183 results about "Batch fermentation" patented technology

The invention belongs to the technical application field of microbial metabolic engineering and synthetic biology, and particularly relates to a pichia pastoris engineering strain for high-yield sclareol as well as a construction method and application of the pichia pastoris engineering strain. The method comprises the following steps: constructing a biosynthetic pathway of sclareol in a host strain, and optimizing an intracellular mevalonic acid metabolic pathway and a central metabolic pathway to obtain an engineering strain A; the host strain is pichia pastoris; or overexpressing or knocking out a metabolism regulation factor in the obtained engineering bacterium A to obtain an engineering bacterium strain B; or, in the obtained engineering bacterium B, a synthetic pathway is targeted to peroxisome and optimized by adopting a cell region compartment strategy, and an engineering bacterium strain C is obtained. The invention provides a pichia pastoris chassis cell for synthesizing a diterpenoid sclareol and an engineering strain for synthesizing sclareol. By introducing and optimizing the expression of a sclareol synthesis pathway, the fermentation yields of sclareol in shake flask batch and bioreactor batch fed-batch fermentation are respectively 631.6 mg / L and 10.5 g / L.

The present invention relates to a recombinant yeast with high squalene production, a construction method and a use thereof, wherein the recombinant yeast is a saccharomyces cerevisiae that expresses or overexpresses HMGR gene, ADA gene, LEU2 gene, MVA pathway gene and squalene synthase gene ERG9. The present invention integrates acetaldehyde dehydrogenase gene ADA and HMG-CoA reductase gene tHMGR in multiple copies in saccharomyces cerevisiae, integrates the screened high-activity phosphoketolase gene PK and phosphotransacetylase gene PTA, weakens epoxysqualene synthesis, strengthens the mevalonate pathway, overexpresses yeast endogenous squalene synthase gene, etc. to obtain a recombinant yeast with high squalene production, so that the production in the shake flask stage of the production process exceeds 5g / L, and the production in the fed batch fermentation stage exceeds 50g / L, breaking through the current bottleneck of microbial synthesis of squalene production, and is suitable for practical promotion and application.

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 technical field of biology, in particular to a recombinant strain for producing nicotinamide mononucleotide and application of the recombinant strain. According to the method, accumulation of the NMN is improved through metabolic flux redirection and gene knockout, a synthesis path of the NMN is successfully established in escherichia coli, and further, the yield of the NMN is effectively improved and reaches 7.0 g / L by optimizing a transport system of NAM-NMN and a PRPP synthesis path. The invention also optimizes the production process of the NMN, realizes the highest yield of the NMN of 28.5 g / L in a fed-batch fermentation system, and obviously improves the yield compared with the traditional batch fermentation.

The invention belongs to the field of application of microbial genetic engineering and metabolic engineering, and particularly relates to a method for improving the biosynthesis efficiency of sclareol. Specifically, in a host strain, gene knockout is realized through a CRISPR / Cas9 technology, or / and a DNA fragment is introduced, and a strain metabolic pathway is adjusted to obtain a corresponding engineering strain; carrying out biosynthesis by utilizing the obtained engineering strain to obtain sclareol; the gene knockout is realized by introducing donor DNA (Deoxyribose Nucleic Acid) containing upstream and downstream homologous arms of a coding frame; the DNA fragment sequentially comprises an upstream homologous arm, a promoter, a gene, a terminator and a downstream homologous arm from upstream to downstream, the host strain is selected from any one of saccharomyces cerevisiae. According to the method, the cell life is associated with product synthesis, and meanwhile, metabolic balance is coupled, so that the synthesis efficiency of a target product is improved, and a foundation is laid for industrial fed-batch fermentation.

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 provides an algae fermentation culture method, which relates to the technical field of bioengineering, and comprises the following steps: selecting microalgae seeds; constructing an index fed-batch model corresponding to the selected microalgae seeds; inoculating the microalgae seeds into a fermentation tank of a Basal culture medium; culturing according to the index fed-batch model under a dark condition; after culturing under a dark condition, adding plant hormones and a nitrogen source to continuously culture to obtain the microalgae. According to the algae fermentation culture method provided by the invention, the growth of microalgae and the accumulation of xanthophyll are facilitated through index fed-batch fermentation, and after the index fed-batch fermentation culture, IAA high-nitrogen combined treatment is matched, so that the xanthophyll yield and yield of the microalgae in a fermentation tank can be effectively improved.

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 relates to the technical field of biology, in particular to an engineering strain for dynamically regulating glucose metabolism to redirect 3-FL production and a construction method of the engineering strain. According to the invention, a gene-coded metabolic switch is adopted and is focused on dynamically regulating and controlling the level of phosphofructokinase (Pfk-I), so that the metabolic state is purposefully converted from a cell growth mode based on a wild strain metabolic pathway to a 3-FL production mode based on an escherichia coli anabolism pathway, and rewriting of a gene regulation network is realized; and the condition of converting the metabolic state from a cell growth mode to a production mode in the batch fermentation of the 3-FL is optimized, so that the production titer and yield of the 3-FL directly fermented from the glucose are improved.

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 present invention provides a plant component hypoglycemic composition and its preparation method and application, belonging to the field of biomedical technology. The pharmaceutical composition is composed of the following raw materials in parts by weight: 2-5 parts of mulberry polypeptide, 5-10 parts of polygonatum and 5-10 parts of eucommia male flower, and its preparation method is to first extract mulberry polypeptide from mulberry, then crush and decoct polygonatum and eucommia male flower, and then add Lactobacillus acidophilus and Bacillus subtilis in sequence for batch fermentation, and take the fermentation supernatant and freeze-dry it and mix it with mulberry polypeptide to obtain the hypoglycemic pharmaceutical composition of the present invention. The pharmaceutical composition prepared by the present invention has a significant inhibitory effect on α-glucosidase in vitro, has a good hypoglycemic effect in vivo, has a high utilization rate of medicinal materials and has no toxic side effects, and has broad market prospects.

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 discloses an automatic treatment device for preparing enzymes through anaerobic fermentation of rotten fruits, and belongs to the technical field of organic waste treatment. The device comprises a smashing assembly, a feeding assembly, a fermentation assembly, a conveying assembly and a filling assembly, the smashing assembly comprises a hopper and a crushing unit, and the crushing unit is connected with the bottom of the hopper; the feeding assembly can lift and turn over the container, and rotten fruits in the container are input into the hopper. The fermentation assembly comprises a fermentation tank and a stirring unit, the stirring unit is arranged in the fermentation tank, and the stirring unit can stir a mixture of pomace, water and syrup; the material conveying assembly is communicated with the fermentation tank and can convey the pomace treated by the crushing unit, water and syrup into the fermentation tank in proportion; the filling assembly comprises a filling unit; the filling unit is communicated with the fermentation tank through a pipeline. The rotten fruit fermentation treatment efficiency can be improved, and batch fermentation industrial rotten fruit treatment is facilitated.

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.