160 results about "Gulonic acid" patented technology

The invention relates to a method for producing mycoprotein and polypeptide organic fertilizer by waste fermented mash and gulonic acid mother solution formed in the vitamin C production technology, which belongs to the field of biotechnology and waste resource utilization technology, and comprises the following steps: adding 5 to 20 parts of gulonic acid mother solution in 100 parts of waste vitamin C fermented mash protein, adjusting pH value to 2.0 to 5.0, and hydrolyzing for 0.5 to 12 hours under the temperature of 40 to 70 DEG C. On the one hand part of protein in fermented mash protein is hydrolyzed to become polypeptide and amino acid to dissolve in water, through adjusting pH value and appropriate concentration, right amount of trace element is added, obtaining polypeptide organic fertilizer, on the other hand the rest protein in mash protein is precipitated, and the precipitated protein is made into mycoprotein through filter-pressing, drying and smashing. The invention recycles the vitamin C fermentation industry waste to produce mycoprotein and polypeptide organic fertilizer, thus not only solving the problem of industrial pollution, but also providing organic fertilizer and feed protein for agricultural production.

The method to extract vitamin C and Gulonic acid from vitamin C mother liquor comprises, using the said mother liquor as material to separate the vitamin C and Gulonic acid completely on analog moving bed with water as eluant at 20~75Deg, and obtaining the said constituents. This invention uses common condensing crystallization separation method.

The invention provides a production method of gulonic acid in the production of vitamin C, which comprises the following steps: (1) ceramic film filtering: filtering and concentrating a fermentation broth through a ceramic-film filtering apparatus, and removing proteins, hypha, suspended substances, colloid, bacteria and other macromolecular organic substances in filter residues to obtain a gulconic sodium dialysate; (2) double-pole film electrodialysis system: processing the filtrate containing gulconic sodium salts by an electrodialysis system to as to convert the gulconic sodium into the gulconic acid solution; (3) nanofiltration concentration: concentrating the gulconic acid solution in a nanofiltration film apparatus to obtain the concentrated gulconic acid solution. The method can reduce the production cost, increase the total yield, enhance the product quality and achieve the aims of environmental protection and simple apparatus operation.

The invention discloses a reconstruction method for producing the Vitamin C precursor 2-KLG with gluconobacter oxydans, and belongs to the field of genetic engineering. According to the invention, a sorbose dehydrogenase (SDH) gene and a sorbosone dehydrogenase (SNDH) gene originated from ketogulonigenium vulgare are expressed in gluconobacter oxydans to obtain a G. oxydans engineering bacteria for production of 2-KLG by using sorbitol. G. oxydans is a frequently used strain in the first step of fermentation in the method of two-step fermentation; in the invention, the SDH and SNDH genes are expressed in G. oxydans, which enables the problem of dependency of ketogulonigenium vulgare on associated fungi to be overcome, direct conversion of D-sorbitol into 2-KLG to be realized and the production process for Vitamin C to be simplified; output of 2-KLG reaches 83 g/L; therefore, the invention has a very good application prospect.

The invention provides a method for producing 2-keto-l-gulonic acid, including the following steps of: feeding nitrogen source nutrient solution from the 20th to the 26th hour in the fermentation process to compensate nutritional demands of microorganism growth in the fermentation process, finishing feeding solution in the 40th to the 45th hour, wherein the dissolved oxygen is controlled to be 30-50% in the nutrient solution feeding process. The invention utilizes a method of feeding nutrient solution at middle and later stages of fermentation, overcomes disadvantage that biomass grows and generates acid slowly, combines control of factors of temperature, pH, dissolved oxygen, low initial glucose, fed-batch glucose compensation and the like, and optimizes the growth relations of biomasses with different sizes in microbial mixed fermentation. The gulonic acid content of the invention is 105-115mg/ml, and the glucose acid convert ratio is 92-95%.

The invention discloses a preparation technology for 2-keto-L-gulonic acid ferment strain, adopting large and small bacteria for mixed culture, wherein, the small bacteria is ordinary ketogenic gulonic acid bacteria, and the large bacteria is Bacillus megatherium. The invention comprises the following steps: firstly, inoculating the small bacteria onto a slant culture medium to cultivate for 16 to 48 hours separately; then, inoculating the large bacteria and continuing to cultivate for 24 to 72 hours with the quantity ratio between the large bacteria and the small bacteria being 1:(20-40); taking sucrose with the concentration of 5-30g/L or fructose with the concentration of 5-20g/L as the main carbon source in the culture medium; introducing the seed mixture of large and small bacteria into the culture medium for cultivation and controlling the bacteria count of seeding liquid to 10-10/ml. The invention changes the proportion between large bacteria and small bacteria, increases the strain density, and improves the strain vitality, thus shortening the fermentation period; the invention is further characterized by unchanging strain, easy operation and widespread use.

A method for improving the stability for the fermentation production of 2-keto-L-gulonic acid belongs to the technical field of bioengineering. The 2-keto-L-gulonic acid is also named as vitamin C. The invention uses three mixture liquids to replace corn pulp, adopts an orthogonal test to investigate the important effects of a metal ion mixture liquid, an amino acid mixture liquid and a vitamin mixture liquid in the fermentation of the vitamin C. Based on the obtaining that the metal ion mixture liquid is the most remarkable affecting factor for cell growing, KGA synthesizing and L-sorbose consumption, the method of the invention further adopts the orthogonal test to investigate the effects of various metal irons to the fermentation of the vitamin C and discovers that increasing the concentration of Mg<2+> can promote the synthesizing of the KGA and relates to the most remarkable affecting factor for the fermentation of the vitamin C. The output and yield of the KGA which adopts optimized combination to carry out the fermentation of the vitamin C are respectively to be 75.2g/L and 0.94g/g. Compared with a common culture medium, the output (67g/L) and the yield (0.84g/g) of the KGA are respectively improved by 12.24 percent and 11.9 percent.

The invention discloses an enhancing method of the production strength of 2-keto-L-gulonic acid (2-KLG), belonging to the field of microbial fermentation engineering. The purpose of enhancing the production strength of 2-KLG is achieved through the addition of the key amino acids, namely L-glycine and L-proline which have significant promotion effect on the production of 2-KLG which adopts the mixed fermentation of the common Ketogulonigeniumvulgare and Bacillusmegaterium. When L-glycine and L-proline are used to enhance the key amino acid components of the corn steep liquor, the fermentationperiod is reduced to 40 hours which is reduced by 21.6%, the concentration of 2-KLG can reach 80.29g.L<-1>, and the acid production rate of 2-KLG can reach 2.00g.(L.h)<-1>, therefore the production strength of 2-KLG is significantly enhanced.

The invention discloses a gulonic acid adsorption separation method utilizing hyper-crosslinked resin. According to the gulonic acid adsorption separation method utilizing the hyper-crosslinked resin, a preprocessed gulonic acid fermentation broth is subjected to the processes of absorption, impurity washing and desorption through an adsorbing column filled with the hyper-crosslinked resin, and finally a gulonic acid solution is obtained. The hyper-crosslinked resin is prepared through a Friedel-Crafts cross-linking reaction and with polystyrene-divinyl benzene as a framework, and is weak in polarity and good in hydrophilia; the average grain diameter is 0.42-0.56mm, the water content is 43wt%, the bore diameter is 1.92nm, the porosity is 30-40%, the wet density 1.09g/cm3, the average specific surface area is 1645.5m2/g, the average pore volume is 0.732cm3/g, and a functional group is a carbanyl group. The adsorption medium adopted by the method has the advantages of being large in adsorption capacity for the gulonic acid, good in adsorptive selectivity, mild in desorption condition and long in service cycle, enabling regeneration to be easy and the like.

The invention discloses a production method and apparatus for high-power concentration of gulonic acid, which comprises delivering a resin ion-exchanged solution or concentrate from a conventional nanofilter to a precise filter through a delivery pump; delivering the material solution to a booster pump, and boosting a pressure to 50-70 bar; delivering the material solution to an online booster pump, mixing a part of concentrate from the outlet of a high-power concentration membrane and the material solution, and boosting the pressure to 55-75 bar; and separating the mixture through the high-power concentration membrane, driving water in the mixture under the high pressure to penetrate a membrane component to enter a dialysate pipe, discharging a part of the concentrated solution, and returning the rest of the concentrated solution to the inlet of the online booster pump. The invention also discloses a production apparatus for high-power concentration of gulonic acid. The invention can increase the concentration of gulonic acid to a higher level, reduce the investment cost of an evaporator, lower energy consumption by 90%, and improve the product quality.

The invention provides a method for synthesizing vitamin C crude product by adopting L-gulonic acid, which comprises the following steps: the L-gulonic acid and a reaction solvent are added to a reaction vessel, and esterification reaction is carried out on the L-gulonic acid and the reaction solvent in the condition of strong acid catalyst so as to generate an L-gulonic acid methyl ester solution; internal esterification reaction is directly carried out on the L-gulonic acid methyl ester solution in alkalinity condition so as to generate a vitamin C sodium salt solution; after the vitamin C sodium salt solution cools down, dilute sulphuric acid is added directly for acidification reaction so as to generate an acidification solution; activated carbon decoloration filtration and concentration filtration are carried out on the acidification solution so as to obtain the vitamin C crude product, wherein esterification reaction, internal esterification reaction and acidification reaction are carried out in the same reaction vessel. The method provided by the invention has simple process, improves production efficiency, reduces labor intensity of workers and improves production environment.

The invention discloses a production technique of 2-keto-L-gulonic acid by high concentration fermentation. In a fermentation medium adopted by the production technique, the initial sorbin concentration is 15mg/ml to 25mg/ml; fermentation is carried out for 4 hours to 8 hours; continuous sorbin feeding is started; fermentation is carried out for 36 hours to 44 hours and then sorbin feeding is finished; during the feeding process, the content of sorbin is controlled to be 15 mg/ml to 30 mg/ml; when fermentation is carried out for 24 hours to 40 hours, pH value is adjusted to be 7.1 to 7.3; when fermentation is carried out for 40 hours to 44 hours, pH value is adjusted to be 7.4 to 7.6 until fermentation is finished; and dissolved oxygen during the whole fermentation process is controlled to be 30 percent to 50 percent. The technique adopted by the invention has total fed sorbin with a concentration of 10.5 percent to 11.5 percent (W/V) at the end of fermentation; the concentration of gulonic acid of fermentation solution is raised by 10 percent to 15 percent; the conversion rates of alcohol and acid are respectively promoted by 3 percentage points; and stability is good; besides, the technique process has simple and convenient operation and is easy for wide application.

The invention discloses a process for refining gulonic acid. The process comprises the following steps of: performing ultrafiltration on fermentation liquor containing sodium gulonate; acidizing the fermentation liquor subjected to the ultrafiltration by using an ion exchange device, wherein the pH value of the fermentation liquor in the ion exchange device is below 2.0; performing nanofiltration preconcentration on the acidized fermentation liquor to obtain preconcentrated liquor; adsorbing the preconcentrated liquor by using active carbon or adsorbent resin; and performing triple effect evaporation on the adsorbed preconcentrated liquor to obtain concentrated liquor, and crystallizing the concentrated liquor to obtain a gulonic acid crystal. In a refining process route, the fermentation liquor is subjected to the ultrafiltration, ion exchange, the nanofiltration preconcentration to remove a large number of macromolecular impurities and soluble protein, and the preconcentrated liquor subjected to the nanofiltration preconcentration is adsorbed to remove impurities such as residual sugar, the soluble protein, pigments and the like further, so the quality of feed liquor before crystallization is optimized, the purity and yield of products are improved, and the discharge capacity of mother liquor is reduced.

The present invention provides a method of synthesizing the vitamin C (V _C ) through the esterification, the pervaporation (PV) membrane separation and integration technology, comprising the process of the methyl esterification between the methanol and the 2-ketone-L-gulonic acid (2-KLG) to produce the 2-ketone-L-gulonic methyl ester (2-Me-KLG). The process comprises the following steps: (1) the solid or liquid 2-KLG is dissolved in the methanol; the temperature is then raised and the catalyst is added; (2) the reaction solution prepared in the first step is pumped into the PV membrane separation device; the reaction solution after the membrane dehydration circulates into the esterification reactor; (3) the product 2-Me-KLG or the reaction solution is directly provided for the later production of the V_C. In the method of the present invention, the area of the used membrane is small; the ratio between the alcohol and acid of the raw material liquid and the reaction temperature are relatively low; the invention greatly improves the purity and collection rate of the 2-Me-KLG; the technology is simple and the process is stable; the present invention is suitable for the industrial production-line.

The invention relates to a method for removing pigment in gulonic acid mother solution, which is characterized in that a simulated moving bed filled with acidic cation resin is adopted, and water is used as a mobile phase. The method has a pigment removing rate of above 70%, is beneficial to enhancing the purity and the yield coefficient of the product and improving the luster of product greatly, and has the advantages of simple operation, low manufacturing cost, environment protection and the like, thus the method is more suitable for large-scale industrial production.

The invention discloses a method for synthesizing vitamin C sodium salt which comprises the steps of: (a) mixing Gulonic acid with methanol solution, and orderly passing through a granular active carbon column and a cation resin column, (b) passing the treating fluid through a dry filter, (c) performing a recycle esterification reaction by passing the treating fluid acquired from the step b through the resin column so as to acquire esterified liquid, (d) pouring the esterified liquid into a constant pressure reactor and performing conversion reaction, and (e) cooling the liquid material B to room temperature after finishing the reaction and centrifuging so as to acquire the vitamin C sodium salt. The method of the invention efficiently promotes the esterifying ratio of products and the yield of the vitamin C sodium salt, shortens the conversion reaction time, simplifies the process, lowers the energy consumption and producing cost, decreases the environmental pollution and prolongs the service life of the device.

The invention discloses a method for producing a 2-keto-L-gulonic acid (2-KLG) vitamin C precursor by modifying Escherichia coli and belongs to the field of genetic engineering. In the method, a sorbitol dehydrogenase (SLDH) gene from gluconobacter oxydans, a pyrroloquinoline quinine (PQQ) gene cluster and a sorbose dehydrogenase (SDH)/ sorbosone dehydrogenase gene (SNDH) from common ketogulonigenium vulgare are expressed in Escherichia coli by a genetic engineering technique to culture an Escherichia coli engineering strain for producing 2-KLG by sorbitol. At present, a two-step fermentationprocess adopted for industrial production of vitamin C in China has the problems of complicated process, many influencing factors, difficult accurate control and large raw material and energy consumption for growing three kinds of microbes at the same time. When the production of the 2-KLG by sorbitol fermentation by the E.coli engineering bacteria is adopted, the production process is simplified, raw materials and energy are saved, and the yield of the 2-KLG can reach 87g/L; and thus, the method has a very bright application prospect.

The invention relates to a preparation method of vitamin C. The preparation method comprises the following steps: by using gulonic acid inorganic salt or gulonic acid as the raw material, adding a 0-38 wt% hydrochloric acid solution, stirring, cooling, introducing HCl gas, heating, and keeping the temperature until the reaction finishes, thereby obtaining the vitamin C. The preparation method has the advantages of simple technique, short reaction time and more environment-friendly production environment, obviously enhances the vitamin C quality and is suitable for vigorous popularization.

The invention provides a method for separating solution rich in gulonic acid and vitamin C by ion exclusion chromatography, which mainly uses different repulsion forces of strong-acidic group of strong-acidic ion exchange resin on the gulonic acid and the vitamin C to separate the gulonic acid and the vitamin C. The method is realized through a moving bed system or a simulated moving bed system, can improve the yield of vitamin C (Vc) products, reduce production cost, and ensure the yield of gulonic acid and vitamin C over 90 percent.

The invention discloses a method for producing gulonic acid (VC precursor). The method comprises the steps as follows: 1) inducing proper amount of Xanthomonas campestris with ray, 2) culturing on a plate at 25-35DEG C for 2-5d, selecting the fine colony which satisfies the requirement of smooth colony, inoculating in a conical flask with culture fluid for culturing, 3) adding into the medium containing glucose for culturing to producegulonic acid. The invention realizes direct production ofgulonic acid from glucose to produce VC, with a converting ratio of 35-50%, has the advantages of simple production, reduces the production cost greatly.

The invention discloses a vitamin C production process improvement method, which comprises: adopting a simulated movement bed chromatographic separation device to separate and purify Vc, gulonic acid and gulonic acid methyl ester in a centrifugation separation mother liquor of Vc production, wherein three materials can be concurrently obtained through separation, the first material is an extraction liquid and is a vitamin C-rich component, the vitamin C purity is more than 90%, the vitamin C-rich component is concentrated and then directly returns to be subjected to crystallization with the original Vc solution, the second material is a raffinate and is a gulonic acid-rich component, the gulonic acid purity is more than 90%, the gulonic acid-rich component is concentrated and then returns to the original process to be esterified, converted and crystallized so as to be recycled, the third material is a raffinate and is a gulonic acid methyl ester-rich component, the gulonic acid methyl ester purity is more than 35%, and the gulonic acid methyl ester-rich component and the second component are combined and concentrated, and returns to the original process to be esterified, converted and crystallized so as to be recycled.

The invention discloses a cleaner production method of Vitamin C. In the method, the steps of converting sodium gulonic acid into gulonic acid are as follows: a. strong acid is added into sodium gulonic acid solution and then the pH of the solution is regulated to be 3.0 to 4.5; b. the solution film with the regulated pH is ultrafiltered; c. the strong acid is added into the ultrafiltered solution so as to regulate the pH to be 1.6 to 1.7; d. the solution with regulated pH in step c is subjected to concentration, crystallization and centrifugation so as to prepare mixed dry products; e. the mixed dry products are dissolved in methanol and then filtered so as to prepare the methanol solution of the gulonic acid. The production method of the invention has the advantages that a large amount of water, caustic soda liquid and hydrochloric acids are saved, and basically no sewage is discharged in the process of conversion, thereby solving the problem of environmental contamination, and meeting the discharge standard prescribed by the state. The production method has basically no loss on yield in the process of conversion, thereby greatly reducing investment and production costs.

The invention relates to a gene capable of regulating and controlling inhibition of vitamin C fermented strain acid production during acid stress and belongs to the technical fields of basic research on vitamin C prepared by fermentation and biological engineering. The key gene for regulating and controlling yield reduction of acid stress induced 2-Keto-L-Gulonic acid (2-KLG) in the production strains is determined, so a metabolic engineering strategy with high pertinence is generated. The expression condition of genes, in particular membrane transport protein genes, of acid-forming bacteria in the vitamin C mixed bacteria fermented system is analyzed by an expression profile chip technology. Compared with ketogenic gulonic acid bacteria subjected to single cultivation, the ketogenic gulonic acid bacteria subjected to mixed bacteria cultivation have 45 upregulated membrane transport protein genes and 15 down regulated membrane transport protein genes. The notable thing is that: 7 iron ion transport protein genes are inhibited to express. After acid stress, the iron ion transport protein genes are over expressed. During fermentation, the iron ions are added to inhibit mixed bacteria to ferment and generate acid. The result shows that the iron ion transport protein exerts reverse action in the process of yield reduction of 2-KLG generated by the acid stress induced mixed bacteria fermentation.

The invention relates to a synthetic process for vitamin C and sodium salts, which pertains to the technical field of chemistry. Gulonic acid and methanol carry out esterification reaction and then react with the sodium salts. The synthetic process is characterized by be executed according to the following steps: (1) the gulonic acid is dissolved in methanol and then goes through a filter with the aperture ranging from 1 Mum to 50 Mum and goes through a filter with the aperture ranging from 0.01 Mum to 1 Mum and a powdered activated carbon layer for gradient decolorization, thus obtaining a destaining solution for further use; (2) the destaining solution circularly reacts for 1.2 hours to 1.8 hours in macroporous hydrogenous resin, thus obtaining esterification solution; (3) the esterification solution is injected into a normal pressure container, kept being stirred at the temperature of 70 DEG C plus or minus 2 DEG C and uniformly added with suspending liquid obtained by mixing sodium bicarbonate with the mole number of 1.01 times to 1.05 times of the gulonic acid or the sodium carbonate with the mole number of 0.505 time to 0.51 time of the gulonic acid with methanol, and then is cooled to the room temperature after thermal insulation is executed continuously, thus obtaining the vitamin C sodium salts after centrifugal dehydration. The process has short synthesis time, products with high purity and good quality, and low energy consumption.

The invention relates to treatment of a gulonic acid mother liquor in VC production, and a method for preparing a fertilizer by using a product obtained by the treatment, wherein the treatment of thegulonic acid mother liquor comprises: hydrolyzing original bacteria or externally added other bacterial residue in a waste liquid at a temperature of 105-121 DEG C by using hydrochloric acid containedin a gulonic acid mother liquor, and the hydrolyzate is neutralized with potassium hydroxide to achieve a pH value of 4-9 to produce an amino acid potassium fertilizer, or the amino acid potassium fertilizer as a culture medium is inoculated with 0.5-2% of Bacillus, and fermentation culture is performed to produce a microbial potassium fertilizer. According to the present invention, the method using the hydrochloric acid contained in the gulonic acid mother liquor or the method for treating the waste with the waste develops the new resource utilization way for the gulonic acid mother liquor,and further develops the new way for treating other waste bacterial residue.