109 results about "Pantoic acid" patented technology

The invention relates to a method used microorganism enzyme resolution DL-pantoic acid lactone to produce D-pantoic acid. It uses the D-pantoic acid lactone hydrolase strain of the fusarium, gibberella, aspergillus, penicillium, rhizopus, gliocladium, aureobasidium to ferment and culture, uses wet thallus as coarse enzyme, DL-pantoic acid lactone as substrate to produce D-pantoic acid. L- pantoic acid lactone can be reclaimed. The DL-pantoic acid lactone gained by racemization reaction can newly be used to do resolution.

The invention relates to the technical field of biosynthesis, and concretely discloses a preparation method of D-pantoic acid inner ester. The preparation method at least comprises the following stepsthat 1, valine is subjected to enzymatic conversion under the effects of catalase and alpha-ketoisovaleric acid reductase to obtain alpha-ketoisovaleric acid; 2, the alpha-ketoisovaleric acid and tetrahydrofolic acid react under the effects of hydroxylmethyl transferase and magnesium chloride to prepare ketopantoic acid; 3, ketopantoic acid is subjected to enzymatic conversion under the effects of alpha-ketoisovaleric acid reductase to obtain pantoic acid; 4, pantoic acid is prepared into the pantoic acid inner ester. The operation process is simple; the conversion condition is mild; the production efficiency is high; the product quality is high; the enzyme used in the process can be cyclically used; the preparation method belongs to an energy-saving, environment-friendly, green and efficient production technology; the industrial production is convenient.

The present invention features improved methods for the enhanced production of pantoate and pantothenate utilizing microorganisms having modified pantothenate biosynthetic enzyme activities and having modified methylenetetrahydrofolate (MTF) biosynthetic enzyme activities. In particular, the invention features methods for enhancing production of desired products by increasing levels of a key intermediate, ketopantoate by enzymes that contribute to its synthesis. Recombinant microorganisms and conditions for culturing same are also are featured. Also featured are compositions produced by such microorganisms.

The invention relates to a method for acquiring genetically engineered bacteria capable of efficiently producing pantothenic acid by introducing the pantothenic acid synthetase genes from different sources into escherichia coli. An efficient escherichia coli protein expression system is adopted for heterologously expressing the pantothenic acid synthetase from different sources, thereby acquiring high-activity pantothenic acid synthetase strain. The strain can efficiently express the pantothenic acid synthetase and convert substrate pantoic acid and beta-alanine into pantothenic acid; the enzyme activity reaches up to 33.52U / mL; the engineering bacteria are fermented for 38h; the generated pantothenic acid is 101.2g / L. The strain has the characteristics of high activity, short fermentation time, high yield, and the like.

The invention provides a method for preparing D-pantoyl lactone by fermentation. The method comprises the following steps: culturing fusarium oxysporum in a culture medium; (2) mixing the cultured fusarium oxysporum with a substrate and hydrolyzing to obtain D-pantoic acid; (3) carrying out lactonization on the D-pantoic acid in the step (2) to obtain the D-pantoyl lactone, wherein the culture medium is prepared from 1 percent to 10 percent of glycerol, 0.2 percent to 2 percent of peptone, 0.2 percent to 2 percent of yeast extract, 0.5 percent to 5 percent of maize slurry, 0.05 percent to 0.2 percent of spirulina polysaccharide, 0.1 percent to 1 percent of glycine and the balance of water.

The invention relates to a recovery method for D-calcium pantothenate mother liquor. The recovery method comprises the following steps: subjecting D-calcium pantothenate mother liquor to recovery of methanol; heating a product obtained after recovery of methanol, adjusting a pH value to 1-3, carrying out sufficient stirring, then performing cooling to room temperature, adjusting a pH value to 4-6,carrying out filtering, and respectively recovering precipitated calcium sulfate and a filtrate; adding ethyl acetate into the obtained filtrate, carrying out sufficient extracting, respectively collecting an ethyl acetate phase and a water phase, and recovering D-pantoic acid lactone in the ethyl acetate phase; and concentrating, cooling and crystallizing the obtained water phase, carrying out filtering and respectively recovering crystallized beta-aminopropionic acid and a filtrate containing D-calcium pantothenate. According to the method provided by the invention, the residual target products and a small amount of unreacted raw materials and auxiliary materials in the D-calcium pantothenate mother liquor are efficiently and fully recovered according to a specific sequence, so resourcewaste and environmental pollution are avoided; and the method has the advantages of easy availability of raw materials, environment-friendly and safe conditions, short reaction time, recyclable solvents, simple steps, high purity of recovered products and the like.

The invention relates to the field of chemical synthesis and discloses a method for preparing D-calcium pantothenate. The method comprises the following steps of: reacting 3-aminopropionitrile with aqueous solution of sodium hydroxide or potassium hydroxide at the temperature of between 96 and 100 DEG C, adding hydrochloric acid to regulate the pH value, and concentrating and crystallizing; dissolving the prepared crystal by using methanol, and filtering to remove salt; and performing ion exchange with 732-type calcium ion exchange resin, carrying out acylation reaction between reaction liquid and D-pantolactone, crystallizing at the temperature of between 15 DEG C below zero and 10 DEG C below zero, and drying to prepare the D-calcium pantothenate, wherein the molar ratio of the 3-aminopropionitrile to the sodium hydroxide and the potassium hydroxide is 1:1.1-1.4, and the molar ratio of the 3-aminopropionitrile to the D-pantolactone 1:1-1.1. The method for preparing the D-calcium pantothenate has the advantages of not using calcium oxide or calcium hydroxide as a calcification agent, avoiding water generation in the reaction process so as to ensure the smooth acylation process, along with simple process, high yield, recycled 732-type calcium ion exchange resin, no generation of three wastes, and contribution to industrial production.

The invention relates to laevorotation lactone specificity hydrolytic enzyme producing strain and the method used it to make chirality oxyacid. The enzyme producing is Fusarium proliferatum Nirenberg ECU2002 with storage number CGMCC 1494. The chirality oxyacid preparing method includes the following steps: using the fungous mycelium, rough enzyme extract or their immobilization derivative as biocatalyst; processing antipode selectivity hydrolysis resolution for a series of racemic chirality lactone to gain many optically active (+)-oxyacid and (+)-lactone which can be hydrolyzed into (-)-oxyacid; (+)-alpha-hydroxyl-beta, beta-dimethyl-gamma-butyric acid which are D-(+)-pantoic acid; simple acidizing to gain chirality intermediate D-(-)-pantoic acid lactone widely used in preparing feed and daily chemical engineering industry.

The present invention relates to a method for producing D-panalcohol by using D-pantoic acid lactone prepared by microbial enzymic hydrolysis anid beta-aminopropanol and making them produce reaction.This invention utilizes the microbial strain fusarium monoliforme SW-902 (i,e, CGMCC No.0536) which can high-produce D-pantoic acid lactone hydrolase and screened and preserved by the inventor to make fermentation and enzyme production, and then to make hydrolysis of DL-pantoic acid lactone, after lactonization of D-pantoic acid, so as to obtain pure product D-pantoic acid lactone, and the D-pantoic acid lactone and beta-aminopropanal are reacted to obtain the invented D-panalcohol.

The invention relates to a preparation method of a vitamin B5 crude product. The preparation method comprises the process steps of carrying out adsorption separation on DL-pantolactone enzymatic hydrolysate by virtue of macroporous adsorption resin, and carrying out purification, filtration, lactonization, solvent extraction, D-calcium pantothenate synthesis and vacuum drying, so as to obtain theVB5 crude product. The method for separating and extracting D-pantoic acid and L-pantolactone by virtue of the macroporous adsorption resin has beneficial effects that the problems that the quality and yield are reduced due to the emulsion easily occurring in an extraction process are overcome; meanwhile, VB5 is separated out by virtue of a solvent-method crystallization technique, so that the crystallization time is shortened, the extraction yield is increased, the product quality is improved, and the economic benefit is increased; and the method is simple and convenient, low in solvent loss,strong in operability and suitable for industrial production, and the environmental pollution can be reduced.

The invention relates to the technical field of genetic engineering, particularly relates to L-pantoyl lactone dehydrogenase and application thereof, and discloses L-pantoyl lactone dehydrogenase derived from Cnuibacter physcomitrellae for the first time. The L-pantoyl lactone dehydrogenase has excellent enzyme activity on L-pantoyl lactone and can be applied to catalytic synthesis of D-pantoyl lactone. A system for chiral inversion synthesis of D-pantoyl lactone with L-pantoyl lactone by multi-enzyme cascade catalysis is constructed, and a genetically engineered bacterial cell co-expressing L-pantoyl lactone dehydrogenase, D-ketopantolactone reductase and glucose dehydrogenase is used as a catalyst, accumulation and spontaneous hydrolysis of the intermediate product ketopantolactone is avoided, and the steps of separating the intermediate product, racemizing the L-pantoyl lactone, lactonizing D-pantoic acid under an acidic condition are omitted, so that the reacting process is simplified, use of acid and alkali is reduced, and the reaction efficiency is improved.

The invention provides a preparation method of dianhydrol. The preparation method comprises the following steps: carrying out acylation reaction on D-pantoic acid lactone and 3-aminopropanol in a solvent to obtain a mixed solution containing D-panthenol, wherein the reaction temperature of the acylation reaction is 10-35 DEG C. The yield of the D-panthenol obtained by the preparation method is high, and the purity is high; meanwhile, the recovered solvent can be continuously used, and is safe and environment-friendly.

The invention relates to an immobilization method of D-pantoic acid lactone hydrolase, comprising the steps: (1) mycelium is pretreated to obtain the target enzyme liquid; (2) macroporous ion exchange resin is pretreated and added with glutaric dialdehyde solution for cross linking; (3) after that, the obtained macroporous ion exchange resin is suspended the D-pantoic acid lactone hydrolase solution and washed by de-ionized water to remove resolvase, so that the immobilized D-pantoic acid lactone hydrolase is obtained. The invention takes the macroporous ion exchange resin D201GF as a carrier and immobilizes the D-pantoic acid lactone hydrolase by two-step reaction of glutaric dialdehyde, thus overcoming the defect of the traditional direct reaction of mycelium. The immobilized enzyme granules have uniform shapes, good mechanical strength and large superficial area, so as to greatly improve the load capacity of enzyme and enhance enzyme activity; the recovery rate of enzyme activity reaches 85% in average, and the recycling times of the immobilized enzyme reaches above 50, so that the reaction cost of the resolvase is reduced. Furthermore, the method has simple technique and little investment for production equipment.

The invention provides a pantolactone hydrolase mutant strain and application thereof. Specifically, the invention provides a mutant pantolactone hydrolase with significantly improved enzyme activity and a host cell for expressing the mutant pantolactone hydrolase. The invention further provides an enzyme preparation containing the mutant pantolactone hydrolase, D-pantolactone in a substrate DL-pantolactone can be completely hydrolyzed into D-pantoic acid, and the mutant pantolactone hydrolase is high in conversion rate, stable in property, capable of being repeatedly used and suitable for industrial production.

The invention relates to the technical field of genetic engineering, in particular to L-panolactone dehydrogenase and application thereof. The L-panolactone dehydrogenase derived from Nocardia asteroides is disclosed for the first time, and the L-panolactone dehydrogenase has good enzymatic activity to L-panolactone, and can be applied to catalytic synthesis of D-panolactone. A system of multi-enzyme cascade catalysis for chiral overturning of the L-panolactone to synthesize the D-panolactone is established, genetically engineered bacterium cells for co-expression of the L-panolactone dehydrogenase, D-ketopantolactone reductase and glucose dehydrogenase are taken as a catalyst, accumulation and spontaneous hydrolysis of an intermediate product, namely ketopantolactone are avoided, the steps of separation of the intermediate product, racemization of the L-panolactone, lactonization of D-pantoic acid under the acid condition and the like are omitted, the reaction process is simplified, using of acid and base is reduced, and the reaction efficiency is improved.

The invention relates to a refining method of D-pantolactone. The method comprises the following steps: mixing a D-pantolactone crude product with the specific rotation of more than -45.0 degrees withethyl acetate, heating and fully dissolving, controlling the temperature at 20-30 DEG C, adding a D-pantolactone seed crystal with the specific rotation of -45.0 degrees to -50.7 degrees, slowly cooling to 5-10 DEG C, carrying out heat preservation crystallization, filtering, and collecting the crystal. The refining method of the D-pantolactone has the advantages that the raw materials are easy to obtain, the conditions are environmentally friendly and safe, the reaction time is short, the solvent can be recycled, the steps are simple, the obtained product is large and uniform in crystal, thespecific rotation can reach -49.5 degrees to -50.5 degrees, the purity can reach 98.0%, and the refining method has extremely high industrial utilization value.

The invention relates to the technical field of gene engineering, in particular to L-pantoyl lactone dehydrogenase and application of the L-pantoyl lactone dehydrogenase. The L-pantoyl lactone dehydrogenase derived from Nocardia farcinica is disclosed for the first time, has good enzymatic activity to L-pantolactone and can be applied to catalytic synthesis of D-pantolactone. A system for multi-enzyme cascade catalysis of chiral overturning of the L-pantolactone to synthesize the D-pantolactone is constructed, genetically engineered bacterium cells for co-expression of the L-pantoyl lactone dehydrogenase, D-ketopantolactone reductase and glucose dehydrogenase are taken as a catalyst, accumulation and spontaneous hydrolysis of an intermediate product, namely ketopantolactone, are avoided, the steps such as separation of the intermediate product, racemization of the L-pantolactone and lactonization of D-pantoic acid under the acid condition are omitted, the reaction process is simplified, using of acid and alkali is reduced, and the reaction efficiency is improved.

The invention relates to the technical field of genetic engineering, in particular to L-pantolactone dehydrogenase and application thereof. The L-pantolactone dehydrogenase derived from Nocardia cyriacigeorgica is disclosed for the first time, the L-pantolactone dehydrogenase has good enzymatic activity for L-pantolactone and can be applied to the catalytic synthesis of D-pantolactone. A system for synthesizing D-pantolactone through multi-enzyme cascade catalyzed L-pantolactone chiral inversion is constructed, genetically engineered bacterial cells which co-express L-pantolactone dehydrogenase, D-keto-pantolactone reducase and glucose dehydrogenase are used as catalysts, the accumulation and spontaneous hydrolysis of the intermediate keto-pantolactone are avoided, the separation of intermediates, racemization of L-pantolactone and D-pantoic acid lactonization under acidic conditions are omitted, the reaction process is simplified, the use of acid-base is reduced, and the reaction efficiency is improved.

The invention discloses a lactone hydrolase mutant in the fields of genetic engineering technology and biocatalysis, and application of the lactone hydrolase mutant as a biocatalyst to resolution of D, L-pantolactone to prepare optically pure D-pantoic acid. The lactone hydrolase mutant is obtained by carrying out single mutation or combined mutation on 1-14 amino acid sites in an amino acid sequence SEQ NO: 2 by taking fusarium oxysporum lactone hydrolase as a parent enzyme. The hydrolysis activity of the lactone hydrolase mutant on D, L-pantolactone is improved by 2.3-128 times compared withthat of parent enzyme. The high-activity lactone hydrolase provided by the invention can be used for large-scale production of preparing optically pure D-pantoic acid by biologically splitting D, L-pantolactone.

The invention discloses a strain of agrobacterium radiobacter of specificity hydrolytic dextral lactone and the application of the agrobacterium radiobacter in preparing dextral lactone compound such as D-(-)-pantoic acid lactone and the like. The enzyme-producing bacterium is the agrobacterium radiobacter and the preservation number is CGMCC2371. Thalli of the bacterium and an immobilized derivative of the bacterium are taken as biocatalyst, dexpazufloxacin which are not needed in pantoic acid lactone racemic mixture is hydrolyzed into (-)-hydroxy acid directly, useful D-(-)-pantoic acid lactone is preserved, and the latter is widely used in producing feedstuff and D-calcium pantothenate and D-panthenol which are useful in the daily chemical industry.

The invention belongs to the technical field of enzyme engineering, and specifically relates to a method for efficiently extracting D-pantoic acid from an enzymatic hydrolysate. According to the method for efficiently extracting D-pantoic acid from the enzymatic hydrolysate, on the basis of the prior extraction method of D-pantoic acid, the extraction of D-pantoic acid from the enzymatic hydrolysate by using existing ethyl acetate as an extracting agent is more thorough through further optimization of the extracting agent. For the technical process of taking D-pantoic acid as a target extract,the interference of other substances in the water phase after the extraction is effectively eliminated, so that the subsequent refining treatment process is simpler.

The invention discloses a D-pantolactone hydrolase producing bacteria in the technical field of bioengineering, and relates to a Thielavia sp. Zmu20201 serving as a microorganism, and the preservationnumber of the strain is CCTCC: M2020062. Meanwhile, the invention further discloses the D-pantolactone hydrolase from the Thielavia sp. Zmu20201 strain and an engineering bacterium strain of the D-pantolactone hydrolase, wherein the amino acid sequence number of the D-pantolactone hydrolase is as shown in SEQ ID NO.2. In addition, the invention also discloses an application of the Thielavia sp. Zmu20201 fermentation thallus or the lactone hydrolase gene engineering strain fermentation thallus as a biocatalyst in resolution of D, L-pantolactone to prepare optically pure D-pantoic acid.

The invention discloses a chiral 2-hydroxy-1, 4-dicarbonyl compound synthesized by catalyzing asymmetric Aldol reaction of fatty aldehyde and glyoxylate or fatty aldehyde and acylformaldehyde monohydrate by taking tetrapeptide TP or an enantiomer ent-TP thereof as a chiral catalyst, and application of a synthetic product. The method for synthesizing the chiral 2-hydroxy-1, 4-dicarbonyl compound through the asymmetric Aldol reaction is shown as a formula 1 and a formula 2. The asymmetric Aldol reaction of fatty aldehyde and glyoxylate or fatty aldehyde and acylformaldehyde monohydrate is catalyzed to synthesize the optically active 2-hydroxy-1, 4-dicarbonyl compound, then the optically active pantoic acid lactone can be further synthesized, and the method has advantages of mild reaction conditions, easy operation, low catalyst consumption, high yield and the like, and can synthesize the 2-hydroxy-1, 4-dicarbonyl compounds with two configurations by using tetrapeptide and the enantiomerthereof.

The invention discloses a strain of agrobacterium radiobacter of specificity hydrolytic dextral lactone and the application of the agrobacterium radiobacter in preparing dextral lactone compound suchas D-(-)-pantoic acid lactone and the like. The enzyme-producing bacterium is the agrobacterium radiobacter and the preservation number is CGMCC2371. Thalli of the bacterium and an immobilized derivative of the bacterium are taken as biocatalyst, dexpazufloxacin which are not needed in pantoic acid lactone racemic mixture is hydrolyzed into (-)-hydroxy acid directly, useful D-(-)-pantoic acid lactone is preserved, and the latter is widely used in producing feedstuff and D-calcium pantothenate and D-panthenol which are useful in the daily chemical industry.

The invention relates to L-pantoic acid lactone dehydrogenase TpLPLDH with good water solubility, a mutant and a coding gene thereof, and application of the L-pantoic acid lactone dehydrogenase TpLPLDH in preparation of a D-pantoic acid precursor intermediate ketone pantoic acid lactone through biological catalysis. The amino acid sequence of the L-pantoic acid lactone dehydrogenase TpLPLDH is as shown in SEQ ID NO. 1 (sequence identifier number 1). The novel protein with the activity of the L-pantoic acid lactone dehydrogenase has the advantages that the protein has the effect of the L-pantoic acid lactone dehydrogenase, L-pantoic acid lactone can be catalyzed to be dehydrogenated to generate keto-pantoic acid lactone, the solubility of the protein is good, and the protein can be used for preparing keto-pantoic acid lactone. The compound is almost completely dissolved in an aqueous solvent (such as a phosphate buffer). According to the mutant with the L-pantoic acid lactone dehydrogenase activity, after the mutant reacts in a system of a substrate L pantoic acid lactone with the final concentration of 1 mM for 30 min at the temperature of 30 DEG C and the speed of 1200 rpm, compared with a wild type, the mutant has the advantages that the substrate conversion rate is increased by 1.84 times, and the enzyme activity is increased by 1.12 times compared with the wild type.

The invention provides a novel process for preparing a D-pantothenic acid calcium salt and relates to a novel process for preparing a D-pantothenic acid calcium salt. According to the basic scheme ofthe invention, the novel process comprises the following steps of: preparing a water solution with the content of 16-26% from DL-pantolactone, adding a biological enzyme which is 5-15% of mass of thewater solution, performing uniform mixing, and then slowly dropwise adding a calcium di-beta-alaninate water solution in which the content of calcium di-beta-alaninate is 20-30% in a molar amount of 0.49-0.51 fold that of D-pantolactone contained in the water solution at 23-30 DEG C; performing reaction all the time until complete hydrolysis of the D-pantolactone, filtering out a clear liquid andrecycling the biological enzyme in a filter cake; keeping a pH value of a reaction system between 6.0 and 7.3 during dropwise adding; concentrating the clear liquid in vacuum of -0.085MPa or above andat 115-120 DEG C after reaction is completed, performing cooling to 65 DEG C or below, adding methanol of which the weight is 0.5-0.8 fold that of D-pantothenic acid calcium di-beta-alaninate, performing mixing and cooling to 20-40 DEG C, performing filtering, and washing, pulping and rewashing the filter cake with the methanol to achieve purification; collecting filtrate of a few times, recovering the methanol and then forming the DL-pantolactone in a racemization manner to reuse; and drying the filter cake through two levels of temperature zones of 105-125 DEG C and 150-160 DEG C, and performing cooling and discharging to obtain the D-pantothenic acid calcium salt. According to the novel process, a precursor substance D-pantothenic acid calcium di-beta-alaninate of the D-pantothenic acid calcium salt is generated in one step during resolution reaction, so that the process of extracting and separating the D-pantolactone is avoided.

The invention relates to a preparation method of D pantoic acid lactone with high conversion rate, which comprises the following steps: an enzyme preparation is added into a substrate solution to prepare D pantoic acid lactone through conversion, the substrate is DL pantoic acid lactone and ammonium formate, and the enzyme preparation is a compound enzyme composed of L pantoic acid lactone dehydrogenase, ketone pantoic acid lactone reductase and formate dehydrogenase. The preparation method has the advantages of simplicity and convenience in operation, environment friendliness and the like.