124 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 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 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 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 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 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 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 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.

The invention relates to the technical field of fermentation of medicinal material production, and discloses a preservation method of a pseudo-ginseng cell strain, which comprises the following steps: acquiring a working cell strain; obtaining a liquid culture medium; carrying out mixed culture on the working cell strain and the liquid culture medium, and dehydrating; and soaking the dehydrated biomass in a cryoprotectant, and then carrying out cryopreservation or refrigerated preservation. Through cryopreservation or refrigerated preservation of the mixed solution, the preserved working cell strain can be adopted for fermentation in real time according to the requirements of a user, the richness of the strain can be guaranteed, the preparation period of the seed solution is saved, multi-batch fermentation can be realized, and the yield is stable.

This invention discloses a method for constructing a high-succinic acid-producing Pichia pastoris cell factory, using Pichia pastoris (… Komagataella phaffii Using GS115 as the chassis, a reductive TCA cycle enhancement strategy and an sdh5 knockout strategy were first constructed, respectively. It was found that a single strategy was insufficient to effectively achieve succinic acid accumulation. Further, the two strategies were combined to obtain the engineered strain SAOG, which achieved succinic acid accumulation. Based on this, the malate transporter SpMAE was introduced to enhance the transmembrane transport capacity of succinic acid, and histidine auxotrophic recovery was combined to improve the growth and fermentation performance of the strain, resulting in the engineered strain SH, whose succinic acid accumulation was 25.1% higher than that of SAOG. Finally, the engineered strain SH achieved a succinic acid yield of 10.47 g / L in shake-flask fermentation and 112.6 g / L after 120 h of continuous fed-batch fermentation in a 5 L fermenter. This invention achieves efficient succinic acid synthesis in the Pichia pastoris chassis through the synergistic regulation of reductive TCA pathway enhancement, succinic acid oxidation blocking, enhanced transmembrane transport, and auxotrophic recovery, exhibiting stable fermentation performance, high yield, and good industrial application potential.

The invention belongs to the field of synthetic biology, and particularly relates to an engineering bacterium for de novo biosynthesis of mendalin as well as a construction method and application of the engineering bacterium. Glucose is taken as a substrate, key enzyme genes are expressed in saccharomyces cerevisiae cells, and the mendalin is de novo synthesized through a microbial fermentation process; according to the constructed engineering strain Med21, the yield of the mediodarin is 180 mg / L; the yield of the constructed optimized strain Med25 can be increased to 450 mg / L after fed-batch fermentation; compared with a traditional chemical synthesis method and a method for extracting from natural plants, the method for synthesizing the medioren does not depend on exogenously added precursor substances, has the advantages of being low in cost, high in production efficiency, small in environmental pollution, high in product yield and purity and the like, provides possibility for large-scale industrial production of the medioren, and has a wide application prospect. The method has important application value and market prospect, and opens up a new way for production and application of the mendalin.

This invention discloses a high-yielding erythritol-producing *Yarrowia lipophila* strain (… Yarrowia lipolytica This invention relates to the field of biotechnology, specifically to *Yarrowia lipophila* CA20 as the starting strain, and its applications. A000039 A gene knockout method was used to construct a high-yield erythritol-producing *Yarrowia lipolytica* strain. The construction method provided by this invention is simple and of significant value in simplifying the construction process of high-yield strains. Furthermore, under the fermentation medium and conditions provided by this invention, the *Yarrowia lipolytica* strain producing high-yield erythritol was fed-batch fermented with glucose as a substrate, achieving a yield of 280.50 g / L and a yield of 0.71 g / g, demonstrating good economic benefits.

The invention discloses a glycosylation-free human enterokinase mutant as well as recombinant expression and application thereof, belongs to the field of genetic engineering and protein technical engineering, and solves the problems of disordered glycosylation modification, low enzymatic activity and low protein expression quantity during recombinant expression of enterokinase in an enterokinase microbiological cell factory. The method comprises the following steps: firstly, carrying out single-site mutation, double-site mutation, three-site mutation and four-site mutation on the basis of a human enterokinase wild type, and constructing a pichia pastoris cell factory to obtain a series of human enterokinase mutants without glycosylation modification. Through calculation design and rational transformation, partial or non-glycosylation is subjected to deep fixed-point transformation, and pichia pastoris cell factory expression is constructed, so that the expression quantity and the total enzyme activity of the mutant are greatly improved. The highest enzyme activity of the glycosylation-free enterokinase mutant generated by fed-batch fermentation in a pichia pastoris cell factory reaches 7.8 * 10 < 4 > U / L, and the enterokinase mutant has the capacity of large-scale industrial fermentation generation and application.

The utility model relates to the technical field of pickles, in particular to a pickle jar which comprises a first jar body, a second jar body and a jar cover, the second jar body is covered on the first jar body, the jar cover is covered on the second jar body, a first pickle storage space is arranged in the first jar body, and the second jar body is provided with a second pickle storage space. Pickled vegetables can be fermented in batches, so that the convenience for people to eat the pickled vegetables is improved.

The invention provides a Trichoderma reesei engineering bacterium for high yield of beta-glucosidase, which is characterized in that a Trichoderma reesei SN1 strain is used as a starting strain, and an expression cassette for knocking out a cre1 gene, a constitutive expression cassette of an aspergillus niger source bglA gene expressed by a promoter cbh1 and a constitutive expression cassette of a 130442 gene expressed by the promoter cbh1 are constructed in a genome of the Trichoderma reesei SN1 strain. Through the knockout of the cre1 gene, the overexpression of the aspergillus niger source bglA gene and the overexpression of the 130442 gene, the yield of the beta-glucosidase in the trichoderma reesei is improved synergistically. Glucose is taken as a raw material, a transglycosylation reaction can be catalyzed by directly utilizing a crude enzyme solution of trichoderma reesei engineering bacteria with high yield of beta-glucosidase to prepare beta-disaccharide, and the beta-disaccharide can be taken as an inducer for cellulase production; a crude enzyme liquid with higher cellulase activity and beta-glucosidase activity is obtained by supplementing an inducer and carrying out fed-batch fermentation.

The invention discloses a preparation, separation and purification method of glucan oligosaccharide, and belongs to the technical field of active oligosaccharide preparation. The preparation, separation and purification method comprises the following steps: adding alcaligenes faecalis into a fermentation culture medium, fermenting to prepare beta-1, 3-glucan, adding beta-1, 3-glucanase when the viscosity of fermentation liquor reaches a certain value, and simultaneously controlling the temperature to promote enzymolysis, so that the beta-1, 3-glucan with high polymerization degree is hydrolyzed into oligosaccharide, and separating and purifying the oligosaccharide. The material liquid viscosity is reduced, and the mass transfer is improved, so that the single-batch fermentation yield is improved. Meanwhile, sugars with different polymerization degrees are separated through a graded alcohol precipitation process, part of impurities are removed, and the product purity is improved. According to the method, the enzymolysis reaction is directly carried out in the fermentation process, so that the steps of polysaccharide extraction and enzymolysis are simplified, the production energy consumption is reduced, and the yield and the production benefit of the final product are improved.

The invention relates to the field of gibberellin production, and discloses a genetically engineered bacterium for producing gibberellin GA3 at high yield as well as a construction method and application thereof. According to the invention, gibberellin GA3 high-yield genetically engineered bacteria are obtained by taking gibberellin as a chassis bacterium and enhancing expression of an AGP3 gene for coding amino acid permease, an FFUJ12592 gene for coding a transcription factor and a Lae1 gene for coding a global regulation factor in a genome of the chassis bacterium. Experiments prove that the GA3 yield of the constructed genetically engineered bacterium can reach 3.36 g / L after the culture medium is fermented for 168 hours and methyl linolenate fed-batch fermentation is carried out, and compared with the GA3 yield without gene modification or methyl linolenate fed-batch fermentation, the GA3 production capacity is remarkably improved.

A fermented natural substance having nitric oxide metabolites fixated and stabilized therein is produced by a process including adding a fermentative strain to nitrogen-containing natural substance and carrying out fermentation by a batch fermentation process. The nitrogen-containing natural substance is at least one selected from the group consisting of lettuce, Sedum sarmentosum, spinach, blueberry, dandelion, pomegranate, cabbage, garlic, Morinda citrifolia, onion, soybean, bean sprout, mulberry leaf, Momordica charantia, Rubus coreanus, Houttuynia cortada, aronia, Humulus japonicus, Coptis chinensis, hijiki, hooker chive, mushrooms, calamus, wild spinach (Seomcho), Chinese cabbage, kelp, apple, mugwort, orange, salmon testis, abalone shell, shellfish shell, cricket, silkworm, Rehmannia glutinosa, and Shipjeondaebo-tang. to Bacillus sp., Bifidobacterium sp., Enterococcus sp., or Lactobacillus sp.