacterial active ingredient; and the fermentation broth of the biocontrol bacteria realizes certain growth promoting effect on a plant.

The invention discloses a bacillus amyloliquefaciens YJ01 which is preserved in China General Microbiological Culture Collection (CGMCC) center of China Committee for Culture Collection of Microorganisms and the preservation number of the bacillus amyloliquefaciens is CGMCCNO.7328. The bacillus amyloliquefaciens is mainly used for preventing and treating a fungal disease of a wine-brewing grape. The bacillus amyloliquefaciens YJ01 can metabolize out antibacterial substances iturin A and surfactin, thereby playing an obvious role in inhibiting various pathogenic fungi of the wine-brewing grape. The bacillus amyloliquefaciens has the advantages of being high in culture proliferation speed, capable of being cultured artificially, easy to operate, convenient to produce and apply, strong in adversity resisting capability, and the like; and the bacillus amyloliquefaciens has significance for preventing and treating diseases such as gray mold, white rot and anthracnose of wine-brewing grape caused by fungi, and is simple in culturing conditions, easy to preserve and easy to produce industrially, thereby having good development and application prospect.

The related preparation method for antibiotic peptide comprises: with ES-2 strain of Bacillus amyloliquefaciens, amplifying culture with medium, taking 2% inoculation, and deep fermenting for 30-48h at 28-35h, or taking inoculation with solid fermentation medium for culturing 2-4day at 28-37Deg; extracting, concentrating, adsorbing, and drying to obtain the product with molecular weight as 994, 1008, 1022, 1036 and 1449, 1463, 1477, 1491 Da as the mixture of surfactin and fangycin. This product can bear heating at 100Deg for 30min, keeps activity in pH 2-12, and can be used as food preservative and additive.

The invention relates to an extracellular antibacterial lipopeptide of plant endophytic Bacillus subtilis Jaasedl and a separation and purification method. The distribution range of the molecular weight of the extracellular antibacterial lipopeptide is mainly between 1,000Da and 2,200Da. The extracellular antibacterial lipopeptide contains an Iturin homologue, a Fengycin homologue and a Surfactin-like Compound homologue. The extracellular antibacterial lipopeptide is a group of uncommon antibacterial lipopeptide mixture produced by a single bacterial strain. The separation and purification method of the extracellular antibacterial lipopeptide comprises the following steps of: after salting out to obtain the crude extract of the antibacterial lipopeptide by using ammonium sulfate from the fermentation liquor of the endophytic Bacillus subtilis Jaasedl, performing Sephedex G-25 molecular sieve chromatography, Cellulose DEAE-52 anion exchange chromatography and FPLC300SB-C18 column chromatography successively, wherein 34 to 37 min of collecting peak has bacteriostatic activity; detecting by Tricine-SDS-PAGE after concentrating; and achieving electrophoretically pure at only one strip to obtain pure extracellular antibacterial lipopeptide. The antibacterial lipopeptide has extremely high research and application value for the development of broad-spectrum antifungal medicaments, and has wide application prospect for natural quality protection of crops and pesticide residue reduction.

The invention relates to a composite antibacterial peptide produced by bacillus natto NT-6 and a production method thereof. The composite antibacterial peptide product is a mixture of three kinds of homologs of antibacterial lipopeptide substances iturins, fengycins and surfactins in the proportions of 10-20 percent, 25-35 percent and 50-60 percent respectively; the antibacterial peptide pure product is a yellow viscous substance which can resist heating of 30 minutes at the temperature of 100 DEG C, can keep activity at the pH of 2-12, has broad-spectrum antibacterial activity for bacteria and fungi and has strong killing effect on protozoa and cancer cells; and experiments prove that the peptide has high safety. The invention also discloses the production method of the composite antibacterial peptide. The peptide can be easily produced by using the process, is relatively low in cost, and can be used as a food preservative, a feed additive, an agricultural biological control agent, a medicament, an animal disease preventing agent and the like.

The invention belongs to the petroleum exploitation field, and discloses an application of iturins sodium surfactin as an oil displacement agent and the oil displacement agent containing iturins sodium surfactin. According to the invention, iturins sodium surfactin as the oil displacement agent has good thermostability and no chromatogram separating phenomenon, alkali and an organic solvent are not added, so that low oil / water interfacial tension can be reached in a small concentration scope, crude oil can be cleaned from the stratum surface, oil displacement efficiency is increased, crude oil recovery efficiency is increased, damage of an alkalescence body to an oil layer can be avoided, and the continuous development for petroleum exploitation can be realized. The composite oil displacement agent comprises iturins sodium surfactin, iturins sodium surfactin has good compatibleness with the chemical oil displacement agent, amount of the chemical oil displacement agent can be obviously reduced when the interfacial tension can reach low level, recovery efficiency of three-time oil extraction of an oil field can be increased, and the cost for three-time oil extraction can be simultaneously and effectively increased.

The invention discloses a production method of bio-surfactin. The method comprises the following steps of: sequentially inoculating bacillus amyloliquefaciens fmb50 to a PDA agar slant culture medium for culture, then inoculating the bacillus amyloliquefaciens fmb50 to a BPY seed culture medium for further culture, and finally inoculating the bacillus amyloliquefaciens fmb50 to a fermentation culture medium, thereby obtaining a culture solution containing bio-surfactin; then diluting the culture solution with water, then regulating the pH, further adding chitosan, sodium alga acid and kieselguhr, performing flocculation, and then filtering; finally dissolving and extracting a filter cake with low alcohols, then filtering, and performing reduced-pressure distillation on the filter liquor, thereby obtaining a bio-surfactin crude product. The production method of bio-surfactin, provided by the invention, is capable of realizing industrial preparation of bio-surfactin; the prepared bio-surfactin is high in purity, so that the method is capable of overcoming the defect that large-scale production of bio-surfactin can not be realized in the prior art.

The invention discloses a method for producing surfactin by applying bacillus amyloliquefaciens, and belongs to the technical field of industrial microbiology. The yield of the surfactin synthesized by applying the bacillus amyloliquefaciens with the collection number of CGMCC No.12593 and obtained by a screening method reaches up to 9.43g / L, and the method for producing the surfactin by applying the bacillus amyloliquefaciens has the characteristics of synchronization of surfactin synthesis and strain growth and no lag phase in a fermentation process. Strains and a fermentation method provided by the invention play an important role in industrial production of the surfactin and have broad application prospects.

The invention provides a kit for detecting chlamydia trachomatis (CT). The kit comprises a nucleic acid releaser and a PCR (Polymerase Chain Reaction) solution, wherein the nucleic acid releaser comprises 0.01-0.5mM / L of surfactin, 20-300mM / L of potassium chloride, 0.01-2% of sodium dodecyl sulfate and 0.05-1% of ethanol; and the PCR solution comprises a forward primer and a reverse primer which are used for amplifying targeted polynucleotide, and a probe for detecting the targeted polynucleotide. A method of releasing nucleic acid by using the nucleic acid releaser in the kit provided by the invention is not obviously different from a boiling method in the detection result, and a violent protein denaturant adopted for the nucleic acid extraction in the method provided by the invention quickly breaks a coat protein structure of a pathogen to release pathogen nucleic acid, so the release and extraction of DNA (Deoxyribonucleic Acid) can be realized without heating; besides, the sensitivity of the provided kit for detecting the CT can be 400 copies / ml, the linearity region of the detection is 400-4.00E+10 copies / ml; and moreover, CT-DNA in an unknown sample such as genital secretions can be quickly and precisely detected by using the kit, so a reliable experiment basis is provided for diagnosing CT infection.

The invention discloses a remediation method of soil polycyclic aromatic hydrocarbon by combining surfactant producing bacteria and mycorrhiza, belonging to the field of environmental modification and pollutant disposal. Based on the actual bottleneck of current phyto-remediation or mycorrhiza remediation, by combining the high-production strain B.subtilis BS1 of the biosurfactant surfactin with AMF, the functions and characteristics of the two are sufficiently realized, wherein the B.subtilis BS1 is used as the donor of the biosurfactant and can enhance the migration of the organic pollutant toward the rhizosphere; and AMF enhances the affinity and absorption effect of the plant against the organic pollutant, improves the tolerance of the plant against the toxicity stress of the organic pollutant, and enhances the metabolic activity of the rhizospheric microorganism. According to the invention, a novel efficient integrated remediation technology is created, the remediation efficiency of PAHs in soil is improved, the remediation time is shortened, and the technology has great practical application values.

The invention relates to a bio-control bacillus subtilis Bs916 extracellular antibiotic lipopeptide surfactin and a separation and purification method thereof. The molecular weight of surfactin is 1007.6, 1021.7 and 1035.7, and belongs to homologues with difference of one methylene. The separation and purification method comprises the following steps: constructing a mutant strain BBFM without a performance of synthesizing other lipopeptide antibiotics of Bs916 through a gene knock-out technology; then extracting coarse product of surfactin from the fermentation broth of BBFM through steps of acid precipitation and methanol extraction, and finally separating and purifying surfactin through NH2 and C18 column chromatography. Through experiments of polyacrylamide gel electrophoresis and high performance liquid chromatography (HPLC) analysis, the experiment results show that the prepared surfactin reaches the electrophoresis purity grade and the chromatography purity grade. The purified surfactin has high a research and application value for development of antifungal and antibacterial drugs, and can also be used as a standard substance in biochemistry.

The invention discloses an engineering bacterium for highly expressing a lipopeptide biosurfactant and application thereof, belonging to the fields of biotechnology and biochemical engineering. The engineering bacterium over-expressing transmembrane protein YcxA is constructed by adopting a gene engineering technology, and transmembrane transport of lipopeptide from the inner part of a cell to the outside of the cell is enhanced, so that the yield of the lipopeptide is significantly improved. Compared with an original strain, the obtained gene engineering bacterium over-expressing lipopeptide carrier protein YcxA has the advantages that the yield of surfactin is improved by 97%, the engineering bacterium can be used for producing the lipopeptide biosurfactant and has a good industrial application prospect, and the yield of lipopeptide in a fermentation broth obtained by fermentation is averagely 1-5 g / L.

The present invention discloses a double-dosage-form moisturizing essence. The double-dosage-form moisturizing essence comprises oil gel and hydrogel. The oil gel and the hydrogel are mixed accordingto a preset ratio when in use; the oil gel comprises the following components in parts by weight: 0.1-6 parts of an active ingredient, 15-35 parts of polyol, 0.1-1 part of sodium surfactin, 10-70 parts of oil and 1-10 parts of water. The double-dosage-form essence is mixed according to the preset ratio when in the use, and the oil gel and the hydrogel have a transparent appearance and good stability. The present invention also discloses a preparation method of the double-dosage-form essence, the oil gel and the hydrogel are prepared separately, a D-phase emulsification method is used to prepare the oil gel, and an emulsifier is low in use amount and can emulsify oil phase with high oil content.

The invention provides an extraction-free direct amplification reagent for real-time fluorescent quantitation PCR and application thereof. The direct amplification reagent comprises the following components: sodium hydroxide (NaOH), Surfactin, dimethyl sulfoxide (DMSO), sucrose and chelex-100. When the reagent is used, there is no need to extract and purify nucleic acid from a sample, only the sample and the reagent need to be mixed according to the equal volume ratio of 1:1. The mixture can be directly used for fluorescent PCR detection. The extraction-free direct amplification reagent is applicable to nucleic acid extraction of human genome, bacteria and viruses in mouth swabs, nasopharyngeal swabs, serum, plasma, genital tract secretion and other samples. The reagent is suitable for both the fluorescence PCR of a probe method and the PCR of a dying method, is suitable for a variety of clinical applications and has the advantages of low cost, simple operation, accurate results and the like, and detection can be simple and quickly completed.

Surfactin is a biosurfactant produced by Bacillus subtilis and is a natural cycloaliphatic peptide having a ring structure made of 7 amino acids. The surfactin has various functions including anti-aging, anti-wrinkle, increasing skin penetration of cosmetic products (skin penetration agent), foaming agent, and emulsifier.

The invention relates to a plant cosmetic preservative composition and a preparation method thereof. The plant cosmetic preservative composition includes a preservative mixture composed of a grapefruit seed extract, an origanum vulgare herb extract and a magnolia officinalis bark extract. The method is characterized in that a raspberry fruit extract and a sophora flower extract are employed to conduct preservative synergy on the preservative mixture. The preservative mixture, the raspberry fruit extract and the sophora flower extract are coated by a lipid material to form a sub-microparticle emulsion, and the lipid material is composed of mannosylerythritol lipid, sodium surfactin, hydrogenated lysolecithin and glycerin in a mass ratio of 5-10:1-3:3-5:20-50. The plant cosmetic preservative composition provided by the invention has low irritation, good preservative effect and effective components from natural plants, and is suitable for cosmetics.

The invention provides a herpes virus EBV (Epstein-Barr Virus) detection kit. The kit comprises a nucleic acid releasing agent and PCR (polymerase chain reaction) reaction solution, wherein the nucleic acid releasing agent comprises 0.01-0.5 mM / L of surfactin, 20-300 mM / L of potassium chloride, 0.01-2% of sodium dodecyl sulphate and 0.05-1% of ethanol; and the PCR reaction solution comprises an upstream primer and a downstream primer used for target polynucleotide amplification, and a probe used for target polynucleotide detection. The detection result of the method for releasing nucleic acid by the nucleic acid releasing agent in the kit disclosed by the invention has no obvious difference with the detection result of a boiling method, a strong protein denaturing agent is used during nucleic acid extraction in the kit disclosed by the invention for rapidly breaking the coat protein structure of a pathogen and releasing the nucleic acid of the pathogen, and release and extraction for DNA (deoxyribonucleic acid) can be rapidly finished without heating; the sensitivity of the EBV detection of the kit disclosed by the invention can achieve 400 copies / ml, and the quantitative linear range is 400-4.00E+09 copies / ml; by applying the kit, rapid and accurate detection can be performed on EBV-DNA in the unknown samples of blood plasma, throat swab, peripheral blood and the like, and reliable experimental basis is provided for diagnosing EBV infection.

The invention provides a mycobacterium tuberculosis TB detection kit. The kit comprises a nucleic acid releasing agent and polymerase chain reaction (PCR) liquid, wherein the nucleic acid releasing agent comprises surfactin of 0.01-0.05 mM / L, potassium chloride of 20-300 mM / L, 0.01 to 2 percent of sodium dodecyl sulfate and 0.05 to 1 percent of ethanol; the PCR liquid comprises an upstream primer, a downstream primer and a probe; the upstream primer and the downstream primer are applied to amplification of target polynucleotide; and the probe is applied to detection of target polynucleotide. No significant difference exists between the detection result of the method for releasing nucleic acid by using the nucleic acid releasing agent in the kit and that of a boiling method. The nucleic acid is extracted by a strong protein denaturing agent, the shell protein structure of the pathogene is quickly damaged, and the nucleic acid of the pathogene is released, release and extraction of DNA can be finished without heating. According to the kit, the TB detection sensitivity can reach 400 copies / ml and the quantitative linear range is 400-4.00E+09 copies / ml. With the application of the kit, the TB-DNA of unknown samples such as sputum can be detected quickly and accurately, and reliable test basis is provided for diagnosis of TB infection.

The invention relates to a promoter replacement method for improving the yield of Bacillus subtilis surfactin, belonging to the biotechnology field. The DNA homologous integration technology is adopted for allowing a promoter Pspac to replace the promoter of a Bacillus subtilis fmbR surfactin synthase gene. The homologous sequence of 1081bp is obtained by amplification from an fmbR strain genome through the PCR method and connected to HindIII and BamH I restriction enzyme cutting sites of a plasmid pMUTIN4, a well constructed vector is converted into wild Bacillus subtilis fmbR, thereby realizing the replacement through the screening by an antibiotic culture medium and the molecular biology method. The method can directly improve an antimicrobial peptide production strain on genetics, thereby having potential application value in industry. The yield of the strain surfactin is improved by about 4.85 times, and the yield of the surfactin can be improved by about 10 times under the IPTG induction.