41 results about "Dehydrocholesterols" patented technology

The invention discloses a process and a device for continuously synthesizing vitamin D3 through photocatalysis, and relates to the technical field of process on-line monitoring and intelligent control. Photochemical reaction requirements under raw materials with different concentrations are matched through multi-band illumination weight dynamic distribution; when the concentration of the 7-dehydrocholesterol fluctuates, the illumination ratio of 290nm to 305nm wavebands is adjusted in real time by a reinforcement learning algorithm, so that the conversion of a pre-vitamin D3 intermediate is more efficient, and the competitive consumption of a by-product path is reduced; the reaction retention time is further optimized through synchronous flow rate adjustment, and local excessive irradiation caused by sudden concentration increase is avoided; the environment temperature and humidity change is incorporated into a state vector, and the control module generates a compensation instruction accordingly; and if the humidity changes suddenly, pulse type flow compensation is triggered to counteract reaction efficiency attenuation. A closed-loop response mechanism reduces the dependence on physical temperature control equipment, and is especially suitable for mobile production scenes.

The invention discloses a synthesis method of 7-dehydrocholesterol and derivatives thereof, which comprises the following steps: cooling a compound (I), cuprous bromide and dimethyl sulfide compound under the protection of inert gas to obtain a reaction system, adding a Grignard reagent (II) or a Grignard reagent (III), stirring, reacting at room temperature, carrying out heating reaction, and cooling to obtain the 7-dehydrocholesterol. Performing post-treatment to obtain a product; if the product is the Grignard reagent (II), the product is 7-dehydrocholesterol and the derivative thereof; if the reagent is the Grignard reagent (III), the product is 25-hydroxy-7-dehydrocholesterol and the derivative thereof. The method is high in total yield, mild in reaction condition, simple in process route, high in product purity, low in three-waste generation amount, simple in post-treatment, environment-friendly and low in equipment requirement, the purity of 95% or above can be obtained through simple recrystallization, and conventional chemical equipment can be used for production.

The invention discloses a recombinant saccharomyces cerevisiae engineering bacterium for producing 7-dehydrocholesterol and application, and belongs to the technical field of genetic engineering and biological engineering. On the basis of an original strain, an exogenous gene DHCR24 and key endogenous genes ERG2 and ERG3 are integrated, and a non-path gene ROX1 is knocked out, so that the accumulation of 7-dehydrocholesterol in a saccharomyces cerevisiae body is further improved; the accumulation amount of the 7-dehydrocholesterol is further increased by adding integration of a speed limiting step gene and cell growth promoting genes SET2 and HAP4 and knockout of an RPH1 gene, and the ability of synthesizing the 7-dehydrocholesterol from the beginning in a saccharomyces cerevisiae body is further improved by completing single-copy integration of TPS1, ERG25, ERG26 and ERG27 genes at a saccharomyces cerevisiae GAL80 site. And the yield of 7-dehydrocholesterol reaches 896.2 mg / L after the constructed strain is subjected to shake flask fermentation for 100 hours.

The invention relates to the technical field of biological medicine, in particular to 7-dehydrocholesterol liposome and application thereof. According to the invention, a biological membrane modified liposome is constructed, and the biological membrane modified liposome comprises an ROS responsive liposome, a neutrophile granulocyte membrane and a renal tubular epithelial cell membrane; the biofilm modified liposome has an active oxygen species (ROS) responsive release function and a dual targeting capability, has the advantages of an integrated drug effect carrier, ROS triggered release, dual bionic targeting, strong target cell uptake and multi-mechanism ferroptosis resistance, and can be used for preparing the biofilm modified liposome. The compound can be used for treating ferroptosis related diseases such as acute kidney injury, myocardial ischemia-reperfusion injury and cerebral ischemia-reperfusion injury, and has wide clinical popularization value.

The present disclosure provides a method for preventing or treating pulmonary arterial hypertension in a subject in need thereof, the method comprising administering to the subject an effective amount of 7-dehydrocholesterol, a salt, oxide, metabolite, or derivative thereof.

The invention relates to the technical field of biochemical engineering, in particular to a method for efficiently converting 7-dehydrocholesterol into vitamin D3. The method comprises two steps of photoreaction and thermal isomerization. Wherein in the photoreaction stage, the reaction selectivity and the yield are remarkably improved by adopting 285 nm ultraviolet irradiation and controlling the energy density within the range of 5-8 W / mL, and the photoreaction yield can reach 60%-70%; a high-temperature condition of 70 + / -2 DEG C is adopted in the thermal isomerization stage, the reaction is carried out in a short time, the reaction period is greatly shortened, the yield is improved, and the thermal isomerization yield can reach 80%-90%; the total yield is superior to that of the traditional process. The method has the advantages of high reaction selectivity, few impurities, high yield, short period and the like, is suitable for industrial production of vitamin D3, and has a wide application prospect.

The invention belongs to the field of bioengineering, and particularly relates to cholesterol 7-dehydrogenase and a method for synthesizing 7-dehydrocholesterol by using the cholesterol 7-dehydrogenase. The invention provides a cholesterol dehydrogenase, and also provides a method for synthesizing 7-dehydrocholesterol by using the cholesterol dehydrogenase. By means of high-efficiency cholesterol 7-dehydrogenase evolved by using enzyme engineering and optimized reaction conditions, dependence on expensive chemical reagents is reduced, meanwhile, energy consumption is reduced, the production cycle is shortened, and the overall production efficiency is improved.

The present invention discloses a method for promoting the synthesis of 7-dehydrocholesterol by Saccharomyces cerevisiae. By mutating phosphomevalonate kinase ERG8 and squalene synthase ERG9 and integrating them into the Saccharomyces cerevisiae genome separately or simultaneously, several mutant strains with significantly increased 7-dehydrocholesterol production are obtained. Among them, phosphomevalonate kinase ERG8 involves mutations at one or more sites such as 75, 187, 192, 205, 206, 314, 322, 349, etc., and squalene synthase ERG9 involves mutations at one or more sites such as 96, 108, 146, 208, 209, 251, 252, 258, 286, 345, 355, 388, 389, 404, 426, 434, etc. The Saccharomyces cerevisiae mutants provided by the present invention can achieve the production of 7-DHC, laying a foundation for metabolic engineering transformation of Saccharomyces cerevisiae to promote 7-DHC synthesis.

The invention is applicable to the technical field of biological medicines, and provides application of lncRNA MCM3AP-AS1 as a diabetic cardiomyopathy biomarker and a therapeutic target. The invention discloses an action mechanism of long-chain non-coding RNA (lncRNA) MCM3AP-AS1 in diabetic cardiomyopathy (DCM). The invention further discloses a preparation method of the long-chain non-coding RNA MCM3AP-AS1. Bioinformatics screening finds that MCM3AP-AS1 is significantly down-regulated in DCM, and the MCM3AP-AS1 can serve as competitive endogenous RNA (ceRNA) to be competitively combined with miR-155-5p, target miR-155-5p / DHCR24 axis and promote expression of DHCR24 (24-dehydrocholesterol reductase), so that HO is subjected to enzymolysis, and oxidative stress and apoptosis of myocardial cells are relieved. By means of the mechanism, the MCM3AP-AS1 becomes a biomarker with diagnosis and treatment potential, and a new target spot and a theoretical basis are provided for clinical diagnosis and treatment of DCM.

The application discloses a method for preparing vitamin D3 by visible light catalysis and belongs to the field of biological medicine product manufacturing. The method uses 7-dehydrocholesterol (7-DHC) as raw material, selects tetrabromofluorescein as a catalyst under visible light with a wavelength of 400-780 nm, and 7-DHC is selectively opened to form previtamin D3 (P3) and then isomerized to vitamin D3 by heat. The method can obtain a high-yield (84%) vitamin D3 product by irradiation for only 90 seconds. The method for preparing vitamin D3 by visible light catalysis overcomes the technical bottleneck of the previous ultraviolet light catalysis, avoids the damage to the human body and the environment caused by ultraviolet light irradiation, is more green and environmentally friendly, has high selectivity, short reaction time and high efficiency, and has a wide industrialization prospect.

The invention provides a 7-dehydrocholesterol solvent-free crystal form as well as a preparation method and application thereof. The invention provides a crystal form A of a compound as shown in a formula I. The crystal form A is radiated by CuK alpha 1, and an X-ray powder diffraction pattern expressed by a 2 theta angle has diffraction peaks at 7.57 + / -0.2 degrees, 7.96 + / -0.2 degrees, 10.56 + / -0.2 degrees and 15.21 + / -0.2 degrees. The crystal form A of the compound shown in the formula I is a high-purity anhydrous solvent-free crystal form, crystal form conversion is not generated in the storage process, the purity of the crystal form A reaches up to 99% or above, and the crystal form A has good application prospects. # imgabs0 #

The invention discloses an improved process for production of vitamin D3 from 7-dehydrocholesterol (7-DHC) and to a simple process for recovery unreacted 7-DHC for further reuse. The invention further describes a process for isolation and purification of Vitamin D3.

The invention discloses a biosensor responding to steroid hormones. The nucleotide sequence of the biosensor is as shown in SEQ ID No. 52. According to the invention, transcriptome data under a steroid hormone stress condition is deeply excavated, a promoter capable of specifically responding to a specific steroid compound is screened out, and the response sensitivity and the dynamic range of the promoter are systematically optimized; the invention develops a biosensor capable of responding to steroid hormones, which can accurately and efficiently screen and characterize specific metabolites. Experimental results show that the biosensor not only can specifically respond to progesterone, but also has different degrees of responsiveness to various steroid hormones such as dehydroepiandrosterone, androstenedione, estradiol, 7-dehydrocholesterol and the like. The characteristic provides more possibilities for wide application of the biosensor in the fields of steroid hormone metabolism research, drug screening, environmental monitoring, industrial production and the like.

The present invention is applicable to the field of biomedicine and provides the use of lncRNA MCM3AP-AS1 as a biomarker and therapeutic target for diabetic cardiomyopathy. The present invention reveals the mechanism of action of the long noncoding RNA (lncRNA) MCM3AP-AS1 in diabetic cardiomyopathy (DCM). Bioinformatics screening revealed that MCM3AP-AS1 is significantly downregulated in DCM. It can act as a competitive endogenous RNA (ceRNA) to competitively bind to miR-155-5p, targeting the miR-155-5p / DHCR24 axis and promoting the expression of DHCR24 (24-dehydrocholesterol reductase), thereby enzymatically hydrolyzing H₂O₂ and alleviating oxidative stress and apoptosis in cardiomyocytes. This mechanism makes MCM3AP-AS1 a biomarker with diagnostic and therapeutic potential, providing a new target and theoretical basis for the clinical diagnosis and treatment of DCM.

The present application relates to the technical field of biological chemical industry, and particularly relates to a method for efficiently obtaining 7-dehydrocholesterol from yeast fermentation liquor. The method of the present application is to treat the yeast fermentation liquor with ionic liquid microemulsion, in which the ionic liquid dissolves polysaccharide on the surface of cell wall, and is used for assisting ultrasonic cell wall breaking, so as to shorten the ultrasonic cell wall breaking time, reduce the cell wall breaking power, and improve the cell wall breaking capacity. In addition, the ionic liquid microemulsion contains oil phase, which is a new process integrating the two-step traditional process of cell wall breaking and solvent extraction into one step, so as to shorten the overall process time, reduce the use of process equipment, simplify the process operation process, and can be completely applied to the environment-friendly industrial production, with low cost and wide application prospect.

The invention belongs to the technical field of prostate cancer diagnosis, and particularly relates to serum 7-dehydrocholesterol as a prostate cancer metastasis and endocrine treatment drug resistance diagnosis marker and application thereof. 7-dehydrocholesterol provided by the invention has different levels in human prostate cancer cell lines such as LNCaP, Du145, PC3 and the like, and is abnormally increased in PC3, so that the 7-dehydrocholesterol can be used as a drug resistance diagnosis marker for prostate cancer metastasis and endocrine therapy; the reagent composition capable of specifically recognizing 7-dehydrocholesterol and generating detectable signals can be used for preparing a prostate cancer detection kit; the 7-dehydrocholesterol provided by the invention has non-invasive, non-invasive and efficient effects as a diagnostic marker, and solves the technical problem of lack of a prostate cancer diagnostic marker in the prior art.

The invention discloses a preparation method of vitamin D3. The preparation method comprises the following steps: S1, obtaining 7-dehydrocholesterol; s2, taking 7-dehydrocholesterol as a raw material, and carrying out primary photochemical reaction under the irradiation of an LED ultraviolet lamp, so that the 7-dehydrocholesterol is converted into pre-vitamin D3 and tachysterol to obtain a reaction solution; s3, adding a triplet-state photosensitizer into the reaction liquid, and carrying out secondary photochemical reaction under the irradiation of an LED (Light Emitting Diode) ultraviolet lamp, so that the tachysterol is converted into pre-vitamin D3, thereby obtaining a mixture containing the pre-vitamin D3; and S4, carrying out a thermal isomerization reaction on the mixture containing the pre-vitamin D3 to obtain the vitamin D3. Raw materials do not need to be recycled after the photochemical reaction, the process is simple, the process operation steps are reduced, the cost is reduced, and the process stability is good.

The application discloses a sterol C24-reductase mutant for reducing by-products and a Saccharomyces cerevisiae for producing 7-dehydrocholesterol, and belongs to the technical field of biology. In order to solve the problems of by-product production and low catalytic activity caused by the wide substrate spectrum of the key enzyme DHCR24 in the current 7-DHC synthesis process, the leucine at the 162th position and the aspartic acid at the 386th position of DHCR24 are mutated through enzyme engineering modification, and a sterol C24-reductase mutant DHCR24M is obtained. The mutant is introduced into a Saccharomyces cerevisiae starting strain, and it is found through fermentation verification that, compared with the unmutated strain, the by-products cholest-7-en-3-ol and cholesta-5,7-dien-3-ol of the mutant strain are reduced by 31.5% and 46.1% respectively, and the target product is increased by 90.2%, which provides a new idea and method for industrial production of 7-dehydrocholesterol and active vitamin D3, and has a very wide application prospect.

The invention relates to a method for simultaneously extracting 7-dehydrocholesterol and vitamin D3, which comprises the following steps: carrying out light conversion on a sample containing 7-dehydrocholesterol to obtain a light conversion solution; performing thermal conversion on the light conversion liquid to obtain thermal conversion liquid; concentrating the thermal conversion solution to obtain a concentrated solution, and stirring to obtain a first precipitate; mixing and dissolving the first precipitate and a first polar solvent, cooling to obtain a precipitate, namely 7-dehydrocholesterol, and drying supernate to obtain the vitamin D3. The sample containing the 7-dehydrocholesterol can be directly actinized without any treatment, the vitamin D3 can be obtained through saponification thermal conversion of the actinized reaction liquid, the vitamin D3 and the 7-dehydrocholesterol are jointly separated out through low-temperature crystallization and separated from other impurities, then the 7-dehydrocholesterol and the vitamin D3 are separated, and the 7-dehydrocholesterol and the vitamin D3 are separated. The purity and the yield of the two are greatly improved.

The invention discloses a P450 enzyme, an enzyme composition and application thereof. The amino acid sequence of the P450 enzyme is as shown in SEQ ID NO: 4. According to the P450 enzyme or enzyme composition provided by the invention, 7-dehydrocholesterol is used as a substrate to synthesize an intermediate 25-hydroxy-7-dehydrocholesterol of 25-hydroxyvitamin D3, and then the 25-hydroxyvitamin D3 is generated through illumination. The 7-dehydrocholesterol is low in price, so that the cost can be effectively reduced. Moreover, by further optimizing reaction conditions, when the concentration of 7-dehydrocholesterol is 13 g / L, the conversion rate can reach 95.5%. The P450 enzyme or enzyme composition provided by the invention has a wide application prospect in the technical field of enzyme catalysis.

The present disclosure relates to fusion proteins including an iron-sulfur protein (e.g., cholesterol 7-desaturase (C7D)) and a ferredoxin reductase (e.g., KshB) that can efficiently convert cholesterol to 7-dehydrocholesterol (7-DHC) in vivo. Also disclosed herein are engineered microbes (e.g., E. coli) expressing the fusion proteins and methods of use thereof.

The present invention relates to the field of pharmaceutical chemistry, and specifically to a method for preparing cholesterol, a derivative thereof, and an analog thereof. The derivative of cholesterol comprises, but is not limited to, 7-dehydrocholesterol, 25-hydroxycholesterol, 25-hydroxy-7-dehydrocholesterol, and ergosterol. In the present invention, a compound represented by formula I can be prepared by means of a microbial transformation using phytosterols as a raw material, followed by preparing cholesterol, the derivative thereof, and the analog thereof.

The invention relates to the technical field of organic synthetic chemistry, in particular to a method for synthesizing plant-derived 7-dehydrocholesterol from dinorhydrin. Comprising the following steps: 1, carrying out sulfonic acid esterification reaction on dinorhydrin to obtain an intermediate 1 or a solution containing the intermediate 1; 2, carrying out first enol esterification or enol etherification on the intermediate 1 to obtain an intermediate 2; 3, carrying out coupling hydrocarbylation on the intermediate 2 to obtain an intermediate 3; 4, carrying out upper debromination and deenolization on the intermediate 3 at the same time to obtain an intermediate 4; 5, carrying out second carbonyl enol esterification on the intermediate 4 to obtain an intermediate 5; and 6, carrying out hydrolysis reduction on the intermediate 5 to obtain the 7-dehydrocholesterol. The botanical 7-dehydrocholesterol is synthesized by taking dinorhydrin as a starting raw material, the total weight yield is 36%, the purity is 98% or above, and industrial synthesis is easy to realize.

The invention discloses a yeast engineering strain for synthesizing 7-dehydrocholesterol as well as a construction method and application of the yeast engineering strain, and belongs to the technical field of metabolic engineering. The invention provides a recombinant Candida tropicalis, which integrates a human-derived C 24-dehydrocholesterol reductase gene HsDHCR24, and can be used for preparing the recombinant Candida tropicalis by overexpressing all genes in a front squalene synthesis route and genes in a rear squalene synthesis route. Meanwhile, by blocking a branch metabolic pathway of ergosterol synthesis and weakening the consumption of ergosterol and by virtue of an endoplasmic reticulum engineering strategy, the yield of 7-DHC obtained by shake-flask fermentation of the recombinant strain is increased from 10.1 mg / L to 432.3 mg / L, and by virtue of culture medium optimization and fed-batch fermentation of a 5L fermentation tank, the final yield reaches 3.65 g / L, so that the yield of 7-DHC is greatly improved. While the yield is increased, a thought is provided for synthesizing other sterol products from chassis cells by the candida tropicalis.

The invention relates to a method for optimizing 7-dehydrocholesterol fermentation by using data fitting and a fermentation method, and the method comprises the following steps: constructing fermentation influence parameters including at least one of fermentation strain activity, fermentation temperature, inoculum size and culture medium components; adjusting the numerical values of the fermentation influence parameters, obtaining an optimal fermentation effect through an experiment, and taking the fermentation influence parameters with the optimal fermentation effect as optimized 7-dehydrocholesterol fermentation parameters; wherein when the fermentation influence parameters are culture medium components, the importance of each component in the culture medium components is analyzed by adopting a random forest model, and a component importance sequence is obtained; performing multiple linear regression modeling by taking at least two components with the top importance sequence as input to obtain a regression equation, and constructing fermentation culture medium components by using the regression equation to obtain optimized culture medium components; the fermentation titer can be improved, and the production cost can be reduced.

The invention relates to the technical field of organic synthetic chemistry, in particular to a method for synthesizing 7-dehydrocholesterol from cholesterol. Comprising the following steps: dissolving cholesterol in a first solvent, adding cyclohexanone and aluminum isopropoxide, and carrying out an Oppenauer oxidation reaction to obtain an intermediate 1; dissolving the intermediate 1 in a second solvent, sequentially adding isopropenyl acetate and p-toluenesulfonic acid, and carrying out first acetylation reaction to obtain an intermediate 2; dissolving the intermediate 2 in a third solvent, and carrying out an upper debromination reaction to obtain an intermediate 3; and dissolving the intermediate 3 in a fourth solvent, then adding 2, 4, 6-trimethylpyridine and acetyl chloride as reaction accelerators, carrying out a second acetylation reaction to obtain an intermediate 4, and carrying out a reduction reaction and purification on the intermediate 4 to obtain the 7-dehydrocholesterol. The 7-dehydrocholesterol is synthesized by the method, the total synthesis yield exceeds 60%, the product purity is 98% or above, and the method is easy to amplify for industrial synthesis.

The invention relates to application of DHCR24 related to diagnosis, treatment and prognosis of cervical cancer. The application comprises the application of the DHCR24 gene expression inhibitor in preparation of drugs for preventing, relieving and / or treating cervical cancer, the application of a reagent for detecting the DHCR24 gene expression level in preparation of products for diagnosis or prognosis of cervical cancer, a cervical cancer risk assessment or prognosis device and a cervical cancer risk assessment or prognosis instrument. The invention finds that 24-dehydrocholesterol reductase (DHCR24) is highly expressed in cervical cancer cells and is related to poor prognosis of cervical cancer; proliferation, migration and invasion of cervical cancer cells and dryness of tumor cells can be inhibited by inhibiting expression of DHCR24, and apoptosis of the cervical cancer cells is promoted. Therefore, DHCR24 can be used as a diagnosis and prognosis marker and a treatment target of cervical cancer, and has a huge application prospect.

The invention discloses a strain for synthesizing 7-dehydrocholesterol as well as a preparation method and application thereof, and belongs to the technical field of biology. Saccharomyces cerevisiae is used as a chassis strain for synthesizing 7-dehydrocholesterol (7-DHC) by a microbiological method, an efficient production strain is constructed by exploring a new gene influencing synthesis of a target product, and the related modification comprises the following steps: firstly, integrating 24-dehydrocholesterol reductase DHCR24, and synthesizing 7-DHC from the beginning in the saccharomyces cerevisiae strain; and then overexpressing key factors SUR7 which are constituent parts of glycosyl phosphatidylinositol anchoring protein ECM33, 1, 3-beta-D-glucan synthase FKS1 and Can1 compound membrane compartments, expanding plasma membrane area of the saccharomyces cerevisiae, activating cell wall integrity to restore membrane fluidity of strains, regulating secretion of cell EV, enriching storage space of 7-DHC, promoting synthesis of 7-DHC, and improving the yield of 7-DHC. Finally, the extracellular yield is increased by 42% compared with that before transformation.

The present invention discloses a P450 enzyme, an enzyme composition, and applications thereof. The amino acid sequence of the P450 enzyme is shown in SEQ ID NO: 4. The P450 enzyme or enzyme composition provided by the present invention uses 7-dehydrocholesterol as a substrate to synthesize 25-hydroxy-7-dehydrocholesterol, an intermediate of 25-hydroxyvitamin D3, and then generates 25-hydroxyvitamin D3 by light. 7-dehydrocholesterol is inexpensive, which can effectively reduce costs. In addition, by further optimizing the reaction conditions, the present invention can achieve a conversion rate of 95.5% when the concentration of 7-dehydrocholesterol is 13 g / L. The P450 enzyme or enzyme composition provided by the present invention has broad application prospects in the field of enzyme catalysis technology.