292 results about "Biomanufacturing" patented technology

The present invention relates to an optimization solving method and system for metabolic flux in a biomanufacturing process. The method comprises: constructing a flux balance equation of a microbial fermentation process; taking metabolic flux as a control variable to be optimized, on the basis of the flux balance equation, constructing a kinetic equation system, and taking into account a constraint condition and a target function in the fermentation process to construct a dynamic flux balance analysis model based on a dynamic optimization problem; on the basis of a control time domain of a production cycle in the fermentation process, constructing a new time domain, and reconstructing the metabolic flux in the new time domain; parameterizing the reconstructed metabolic flux by means of a piecewise constant approximation strategy; on the basis of the parameterized metabolic flux, converting the dynamic flux balance analysis model from the dynamic optimization problem into a static optimization problem; and solving the optimal metabolic flux of the dynamic flux balance analysis model based on the static optimization problem. The present invention effectively improves the product quality by solving the optimal metabolic flux in the microbial fermentation process.

The invention provides a construction method and application of a pichia pastoris strain capable of efficiently secreting and expressing recombinant human-derived protein, and relates to the technical field of synthetic biology. According to the invention, pichia pastoris is used as a chassis strain, a ku70 gene and / or a pep4 gene in a pichia pastoris genome are / is knocked out through a gene editing technology, and a new signal peptide O factor signal is introduced, so that the secretion efficiency of the pichia pastoris for secreting and expressing recombinant human-derived protein can be remarkably improved, the problem of protein degradation is effectively improved, and the recombinant human-derived protein can be obtained. And meanwhile, a high-copy pichia pastoris strain is efficiently obtained, so that the expression quantity of recombinant human protein is improved, and powerful support is provided for research and development of related drugs and biological manufacturing.

The invention provides a cell culture medium aided design system and method based on a large oracle model, and belongs to the technical field of artificial intelligence. A plurality of candidate culture medium formula schemes meeting user requirements are generated by obtaining culture medium design parameters and requirements and designing structured cues and inputting the designed structured cues into a large language model; a substance matched with the structured cue word is retrieved by utilizing a retrieval enhancement technology, a replacement substance is recommended for a user to refer, the user selects and purchases a required experimental material according to own requirements, a candidate culture medium formula scheme is fed back and evaluated in an actual experiment, and the system performs iterative optimization based on the feedback and evaluation, so that the accuracy of the experiment is improved. And a more appropriate candidate culture medium formula scheme is provided. According to the cell culture medium aided design system and method, intelligence, scientization and individuation of culture medium formula scheme design are achieved, experiment cost is reduced, repeatability and stability of cell culture experiments are improved at the same time, and therefore wide application of culture medium design in the field of biological manufacturing is promoted.

The invention relates to the technical field of enzyme engineering, in particular to an L-glutamate oxidase mutant as well as preparation and application thereof. Through mutation of the 77th amino acid and the 171st amino acid, the active center configuration of the enzyme is optimized, the binding capacity with a substrate and the catalytic conversion efficiency are enhanced, and the enzyme activity is remarkably improved. The mutant disclosed by the invention shows stronger specificity to L-glutamic acid (salt), effectively reduces non-specific reaction and improves the product purity. And in the presence of a high-concentration substrate, the enzyme activity is not inhibited by the substrate. The prepared L-glutamate oxidase mutant with high enzymatic activity can be efficiently expressed in escherichia coli, the problem that the expression quantity of natural enzymes in streptomyces is limited is solved, the acquisition efficiency of the enzymes is greatly improved, and the production cost is reduced. The mutants can be directly applied to biological manufacturing of alpha-ketoglutaric acid, so that the production efficiency of products is remarkably improved.

The invention discloses a high-performance carbonyl reductase mutant and application thereof in synthesis of a series of chiral alcohols. The high-performance carbonyl reductase mutant is obtained by performing the following mutations on an amino acid sequence as shown in SEQ ID NO.2: the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine, and the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine; and / or the 100th glycine is mutated into any one of glutamic acid, serine, lysine or asparagine. Compared with a wild type enzyme, the thermal stability and the catalytic activity of the obtained mutant are remarkably improved, particularly, single point mutants Mut-A69N, Mut-G100E and Mut-G100N and a combined mutant Mut-A69N-G100N show excellent relative enzyme activity, thermal stability and substrate tolerance. The mutant has high catalytic activity and stability to various chiral alcohols, and the contradiction that the traditional carbonyl reductase is easy to inactivate at high temperature and the catalytic efficiency is difficult to achieve at the same time is solved. The invention provides an efficient and stable enzyme catalysis tool for green biological preparation of chiral drug intermediates.

The invention belongs to the technical field of microbial genetic engineering, and particularly relates to an alpha-2, 3-sialyltransferase mutant and application thereof. The a-2, 3-sialyltransferase mutants V175N, P198A, G173D, D174Q, F196K and A315H are obtained by performing site-specific mutagenesis on six amino acids in an a-2, 3-sialyltransferase amino acid sequence coded by a gene derived from Campylobacter jejuni (OH4384), the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3'-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the P198A, the G173D, the D174Q, the F196K and the A315H, the optimal recombinant microbial strain 3 '-SL-09 (V175N / P198A / D174Q) is obtained through combination of multiple mutation sites, the yield can reach 5.91 g / L, the bottleneck problem of low catalytic efficiency of sialyltransferase is effectively solved, and an enzyme element with excellent performance is provided for high-efficiency and low-cost biological manufacturing of 3'-SL.

The invention discloses a method for constructing a multicellular living body material by adopting a droplet bioreactor, which belongs to the crossing field of biological materials and microbial engineering, and comprises the following steps: (1) resuspending different cells in different polymer solutions to prepare degradable or phase-separable hydrogel particles containing cells; (2) dispersing the hydrogel particles in a cross-linkable hydrogel matrix to form a composite hydrogel; and (3) curing the composite hydrogel, and then carrying out degradation treatment to obtain the liquid cavity structure multicellular system bioreactor with spatial partition and dynamic interaction characteristics. According to the invention, ordered distribution and metabolism synergy of different microorganisms in space can be realized, controllable degradability and excellent mass transfer performance are realized, bacterial slow release and metabolite exchange can be maintained, and the method is suitable for long-term stable operation of a continuous flow reactor. The method has wide application prospects in the fields of green biological manufacturing, renewable energy source production, environmental restoration, biomedical engineering and the like.

The invention discloses a recombinant VI-type collagen as well as a preparation method and application thereof. The recombinant VI-type collagen comprises a protein gene segment derived from a natural VI-type collagen alpha2 chain, and proteins P0607, P060702 and / or P060703 capable of promoting cartilage cell proliferation activity are expressed. Gene segments derived from a natural type VI collagen alpha2 chain are integrated through a genetic engineering means, sequence optimization and repetition are carried out on the gene segments to enhance the biological activity of the collagen, the recombinant type VI collagen P0607, P060702 and P060703 can be stably and efficiently produced, the recombinant collagen has the effect of promoting chondrocyte proliferation, and the recombinant collagen can be applied to the field of cartilage cell proliferation. An efficient and reliable biological manufacturing way is provided for research, development and production of osteoarthritis medicines.

A semi-carbonized carbonized material is produced from the raw wood material to be treated, a highly energy semi-carbonized carbonized material is produced from the raw wood material to be treated, and a highly energy wood biogas is produced from the highly energy semi-carbonized carbonized material. [Solution] The system includes a subcritical water reaction treatment device, a wood biopellet generation device, and a wood biogasification furnace. The subcritical water reaction treatment device forms a semi-carbonized wood biomaterial consisting mainly of low molecular weight cellulose components and low molecular weight lignin components from wood material, and multiple hemicellulose-based monosaccharides that are solidified and fixed to these cellulose components and lignin components. The wood biopellet generation device produces wood biopellets from the semi-carbonized wood biomaterial, and the wood biogasification furnace produces wood biogas from the wood biopellets.

The invention relates to bacillus megatherium XW-02 (the preservation number is CCTCC NO: M 2025185), a microbial preparation of the bacillus megatherium XW-02, and application of the microbial preparation. The bacillus megatherium XW-02 has the advantages that the preservation number is CCTCC NO: M 2025185; the strain can efficiently express xylanase through xylan induction, hemicellulose in tobacco leaves is remarkably degraded, chemical composition is optimized, and combustion harmful substances are reduced. The treated cigar has the advantages that the aroma richness is improved, the purity is improved, the taste mellowness is improved, and the overall quality is improved by 1-2 grades. The xylan extracted from agricultural wastes is used as an inducer to construct a green biological manufacturing system, the cost is reduced by more than 80% compared with the traditional process, and the emission of three wastes is reduced. The process compatibility is high, the method can be matched with an existing production line only by additionally arranging a strain culture module, the single-line annual tobacco leaf treatment amount reaches 500 tons, and economic benefits are remarkable. The invention provides a safe, efficient and environment-friendly quality improvement solution for the tobacco industry, and has great industrialization value.

The invention relates to the technical field of textile bio-manufacturing and industrial process automation control, in particular to a bio-based fabric production temperature and humidity intelligent regulation and control system which comprises a data acquisition unit used for acquiring temperature and humidity sensor values in a fermentation curing workshop in real time and reading current heating power of an actuator and a fan rotating speed instruction; the reference state reconstruction unit is used for receiving the input state vector and generating an interference-free reference vector; the disturbance simulation unit is used for generating a theoretical damaged state vector representing a specific fault mode; the double-track difference unit is used for generating a real difference feature vector and a theoretical difference feature vector; the coupling regulation and control unit is used for calculating a similarity value of the real difference feature vector and the theoretical difference feature vector in a number domain space; according to the method, non-ideal factor interference is effectively eliminated, and the physical interpretability and numerical traceability of the reference data are ensured.

The present disclosure relates to improved biomanufacturing systems and methods with an ability to effectively control process variability to consistently generate high-quality cell-based therapies. In particular, disclosed herein is a using model-based control for determining process parameters in a bioreactor in real time. Additionally disclosed herein is a system for generating an AI digital twin model for a commercial-scale bioreactor.

The invention discloses a transporter T6417 and application thereof in increasing the yield of natural products of streptomyces, and belongs to the technical field of genetic engineering. In order to improve the yield of a streptomyces natural product, the method comprises the following steps: screening transporters related to the yield and a production time sequence on the basis of analysis on transcriptome data of streptomyces cerevisiae NEAU6, and carrying out function annotation analysis to obtain a candidate transporter T6417; then, different promoters are used for carrying out adaptation on the expression of the T6417, and it is found that the yield of natural products can be increased by overexpressing the transporter T6417 in streptomyces obscurvius, streptomyces bingchengensis or streptomyces avermitilis. The invention provides a valuable biosynthetic element for efficient biomanufacturing of streptomyces.

The invention belongs to the technical field of metabolic engineering and biological manufacturing, and particularly relates to an engineering bacterium for producing gallic acid (GA) and application of the engineering bacterium in production of gallic acid. According to the method, key metabolic nodes in the gallic acid production process are systematically regulated, for example, the hydroxylase activity is improved, the pyruvic acid flow direction is regulated, the aromatic amino acid precursor supply is improved, and the dynamic balance between the engineering bacterium growth and the gallic acid product synthesis is realized, so that the gallic acid yield and the carbon yield are remarkably improved. Experiments show that the gallic acid yield of the engineering bacteria prepared by the invention reaches 12.5 g / L in a shake flask experiment, and reaches 109.8 g / L in a 3 L fermentation tank; and the carbon yield is increased to 0.64 g / g glucose, which is the highest level reported in the current literature.

The invention discloses a method and application for enhancing cytidine production, and belongs to the field of genetic engineering and microbial engineering. The present invention knocks out cytidine deaminase gene cdd, uridine kinase gene udk, pyrimidine-specific ribonucleoside hydrolase gene rihB, nucleotide 5'-monophosphate nucleosidase gene ppnN, pyruvate oxidase gene poxB and integrates and expresses 5'-CTP diphosphohydrolase, orotate phosphoribosyltransferase, uridylate kinase mutant gene PyrH (R92G / D93G), as well as phosphoribosyl pyrophosphate synthetase gene prs, glucose-6-phosphate dehydrogenase gene zwf and 6-phosphogluconate dehydrogenase gene gnd, and constructs a recombinant Escherichia coli with enhanced cytidine synthesis ability, and the cytidine yield reaches 61.9g / L in a 30L fermenter without adding antibiotics and inducers, which can be used to realize the industrial production of cytidine and realize green biomanufacturing.

The invention discloses an SAM hydrolase mutant and application thereof, the specific activation effect of halogen anions (F <->, Cl <->, Br <-> and I <->) on SAM hydrolase (containing MJ1651 wild type and mutant) is found for the first time, and the active sequence of the SAM hydrolase is F <-> > I <-> > Br <-> = Cl <->. According to the method, 400 mM of halide salt is added into a reaction system, so that the catalytic efficiency is improved in a breakthrough manner; the yield of wild type enzyme adenosine is improved by 20-300% (optimally, an F-system); the yield of the mutant R89C is soaring up by nearly 6 times under the action of KF; the activity of the immobilized enzyme is still more than 90% after the immobilized enzyme is repeatedly used for 5 times. The method is easy and convenient to operate, green production is achieved with water as the solvent (three wastes are reduced by 90%), the comprehensive cost is reduced by 41%-80 / kg, and an efficient solution is provided for industrial biological manufacturing of antiviral drug intermediates and the like.

The invention discloses an escherichia coli engineering bacterium for producing 5-ALA by an anaerobic fermentation method as well as a construction method and a fermentation method of the escherichia coli engineering bacterium. The engineering bacterium is constructed by taking escherichia coli (E. coli) ZX11 as a host and performing heterologous expression on 5-ALA synthetase (HemA) from rhodobacter capsulatus. In order to improve the soluble expression level of the HemA, the HemA is subjected to fusion expression with a maltose binding protein (MBP) tag. Furthermore, the engineering bacterium can also heterologously express a visual purple protein (PR) fused with the MBP tag. The construction method of the engineering bacteria comprises the following steps: sequentially cloning the MBP-HemA fusion gene and the PR gene to an expression vector and transferring into host bacteria. By utilizing the engineering bacteria provided by the invention, anaerobic fermentation can be carried out in a culture medium taking glucose as a carbon source, and 5-ALA can be efficiently synthesized. The invention provides an effective way for realizing green and energy-saving anaerobic biological manufacturing of 5-ALA.

A method for monitoring a process of producing a multicellular structure from a population of cells in a 3D cell culture is described. The method comprises obtaining one or more 3D images of the 3D cell culture obtained by imaging the cell culture at different depths and / or from different angles, processing the one or more images to obtain a plurality of image-derived features, and determining values of one or more indicators indicative of progress or outcome of the maturation process using a statistical model, the statistical model is adapted to predict values of one or more indicators indicative of progress or outcome of the maturation process using an input comprising a plurality of image-derived features.

A biofabrication method for producing cellulose-based materials. The method includes providing active cellulose-producing bacteria such as Gluconacetobacter and culturing media in a container; combining organic additives or inorganic additives with the active cellulose-producing bacteria in the container to produce a cellulose hydrogel matrix composed of entangled bacteria-produced cellulose nanofibers; controlling a concentration of the additives in the cellulose hydrogel matrix; and exposing the cellulose hydrogel matrix in selected thermal environment to create a biofabricated functional material.

The invention discloses a high-industrial-property yarrowia lipolytica strain and application thereof, and belongs to the technical field of microbial engineering and fermentation. The mutant is derived from a natural bacterial strain of fermented vegetables, is obtained through high-throughput cell screening, has the advantages and characteristics of high salt resistance, low pH resistance, efficient utilization of an acetic acid substrate, high protein content, no need of vitamins for growth and the like, and is suitable for industrial production of microorganisms; the method can support efficient production of bio-manufactured products under strict conditions of acetic acid carbon source, no sterilization, no seawater, no vitamin and the like.

The invention discloses a genetically engineered bacterium, a method for constructing the genetically engineered bacterium and a method for preparing emodin and physcion by using the genetically engineered bacterium. The genetically engineered bacterium carries an exogenous gene; the exogenous gene comprises a gene npgA for coding phosphopantetheinyl transferase and a gene gedC for coding polyketide synthase, further comprises at least one of a gene HyTE and a gene gedB for coding beta-lactamase thioesterase, and further comprises at least one of a decarboxylase gene TpCK and a gene AfDC. According to the invention, a complete synthetic route of the physcion is constructed from the beginning in a heterologous host microorganism for the first time; a novel microbial fermentation mode is used for replacing traditional plant extraction, and novel biological manufacturing of the emodin and the emodin monomethyl ether is developed.

The present invention provides methods for glycosylation and compound preparation. The compounds include galactosylated, sialylated, fucosylated, and N-acetylglucosaminated compounds from simple oligosaccharides and sugars of animal, plant, or microbial origin. In certain embodiments, the invention provides for trinucleotide-free enzymatic production of oligosaccharides starting from simple sugars, including plant-based sugars. The present invention also provides methods of enzymatic production of fucosylated oligosaccharides and fucosylated antibody-glycan conjugates from ordinary enzymes. The production may be a cell-free, one-pot synthesis using an enzyme and, in some embodiments, an immobilized enzyme. The synthesis is a highly customizable and efficient cell-free manufacturing process. In some embodiments, lactose derivatives and human milk oligosaccharides (HMOs) are produced.

The invention belongs to the technical field of microbial genetic engineering, and particularly relates to an alpha-2, 6-sialyltransferase mutant and application thereof. The a-2, 6-sialyltransferase mutants M133L, E140D, D228H, N289D and G322Q are obtained by carrying out site-specific mutagenesis on five amino acids in an a-2, 6-sialyltransferase amino acid sequence coded by a gene derived from Photobacterium sp.JT-ISH-224, and compared with wild type alpha-2, 6-sialyltransferase, the yield of the single-point mutant 6 '-SL of the E140D and the yield of the single-point mutant 6'-SL of the G322Q are increased by 87.80% and 78.01% respectively. The yield of the optimal recombinant microbial strain 6 '-SL-7 (E140D / G322Q) obtained by mutation site combination can reach 1.471 g / L, the bottleneck problem of low catalytic efficiency of sialyltransferase is effectively solved, and an enzyme element with excellent performance is provided for high-efficiency and low-cost biological manufacturing of 6'-SL.

The disclosure features methods that include obtaining a vibrational spectrum of a solution in a biological manufacturing system, analyzing the vibrational spectrum using a first chemometrics model to determine a value of a first quality attribute associated with the solution, analyzing the vibrational spectrum using a second chemometrics model to determine a value of a second quality attribute associated with the solution, and adjusting at least one parameter of a purification unit of the biological manufacturing system based on at least one of the values of the first and second quality attributes.

A microbial cellulose material is provided comprising an acid, a metal ion, and microbial cellulose. The acid can be a polyphenol. The metal ion can be selected from a group consisting of iron ion (Fe2+), iron ion (Fe3+), copper ion (Cu2+), aluminum ion (Al3+), magnesium ion (Mg2+), zinc ion (Zn2+), nickel ion (Ni2+), germanium ion (Ge4+), titanium (Ti4+), molybdenum (Mo6+) and tungsten ion (W6+). The microbial cellulose can be a bioplasticized microbial cellulose.

The invention relates to a method for efficiently preparing maytansinol by using heterologous alanyl enzyme. The invention provides a genetically engineered bacterium HGFS10 for efficiently preparing maytansinol by using heterologous alanyl enzyme. The genetically engineered bacterium HGFS10 is used for heterologous expression of D-alanyl enzyme, overexpression of a positive regulatory factor gene asm18, overexpression of a C-4 / 5 epoxidase gene asm11 and overexpression of an amide N-methylase gene asm10. The coding gene of the D-alanyl enzyme is smast C, the nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the coding gene is as shown in SEQ ID NO. 2. The in-vitro catalytic efficiency of the D-alanyl enzyme SmAstC on N-demethyl-4, 5-deepoxymaytansinol is 8 times that of a homologous protein AstC of the D-alanyl enzyme SmAstC, and a good tool enzyme is provided for biological manufacturing of maytansinol. D-alanine is added into the culture medium, a direct acyl donor of the enzyme SmAstC is optimized, and the yield of maytansinol reaches 16.18 mg / L and is increased by 15.7% compared with the yield of maytansinol without adding D-alanine into the culture medium.

This invention relates to the fields of microbial genetic engineering and fermentation biomanufacturing technology, specifically disclosing a recombinant Corynebacterium glutamicum with high 5-aminolevulinic acid (ALA) production and its applications. Addressing the problems of existing strains such as crude enzyme activity regulation, inhibited cell growth, the need for external inhibitors, and low yield, this invention uses Corynebacterium glutamicum as a host and removes the gdhA gene without scarring to reduce precursor competitive metabolism. Site-directed mutations of the endogenous hemB gene (S288A, D128E, R231K) are performed to moderately reduce dehydratase activity. A high-expression plasmid pXMJ19-hemA containing the hemA gene from Rhodopsinus capsulatum is constructed and introduced into the modified strain to obtain a recombinant strain with optimized triple metabolism. This strain reduces product degradation without the need for external dehydratase inhibitors, does not affect cell growth, and achieves a yield up to 4.30 times higher than the control after 42 hours of fermentation, with ALA accumulation exceeding 8.9 g / L. The strain exhibits stable characteristics, the process is simple, and it has good industrial application value.

To provide a system and a method for measuring the product quality attribute of a sample including a recovered sample from a continuous biological manufacturing system.SOLUTION: A system 100 for measuring the product quality attribute of an analyte of a biological sample includes a first flow control device 106, a sample purification device 108, a first sample analyzer 114a, second sample analyzers 114b, 114c, 114d, a second flow control device 110, a detector 116, and a control unit 122. The first sample analyzer 114a includes a first chromatography column. During operation of the system, the control unit 122 adjusts the arrangement of the second flow control device 110 to direct a portion of the biological sample to one of the first and second sample analyzers. The product quality attribute of the analyte of the biological sample is determined on the basis of the analysis of the portion of the biological sample conducted by one of the first and second sample analyzers.SELECTED DRAWING: Figure 1

The present invention relates to a device for a scalable biomanufacturing platform for the production of modified cells such as CAR-modified T-cells while eliminating on-target / off-tumor toxicity and decreasing the current production cost by 500 times (per treatment), and to attendant methods. The includes a first chamber for proliferating a population of cells and a second chamber for modifying the cells to express a desired T-cell receptor antigen.

This invention discloses a recombinant vector, recombinant bacteria, and fermentation method for high-yield GHK production, belonging to the field of biomedical technology. The recombinant expression vector of this application comprises a bicistronic expression cassette controlled by a T7 / lac heterozygous promoter, sequentially containing the encoding (GHK). n The vector encodes the first cistron of the polypeptide and the second cistron, where n is an integer from 2 to 200. When this vector is transformed into *E. coli* BL21, the resulting recombinant bacteria can simultaneously express (GHK) after IPTG induction. n Polypeptides and trypsinogen. During fermentation, trypsinogen is activated and enables the processing of (GHK). n In situ enzymatic cleavage of peptides efficiently releases GHK. This invention integrates the traditional multi-step process into a "one-step fermentation," simplifying the production process and reducing purification difficulty. Examples show that the GHK yield can reach up to 3.9 g / L, providing an efficient and convenient new strategy for the large-scale biomanufacturing of GHK.