39 results about "Schizochytrium sp." patented technology

The invention discloses a method for producing docosahexaenoic acid by strengthening schizochytrium limacinum, which comprises the following steps: carrying out fermentation culture on a schizochytrium limacinum strain until the growth period is ended (a nitrogen source is used up); when the growth period of the schizochytrium limacinum is ended, exogenously adding hydrogen peroxide, culturing, exogenously adding para aminobenzoic acid, and culturing; and repeating for several times, and ending the fermentation to obtain the DHA-containing fermentation liquor. Compared with execution of a blank group without addition of exogenous additives, an independent hydrogen peroxide intermittent feeding control strategy and an independent aminobenzoic acid intermittent feeding control strategy, the DHA yield is increased by 25.21% or above when the hydrogen peroxide-p-aminobenzoic acid periodic control method is adopted, and the highest level of 1.49 g / L is achieved.

The invention relates to the technical field of microbial fermentation, and discloses application of a regulatory factor in promoting schizochytrium limacinum to produce DHA (docosahexaenoic acid), and the regulatory factor contains beta-carotene and reduced glutathione. In addition, the invention also provides a method for improving DHA production of schizochytrium limacinum and a preparation method of microbial oil. The method comprises the following steps: fermenting schizochytrium limacinum in a culture medium, and adding beta-carotene and reduced glutathione into the culture medium in the fermentation process; in the culture medium, the content of the beta-carotene is 1.5 to 2g / L. The beta-carotene and the reduced glutathione can effectively improve the content of grease and DHA prepared by fermentation of schizochytrium limacinum, the content of DHA can reach 63.9%, and the beta-carotene and the reduced glutathione are non-toxic and high in safety.

The invention belongs to the technical field of biology, and discloses a yarrowia lipolytica engineering strain for efficiently producing eicosapentaenoic acid based on citric acid metabolism regulation, a method and application of the yarrowia lipolytica engineering strain. Delta9 elongase, Delta8 desaturase, Delta5 desaturase and Delta17 desaturase genes in a fatty acid synthesis route, heterologous ATP-citrate lyase and a mitochondrial citric acid vector are overexpressed. According to the invention, ATP-citric acid lyase and a mitochondrial citric acid carrier gene which are respectively derived from Schizochylene sp. And Saccharomyces cerevisiae are jointly expressed in the yarrowia lipolytica engineering strain, so that the yields of EPA and grease in the yarrowia lipolytica engineering strain are remarkably improved, and compared with a control strain, the yields of the grease and the EPA are respectively improved by 243.35% and 161.85%.

The invention relates to the technical field of genetic engineering, in particular to a recombinant vector, an expression plasmid, a recombinant schizochytrium limacinum mutant and a method for increasing the DHA content of microorganisms. The LIP1 gene for coding lipase in the microbial lipid droplet protein is knocked out through a homologous recombination technology, and the production of lipase in the microbial lipid droplet can be effectively inhibited, so that the hydrolysis process of triglyceride of polyunsaturated fatty acid stored in the lipid droplet to glycerol and free fatty acid in the later stage of microbial culture is blocked, and DHA is prevented from being lost from a TAG repository; experimental verification shows that in microorganisms represented by schizochytrium limacinum, the DHA content can be increased by 21.8% compared with that of a wild type, the yield and purity of DHA are increased, the stability of the microbial fermentation process is guaranteed, and application and popularization are facilitated.

The invention belongs to the technical field of genetic engineering, and discloses a schizochytrium limacinum genetic engineering strain capable of reducing the proportion of DPA in grease and correspondingly increasing the content of DHA, a method and application of the schizochytrium limacinum genetic engineering strain. The strain is characterized in that an omega-3 desaturase gene derived from Platyneria dumeriii and / or Lepeophtheria salmonis is subjected to heterologous expression, so that the proportion of DPA (docosahexaenoic acid) in grease is reduced, and the content of DHA (docosahexaenoic acid) is correspondingly increased. According to the invention, by heterologous expression of Pd delta 19Des and Ls delta 19Des genes in schizochytrium limacinum, desaturation activity of two desaturases to a delta 19 position is verified, and conversion from omega-6 type DPA to omega-3 type DHA is effectively promoted.

The invention belongs to the technical field of biochemical engineering, and discloses a production method for improving the content of odd-chain fatty acid in grease by modifying schizochytrium limacinum and optimizing a substrate and application of the method. According to the method, schizochytrium limacinum HX-308 serves as a receptor strain, high-yield odd-chain fatty acid engineering bacteria are obtained through gene overexpression and inhibition, and the high-yield odd-chain fatty acid is obtained. Accumulation of a propionyl CoA precursor pool for synthesizing odd-chain fatty acid is improved, endogenous methylmalonyl-CoA mutase coding gene MUT is inhibited, so that a competitive pathway of the odd-chain fatty acid is inhibited, and then through substrate optimization, the content of fermented grease and the content of the odd-chain fatty acid are obviously improved. The expression of related enzymes in schizochytrium limacinum is adjusted through a genetic engineering strategy to improve the odd-chain fatty acid synthesis, the synthesis and utilization of the key intermediate propionyl coenzyme A in the odd-chain fatty acid synthesis process are increased, the metabolic pathway of the propionyl coenzyme A is inhibited, and substrate optimization is assisted to improve the odd-chain fatty acid yield.

The invention belongs to the technical field of bioengineering, and discloses a genetic engineering strain for enhancing schizochytrium limacinum DHA oil production based on carbon flux guidance and fatty acid transport, a method and application thereof. The gene engineering strain is obtained by introducing an optimized coding gene ACLY and / or Fat1p coding gene related to guiding carbon flux and fatty acid transport into a schizochytrium limacinum body; wherein the gene sequence of the gene ACLY is SEQ ID No. 9, and the gene sequence of the gene Fat1p is SEQ ID No. 10. According to the invention, a schizochytrium limacinum engineering bacterium for enhancing DHA oil production based on carbon flux guidance and fatty acid transfer is constructed by means of metabolic engineering. The schizochytrium limacinum engineering bacterium is combined with fatty acid transfer based on carbon flux guiding, can be used for grease synthesis by guiding carbon flux, and can prevent grease from being degraded through fatty acid transfer.

The invention provides a method for producing polyunsaturated fatty acid, and belongs to the technical field of biological fermentation. The method for producing polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid from schizochytrium limacinum comprises the following steps: activating schizochytrium limacinum in a seed culture medium to obtain a schizochytrium limacinum seed culture solution; inoculating the schizochytrium limacinum seed culture solution into a fermentation culture medium for culturing, wherein the fermentation culture medium comprises rapeseed protein powder; and extracting the culture solution in the cultured fermentation culture medium to obtain the polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid. According to the present invention, the yield of polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid can be significantly improved.

The invention relates to the technical field of grease processing, and discloses a method for increasing the sn-2 DHA content in schizochytrium limacinum triglyceride. The method comprises the following steps: adding a metabolic regulator into a fermentation culture solution containing schizochytrium limacinum, carrying out fermentation culture until the total fermentation time is more than 120 hours, and collecting a grease product, the metabolic regulator is selected from at least one of choline chloride, inositol and lysophosphatidic acid. According to the method provided by the invention, the lipid metabolic flux of schizochytrium limacinum can be effectively regulated and controlled, and directional enrichment of DHA to the sn-2 site of triglyceride is remarkably promoted, so that the yield and content of sn-2 DHA are improved.

The invention belongs to the technical field of biochemical engineering, and discloses a method for promoting schizochytrium limacinum to quickly accumulate Omega-3 fatty acid by using a natural antioxidant and application of the schizochytrium limacinum, the method comprises the following steps: performing fermentation culture on the schizochytrium limacinum, adding a mixed exogenous antioxidant melatonin and a rosemary extract in the fermentation culture process, and performing fermentation culture on the mixed exogenous antioxidant melatonin and the rosemary extract to obtain the Omega-3 fatty acid-containing schizochytrium limacinum. The melatonin and rosemary extract mixed antioxidant can interact to regulate fermentation in schizochytrium limacinum to increase fatty acid synthesis and oxidation stability. According to the present invention, in the experiment of schizochytrium limacinum fermentation oil production, the addition of the specific amount of the mixed antioxidant composition during the fermentation results show that the oil yield can be increased, the PUFA ratio in the specific concentration range can be significantly increased by 32%, the PUFA use amount can be reduced by 40% through the compounding use, and the method can save manpower, material resources and cost, and can provide the good application prospect. The grease production efficiency is greatly improved, and the method is suitable for industrial production.

This invention discloses a fermentation method for increasing the yield of odd-chain fatty acids from *Schizochytrium*. *Schizochytrium* is inoculated into a fermentation medium containing assimilated carbon and nitrogen sources for fermentation. The carbon sources include carbon source a (a common carbon source) and carbon source b (a precursor that can be converted into propionyl-CoA intracellularly). The fermentation is carried out in a shake flask or fermenter. When fermentation is carried out in a fermenter, carbon source b is continuously added to the fermentation system at a constant flow rate. This invention, by establishing a mixed carbon source culture system and continuously supplementing propionyl-CoA precursors during fermentation in a fermenter, reduces the transient toxicity of propionic acid, maintains cellular energy metabolism and redox homeostasis, and improves the effective supply of intracellular propionyl-CoA, thereby significantly increasing the yield of odd-chain fatty acids.

Provided herein are microorganism of a Thraustochytrid and a method for preparing bio-oil using the same, and more particularly, Schizochytrium sp. PB31 (PTA-123692) having bio-oil producibility, and a method of preparing bio-oil, particularly bio-oil having a content of omega-3 and / or 6 long-chain polyunsaturated fatty acids of 30% or more by weight or more based on total fatty acids, characterized by culturing the microorganism.

The application belongs to the technical field of biology, and discloses a Yarrowia lipolytica engineering strain for efficiently producing eicosapentaenoic acid based on citric acid metabolism regulation, a method and application, wherein a chassis strain of the engineering strain is Yarrowia lipolytica Po1f, and genes of Δ9 elongase, Δ8 desaturase, Δ5 desaturase and Δ17 desaturase in a fatty acid synthesis pathway, a heterologous ATP-citrate lyase and a mitochondrial citrate carrier are overexpressed. ATP-citrate lyases and mitochondrial citrate carrier genes respectively derived from Schizochytrium sp. and Saccharomyces cerevisiae are combined to be expressed in the Yarrowia lipolytica engineering strain, so that the production of EPA and oil in the Yarrowia lipolytica engineering strain is significantly improved, and compared with a control strain, the production of oil and EPA is respectively increased by 243.35% and 161.85%.

The invention belongs to the technical field of biochemical engineering, and discloses a method for producing squalene by inducing schizochytrium limacinum through a salt stress strategy and application, the method comprises the following steps: fermenting a schizochytrium limacinum strain for fermentation for 0-72 hours, adding 3g / L of NaCl into a fermentation medium, after 72 hours, adding NaCl into the fermentation liquid to 5g / L, continuing to ferment for 120 hours, measuring the sugar consumption every 12 hours in the fermentation process, and supplementing glucose to 80g / L. According to the method, the squalene yield of schizochytrium limacinum is remarkably increased through a salt stress strategy, an expensive inducer is prevented from being used, and the production cost is effectively reduced. 3 g / L of low-concentration salt is added in 0-72 h of the early stage of fermentation to serve as an adaptation period, so that schizochytrium limacinum grows rapidly and is induced to start an osmotic regulation mechanism, the concentration of NaCl is increased to 5 g / L in 72 h, at the moment, the schizochytrium limacinum enters a lipid accumulation period, and high-concentration NaCl is utilized to stress the schizochytrium limacinum to efficiently accumulate squalene.

The invention relates to the technical field of gene engineering, and discloses a high-strength promoter, a schizochytrium limacinum engineering bacterium based on the promoter and application of the schizochytrium limacinum engineering bacterium. Phosphoric acid pantetheinyl transferase (PPTase) is a key rate-limiting enzyme for biosynthesis of docosahexaenoic acid. Four high-strength promoters which can be strongly expressed in a schizochytrium limacinum chassis are obtained through screening and comprise a P3431 promoter, a P5659 promoter, a P6631 promoter and a P4862 promoter, and the nucleotide sequences of the promoter are shown as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4 respectively; furthermore, the high-strength promoter is connected with PPTase and is integrated into a schizochytrium limacinum genome, so that the yield of the docosahexaenoic acid is remarkably increased. After a P3431 promoter, a P5659 promoter, a P6631 promoter and a P4862 promoter are respectively connected with PPTase and are transferred into schizochytrium limacinum for expression, the docosahexaenoic acid yield is respectively increased by 25%, 37.5%, 43% and 36.5% compared with that of a wild type.

The present application relates to a novel strain of Schizochytrium sp. with easy intracellular oil extraction and a method for producing oil containing omega3 using same. The novel microalgae of Schizochytrium sp. according to the present invention has a high content of fat in biomass, specially a high content of unsaturated fatty acids such as docosahexaenoic acid and eicosapentaenoic acid, and thus it is very easy to extract biomass produced by itself or by cultivation and fermentation and fat components including unsaturated fatty acids from the biomass. Accordingly, the microalgae, and dried biomass and bio-oil prepared therefrom can be helpfully used as a composition for feed or a composition for food.

The present invention relates to a novel Schizochytrium sp. strain containing a high protein content, antioxidant pigments and omega-3 fatty acids, and a method for producing a biomass containing a high protein content, antioxidant pigments and omega-3 fatty acids by using the strain. The novel Schizochytrium sp. strain of the present invention simultaneously contains, in the cells thereof, a high content of protein ingredients including amino acids such as glutamic acid, arginine and aspartic acid, antioxidant pigments of beta-carotene and canthaxanthin, and omega-3 unsaturated fatty acid ingredients including eicosapentaenoic acid and docosahexaenoic acid, and thus the strain itself or the biomass and bio-oil produced by culturing and fermenting the strain can be effectively used as a feed composition or a food composition and the like.

The invention belongs to the technical field of microbial engineering and synthetic biology, and particularly relates to a schizochytrium limacinum mutant strain A1-11 and application thereof in synthesis of odd-carbon-chain fatty acid. According to the invention, ARTP plasma mutagenesis is carried out on wild-type Aurantiochytrium sp. Sdu080, a mutagenic strain A1-11 with genetic stability is successfully screened, and compared with an original strain, the total proportion of C15: 0 and C17: 0 of the mutagenic strain under a conventional shake-flask culture condition is increased from 7.17% to 33.13%. Furthermore, after 20 mM of L-valine is added into the fermentation liquid culture medium, the proportion of odd fatty acid of A1-11 can be further increased to 52.14%. The schizochytrium limacinum A1-11 provided by the invention not only remarkably breaks through the limitation of the odd fatty acid yield and proportion in the prior art, but also has the advantages of simple process, easiness in large-scale fermentation and the like.

The present invention relates to the technical field of microbial preparation, specifically to Schizochytrium sp. and its applications, as well as methods and products for producing EPA, DHA, and heptadecanoic acid. Schizochytrium sp. SK-3 was deposited at the China Center for Type Culture Collection on October 23, 2024, with the deposit number CCTCC NO: M 20242315. The application of Schizochytrium sp. SK-3 in the production of EPA, DHA, and high-yield heptadecanoic acid. This strain is cultured in a fermentation medium containing a carbon source, a nitrogen source, and inorganic salts at 20-30 °C for 72-96 hours to obtain oils containing long-chain fatty acids such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and heptadecanoic acid, which can be used to prepare food products, cosmetics, or pharmaceutical compositions for animals or humans.

The present invention discloses a genetically engineered Schizochytrium strain for efficient DHA production, as well as a construction method and application. The genetically engineered strain is obtained by knocking out the peroxisome matrix protein gene PEX10 in the wild-type Schizochytrium, which disrupts the β-oxidation pathway and blocks the degradation of fatty acyl-CoA; overexpressing the acetyl-CoA carboxylase gene ACC1 to provide more malonyl-CoA precursors, thereby promoting fatty acid biosynthesis; and overexpressing the diacylglycerol acyltransferase gene DGAT to promote the accumulation of fatty acids in the form of triglycerides. Under shake flask conditions, after fermentation, the DHA and PUFA produced reached 55.10% and 70.47% of the total oil content, respectively. Compared with the wild-type strain, DHA and PUFA increased by 21.6% and 24.77%, respectively, without a significant change in biomass, showing extremely high application potential.

The present invention discloses a Schizochytrium genetic engineering strain that reshapes the FAS pathway, a construction method and an application. The Schizochytrium genetic engineering strain of the present invention is obtained by heterologously expressing the C16 elongase gene E16, the C18 elongase gene E18, the Δ9 desaturase gene D9, the Δ5 desaturase gene D5, the Δ6 desaturase gene D6, and the Δ15 desaturase gene D15 in the wild-type Schizochytrium. The recombinant Schizochytrium described in the present invention can produce EPA accounting for 2.08% of the total fatty acids, and can also produce at least DHA accounting for more than 41% of the total fatty acids. In addition, the present invention claims a method for producing unsaturated fatty acids through the dual-pathway channels of PKS and FAS, which is beneficial to promoting the popularization and application of the technology for Schizochytrium to produce EPA and DHA.

The present invention relates to a novel strain of Schizochytrium sp. with easy intracellular oil extraction and a method for producing oil containing omega-3 using same. The novel microalgae of Schizochytrium sp. according to the present invention have a high fat content compared to other biomass and especially have a high content of unsaturated fatty acids such as docosahexaenoic acid and eicosapentaenoic acid, and thus allow easy extraction of biomass produced from the microalgae or by culturing and fermenting, and easy extraction of fat components including unsaturated fatty acid from the biomass. Therefore, the microalgae, and a dried biomass product and bio-oil produced therefrom may be utilized as a composition for feed or a composition for food.

The invention discloses a fermentation process and application of algal oil containing specific DHA distribution characteristics, and belongs to the technical field of microbial fermentation and functional algal oil preparation. According to the method, sodium citrate participating in tricarboxylic acid circulation is utilized to supplement and enhance central carbon metabolic flux, and more sufficient acetyl coenzyme A and precursor substances are provided for fatty acid synthesis, so that the fatty acid synthesis flux is up-regulated; the phytosterol is used as a cell membrane fluidity regulator, so that the activity of the Sn-2 site acyltransferase in the schizochytrium limacinum is favorably activated, and the DHA is directionally guided to be combined to the Sn-2 site of the TAG. According to the invention, the two substances are synergistically added to a schizochytrium limacinum fermentation system, so that the three targets that the total DHA content gt is 40%, the ratio of Sn-2DHA to the total DHA gt is 50% and the specific triglyceride molecule content reaches the standard can be simultaneously realized accidentally, and the problem that the DHA content and distribution are difficult to regulate and control synchronously in the prior art is effectively solved.

A microbial oil includes a triglyceride. Sn-2 fatty acids of the triglyceride in the microbial oil contain 23% or more by weight of DHA. The microbial oil is prepared by fermentation using a Schizochytrium strain, where the Schizochytrium strain is Schizochytrium sp. with an accession number of GDMCC No. 60733. A food including the microbial oil is also provided.

The invention discloses a method for increasing the yield of stearic acid in schizochytrium limacinum based on lipid droplet targeting and application, and belongs to the technical field of microbial metabolism engineering. The invention aims to solve the technical problems of non-specific positioning of single lipid droplet protein signal peptide and difficulty in efficiently producing stearic acid in schizochytrium limacinum. The invention provides a fusion protein containing a lipid droplet targeting signal peptide and palmitic acid elongase, wherein the amino acid sequence of the lipid droplet targeting signal peptide is shown as SEQ ID NO: 5, and the lipid droplet targeting signal peptide is connected to the N-terminal of the palmitic acid elongase. The fusion protein is expressed in a schizochytrium limacinum genetic engineering strain and is subjected to fermentation culture, so that the yield of stearic acid is effectively increased. The fusion protein and the method are mainly used for improving the stearic acid yield of schizochytrium limacinum, can be used for plant subcellular localization research, provide a favorable method for subcellular localization research of microalgae such as schizochytrium limacinum and have important significance.

The invention belongs to the technical field of bioengineering, and discloses recombinant schizochytrium limacinum for producing DHA (docosahexaenoic acid) lipid by using carbon dioxide as well as a construction method and application of the recombinant schizochytrium limacinum, and the recombinant schizochytrium limacinum is obtained by introducing optimized encoding genes FDH, FK, FPR and Ti related to CO2 into schizochytrium limacinum; wherein the gene sequence of the gene FDH is SEQ ID No. 17, the gene sequence of the gene FK is SEQ ID No. 18, the gene sequence of the gene FPR is SEQ ID No. 19, and the gene sequence of the gene Ti is SEQ ID No. 20. The schizochytrium limacinum engineering bacterium capable of efficiently and rapidly utilizing the CO2 is constructed through a metabolic engineering means, and the schizochytrium limacinum engineering bacterium can utilize the CO2 as a carbon source to produce the DHA lipid, so that the production cost of the DHA lipid can be reduced, and the production economy of the DHA lipid can be improved.

The present invention belongs to the field of bioengineering technology and discloses an engineered Schizochytrium strain that co-expresses the PDC-ACC genes, as well as a construction method and application. The engineered strain is derived from Schizochytrium by expressing the pyruvate decarboxylase complex gene PDC and the acetyl-CoA carboxylase gene ACC, both found in Yarrowia lipolytica. The PDC and ACC genes in the engineered Schizochytrium strain are cloned from Yarrowia lipolytica as Po1f. Traditional fermentation yields 43.90 g / L of oil. Overexpressing the PDC and ACC genes in this engineered strain increases the oil content of the Schizochytrium strain by 53.13% to 67.22 g / L, and also increases the DHA content by 36.97%, from 40.71% to 55.76%.

This invention relates to a method for preparing and applying a fermented product of Bifida Ferment Lysate and Dioscorea Oolong with enhanced moisturizing and anti-aging effects, belonging to the field of microbial fermentation technology. This invention discloses a *Bifidobacterium longum*, the nucleotide sequence of which is shown in SEQ ID NO: 1. This invention utilizes *Bifidobacterium longum* HJ-005 to ferment *Dioscorea opposita* and *Dioscorea opposita* to prepare a *Bifida Ferment Lysate* fermented product of *Dioscorea opposita*, simultaneously increasing the content of absorbable diosgenin, anthocyanins, arginine, and organic selenium, with better effects compared to other lactic acid bacteria. It can significantly reduce the water loss and shrinkage of zebrafish embryos, exhibiting a moisturizing effect, and can significantly promote the expression of zebrafish type I collagen and Elna genes, promoting collagen and elastin regeneration, thus possessing anti-aging effects and can be used as a new raw material for cosmetics.

Disclosed are a ruminant methane emission inhibitor, a preparation method thereof, and use thereof. The ruminant methane emission inhibitor of the present application is prepared from components including 1-5 parts by weight of Schizochytrium sp. powder and 0.05-0.5 parts by weight of cellulase. The ruminant methane emission inhibitor of the present application can reduce the amount of methane generated by ruminants while increasing the total fermentation activity of the rumen.