118 results about "Mortierella alpina" patented technology

The present invention relates to a process for preparing a biomass, such as from a microbial fermentation, for an extraction process to separate desired chemicals, nutritional products, bioactive components, proteins, carbohydrates, and lipids, from the biomass. Particularly preferred substances to extract include docosahexaenoic acid, docosapentaenoic acid, and arachidonic acid. The present invention also includes extracting the prepared biomass. Biomasses to be treated in accordance with the methods of the invention include plant, animal, and microbial biomass, particularly a microorganism such as Crypthecodinium cohnii and a fungus such as Mortierella alpina.

An acyltransferase is provided, suitable for use in the manufacture of microbial oils enriched in omega fatty acids in oleaginous organisms. Specifically, the gene encoding diacylglycerol acyltransferase (DGAT2) has been isolated from Mortierella alpina. This gene encodes an enzyme that participates in the terminal step in oil biosynthesis in fungi and yeast and is expected to play a key role in altering the quantity of long-chain polyunsaturated fatty acids produced in oils of oleaginous organisms. Most desirably, the substrate specificity of the instant DGAT2 will be particularly useful to enable accumulation of long-chain PUFAs having chain lengths equal to or greater than C20 in oleaginous yeast, such as Yarrowia lipolytica.

The present invention relates to a fungal C16 / 18 fatty acid elongase that is able to catalyze the conversion of palmitate (16:0) to stearic acid (18:0). Specifically, the nucleotide sequence of a Mortierella alpina C16 / 18 fatty acid elongase is provided (designated as “ELO3”). Methods of increasing microbial oil production, increasing carbon flux into the polyunsaturated fatty acid biosynthetic pathway and increasing the content of polyunsaturated fatty acids by over-expression of the C16 / 18 fatty acid elongase are described herein. Most desirably, the substrate specificity of the instant ELO3 will be particularly useful to enable accumulation of long-chain polyunsaturated fatty acids in oleaginous yeast, such as Yarrowia lipolytica.

Glycerol-3-phosphate o-acyltransferase (GPAT) participates in the first step of oil biosynthesis and is expected to play a key role in altering the quantity of long-chain polyunsaturated fatty acids (PUFAs) produced in oils of oleaginous organisms. The present application provides a nucleic acid fragment isolated from Mortierella alpina encoding a GPAT that is suitable for use in the manufacture of oils enriched in omega fatty acids in oleaginous organisms. Most desirably, the substrate specificity of the instant GPAT will be particularly useful to enable accumulation of long-chain PUFAs having chain lengths equal to or greater than C20 in oleaginous yeast, such as Yarrowia lipolytica.

The present invention relates to a fungal C16 / 18 fatty acid elongase that is able to catalyze the conversion of palmitate (16:0) to stearic acid (18:0). Specifically, the nucleotide sequence of a Mortierella alpina C16 / 18 fatty acid elongase is provided (designated as “ELO3”). Methods of increasing microbial oil production, increasing carbon flux into the polyunsaturated fatty acid biosynthetic pathway and increasing the content of polyunsaturated fatty acids by over-expression of the C16 / 18 fatty acid elongase are described herein. Most desirably, the substrate specificity of the instant ELO3 will be particularly useful to enable accumulation of long-chain polyunsaturated fatty acids in oleaginous yeast, such as Yarrowia lipolytica.

A process for preparing arachidonic acid oil by microbial fermenting includes culturing and activating Mortierella alpina (M0223), culturing seed, inoculating the seed in shake flask or fermentor, fermenting, collecting wet thallus, baking and extracting oil. Its advantages are high purity and safety.

The invention relates to a method for producing arachidonic acid oil by Mortierella alpina fermentation on the basis of dissolved oxygen control. The method comprises the following steps: carrying out progressive amplification culture on the Mortierella alpina strain in a test tube slant culture medium and a shake culture medium to prepare a seed solution, and inoculating in a fermentation tank to perform fermentation, wherein the whole fermentation process is divided into an early fermentation stage, a medium fermentation stage and a later fermentation stage. The initial rotation speed is set at 0 rpm, and the initial dissolved oxygen is set at 100%; in the early fermentation stage, the dissolved oxygen is lowered and stable at 40-70%; in the medium fermentation stage, the dissolved oxygen is controlled at 20-40%; and in the later fermentation stage, the dissolved oxygen is controlled at 40-70%. After the fermentation is finished, the Mortierella alpina cell dry weight, oil content, percent of arachidonic acid in total oil, and unit yield of arachidonic acid can respectively reach 55g / L, 54%, 65% and 20.628g / L, and the arachidonic acid production intensity is up to 2.292g / (L*d).

The invention relates to a method for producing arachidonic acid grease by utilizing a mortierella alpina mutant strain, and the arachidonic acid grease produced by the method. The method comprises the steps that the mortierella alpina mutant strain is fermented; a fermentation product is collected; aftertreatment is performed; the arachidonic acid grease is obtained. The mortierella alpina mutant strain (mortierella alpina ZR36) is preserved in China Center For Type Culture Collection (CCTCC) on September 13, 2013; the preservation address is Wuhan University, Wuhan City, China; the preservation number is CCTCC NO:M2013419. The total content of saturated fatty acid, particularly above eicosanoic long-chain saturated fatty acid in the arachidonic acid grease produced by the mortierella alpina mutant strain is relatively low (lower than 15wt%); a freezing point of the arachidonic acid grease is relatively low (low to 4 DEG C); the arachidonic acid grease cannot be frozen or crystallized even if at a lower temperature; saturated fatty acid cannot be separated out; the arachidonic acid grease can keep clear and transparent, and has higher quality.

The present invention relates to a microbial agent for treating heavy metal polluted soil, a preparation method and applications thereof. The microbial agent comprises the following raw materials: rice husk, pig manure, chicken manure, soybean powder, single cell protein, a Mortierella alpina fermentation broth, a Trichoderma asperellum fermentation broth, and a Mucor circinelloides fermentation broth. According to the present invention, the microbial agent prepared by mixing Mortierella alpina, Trichoderma asperellum and Mucor circinelloides according to a certain ratio is firstly used, can be used in the complex and changing environment, and can provide effects in the poor soil environment, wherein the functional microbes provide high tolerance to heavy metals, and can maintain activity and perform efficient repair in the heavily heavy metal polluted soil.

The invention belongs to the field of fermentation engineering, relates to a method for producing ARA (arachidonic acid) through microbial fermentation, and particularly relates to a method for producing compound fat containing the ARA through industrial fermentation of a mortierella alpine strain. More particularly, the invention relates to a method for culturing microorganisms for producing the ARA, wherein the method for culturing the microorganisms for producing the ARA comprises the steps of controlling the fermentation temperature to be 20 to 25 DEG C after culturing in a fermentation tank for 100 to 140h; and / or, stopping ventilation or reducing the ventilatory capacity by not less than 50 percent at the 8th to 10th day of the culturing in the fermentation tank. The method for preparing the ARA has the advantages that the yield is high, the purity is high, and the large-scale industrial production of the ARA is facilitated.

The invention discloses a technology for extracting arachidonic acid oil by an aqueous enzymatic extraction. The technology comprises the following steps: using Mortierella alpina as an original strain, fermenting and culturing, diluting by adding distilled water to obtain a thallus suspension, regulating the pH value of the suspension, adding a biological enzyme, oscillating at constant temperature for 6-10 hours, adding ethanol and normal hexane to carry out oil extraction, centrifugating to delaminate, taking out an organic solvent phase, adding normal hexane into the left water phase and solid phase to carry out secondary extraction, centrifugating to delaminate, combining with the organic solvent phase, and carrying out rotary evaporation on the organic solvent phase to obtain the oil and an organic solvent. The invention has the advantages of low energy consumption, low cost, high yield of arachidonic acid oil, high quality, simple reactor, easy batch large-scale application and the like.

The invention discloses a method for preparing an ARA (Arachidonic Acid) by culturing a mortierella alpina by utilizing a solid material culture medium. The method comprises the following steps of using rice bran and niblet as material, adding inorganic salt in defined amount, using the inorganic salt as the solid material culture medium, collecting and smashing thallus after culturing strain in liquid, inoculating smashed thallus to the solid material culture medium, controlling culture temperature, mosturizing and controlling relative humidity during a culture process and appropriately stirring, and generating a large amount of spores. pH (Potential Of Hydrogen) segmentation regulation and control can be carried out during a fermentation process by using a citric acid and citrate. According to the method disclosed by the invention, the amount of seed spores of is large, the sunchronism is good, the activity is strong, the fermentation period of the mortierella alpina can be shortened, the growth of the mortierella alpine and the accumulation of ARA grease can be facilitated through the pH segmentation regulation and control, and the production and the quality of the ARA can be increased.

The invention discloses a mortierella alpine and the application thereof. Bacterial strains are conserved in China Center for Type Culture Collection (CCTCC) with a CCTCC NO. M 207067. The output of arachidonic acid produced by the bacterial strains upon fermentation can be up to 10g / l and the yield is up to 1.54g / l / d, thereby, the bacterial strains is applicable to industrial production of the arachidonic acid.

The invention provides a technology for preparing grease containing rich polyunsaturated fatty acid by taking lignocellulose type agricultural and forest wastes as raw materials through fermenting mortierella alpine, aiming at the problems in the prior art. The technology comprises the steps of pre-treating, saccharifying, carrying out mortierella alpine fermentation, extracting the grease and thelike. According to the technology provided by the invention, a fermentation phase of the mortierella alpine adopts a temperature gradual reduction method and the yield of ARA (Arachidonic Acid) can be remarkably improved; the ratio of the ARA in the total grease can reach 48 percent. Compared with the prior art, the technology adopts a cellulose preparation based on fiber-production globule bacteria to realize the saccharifying of lignocellulose, so that the cost of a carbon source and the enzyme utilization cost of a saccharifying phase are remarkably reduced; the cellulose saccharifying efficiency is 80 to 90 percent. Furthermore, a culture medium at a lignocellulose saccharifying phase and a fermentation culture medium in the technology can be circularly utilized, so that water can beremarkably reduced and the dosage of chemicals also can be remarkably reduced; the technology has remarkable effects of reducing wastewater emission and reducing the cost.

The invention provides restructured mortierella alpina strain MA-oPpFADS17-4 with heterologous experession from omega-3 desaturase genes of phytophthora parasitica, and provides a method for establishing the restructured mortierella alpina. The mortierella alpina uracil heterotrophia type strain is used as materials and produces a constant-temperature high-yield EPA restructured strain through the genetic operation technology of agrobaterium tumefaciens mediate. Production of EPA reaches 1197.3 mg / L and accounts for 31.5% of TFA content, and conversion rate for AA is 77.6%. The restructured mortierella alpina strain is of great significance for basic theory research and product development of the oil-producing fungus, mortierella alpina.

The invention relates to microbe fermentation, in particular to a method for preparing arachidonic acid through fermentation with addition of a product accelerating agent. The method comprises the following steps: inoculating mortierella alpina in a sterile fermentation culture medium containing a silicon source, a nitrogen source and inorganic salts for aerobic culture, collecting thalluses after culture, and extracting arachidonic acid-containing grease from the thalluses, wherein each liter of the sterile fermentation culture medium contains the following raw materials of the product accelerating agent: 1 to 5 mg / l of biotin, 0.5 to 2 g / l of glutamic acid, 1 to 3 g / l of citric acid, 1 to 5 [mu]mol / l of hydrogen peroxide and 1-5% of n-dodecane according to concentration in the fermentation culture medium. The method has the advantages that the technical problems of low yield, relatively high fermentation culture medium cost and the like in the prior art are solved; the yield is high; the fermentation process is simple; the operation is easy; the industrialized prospect is good.

The invention provides a recombinant mortierella alpina strain Ma-oPaFADS17 with heterologous expression coming from pythium aphanidermatum Omega-3 desaturase genes and a method for constructing the mortierella alpina strain with heterologous expression coming from pythium aphanidermatum Omega-3 desaturase genes. The method adopts mortierella alpine uracil auxotrophic strain CCFM 501 as materials and uses an agrobacterium tumefaciens mediated genetic manipulation technology to obtain a recombinant strain with a high yield of EPA in the environment of normal temperature, wherein the yield of EPA reaches 617.1 mg / L, and the content of the EPA accounts for 18.7 percent of the total amount of fatty acid, which has an important meaning for the basic research and product development of oil-producing fungus mortierella alpine.

The invention provides a recombinant mortierella alpina strain for heterologous expression of an MpFADS6 gene by virtue of a delta-6 desaturase gene MpFADS6 from tiny microcystis. The invention also provides an application of the recombinant mortierella alpina strain in production of EPA. By adding 1% of peony seed oil or 50g / L of peony seed pulp juice into a culture medium, the yield of the EPA is effectively increased, which has important significance for basic theory research of metabolic regulation in synthesis of fatty acid of an oil-producing fungus namely mortierella alpine and production of PUFAs. Compared with over-expression of an omega 3 fatty acid desaturase gene in the mortierella alpine, the yield of the EPA is increased by 5 times and can reach 588.5+ / -29.6mg / L.

A preparation method of arachidonic acid relates to the arachidonic acid. The method is as follows: optimizing culturing conditions of mortierella alpina arachidonic (ARA), fermenting in batches on basal media, then fermenting in batches on the optimized media by means of using yeast powder and corn steep liquor as nitrogen source, performing a fermenter enlarging experiment by using the yeast powder as the nitrogen source, culturing the mortierella alpina on the optimized media by using the corn steep liquor and the yeast powder as the mixed nitrogen source, and obtaining the mixed nitrogen source optimized media by replacing the nitrogen source on the optimized media into the corn steep liquor and the yeast powder; fermenting the mortierella alpina in batches on the mixed nitrogen source optimized media; performing high-glucose acclimatization to the mortierella alpina and fermenting the acclimated strains in batches on the mixed nitrogen source media; when the concentration of the glucose is reduced to 10g / L, performing fed-batch operation by 600 g / L of glucose so as to control the residual glucose within about 10 g / L, and starting to harvest dry strains, grease and the ARA after culturing for 168 h.

The invention discloses a single-carrier polycyclic aromatic hydrocarbon mixed bacterial agent with low temperature degradation-resistant effect and a preparation method and application thereof. The single-carrier polycyclic aromatic hydrocarbon mixed bacterial agent is prepared from a mixed bacterial agent and a corn cob which is used as a carrier, wherein the mixed bacterial agent is obtained bymixing pseudomonas SDR4 and mortierella alpine JDR4 which are screened from frozen and thawed soil according to a volume ratio of 1:1. The single-carrier polycyclic aromatic hydrocarbon mixed bacterial agent with low temperature degradation-resistant effect has the advantages that the biological activity is good, the cost is low, the preparation technology is simple, the secondary pollution is avoided, the recycling is not needed, the malicious competition, devouring and poisoning to microorganisms by indigenous flora, bacteriophage and toxic matters can be effectively shielded, and the highefficiency can be stably realized in the complicated environment; the effect of restoring the frozen and thawed soil which is polluted by polycyclic aromatic hydrocarbon in cold areas is obvious.

The invention discloses a decomposition maturing agent and the application thereof to green waste recycling. A fermentation broth is combined by adopting 20 to 50 parts of streptomyces griseus, 5 to 15 parts of pholiota adipose, 5 to 15 parts of mortierella alpine, 5 to 15 parts of flavobacterium columnare, 5 to 15 parts of pseudomonas stutzeri and 20 to 40 parts of rhodopseudomonas palustris; the bacterial broth is fully adsorbed on a carrier, namely crushed green waste of which the particle size is 1 to 30mm; water is added to the green waste until the water content reaches 40 percent to 70 percent, and then 100 to 300g of decomposition maturing agent are added to the green waste per cubic meter according to a volume ratio; auxiliary materials are urea and calcium superphosphate, and the weight ratio of the urea to the calcium superphosphate is 1:3; the weight ratio of the carrier to the auxiliary materials is 1:2; the decomposition maturing agent is obtained after the fermentation broth, the carrier and the auxiliary materials are evenly mixed; the decomposition maturing agent and the green waste are composted and fermented in a sealed manner at a temperature of 20 to 40 DEG; compost is turned once every 30 days, and water is supplemented to the compost until the water content reaches 40 to 70 percent when the compost is turned; and after being composted for 120 days, the green waste is spread and aired to be dried, and then organic fertilizer is obtained. The decomposition maturing agent has a wide application value.

The invention relates to a new separated bacterial strain of mortierella alpine (Mortierella Alpina) and an application thereof. Particularly, the invention relates to CGMCC No.6254, offspring thereof, culture and / or mutant strain and a method and application in preparing arachidonic acid by using the bacterial strain, offspring thereof, culture and / or mutant strain. The separated bacterial strain is a natural bacterial strain, and can stably produce arachidonic acid with a yield as high as 14.32g / L.

The invention discloses a method for promoting the high-efficiency accumulation of arachidonic acid grease. After the mortierella alpina strain is cultured to be activated, rapid mycelium multiplication culture is carried out on culture medium rich in the carbon source and other nutrient substances, mycelia are collected, rapid grease accumulation culture is then carried out on culture medium rich in the carbon source but lack in other nutrient substances, the mycelia are collected, rapid arachidonic acid accumulation culture is carried out on culture medium lacking carbon in the end, and the mycelia are collected. The technique can accurately control each stage of the arachidonic acid grease fermentation process, has the characteristics of short production period, high arachidonic acid production intensity, good grease quality and the like, and is easy to operate and industrially apply.

The invention relates to a method for promoting production of mortierella alpina and oil yield of arachidonic acid. The method comprises the following steps that fermented clear liquid of the mortierella alpina is collected, and through extraction and concentration of hydrophilic organic solvent, an accelerating agent which can adjust and control the production of the mortierella alpine and the oil yield of the arachidonic aicd is obtained by screening; and in the 0-48 hours at the early stage of fermentation, the accelerating agent is added to a normal medium according to the ratio of 0.1-0.5%, and a conditioned medium is prepared to inoculate and cultured at the temperature of 28 DEG C. After the fermentation, in fermentation results of some embodiments, biomass is 43.2 g / L, oil content is 61.3%, ARA content is 60.7%, ARA yield is 16.1 g / L, and the fermentation period is shortened from 192 hours to 168 hours. The method for promoting the production of the mortierella alpina and the oil yield of the arachidonic acid is simple and effective, promotes the production of the mortierella alpine, improves the oil yield of the arachidonic acid, and is of great significance to industrial application.

The invention discloses a mortierella alpine mutant strain, a method for producing arachidonic acid oil by using the same and arachidonic acid oil. The mortierella alpine mutant strain (Mortierella alpine Y16) is preserved in the China Center for Type Culture Collection (CCTCC) on July 2, 2015, the preservation address is Wuhan University in Wuhan City of China, and the preservation number is CCTCC NO: M2015421. In the arachidonic acid oil produced by using the mortierella alpine mutant strain, the content of triglyceride is more than or equal to 92wt%, the content of arachidonic acid is at least 48wt%, and the content of long-chain saturated fatty acid with more than 20 carbons is lower than 6.3wt%.

The present invention relates to processes for the production of arachidonic acid containing oils, which preferably are substantially free of eicosapentaneoic acid. The invention also relates to compositions containing such oils, in an unmodified form, and to uses of such oils. In a preferred embodiment, Pythium insidiosum is cultivated, harvested and the oil is extracted, recovered, and used as an additive for infant formula. In an alternative embodiment, Mortierella alpina is cultivated, harvested and the oil is extracted, recovered, and used as an additive for infant formula.

The invention discloses a Mortierella alpina strain capable of realizing high-yield production of arachidonic acid oil and application thereof. The Mortierella alpina strain is classified and named asMortierella alpine GM712 and has been preserved in China General Microbiological Culture Collection Center (CGMCC), with an accession number of CGMCC No. 15272. When the strain is used for fermentation production of arachidonic acid, the output of arachidonic acid can reach as high as 14.0 g / L, and the yield of arachidonic acid can reach 1.76 g / L / d. The strain of the invention can be used for industrial production of arachidonic acid.