62 results about "Nannochloropsis oculata" patented technology

The invention relates to genetic engineering transformation of nannochloropsis oculata and particularly relates to a method for carrying out genetic transformation by using Cas9/CRISPR at a fixed point. A compound is formed by overexpressed Cas9 protein in the nannochloropsis oculata and gRNA, the Cas9 protein is used for carrying out site-specific cleavage on a genome double-stranded DNA under the guidance of the gRNA to cause gene mutation, and mutation is effectively screened by combining a sequencing technique. Fixed-point genetic engineering transformation is carried out on the nannochloropsis oculata by using a CRISPR/Cas technology, so that the algae species improvement efficiency of the nannochloropsis oculata can be significantly improved.

The invention belongs to the field of microbial fermentation engineering, and particularly relates to a technology of consolidated bioprocessing (consolidated bioprocessing, CBP). Cellulose is fermented and degraded by a cellulose degradation strain (such as clostridium thermal fiber) to generate saccharides and organic acids, such as glucose, lactic acid and acetic acid; and biodiesel and co-produced products with high additional values, such as timnodonic acid, docosahexenoic acid and protein feed, are produced by serial fermentation combined with large-scale heterotrophism or raising cultivation of microalgae (such as schizochytrium limacinum, chlorella and nannochloropsis oculata). The method has the characteristics that anaerobic cellulose degradation bacteria are utilized to degrade the preprocessed lignocellulose material, so that the cellulose and hemicelluloses are converted into carbon sources which can be utilized by the microalgae such as soluble saccharide and organic acids; the carbon sources are inoculated into a microalgae seed liquid to be fermented after a certain nitrogen source and other nutrient elements are added, and oil is accumulated to generate the biodiesel and other products with high additional values.

The invention provides a culture method for efficiently inducing accumulation of nannochloropsis oculata fat. The culture method is as follows: preparing a culture medium and daily-added nutrient liquid; carrying out biomass accumulation culture on annochloropsis oculata for 5-8 days; introducing CO2 in annochloropsis oculata culture liquid, and regulating the concentration of the daily added nutrient liquid; and carrying out ultrasonic processing on the annochloropsis oculata culture liquid one to three times at each interval of 10 seconds and for 5-60 seconds per time by using an ultrasonic instrument under a certain frequency, so that the accumulation of the nannochloropsis oculata fat is induced. The biomass of the nannochloropsis oculata cultured by the method is 1.5-2 times as much as that of the prior art, the content of oil in the nannochloropsis oculata cultured by the method is improved by 40%-65% compared with the prior art, and the growing period of the nannochloropsis oculata cultured by the method is shortened by 2-3 days; and in the invention, carbon source is supplemented in a form of introducing CO2, and a large amount of CO2 waste gases generated in the industrial production process are absorbed in the process of the large-scale production of the nannochloropsis oculata for inducing the accumulation of fat, thus the culture method has an important significance for energy conservation and emission reduction as well as environment protection.

The invention provides a formula of a nannochloropsis oculata medium and a three-stage cultivation method. The formula is as follows: 7.5*10 <-2>g of NaNO3, 1*10 <-2>g of KH2PO4, 5*10<-1>g of NaHCO3, 2*10 <-4>g of ferric citrate (FeC6H5O7.5H2O), 5*10 <-7>g of VB12, 1*10<-4>g of VB1, 5*10 <-7>g of VH, 5 IU of 920 plant growth hormone (namely gibberellin), 2mL of human urine, and 1000mL of disinfected seawater. The experimental results show that the optimized medium and three-stage cultivation method for culturing nannochloropsis oculata have the advantages of fast growth, high yield, low cost, easy operation and little pollution.

The invention relates to a method for controlling microbe indexes in microalgae powder by combination of pasteurization and spray drying and a method for producing microalgae powder. The method comprises the following steps: pasteurizing a microalgae concentrated solution to obtain a sterilized concentrated solution, and drying the sterilized concentrated solution to obtain microalgae powder, thereby controlling the microbe indexes in the microalgae powder (including but not limited to total bacterial count, mold quantity and yeast quantity); and producing the microalgae powder. The method can effectively control the microbe indexes (especially total bacterial count) of the algae powder, and can maximally maintain the activity of the active substances in the microalgae. The method is suitable for but not limited to producing Chlorella, Haematococcus pluvialis, spirulina, Dunaliella, Nannochloropsis oculata, Schizochytrium, Crypthecodinium cohnii and various algae powders, and is hopeful to become a universal production method capable of effectively controlling microbe indexes (especially total bacterial count) in the algae powder.

A process for accelerated and sustainable algae growth using a vertical tank photobioreactor, which includes steps involving the sustainable algae growth, preferably of the strain nannochloropsis oculata, the harvest of a portion of the algae in a harvester apparatus, the reduction and recycling of byproducts using recycled waste and conversion of gasses to useable and sustainable recycling within the process, the algae product supplied in wet or dry form for the further production of biofuels through the conversion of the algae to crude oil.

The invention discloses a method for extracting EPA from nannochloropsis oculata by a compound enzyme method. The method comprises the steps: subjecting nannochloropsis oculata fermentation liquor tocentrifugation, and collecting nannochloropsis oculata mud; adding a certain volume of buffer solution into the nannochloropsis oculata mud, then, adding a compound enzyme, and carrying out a reaction, so as to obtain an enzymolysis solution; subjecting the enzymolysis solution to centrifugation so as to obtain first free oil and an emulsion, subjecting the emulsion to further demulsified centrifugation so as to obtain second free oil, merging the first free oil and the second free oil, so as to obtain crude oil; and further purifying the obtained crude oil through a supercritical CO2 purification method, thereby obtaining the EPA. According to the method, the nannochloropsis oculata is treated by the compound enzyme, cell structures of microalgae are subjected to enzymolysis destruction under certain reaction conditions, meanwhile, the degradation action on complexes such as lipopolysaccharides and lipoproteins is strengthened, the fluidity of fat and oil is improved, and the fat andoil can be liberated. According to the method, the extraction purity of the EPA is 95% or more.

The invention relates to a method for controlling microbe indexes in microalgae powder by combination of microwave sterilization and spray drying and a method for producing microalgae powder. The method comprises the following steps: carrying out microwave sterilization on a microalgae concentrated solution to obtain a sterilized concentrated solution, and drying the sterilized concentrated solution to obtain microalgae powder, thereby controlling the microbe indexes in the microalgae powder (including but not limited to total bacterial count, mold quantity and yeast quantity); and producing the microalgae powder. The method can effectively control the microbe indexes (especially total bacterial count) of the algae powder, and can maximally maintain the biological activity of the active substances in the microalgae. The method is suitable for but not limited to producing Chlorella, Haematococcus pluvialis, spirulina, Dunaliella, Nannochloropsis oculata, Schizochytrium, Crypthecodinium cohnii and various algae powders, and is hopeful to become a universal production method capable of effectively controlling microbe indexes (especially total bacterial count) in the algae powder.

The invention discloses a wall-breaking and surfactant-assisted method used for extracting grease from microalgae containing oil. The wall-breaking and surfactant-assisted method comprises following steps: (1) pH value of a suspension liquid of microalgae containing oil is adjusted to be 7.5 to 14, and is subjected to hydro-thermal treatment; (2) after hydro-thermal treatment, enzymatic hydrolysis is carried out in the presence of enzymes; and (3) after enzymatic hydrolysis, a surfactant is added for reaction, and an organic solvent is added for extraction and separation of grease. Grease extraction rate of the wall-breaking and surfactant-assisted method is high; operation conditions are mild; nutritional ingredients of microalgae can be maintained effectively; using amount of the organic solvent is small; the wall-breaking and surfactant-assisted method is friendly to the environment, is low in energy consumption, and is suitable for nannochloropsis oculata, chlorella, diatom, scenedesmus, and monoraphidium komarkova-legnerova; and application range is wide.

The invention relates to the technical field of biological gene engineering, and particularly relates to a method for importing exogenous DNA (deoxyribonucleic acid) into nannochloropsis oculata chloroplast and a related chloroplast genome sequence. The gene sequence comprises a sequence shown by SEQ ID No.1, a sequence shown by SEQ ID No.2 and an exogenous gene inserted between the two sequences. The method comprises the concrete step of importing a chloroplast transformation vector comprising the sequence shown by SEQ ID No.1, the sequence shown by SEQ ID No.2 and the exogenous gene inserted between the two sequences into nannochloropsis oculata cells by an electroporation process. The method provided by the invention can be used for designing and establishing a chloroplast genetic modification system universal in nannochloropsis oculata and a chloroplast genetic modification system universal in multiple species of unicellular microalgae according to the conserved genes among the genomes of seven nannochloropsis oculata chloroplasts and the conserved genes of the nannochloropsis oculata chloroplast and diatom or brown algae chloroplast genome.

The invention relates to the technical field of aquaculture fry breeding, in particular to a special indoor artemia breeding method for seawater fish and shrimp fry breeding. The method specifically comprises the following steps: step 1, indoor artemia breeding; step 2, bait feeding; and step 3, nutrient enrichment. The indoor artemia breeding method is established by optimizing the culture method, the breeding yield is stable, the survival rate is high, in addition, the cultured artemia is subjected to nutrient enrichment through schizochytrium limacinum and nannochloropsis oculata, the nutritive value of artemia is increased, especially the unsaturated fatty acid content is increased, and therefore, high-quality biological bait is provided for aquatic fry breeding; and the method can beapplied to large aquatic fry breeding fields and can also be used for commercial production.

The invention discloses a puffing feed using marine microalgae as main raw materials, for takifugu obscurus, and a making method of the puffing feed, and belongs to the field of feeds. According to the puffing feed and the making method thereof disclosed by the invention, the nutritional requirements of the takifugu obscurus are analyzed in advance, nannochloropsis oculata powder, chlorella powder, and spirulina powder in the feed are rich in EPA and DHA, which are favorable for the health and the rapid growth of the takifugu obscurus, the compounding ratio is scientific, the nutrition is balanced, the nutritional requirements of the takifugu obscurus can be met, and the feed can promote the food intake and the growth of the takifugu obscurus; the feed is rich in fragrance, and good in a food-calling effect, the sizes of particles of the feed conform to the foraging requirement of the takifugu obscurus, the quality guarantee period is long, and the immunity of fish can also be improved; the feed is high in ageing degree, and good in water resistance; few feeds are wasted, the feed coefficient is low, pollution to water is small, the use is convenient, and the feed is convenient for fish breeding in a scaled, industrialized and mechanized manner.

The invention discloses a special compound feed for industrial aquaculture of penaeus monodon. The special compound feed comprises the following components in percentage by weight: 20%-25% of fish meal, 5%-8% of dephenolized cottonseed protein, 6%-10% of artemia powder, 6%-6.7% of degreased silkworm chrysalis meal, 6%-12% of scallop skirt powder, 3%-6% of enzymolysis fish soluble pulp powder, 1%-3% of dried porcine soluble powder, 2%-5% of superfine enteromorpha powder, 3%-6% of schizochytrium limacinum powder, 3%-6% of nannochloropsis oculata powder, 2%-5% of marine red yeast dry powder and the balance of water, 2%-3% of shrimp shell powder, 1%-3% of kelp powder, 7%-10% of flour, 3%-5% of peas, 1.5%-3% of lysolecithin, 1%-1.6% of fish oil, 0.2%-0.4% of taurine, 2%-3.5% of monocalcium phosphate, 0.8075%-1% of vitamin premix and 1.0925%-1.4% of mineral premix. The special compound feed for industrial aquaculture of the penaeus monodon is high in nutritional value, easy to digest, good in curing degree and high in stability, the growth performance, the anti-stress capability, the oxidation resistance and the body color of the penaeus monodon are remarkably improved, and nitrogen and phosphorus emission is reduced.

The invention relates to a nannochloropsis oculata mutant strain and applications thereof. The nannochloropsis oculata mutant strain is taxonomically named Nannochloropsis Sp. The nannochloropsis oculata mutant strain has the advantages of stable properties, strong environment adaptability, and strong resistance to pesticide and a high temperature.

The invention discloses a compound Sargassum, fruit and vegetable beverage capable of stimulating children's intelligence development and a preparation method of the compound Sargassum, fruit and vegetable beverage and belongs to the field of beverage production. The compound Sargassum, fruit and vegetable beverage is prepared from Dunaliella Salina powder, Isochrysis Galbana powder, Nannochloropsis Oculata powder, scallop juice, Concha Ostreae juice, grape juice, pineapple juice, kiwi-fruit juice, tomato juice, orange juice, Spinacia Oleracea juice, carrot juice and distilled water according to a scientific technology. The compound Sargassum, fruit and vegetable beverage capable of stimulating children's intelligence development is scientific and reasonable in formula, available to raw materials, simple in technology, low in cost, green, natural, unique and harmonized in flavor, mild in taste, free from foreign smell, clear, transparent, cool, refreshing, good in fluidity and natural in faint scent, and is capable of tonifying the spleen, reinforcing the stomach, nourishing the heart, moistening the lung and nourishing the liver and kidney, thereby being suitable for people of all ages and particularly helping stimulate brain development and enhance intelligence of children.

The invention discloses a method for excavating and selecting anabolic markers of microalgae oil. In a nitrogen deficiency induction condition, three kinds of algae with extremely poor affinis, including freshwater chlorella, seawater nannochloropsis oculata, and polar coccomyxaceae, are used as subjects, and based on cytomics, metabonomics, andbioinformatics methods, a same anabolic marker synthesized and accumulated by the three kinds of algae cell oil with extremely poor affinis is excavated and selected. Using complementary analysis among different algaes and different omics, a result one-sidedness caused by using a single algae and a single analysis method can be effectively prevented, so as to greatly increase selection identification rate of a key anabolic marker synthesized and accumulated by the microalgae oil and corresponding precision. The method provides theoretical basis for further improving yield of the microalgae oil and revealing an anabolism general mechanism of themicroalgae oil.

The invention discloses a method for simultaneously extracting a nannochloropsis oculata protein and dietary fibers by ultrasonic assistance, and relates to the technical field of extraction of a protein and dietary fibers. The method comprises the steps of degreasing, wall breaking, double-enzyme enzymolysis, enzyme deactivation, solid-liquid separation, protein extraction, soluble dietary fiber separation, heavy metal removal and the like. The method disclosed by the invention is simple to operate, high in protein and dietary fiber extraction rate and low in heavy metal content, and three products including the nannochloropsis oculata protein, the nannochloropsis oculata soluble dietary fiber and the nannochloropsis oculata insoluble dietary fiber are simultaneously obtained in one technological process.

The invention belongs to the technical field of microalgae polysaccharide preparation, and particularly relates to a preparation method of nannochloropsis oculata polysaccharide with antioxidant activity. An ultrasonic-microwave-enzyme method is used for synergistically extracting the polysaccharide in the nannochloropsis oculata, and during extraction, the cavitation crushing effect of ultrasonic waves on nannochloropsis oculata powder, the high-energy effect of microwaves on the nannochloropsis oculata powder and the efficient and specific property of enzymes on cell walls of the nannochloropsis oculata powder are fully combined, so the dissolution of the polysaccharide in the nannochloropsis oculata powder is increased, and the extraction rate of the nannochloropsis oculata polysaccharide is greatly improved; the extracted nannochloropsis oculata polysaccharide is subjected to alpha-amylase modification to change the structure of the polysaccharide, so the groups and the structure of the polysaccharide subjected to shear modification on the space structure of the polysaccharide are changed, the physicochemical properties of the polysaccharide can be changed to different degrees, and the biological activity of the polysaccharide is improved; the free radical scavenging rate (-Sa/%) of the modified nannochloropsis oculata polysaccharide DPPH can be increased to 78.9% from 61.5% of the modified nannochloropsis oculata polysaccharide DPPH, and the increasing rate is 28.29%.

The invention discloses a method for promoting propagation of bait microalgae. The invention provides a method for expanding propagation of bait microalgae. The highest propagation density of the baitmicroalgae is increased and the platform period maintaining time of the bait microalgae is prolonged by adding an algal beneficial bacterium separated from a pseudominiature sea chain algae solutionand a nannochloropsis oculata algae solution. The phycomycetes beneficial bacteria are bacillus marinus with the preservation number of CCTCC M 20200448. According to the method disclosed by the invention, the propagation speed of the nannochloropsis oculata and the nannochloropsis oculata is much higher than the microalgae culture speed of a conventional microalgae nutrient solution, the highestpropagation density and the plateau period maintenance time are much higher than those of a conventional culture method, more microalgae can be supplied under the same water body condition, and meanwhile, the plateau period is relatively often not easy to decay, so the microalgae quantity control and the production management in the actual shellfish large-scale production process are very facilitated.

The invention belongs to the technical field of artificial breeding of coral, and particularly relates to a preparation method of composite liquid bait for artificial breeding of coral. Specifically, the composite liquid bait is prepared through the following steps of by using shrimp meat, mussel sexual gland and fish oil as raw materials, adding compound protein enzymes, and performing enzymolysis to prepare enzymolysis supernate and fish oil mixed liquor for standby application; by using ulva lactuca liquid and nannochloropsis oculata as raw materials, preparing nannnochloropsis gaditana and ulva lactuca mixed liquor for standby application; enabling saccharomyces cerevisiae, candida utilis seed liquid and rhodospirillum rubrum seed liquid to be made into mixed bacterium seed liquid for standby application; adding complex vitamins to all of the liquid for standby application so as to obtain the composite liquid bait for artificial breeding of coral. The composite liquid bait for artificial breeding of coral, prepared by the method disclosed by the invention, is comprehensive in nutrient components and good in balance, is harmless, and has the efficacy of purifying water quality, and the survival rate and the growth rate of the artificially bred coral can be increased.

The invention relates to the technical field of fertilizers for tea leaves, in particular to a microalgae fertilizer suitable for northern tea leaves, the microalgae fertilizer comprises the following components in parts by weight: 5-10 parts of medium trace element fertilizers, 3-5 parts of nannochloropsis oculata liquid, 2-5 parts of trichoderma viride and 50-80 parts of tea residues. According to the microalgae fertilizer, the nannochloropsis oculata solution, the trichoderma viride and the tea residues are jointly used, so that soil can be effectively improved, and the yield of tea leaves can be increased; the nannochloropsis oculata solution has a nitrogen fixation effect, a biological membrane can be formed after the trichoderma viride and the nannochloropsis oculata solution are mixed, tea leaf effective components such as tea polyphenol contained in the tea residues are acidic substances, the pH of soil can be reduced, media are provided for the nannochloropsis oculata solution and the trichoderma viride, and the promotion effect is achieved; the medium trace element fertilizer is matched so that the fertilizer efficiency is improved, and the soil quality is improved.

The invention provides an extraction process of nannochloropsis oculata essential oil. The extraction process comprises the steps of pectinase treatment, thallus collection, juice extraction and distillation. In the pectinase treatment process, the addition amount of pectinase is 3.5-4.3 u/g, the enzymolysis temperature is 35-40 DEG C, and the condition is maintained for 4-5 hours. The extraction process has the advantages of being easy to operate, low in cost and suitable for wide popularization. The essential oil obtained by the extraction process has the advantages of no fishy smell and light and pleasant smell.