124results about How to "Optimize culture conditions" patented technology

The present invention provides methods and devices for the knowledge-based discovery and optimization of differences between cell types. In particular, the present invention provides visual servoing optical microscopy, as well as analysis methods. The present invention provides means for the close monitoring of hundreds of individual, living cells over time: quantification of dynamic physiological responses in multiple channels; real-time digital image segmentation and analysis; intelligent, repetitive computer-applied cell stress and cell stimulation; and the ability to return to the same field of cells for long-term studies and observation. The present invention further provides means to optimize culture conditions for specific subpopulations of cells.

The invention discloses a sinorhizobium meliloti strain capable of being applied to fermentation to generate vitamin B12, wherein the preservation number of the sinorhizobium meliloti strain is CGMCC No.9638. The invention also discloses a production and preparation method of vitamin 12 by the sinorhizobium meliloti strain in a fermentation manner. The method comprises a fermentation culture condition of the strain and an extraction method of the vitamin 12 in fermentation liquor; the method comprises the following steps: inoculating the strain on a solid culture medium into a triangular flask as a seed solution; inoculating the seed solution into a fermentation culture medium at 30 DEG C with 10% of inoculation amount; fermenting and cultivating at 200rpm for 200 hours, and then collecting the fermentation liquor; and adding a transforming agent to transform the fermentation liquor into cyano cobalt amine. The vitamin 12 is widely applied to the industries such as feeds, additives, medicines and cosmetics as a special factor in an adjusting organism growth process; and the requirement of the vitamin 12 rises year by year. The strain fermentation process is easy to control, so that the strain fermentation process is not affected by the factors such as seasons; the requirements on raw materials are simple; and fermentation production is developed into a main production mode.

The invention relates to a Wickerhamomyces anomalus strain for highly yielding ethyl acetate, and a culture method and an application thereof. The strain for highly yielding ethyl acetate is named asa Wickerhamomyces anomalus Y3604 strain, and is preserved in China General Microbiological Culture Collection Center on October 12, 2016 with the preservation number of CGMCC No.13103; and the 26S rDNA D1 / D2 sequence of the strain is similar to the 26S rDNA D1 / D2 sequences of multiple other Wickerhamomyces anomalus strains. The strain has good tolerance to glucose, ethanol and ethyl acetate, and has a wide range in the growth pH value and the temperature; and the Wickerhamomyces anomalus Y3604 strain has a high ethyl acetate yield, and a result of detection shows that the ethyl acetate yield of the strain can reach 19.17 g / L. The strain can be applied to the brewing industry of Chinese liquor, yellow rice wine and soy sauce, demanding for the ethyl acetate.

The present invention relates to the process of promoting haemtococcus pluvialis growth, raising its population density and controlling the synthesis and accumulation of astaxanthin as intracellular secondary catabolite in a photobiological reactor system. The process includes establishing a photobiological reactor system with main reactor made of transparent material, outside controllable lighting facility, speed regulating vane wheel stirrer, water rolling wheel, bacteria-free ventilating and oxygen unit, salty and fresh water nutritious liquid compounding and temperature controlling unit, and cleaning and sterilizing unit; altering environment conditions to induce haemtococcus pulvialis to multiply, raise population density and regulating synthesis, accumulation and collecting haemtococcus pluvialis with rich astaxanthin.

The invention discloses a method for propagating Millettia Speciosa germchit by cuttage, including first-phase preparation and later management: 1) during the first-phase preparation, Millettia Speciosa tresses are selected and cut into stem segments for spare use; river sand and coco bran are evenly mixed according to certain proportion to obtain a substrate for spare use; 2) during the later management, Millettia Speciosa stem segments are dipped with root stimulate at the base parts and then stuck into the substrate; the stem segments are watered to promote the close contact between the base parts thereof and the substrate; the stuck stem segments are shaded for culture, with the moisture of the substrate and air humidity being guaranteed; after taking root, the stem segments are fertilized with urea solution until the germchit grows up. The invention has the advantages of simple process flow and low manufacturing cost, and, effectively utilizes the scrap Millettia Speciosa tresses; by optimizing the root stimulate and the substrate formula, the invention improves the culture conditions and increases the survival rate of the stuck stem segments. The method has high feasibility, simple operation, low cost input and high survival rate and achieves the dual purpose of recycled utilization of resources and batch production of the germchit.

The invention belongs to the technical field of biology engineering and particularly discloses an actinomycete strain KN37 and an application thereof. Through microscopic observation and 16S rDNA sequence analysis, the condition that the separated actinomycete strain KN37 belongs to Streptomyces is identified, and the preservation number of the actinomycete strain KN37 is CGMCC No.13160. A fermentation broth of the strain KN37 has a better inhibition effect on multiple phytopathogens such as Botrytis cinerea, Alternaria solani, Fusarium oxysporum, Fulvia fulva and the like and can be used for preparing microbial agents for controlling crop diseases.

The invention relates to a preparation method and application of a marine fungi aspergillus terreus butyrolactone compound butyrolactone-I. The invention discloses application of the marine fungi aspergillus terreus butyrolactone compound butyrolactone-I as shown in a formula (I) to preparation of medicaments for resisting to peripheral and neurogenic inflammation and resisting to neurodegenerative diseases. The formula is shown in the description. The invention finds that besides a DPPH free radical, butyrolactone-I can also well remove an ABTS free radical and an OH free radical; the marinefungi aspergillus terreus butyrolactone compound butyrolactone-I has better oxidation resistance and inflammation resistance and has better neuroprotection activity; therefore, the marine fungi aspergillus terreus butyrolactone compound butyrolactone-I has a wide application prospect in the aspect of preparing the medicaments for resisting to peripheral and neurogenic inflammation and resisting toneurodegenerative diseases. Meanwhile, by the preparation method of the butyrolactone-I, which is provided by the invention, the butyrolactone-I can be successfully prepared; the method is simple andeasy to realize large-scale production; culture conditions can also be optimized by addition of an inducer, so that yield of the butyrolactone-I is greatly improved.

The invention relates to the technical field of solid fermentation, in particular to a solid fermentation method of lactarius deliciosus mycelium and an application of solid fermentation extractive thereof. The solid fermentation method of lactarius deliciosus mycelium comprises the steps of: performing solid fermentation on the lactarius deliciosus mycelium by a culture medium of the invention under certain culture conditions, wherein the certain culture conditions are as follows: the carbon-nitrogen ratio of the culture medium is 1:5 to 1:8, the temperature is 18-37 DEG C, the pH is 5-8, the day age of strain is 24-120 h, the inoculation amount is 10-30%, and the fermentation period is 2-7 d. The product obtained after the solid fermentation procedure of the solid fermentation method is extracted by such methods as ultramicro pulverization, enzyme treatment and microwave or ultrasonic, and the like so as to obtain an extractive containing polysaccharide and polypeptide. The extractive can be used as a food additive to be applied in the healthcare field and can be also used as a feed additive to be applied in the animal feed field. According to the invention, a proper dosage of plant Chinese medicinal herb additives can be added in the culture medium to improve the yield of the mycelium and the intracellular and extracellular polysaccharide and polypeptide so as to reinforce the pharmacological activity of the extractive.

The invention discloses a preparation method and applications of freeze-dried powder of acetic acid bacteria. The preparation method comprises following steps of: 1) inoculating the acetic acid bacteria into a liquid culture medium and culturing at 28-30 DEG C for 10-30 h to obtain a primary strain; 2) inoculating the primary strain into a fermentation culture medium, culturing at 28-33 DEG C for 10-30 h, and adding an aqueous sodium hydroxide solution having a concentration of 5-25 wt% during the fermentation process to obtain a fermentation liquid mixture; 3) filtering the fermentation liquid mixture or centrifuging and separating to obtain bacterial sludge; 4) uniformly mixing the bacterial sludge and a bacterial sludge protective agent to obtain a bacterial suspension; and 5) freeze-drying the bacterial suspension to obtain the freeze-dried powder of the acetic acid bacteria. After the freeze-dried powder of the acetic acid bacteria is mixed with various additives, alcoholism-relieving tablets and alcoholism-relieving beverages, which have good alcoholism-relieving effects, can be obtained through different processes. The preparation method is simple and the freeze-dried powder of the acetic acid bacteria is high in bacterium content and long in storage time.

The invention discloses a microfluidic device and method for optimal control on microbial fertilizer bacterium culture conditions. The device comprises a case body and a PC (personal computer) positioned outside the case body, wherein a gas pump, a 10-channel constant-pressure sampler, an objective table, a microcontroller and a three-stage microfluidic chip are fixedly arranged in the case body. A complete optimal control method of nutrient solution main component screening, main component concentration proportion optimization and sample introduction parameter and temperature optimization of the optimal proportion nutrient solution is adopted to implement the microfluid intelligent control, temperature control and intelligent monitoring double feedback and also implement microfluid intelligent control, external shape image automatic treatment and interior physiologic metabolite activity electrochemical detection double feedback, thereby directly controlling the multichannel sample introduction mode and sample introduction parameters to perform the microfluidic microbial fertilizer bacterium culture. The method can be used for culture condition optimal control on any microbial fertilizer bacterium under the condition of unknown nutrient solution components.

The invention relates to an asparagus extract bifidobacterium fermentation activity beverage and a preparation method thereof. An asparagus extract, brown sugar and stachyose serve as a culture medium; the following strains as shown in the specification are subjected to anaerobic fermentation to culture a bifidobacterium; after the anaerobic fermentation, a fermented probiotic liquid is subjected to sterile filling; and the bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and saccharomyces cerevisiae are freeze-dried bacteria bought from a culture collection center. The beverage supplements a specific bifidus factor and an exogenous bifidus factor in a plant in order to culture high-density bifidobacterium longum, increase the unit bacterial count of other lactic acid bacteria, conduct enzymolysis of small protein converting peptide in the culture medium, increase the content of amino acid essential for a human body, and improve the activity of asparagus juice, and a health care function of the whole fermentation liquid. The technology has the advantages that a strict anaerobic condition of the bifidobacterium is overcome, an expensive bifidobacterium fermentation device is removed, and the health care function of metabolic products of an active substance and a microorganism containing asparagus, as well as the fermentation liquid of the viable bacteria is enhanced.

The invention discloses a method for preparing a heat-resisting attenuated virus live vaccine for goatpox by using a BHK21-C13 passage cell. Based on the existing attenuated virus AV41 for goatpox, which has excellent immunogenicity, the method comprises the following steps: culturing 25-28th generations of virus solution with a heterologous BHK21 cell, and then adding an appropriate heat-resisting freeze-drying protective additive to obtain the heat-resisting attenuated virus live vaccine for goatpox. The passage cell vaccine used by the method is superior to the primary cell vaccine in the aspects of virus yield, homogeneity, purity and the like, not only guarantees the effective level of the vaccine and has a better heat-resisting protection effect, but also can ensure the safety of a purebred pregnant goat. As the existing BHK21 cell adopts a mature suspension culture process, the exploration of various parameters of adherent culture lays a solid foundation for the next promotion of suspension process.

The invention discloses a method for in vitro regeneration of a Feizixiao litchi variety. The method comprises the following steps: step 1, taking the anther of the Feizixiao litchi variety as an explant for callus induction and culture, and screening to obtain embryogenic callus; the culture medium is added with 2, 4-D with final concentration of 3 mg / l, BA with final concentration of 0.5 mg / l, NAA with final concentration of 0.5 mg / l and sugar with final concentration of 30 g / L on the basis of a culture medium MS; step 2, carrying out somatic embryo induction and culture to the embryogenic callus to obtain a somatic embryo; the culture medium is added with NAA with final concentration of 0.1 mg / l, LKT with final concentration of 5 mg / l, inositol with final concentration of 100 mg / l and sucrose with final concentration of 50 g / L on the basis of the culture medium MS; step 3, carrying out maturation to the somatic embryo to obtain a mature embryo, culture mediums are MS, 50 ml / L coconut milk and 60 g / L sucrose; and step 4, carrying out regeneration culture to the mature embryo, the culture medium is added with sucrose with final concentration of 30 g / L and agar with final concentration of 7 g / L on the culture medium MS. The invention establishes a method for in vitro plant regeneration of a Feizixiao litchi variety, achieves the high induction rate, improves the production efficiency, reduces the cost and lays a good foundation for litchi variety improvement and biotechnology breeding.

The invention discloses a culture method capable of maintaining the self-renewal state of mouse epiblast stem cells. The culture method includes: acquiring, performing passage on and culturing the mouse epiblast stem cells under culture conditions, and observing and detecting the self-renewal state of the cultured mouse epiblast stem cells. The culture method has the advantages that the problems that the state of cells cultured under traditional culture conditions is unstable, passage operation is inconvenient, and existing culture conditions are high in cost, complex in action factors and unfavorable for subsequent researches, and the like are solved; the method is economic, efficient, convenient to operate, stable in effect, beneficial to researches, and the like; the method can provide a clue for the improvement of the culture conditions of current human embryonic stem cells and other kinds of pluripotent stem cells and plays an important role in the promotion of the smooth development of stem cell foundation and application researches.

The invention relates to a liquid strain culturing method and culturing method of boletus aereus. The method comprises the steps that liquid strains of boletus aereus are inoculated to a fermentationtank, and after six stages of culturing of strain mycelium breaking, mycelium restoring, culturing adapting germinating, mycelium growing and mycelium exponential growing period are conducted, fermentation is completed. According to the method, different culturing conditions are made aiming at the growing characteristics of germ in different stages, and the boletus aereus is adapted to growth in the fermentation tank to obtain higher growing rate; culturing conditions of the boletus aereus liquid strains in the fermentation tank are studied, mechanical inoculation feasibility is provided, andthe method has higher production efficiency compared with an existing triangular flask culturing method and has a lower pollution rate compared with a solid grain culturing method.

The invention relates to an asparagus extract bifidobacterium fermentation activity beverage and a preparation method thereof. An asparagus extract, brown sugar and stachyose serve as a culture medium; the following strains as shown in the specification are subjected to anaerobic fermentation to culture a bifidobacterium; after the anaerobic fermentation, a fermented probiotic liquid is subjected to sterile filling; and the bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and saccharomyces cerevisiae are freeze-dried bacteria bought from a culture collection center. The beverage supplements a specific bifidus factor and an exogenous bifidus factor in a plant in order to culture high-density bifidobacterium longum, increase the unit bacterial count of other lactic acid bacteria, conduct enzymolysis of small protein converting peptide in the culture medium, increase the content of amino acid essential for a human body, and improve the activity of asparagus juice, and a health care function of the whole fermentation liquid. The technology has the advantages that a strict anaerobic condition of the bifidobacterium is overcome, an expensive bifidobacterium fermentation device is removed, and the health care function of metabolic products of an active substance and a microorganism containing asparagus, as well as the fermentation liquid of the viable bacteria is enhanced.

The invention relates to a CYP101 enzyme recombinant vector, a construction method thereof and a CYP101 enzyme high-efficiency expression and purification method, and belongs to the field of biotechnology. A PET28a-CYP101 high-efficiency expression vector is constructed and transformed into BL21 plysS strain, and high-purity enzyme with purity higher than 95% is obtained by a simple two-step purification method including affinity chromatography and ion exchange chromatography after inducible expression. Compared with the prior art, CYP101 is simply and efficiently produced, the cost is low, purification is facilitated, and 23mg of target protein with the purity higher than 95% can be obtained by per liter of culture media. The CYP101 enzyme high-efficiency expression and purification method can be used for expressing and purifying CYP101 enzyme, the production efficiency of a laboratory is greatly improved, and experimental cost is greatly reduced. The CYP101 enzyme high-efficiency expression and purification method has an excellent application prospect for large-scale CYP101 enzyme production and purification.

The invention relates to the fields of gene engineering and biosynthesis, and concretely relates to a construction body for efficiently synthesizing ethylene, and a construction method and an application thereof. The construction body comprises an active promoter and an ethylene synthetase gene (efe) controlled by the promoter. Carbon dioxide can be immobilized by above engineering bacterium through using solar energy in order to synthesize ethylene. An ethylene synthesis approach is constructed in cyanobacterial cells in order to realize immobilization of carbon dioxide in photosynthetic microbal cyanobacterial cells through using solar energy and synthesis of ethylene, wherein energy for ethylene synthesis is from the solar energy, and a carbon source is carbon dioxide. The method can be used to prepare bioethylene without restriction of petroleum shortage, and the bioethylene does not increase carbon emission.

The invention relates to a method used for co-culturing of ethanol high yield Saccharomyces cerevisiae YF1914 and an ethyl acetate high yield Wickerhamomyces anomalus YF1503 bacterial strain, and applications thereof. The Saccharomyces cerevisiae YF1914 is preserved in China General Microbiological Culture Collection Center on Oct. 20th 2017, and the preservation number is CGMCC No.14827; the sequence similarity of 26S rDNA D1 / D2 sequence of the Saccharomyces cerevisiae YF1914 and 26S rDNA D1 / D2 sequence of a plurality of other Saccharomyces cerevisiae bacterial strains is relatively high; ina sorghum enzymatic hydrolysate culture medium, co-culturing of the two yeast bacterial strains is benefical for increasing of ethyl acetate content. The double bacterial strain co-culturing method can be applied in brewage industry such as baijiu, yellow rice wine, and soy sauce with requirements on ethyl acetate.

The asymmetrical resolution method for (R)-mandelic acid comprises: screening the (Brevibacterium flavum) AS 1.818 for culture and full cell preparation; with racemic acid as substrate, catalytic converting to obtain the product with optical purity up to 90%e.e. This invention improves product optical purity and has important meaning for enzyme development and resolution.

The invention discloses a piriformospora indica fermentation method. The method comprises the following steps: by taking a PDB culture medium as a culture solution, regulating the pH of the culture solution to be 5.0-5.1 by using a HCl solution with the concentration of 1mol / L, and performing culture fermentation on piriformospora indica at the temperature of 27.5-28.5 DEG C for 14-15 days, so as to obtain the fermentation liquor. The invention also discloses a method for performing response surface optimization in the piriformospora indica fermentation process. The method comprises the following steps: setting the three variables including the pH, the concentration of the PDB culture medium and the culture time; optimizing according to the Box-Behnken design principle by utilizing design expert software. According to the method, the yield of secondary metabolites can be increased, and the consumption of nutritional ingredients and culture media is reduced.

The asymmetrical reduction bio-method for (R)-mandelic acid comprises: screening the Saccharomyces cerevisiae AS 2.150 for culture and full cell preparation; with benzoylformic aicd as substrate, adding assist substrate, catalytic converting to obtain the product with optical purity up to 90%e.e. This invention improves product optical purity and yield, and has important meaning for enzyme development and resolution.

The invention relates to aeribacillus pallidus Jh-7 antagonizing xanthomonas campestris pv. oryzae. The invention aims at providing the aeribacillus pallidus Jh-7 antagonizing the xanthomonas campestris pv. oryzae firstly. The aeribacillus pallidus contains an ingredient for inhibiting the xanthomonas campestris pv. oryzae, and a good approach is provided for biological control of rice diseases caused by the xanthomonas campestris pv. oryzae. The invention aims at providing a metabolism crude extract of the aeribacillus pallidus Jh-7 secondly. The invention aims at providing an application ofthe metabolism crude extract of the aeribacillus pallidus Jh-7 thirdly. The invention aims at providing a biological pesticide containing the aeribacillus pallidus Jh-7 fourthly.

The invention discloses a pepper CaCOI1.2 gene. An open reading frame sequence is composed of the 186th to 1997th nucleotide of the SEQ (sequence) ID NO.1. The invention further discloses a recombinant expression vector containing the pepper CaCOI1.2 gene, a transformant, an application of the pepper CaCOI1.2 gene to large-fruit pepper plant breeding and a specific application method. Research results show that an over-expression vector containing the pepper CaCOI1.2 gene is introduced into a pepper of new Su Jiao Wang strain through agrobacterium mediates, and pepper fruits of the obtained transgenic positive plants are bigger than those of non-transgenic pepper plants, so that the yield of the pepper is greatly increased; the CaCOI1.2 gene is a self gene of the pepper, thereby the transgenic plants obtained by introducing the CaCOI1.2 gene into pepper plants have no influence on the environment safety and food safety; and the pepper transgenic method is high in transduction efficiency.

The invention relates to the technical field of fermentation industry, in particular to a preparation method of selenium-rich yeast. A soil sample from Bama County, Guangxi is selected, the content ofselenium in Bama soil and grains is 10 times or more higher than the national average, a selenium-rich yeast strain is obtained through screening and breeding of yeast strains in the soil sample, theselenium ion concentration and the culture condition are optimized, and the selenium content of the yeast is increased. The selenium-rich yeast obtained through separation and screening of the selenium-containing soil is subjected to domestication with sodium selenite with gradient concentrations, yeast colonies with high selenium resistance are screened, then, ultraviolet-induced mutation is performed, the selenium enrichment capability of the yeast is enhanced, and the yeast with high biomass and selenium enrichment capability is obtained.

The invention belongs to the technical field of microorganisms, and relates to marine-derived Aspergillus terreus M7 with an antibacterial effect, and separation and application of a secondary metabolite of the Aspergillus terreus M7. The strain is classified and named as Aspergillus terreus M7, and is preserved in China Center for Type Culture Collection (CCTCC) on June 7, 2021, with the preservation number of CCTCC NO: M 2021679. The strain is fermented by a soybean solid culture medium, and is subjected to methanol extraction, silica gel G chromatography, Sephadex LH-20 gel column chromatography, silica gel H chromatography and preparative liquid phase separation and purification to obtain a compound terramide A. The terramide A has obvious antibacterial activity on acinetobacter baumannii ATCC 19606, and has the potential of being developed into an antibacterial agent.

The invention relates to a one-step seedling and efficient in-vitro propagation method for Begoniaceae gynura bicolor, which is an endangered plant with homology of medicine and food. The method provided by the invention comprises the following steps of: preparing a culture medium, and adjusting the culture medium through preparing, subpackaging and sterilizing; carrying out sterile inoculating directly on gynura bicolor aseptic seedling leaves which are taken as explants; culturing the inoculated culture medium in a culture chamber for 70-80 days under appropriate conditions, wherein the number of a single explant induced seedlings is 20 above; and decapping, transplanting regenerating plants to appropriate conditions, and culturing without hardening-seedling, wherein the commodity seedling survival rate achieves 100%. According to the method provided by the invention, the gynura bicolor tissue culture seedling regeneration period is short; the original seed property is kept effectively; the vegetative propagation coefficient is improved obviously; the operation is simple and convenient; the culture program is simple; the seedling cost is lowered obviously; and the method can act as an effective technology of industrialized production of high-quality seedlings of Begoniaceae gynura bicolor.

The invention discloses a camellia changii variety breeding method which comprises the following steps of selecting budding flowers by taking fragrant flowers with rich flower colors and flower types as breeding aims in a camellia changii male parent blooming season, poking petals, quickly removing all stamens, smearing pollens to be pollinated on stigmas, folding an adhesive plaster in two to completely cover the stigmas for controlled pollination to realize the controlled pollination of theaceae plants, and performing seed breeding of hybrids under an indoor control condition. The variety of camellia changii is small, the number of female parents is small, and the breeding potential is low. By a tissue culture technology disclosed by the invention, the problems can be well solved; rare and precious resources, namely the camellia changii, can be protectively used; the problem that the camellia changii seedlings are scarce can be alleviated.