157 results about "Batch fermentation" patented technology

The invention discloses a Raman spectrum-based online detection method for glucose and ethanol in a yeast fermentation process. The Raman spectrum-based online detection method specifically comprises the following steps: preparing a yeast seed solution, and inoculating the yeast seed solution into a fermentation tank for fermentation; in the fermentation process, an immersion type Raman optical fiber probe is used for collecting a Raman spectrum in real time, and meanwhile sampling is conducted at regular time to measure the concentration of glucose and ethanol; sequentially carrying out median filtering, adaptive iteration reweighted penalty least square method (airPLS) baseline correction, Savitz-golay (SG) smoothing processing and characteristic peak normalization on the acquired spectrum; respectively establishing concentration prediction models of glucose and ethanol by adopting a PLS1 algorithm; and applying the model to a subsequent batch fermentation process to realize online prediction of glucose and ethanol. According to the method, manual sampling is not needed, prediction is accurate, operation is easy and convenient, and in-situ online detection of glucose and ethyl alcohol in the yeast fermentation engineering can be achieved.

The invention provides a genetically engineered bacterium for producing O-succinyl-L-homoserine as well as a construction method and application of the genetically engineered bacterium. In a chassis bacterium genome, the expression of a 2-ketoglutaric acid decarboxylase encoding gene sucA is enhanced, and the expression of a succinyl-coenzyme A synthetase encoding gene sucD is weakened, so that the supply of succinyl-coenzyme A is increased; the method comprises the following steps: increasing the NADPH (Nicotinamide Adenine Dinucleotide Phosphate) reducing capacity and ATP (Adenosine Triphosphate) energy supply of a chassis bacterium, increasing DNA (Deoxyribose Nucleic Acid) in combination with a transcription dual regulatory factor ompR to improve the stress resistance of escherichia coli under high osmotic pressure, and introducing an overexpression plasmid containing a homoserine transsuccinylase coding gene metA to construct the genetically engineered bacterium for producing O-succinyl-L-homoserine. The engineering strain obtained through a systematic metabolic engineering modification strategy can realize effective accumulation of OSH, the shake flask yield of OSH reaches 19.8 g / L, the fed-batch fermentation yield of a 5L fermentation tank reaches 110.5 g / L, the sugar-acid conversion rate reaches 52.6%, and a foundation is laid for subsequent construction of high-yield OSH engineering bacteria.

The invention discloses a spermidine synthetase mutant and application of the spermidine synthetase mutant in synthesis of spermidine, and belongs to the technical field of genetic engineering and metabolic engineering. According to the method, the spermidine synthetase is mutated firstly, the efficiency of the spermidine synthetase is improved, and on the basis, the dissolved oxygen and fed-batch concentration are optimally controlled through fermentation, so that the yield of spermidine is improved. The corynebacterium glutamicum provided by the invention is an engineering bacterium with plasmids and without defects, and after the corynebacterium glutamicum is subjected to fed-batch fermentation in a 5L fermentation tank for 68 hours, the yield of spermidine reaches 25.1 g / L, and the sugar-acid conversion rate is 17.9%.

The invention relates to the field of bioengineering, in particular to a construction method and application of recombinant yarrowia lipolytica with high succinic acid yield. The recombinant yarrowia lipolytica disclosed by the invention simultaneously expresses a reduced tricarboxylic acid pathway related gene, a glyoxylic acid circulation pathway related group, a heterologous dicarboxylic acid transport related gene and a mitochondrial transport related gene, and an endogenous dicarboxylic acid transport protein coding gene is also knocked out; and a natural promoter of an isocitrate dehydrogenase coding gene IDH1 is replaced. The recombinant yarrowia lipolytica constructed by the invention can produce 112.85 g / L of succinic acid after fed-batch fermentation under the condition of not controlling pH, and efficient synthesis of succinic acid in the yarrowia lipolytica is realized.

The invention provides a method for producing glycosyltransferase or sucrose synthase by fermenting recombinant escherichia coli, and belongs to the technical field of biological fermentation. According to the fermentation method provided by the invention, a three-stage material supplementing control fermentation strategy is adopted, namely after fermentation is carried out for 5 hours, material supplementing operation is started, the material supplementing speed is 6.24-7.20 g / L / h, fermentation is carried out for 8-10 hours, the material supplementing speed is reduced to 4.80 g / L / h, the temperature is reduced to 25 DEG C, and induction is carried out; then maintaining the material supplementing speed until the fermentation OD600 is 65-75, and then reducing the material supplementing speed to 3.84-4.80 g / L / h until the OD600 is not obviously increased any more; the material supplementing mode has remarkable advantages in the aspects of improving fermentation OD600 and fermentation enzyme activity, shortening the fermentation period, reducing the cost and the like.

The present invention relates to the process to prepare recombinant lectin having amino acid sequence of SEQ ID NO:1, wherein the process comprises fed-batch fermentation of a clone in a host cell at specific feeding rate with carbon to Nitrogen ratio of 3:1 to 6:1. The invention also relates to the process of purification of recombinant lectin having amino acid sequence of SEQ ID NO: 1.

The invention provides a construction method and application of recombinant yarrowia lipolytica with high yield of tyrosol, and belongs to the field of bioengineering. The recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain, and the recombinant yarrowia lipolytica with high yield of tyrosol is named as a yarrowia lipolytica XJ-TL strain. The recombinant yarrowia lipolytica disclosed by the invention expresses an ethanol dehydrogenase encoding gene, and simultaneously expresses any one or a combination of more of a DAHP synthase mutant encoding gene, a chorismate mutase mutant encoding gene, a glucose 6-phosphate dehydrogenase encoding gene, a transketolase encoding gene and a transaldolase encoding gene; meanwhile, a natural promoter of a pyruvate kinase coding gene is truncated. After the recombinant yarrowia lipolytica XJ-TL strain is subjected to fed-batch fermentation, tyrosol can be synthesized at a relatively high level. The construction method of the recombinant yarrowia lipolytica is easy to operate, and efficient synthesis of tyrosol can be achieved.

The preparation method comprises the following steps: mixing salvia miltiorrhiza-schisandra chinensis superfine powder and roxburgh rose normal juice in a ratio of 3: 2: 5, carrying out composite enzymolysis and saccharomyces cerevisiae pretreatment, and sequentially carrying out aspergillus oryzae solid-state fermentation, acetobacter pasteurianus liquid-state index fed-batch fermentation and lactobacillus plantarum anaerobic flavor fermentation, so as to obtain the vinegar product with the homology of medicine and food. And parameters are regulated and controlled in real time through the three-layer stacked LSTM neural network. According to the invention, the conversion rate of tanshinone IIA to tanshinol reaches 65.8%, the total acid is greater than or equal to 7.2 g / 100mL, at least 21 flavor substances are contained, the batch difference rate is 4.2%, the energy consumption is reduced by 29.2%, the equipment utilization rate is increased by 50%, the product safety reaches the standard, and the method has a health-care function and palatability, and is suitable for industrial production.

The invention discloses a yeast fed-batch fermentation nutrition addition prediction method based on deep learning, which establishes a bimodal space-time prediction model and can predict the nutrition addition amount of traditional strains. When a new strain needs to be subjected to yeast fed-batch fermentation, network parameters of the dual-mode space-time prediction model after field adaptation are determined through an MMD loss function, the loss function is consolidated through an elastic weight, the network parameters of the dual-mode space-time prediction model after field adaptation are optimized, and the network parameters of the dual-mode space-time prediction model after field adaptation are optimized; an optimized bimodal space-time prediction model is obtained; and on the basis of the optimized bimodal space-time prediction model, obtaining the fused characteristics of the fermentation process parameters of the new strain so as to obtain the predicted nutrition addition amount. The control strategy can be automatically adjusted and optimized for a new strain or a new fermentation process, the adaptability and flexibility of the system are greatly improved, and the development time and cost of the new strain are reduced.

The invention discloses recombinant escherichia coli capable of increasing the yield of L-homoserine and a preparation method of the recombinant escherichia coli, and belongs to the technical field of biology. By constructing a multi-way glucose transport system, L-homoserine synthesis precursor supply is enhanced, high-energy-consumption genes are knocked out, ATP regeneration is enhanced, ATP availability is improved, an acetic acid conversion way is established, and an L-homoserine degradation way is blocked, so that a metabolic flow of a synthesis way is further enhanced; a pyruvate carboxylase mutant and a designed promoter are introduced to weaken expression of citrate synthase and reconstruct key metabolic nodes, and fed-batch fermentation conditions of the strain in a 5L bioreactor are optimized according to metabolic characteristics of the strain; the genetically engineered bacterium which is clear in genetic background, does not carry plasmids, does not need induction and can stably and efficiently produce the L-homoserine is obtained, the yield of the L-homoserine reaches 141.5 g / L after the L-homoserine is fermented for 56h in a 5L fermentation tank, the saccharic acid conversion rate reaches 40%, and the genetically engineered bacterium has a certain application prospect.

The invention discloses a construction method of recombinant corynebacterium glutamicum for fermentation synthesis of poly beta-(1, 6)-N-acetylglucosamine polysaccharide, and belongs to the technical field of bioengineering. The recombinant corynebacterium glutamicum capable of preparing the PNAG through fermentation is constructed by using synthetic biology and genetic engineering technologies for the first time, cheap carbon sources and nitrogen sources can be utilized, the recombinant corynebacterium glutamicum is suitable for high-density fed-batch fermentation, and the generation cost of the PNAG is reduced; the yield after 42 hours of fermentation can reach 6.82 g / L, and the method has industrial application potential.

The present invention belongs to the field of microbial metabolic engineering and synthetic biology technology applications, and particularly relates to a Pichia pastoris engineered strain that produces high sclareol production, as well as its construction method and application. A sclareol biosynthesis pathway is constructed in a host strain, and the intracellular mevalonate metabolic pathway and the central metabolic pathway are optimized, thereby obtaining an engineered bacterial strain A; the host strain is Pichia pastoris; or, metabolic regulatory factors are overexpressed or knocked out in the above-obtained engineered bacterial strain A, thereby obtaining an engineered bacterial strain B; or, in the above-obtained engineered bacterial strain B, a cell compartmentalization strategy is used to target the synthesis pathway to the peroxisome and optimize it, thereby obtaining an engineered bacterial strain C. The present invention provides a Pichia pastoris chassis cell for synthesizing the diterpenoid compound sclareol and an engineered strain for synthesizing sclareol; by introducing and optimizing the expression of the sclareol synthesis pathway, the sclareol yields in shake flask batch fermentation and bioreactor batch fed-batch fermentation reach 631.6 mg / L and 10.5 g / L, respectively.

The invention relates to a method for producing fermented vegetable oil by a semi-continuous fed-batch fermentation process, which comprises the following steps: firstly, inoculating a strain into a fermentation culture medium for thallus propagation under proper conditions, then mixing with crude vegetable oil according to a certain proportion, fermenting at proper temperature, and discharging part of fermentation liquor after the fermentation index reaches a set value, thereby obtaining the fermented vegetable oil. And returning a water phase containing active thalli to the fermentation tank, and meanwhile, supplementing fresh vegetable oil crude oil with the same mass as the water phase to continue fermentation. And repeating the steps until the time of the acid value of the oil phase of the fermentation liquid reaching the set value exceeds 24 hours or is more than two times of the time of the fermentation reaching the set value after the previous oil supplement, ending the fermentation, and centrifugally separating and purifying the separated oil phase to obtain the fermented vegetable oil. The production efficiency of the fermented vegetable oil can be greatly improved, the single-batch production yield is improved, the stability of the quality of the fermented oil is ensured, the production cost is saved, and the method can be widely applied to the cosmetic and skin care field, the food field or the pharmaceutical industry.

The invention relates to a preparation method of microbial exopolysaccharide and application of the microbial exopolysaccharide in a culture medium for tissue culture, and belongs to the technical field of microbial polysaccharide preparation. According to the preparation method of the microbial exopolysaccharide, the microbial exopolysaccharide is prepared in a batch fermentation mode by utilizing different microorganisms, a beet extract and supplementary materials are added into a culture medium of the microbial exopolysaccharide, the microorganisms are promoted to generate the exopolysaccharide, and further purification, enzymolysis and separation are performed to obtain the microbial exopolysaccharide. The finally obtained target microbial exopolysaccharide has good effects of promoting tissue culture seedling development and rooting, and can be applied to preparation of a culture medium for tissue culture.

The invention provides a combined type three-dimensional composting device, and relates to the technical field of waste resourceful treatment. A fermentation bag; a movable stacking rack; the fermentation shed is a telescopic shed body, the movable stacking frame is movably arranged in the fermentation shed, and a temperature detection device is further arranged in the fermentation shed. According to the invention, material loading and unloading can be carried out more conveniently, the space is fully utilized by adopting a stacking composting mode, sequencing batch fermentation can be carried out, air intake in the fermentation process can be controlled through the arrangement of the bidirectional air pipe, compost is fully fermented, the problem that the compost is not fully contacted with oxygen in the aerobic composting process is solved, and the aerobic composting efficiency is improved. And moreover, the device can be used for fermenting wastes on a large scale, the fermentation space is flexible and adaptive, and both the mounting convenience and the operation and maintenance cost are considered.

The present invention relates to a method for preparing microbial exopolysaccharides and their use in tissue culture medium, belonging to the technical field of microbial polysaccharide preparation. The method for preparing microbial exopolysaccharides utilizes batch fermentation of different microorganisms, adds a beet extract and a supplement to the culture medium to promote microbial exopolysaccharide production, and further purifies, enzymatically hydrolyzes, and separates the exopolysaccharides. The resulting target microbial exopolysaccharides have a good effect on promoting the development and rooting of tissue culture seedlings and can be used to prepare tissue culture medium.

The invention discloses a method for constructing a high-yield natamycin engineering strain through overexpression of an extracellular transport protein gene pimA and a pathway specific regulation gene pimR, and the high-yield engineering strain is applied to fermentation production of natamycin. The yield of the natamycin can reach 19.49 g / L through fed-batch fermentation in a fermentation tank of 5L of the high-yield strain S.ilvosporeus pimA-pimR, and the yield of the natamycin is improved by 33.12% compared with the yield of a chassis strain S.ilvosporeus TUST01; the production intensity is improved by 33.33% compared with the production intensity of a chassis strain S.ilvosporeus TUST01, and the strain has a relatively high industrial application value.

The invention discloses a method for non-induction biosynthesis of low molecular weight gamma-polyglutamic acid, which specifically comprises the following steps: cloning a gamma-polyglutamic acid synthetase gene cluster pgsBCA of bacillus licheniformis to corynebacterium glutamicum F343 with high glutamic acid yield for exogenous expression; a gamma-polyglutamic acid degrading enzyme gene pgdS and pgsBCA from Schlaughua Zehnder bacillus are subjected to tandem expression, so that the biosynthesis of the low-molecular-weight gamma-polyglutamic acid (LMW-gamma-PGA) is realized. Furthermore, natural promoters are screened, constitutive promoters with different intensities are introduced, and the optimal promoter Pdep-A16 is used for replacing the original inducible promoter Ptac, so that synthesis of the LMW-gamma-PGA without an inducer is realized. Finally, continuous fed-batch fermentation is carried out in a 15-L fermentation tank, the yield of the LMW-gamma-PGA reaches 37.42 g / L, and the production intensity is 0.78 g / L / h. And the LMW-gamma-PGA production process without adding an inducer is successfully constructed.

The invention relates to a feeding method for improving the fermentation unit of tylosin, which comprises the following steps: inoculating a seed solution into a fermentation tank filled with a liquid fermentation culture medium by taking streptomyces fradiae as a fermentation strain, producing tylosin by adopting a fed-batch fermentation process, fermenting for 12 hours, adding a sodium malonate solution, reacting for 2 hours, adding a VB12 solution, fermenting for 30-130 hours, and then adding a VB12 solution to react for 2 hours. And supplementing the mecobalamin solution every 12 hours. By adopting the material supplementing process, the fermentation unit of tylosin can be effectively increased by 2-4% or above, and the material supplementing process is suitable for industrial production.

The application discloses a construction method and application of a genetically engineered corynebacterium glutamicum for synthesizing a bacteriochlorophyllide amino acid shinorine, and belongs to the field of genetic engineering and fermentation technology. The genetically engineered corynebacterium glutamicum is constructed by taking corynebacterium glutamicum ATCC 13032 as a host strain, knocking out a competitive pathway gene, integrating a key enzyme gene in multiple copies on a genome, knocking out an acid production pathway gene and a serine dehydrogenase gene, and expressing a gene cluster for synthesizing shinorine on a plasmid, so that the synthesis yield of shinorine in a microbial cell is improved. The strain can produce shinorine in a flask, and 1.12 g / L of shinorine can be accumulated in 72 hours. In fed-batch fermentation in a 50 L fermenter, the strain can be cultured to 8.6 g / L.

The invention provides a VniNatt enzyme mutant, nucleic acid, a carrier or a host cell, and application of the VniNatt enzyme mutant, nucleic acid, a carrier or a host cell in fermentation production of NMN. Compared with a wild type, the mutant can catalytically produce NMN with higher efficiency, and has good thermal stability. BL21 engineering bacteria are constructed on the basis of the VniNamt enzyme mutant and are used for producing NMN through fermentation, and the yield of the NMN can reach 54 g / L by adopting a fed-batch fermentation process.

This invention discloses a nicotinamide phosphoribosyltransferase mutant, a recombinant strain, and a method for constructing the same. The mutants are I457V, G344A, Q378A, T283G, or F182A, obtained by single-point mutation of the wild-type nicotinamide phosphoribosyltransferase shown in SEQ ID NO:1. This invention utilizes a method derived from... Schizotequatrovirus By rationally designing and site-directed mutagenesis of KVP40 nicotinamide phosphoribosyltransferase, combined with metabolic engineering to block the NMN degradation pathway and enhance substrate synthesis and product transport, the yield of β-NMN by the recombinant strain during shake-flask fermentation reached 4.82 g / L. Through optimization of fed-batch fermentation conditions, the highest yield of β-NMN during fed-batch fermentation in shake flasks reached 15.60 g / L.

This invention discloses a method for the efficient synthesis of gastrodin using recombinant Corynebacterium glutamicum, belonging to the fields of genetic engineering, fermentation engineering, and synthetic biology. This invention uses Corynebacterium glutamicum as the host bacterium and knocks out… pobA The gene was expressed, and the engineered strain WN01 / pB6 was constructed using the expression plasmid pEC-XK99E to express cladoid-pyruvate lyase, carboxylic acid reductase, and glycosyltransferase, achieving heterologous synthesis of gastrodin. Further overexpression was then performed. aroG Genes, overexpression of protein-coding genes that activate carboxylic acid reductase sfp The engineered strain WN05 / pB6 was able to synthesize 7.37 g / L of gastrodin at the shake flask level, and the gastrodin yield reached 17.77 g / L at 72 h of batch fermentation in a 5-L fermenter, with a yield of 0.247 g / L / h. This provides a brand-new biomanufacturing platform for the industrial production of gastrodin.

The application discloses a kind of high-yield O-succinyl-L-homoserine recombinant escherichia coli and its construction method and application, belong to genetic engineering and fermentation engineering technical field.The recombinant escherichia coli is expressed by expressing anti-feedback inhibition homoserine O-succinyltransferase metA fbr , aspartate ammonia-lyase aspA, aspartate transaminase aspC, 2 ketoglutarate decarboxylase sucA, dihydrothioctic acid succinyltransferase sucB and efflux protein yjeH, knock out metJ, improve intracellular NADPH availability, to enhance OSH synthesis pathway metabolic flow, precursor succinyl coenzyme A supply and product efflux.Using 5 L bioreactor for fed-batch fermentation, after fermentation 60 h, OSH production is as high as 137.4 g / L, and sugar acid conversion rate reaches 50.5%.The application provides engineering strain and technical realization path for the efficient fermentation of OSH.

A method for preparing recombinant human albumin with high expression and low O-glycosylation level, comprising taking at least one measure during fermentation to improve the protein expression rate and reduce the O-glycosylation level of a protein; and specifically comprising: S1, seed solution preparation; S2, batch fermentation; S3, glycerol fed fermentation; S4, methanol fed fermentation; S5, fermentation product collection.

The present invention discloses a method for fermenting and preparing a multi-component short- and medium-chain fatty acids and a bacterial agent, and belongs to the technical field of brewing microorganisms. The present invention utilizes Bacillus lactis and Lithomonas sulfophilus to co-ferment and prepare a functional bacterial liquid, which can improve the production of caproic acid while also improving the biomass of the two microorganisms, and can also produce valeric acid. When the "simplified bacterial community" obtained by screening the caproic acid-producing functional simplified bacterial community screening method of the present invention is used to prepare a functional bacterial liquid, the caproic acid production is high and the butyric acid production is low, and at the same time, large-scale continuous production operation can be achieved through semi-continuous feeding operation, and the production efficiency is significantly better than the batch fermentation method. After the composite short- and medium-chain fatty acids produced by the technology of the present invention are enzymatically esterified, composite esters containing multiple esters can be obtained through conventional distillation extraction, which can be used for flavoring food and beverage wine. In addition, the bacterial agent obtained in the fermentation process of short- and medium-chain fatty acids can be used for cellar mud production and cellar maintenance.

The application discloses a genetically engineered bacterium for efficiently producing itaconic acid by using corn stalk hydrolysate and application thereof, and belongs to the technical field of gene recombination and metabolic engineering. The genetically engineered bacterium for efficiently producing itaconic acid by using corn stalk hydrolysate is characterized by the following aspects: taking a strain BW08 as a starting strain, heterologously expressing a homocentric acid decarboxylase gene CAD, deleting an isocitric acid dehydrogenase gene Icd, heterologously expressing a xylose acid dehydrase gene XD, a 2-keto-3-deoxy-xylose acid dehydrase gene KDXD and an alpha-ketoglutaric acid semialdehyde dehydrogenase gene KGSADH, and heterologously expressing a citric acid synthase gene gltA. In the fed-batch fermentation, the yield of itaconic acid of the genetically engineered bacterium reaches 68.6 g / L, and the conversion rate reaches 0.6 g / g (total sugar). The genetically engineered bacterium can efficiently metabolize corn stalk hydrolysate to produce itaconic acid.

The invention discloses a beta-elemene synthase and an application of the beta-elemene synthase in preparation of beta-elemene. On the basis of EL-Y0 chassis bacteria of high-yield precursor FPP, ERG20 (FPP synthase) and AarTPS72 (beta-elemene synthase) are subjected to fusion expression through flexible linker (GGGS) 2, and the metabolic flux of an MVA pathway is improved by overexpressing genes HIF-1, UPC2-1 and tHMGR, so that the biosynthesis of beta-elemene is improved; the engineering bacterium EL16 for producing the beta-elemene is successfully obtained; the final shake flask yield of the beta-elemene is 748.1 mg / L; in addition, the engineering bacterium EL16 is used for carrying out fed-batch fermentation in a 5L bioreactor, and the final titer of beta-elemene is 14.60 g / L.

The invention discloses a method for improving the yield of L-threonine by enhancing Escherichia coli electronic respiration transfer and energy transfer efficiency, and belongs to the technical field of genetic engineering. According to the invention, a cytochrome bo oxidase subunit CyoA mutant derived from escherichia coli, a cytochrome bo oxidase subunit CyoB mutant derived from pseudomonas aeruginosa, a cytochrome bd oxidase cydAB gene cluster derived from escherichia coli and a polyphosphate kinase PPK mutant derived from radiation-resistant coccus are introduced, amn genes are knocked out, and a cytochrome bo oxidase gene is obtained. The electron transfer efficiency in escherichia coli and the production efficiency of energy ATP (adenosine triphosphate) are enhanced, and the production efficiency of L-threonine and the carbon yield are further improved. Finally, the obtained engineering strain ECTHRS-5 is subjected to fed-batch fermentation in a 5L fermentation tank to accumulate 160.8 g / L of L-threonine, and the sugar-acid conversion rate is 0.62 g / g.

The invention discloses a Kluyveromyces marxianus engineering strain and an application thereof in high-temperature fermentation production of 3-hydracrylic acid, which are characterized in that heat-resistant Kluyveromyces marxianus is used as a platform, a gene editing technology is utilized, a glycerin metabolism way of the Kluyveromyces marxianus is reconstructed, and MCRN940V / K1106W / S1114R mutant gene is over-expressed to finally obtain a YZB521 strain. Under the aerobic condition of 37-42 DEG C, 21.26 g / L of 3-hydracrylic acid and 23.95 g / L of 3-hydracrylic acid can be synthesized through glycerol fed-batch fermentation, and 10.31 g / L of 3-hydracrylic acid and 9.02 g / L of 3-hydracrylic acid can be synthesized through industrial waste glycerol fed-batch fermentation.