35 results about "Microbial host" patented technology

The present disclosure is generally related to the fields of microbial host cells, molecular biology, protein engineering, fermentation, protein production, and the like. Certain aspects of the disclosure are related to novel promoter and 5′-untranslated region a nucleic acid (DNA) sequences.

The present disclosure relates to the use of a switch for the production of heterologous non-catabolic compounds in microbial host cells. In one aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured under aerobic conditions followed by microaerobic conditions, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions. In another aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured in the presence of maltose followed by the reduction or absence of maltose, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions.

The invention discloses a method and system for synchronously detecting host chromatin openness and in-vivo microbiome based on transposase, and belongs to the technical field of biological sequencing data analysis. According to the method, transposase is used for selectively fragmenting an open chromatin region of a host, and a microbial genome is almost randomly cut, so that synchronous enrichment of host and microbial DNA is realized; after high-throughput sequencing library construction and double-end sequencing, sequencing data is split into host source and non-host source reads through bioinformatics analysis, host chromatin state and microorganism composition are analyzed respectively, and a microorganism-host epigenetic regulation network is constructed; the invention further provides a matched DNA sequencing library and an analysis system, multi-scene research of infectious diseases, intestinal microecology, tumor microenvironment and the like is supported, a public database can be reanalyzed, and potential microbial interaction signals are mined. According to the method, the host-microorganism interaction research efficiency is remarkably improved, and a high-sensitivity and integrated technical scheme is provided for epigenetic regulation mechanism analysis.

The present disclosure describes a genetically modified microbial host cell expressing one or more heterologous genes encoding a modified beta-lactoglobulin (BLG), thereby producing the said modified BLG, wherein the modified BLG comprises at least one substitution of a non-essential and / or non-branched amino acid with an essential and / or branched amino acid compared to a parent unmodified BLG.

Disclosed herein are methods for producing interfering RNA biopesticides, such as microbial host organisms engineered to produce interfering RNA (iRNA) molecules which selectively undermine the survival of cockroaches. Such iRNA molecules selectively inhibit the expression of a gene in a specific insect population, such as a cockroach population, by RNA interference. Also disclosed herein are polynucleotides, such as expression cassettes encoding iRNA molecules and facilitating integration, such as stable integration into the genome of a host. Further disclosed herein are compositions including the disclosed nucleotide sequences and host organisms, along with methods of using the same to control cockroach populations.

This invention discloses a plasmid for gene overexpression in non-pattern *Bacillus laterosporus*. Its leader region contains a methyltransferase expression cassette derived from the *Bacillus laterosporus* i83 genome, enabling stable existence in specific hosts containing the RM system. Based on this, a PidR2 promoter is added to obtain the pMCPidR2 shuttle plasmid. The PidR2 promoter effectively drives target gene expression, the pWV01 replicon ensures plasmid replication in non-pattern hosts, and the chloramphenicol resistance gene is used for positive colony selection. The plasmid contains a TraJ element, allowing it to be introduced into non-pattern microbial hosts via conjugation transfer. This invention also provides a target gene expression strategy in non-pattern microorganisms, achieving high transformation efficiency and high strain yield through conjugation transfer transformation of donor bacteria, conjugation transfer helper bacteria, and recipient bacteria, resulting in stable expression of the target protein. This invention provides an effective vector for efficient and stable gene overexpression in specific non-pattern microorganisms, helping to solve the problem of difficult genetic manipulation of industrial production strains.

The present disclosure relates, in part, to microbial hosts capable of synthesizing cis-3-hexenol, cis-3-hexenal, trans-3-hexenol, trans-3-hexenal, trans-2-hexenal, cis-2-hexenal and related compounds from hexanoic acid and methods for the preparation of cis-3-hexenol, cis-3-hexenal, trans-3-hexenol, trans-3-hexenal, trans-2-hexenal, cis-2-hexenal and related compounds.

Described herein are compositions and methods for generating a chassis for high-capacity production of different classes of high-value terpenoids. The chassis is a microorganism host that expresses a first operon comprising a modified ribosomal binding site, a Hmg-CoA reductase (HmgR) polypeptide, a 3-hydroxy-3-methylglutaryl CoA synthase (HmgS) polypeptide, and a β-ketothiolase (PhaA) polypeptide; and a second operon comprising a modified ribosomal binding site, a mevalonate kinase (MvK1) polypeptide, a phospho-mevalonate kinase (MvK2) polypeptide, and a diphosphomevalonate decarboxylase (MvD) polypeptide.

The invention relates to a microbial strain DNA (deoxyribonucleic acid) mixed coding and an alternating Transformer depth prediction method for gene expression of the microbial strain DNA mixed coding, and belongs to the crossing field of bioinformatics and artificial intelligence. The core of the method is to construct a deep learning model capable of efficiently capturing DNA sequence characteristics, and end-to-end accurate prediction of the expression level of a target gene in a specific microbial host is realized. The method comprises the following steps: carrying out binary coding on four conventional basic groups of DNA of a target strain, splicing eight key biological characteristics closely related to gene expression, and carrying out hybrid coding to generate a combined characteristic matrix; a deep learning network containing an alternating Transform coding structure is established, local structure information of a DNA combination feature matrix and a global context dependency relationship are deeply fused, and the analysis precision of microorganism DNA sequence data and the prediction accuracy of a gene expression level are improved. According to the technical scheme, an efficient and accurate computer-aided prediction tool is provided for optimization design of genetic elements (such as promoters) and gene sequences, the expensive experiment trial and error cost and the strain characterization cost can be remarkably reduced, and the research and development process of high-yield strains is accelerated.

The present invention relates to a recombinant microbial host cell comprising an operative biosynthetic metabolic pathway capable of producing one or more compounds selected from the group consisting of L-dopa, dopamine, (S)-Norcoclaurine and derivatives thereof; said pathway comprising a heterologous L-tyrosine hydroxylase (TyrH) converting L-Tyrosine into L-dopa capable of increasing the cell production of the Compound compared to a reference L-tyrosine hydroxylase having the sequence set forth in SEQ ID NO: 58.

The present invention relates to methods and compositions for preventing erroneous incorporation of n-leucine into a protein during the production of a recombinant protein in a bacterium. The invention also provides microbial host cells and nucleic acid molecules for use in the methods and compositions provided herein.

The present invention introduces an innovative approach for the manipulation of microorganisms to generate colored bioplastics. This is achieved by concurrently expressing genes responsible for pigment production and genes involved in bioplastic synthesis within a microbial host. These genes can be synthesized, obtained from a different host through cloning, or naturally occurring within the host organism. The resultant color compounds become encapsulated within the extended bioplastic polymers within the cells, resulting in the formation of naturally pigmented bioplastics.

A recombinant microbial host cell having improved in vivo conversion of reticuline and derivatives thereof (such as thebaine and / or oripavine) to relevant downstream opioids (such as neopinone, oripavine, northebaine, nororipavine or morphinone) and related compounds (such as heroin, morphine, codeine, thebaine, oripavine, oxycodone, hydrocodone, hydromorphone, oxymorphone, buprenorphine, naltrexone, naloxone or nalbuphine), wherein the microbial (such as fungal) host cell is heterologously expressing at least one functional transporter protein capable of transporting reticuline or a derivative thereof (such as thebaine and / or oripavine) and a heterologously expressed enzyme capable of acting upon reticuline or a derivative thereof. The invention also relates to uses of the microbial host cells and methods of making an opioid compound and / or opioid precursor compound and / or opioid derivative of interest.

Embodiments of the present invention provides recombinant systems, methods, and compositions for the biological production of Testosteronan (Testan), a linear α-(1→4)-linked glucuronic acid–N-acetylglucosamine polysaccharide. Microbial hosts engineered to express full-length CtTS, N-terminal truncation variants (including d64-CtTS), or the Pseudomonas CIPTS homolog synthesize high–molecular-weight Testan (≥800 kDa) with low polydispersity. Structural analysis by ¹H NMR and 2D HSQC demonstrates that all recombinant enzymes produce Testan that is chemically indistinguishable from native polymer. Purification using ultrafiltration and strong-anion exchange chromatography yields highly defined molecular-weight fractions, including monodisperse populations (PDI ~1.02). The invention further provides sulfated derivatives of Testan produced via aqueous sulfation methods, as well as Testan and functionalized Testan compositions useful for chromatographic stationary phases, biomaterial scaffolds, and chemically modifiable polymer platforms. These systems provide the first scalable, fermentation-based production methods for Testan and Testan-derived materials.

The present invention relates to polypeptide comprising an immunoglobulin single variable domain (ISVD) that specifically binds and inhibits a urease, which ISVD is modified to improve one or more of its protease resistance, thermostability, producibility in microbial hosts and in situ performance in animal excreta. Alone or in combination, the ISVD's of the invention are able to inhibit a broad range of microbial and plant-based ureases. The urease-inhibiting polypeptides of the invention can be used in compositions such as animal feed, animal feed premixes and supplements, to be fed to animals, for reducing the amount of ammonia released from the animal and / or for reducing the amount of ammonia released from the animal's excreta, with the aim to reduce ammonia emissions from agriculture into atmosphere and / or ecosystems. In addition, in the polypeptides can be used in an environmental application like stable floors, slurry pits and urea-based fertilizers for reducing the emission of ammonia.

The invention relates to the technical field of biological medicines, in particular to an ACT structural domain protein derived from coprobacterium rodent and application of the ACT structural domain protein in preparation of anti-tumor and anti-radiation medicines. The invention provides ACT structural domain protein. The amino acid sequence of the ACT structural domain protein is shown as SEQ ID NO: 2. As a functional protein derived from intestinal probiotics, the ACT structural domain protein provided by the invention provides a new molecular probe and a research object for revealing an action mechanism of intestinal flora in tumor generation and development. The protein not only provides an entry point for research on interaction of intestinal microecology and tumor immunity, but also lays a foundation for development of novel biomarkers and drug targets based on microbiome, and has important scientific value in the research field of tumor biology and microorganism-host interaction.

he present disclosure describes a genetically modified microbial host cell expressing and secreting a compound of interest, wherein the cell comprises a native gene involved in the regulation of secretion from the cell of the compound of interest with a signal peptide and / or without a signal peptide wherein the native gene has been modified to alter its functionality in the cell whereby the secretion and yield of the compound of interest from the cell is increased compared to a cell where the gene has not been modified.

Disclosed herein are methods for producing interfering RNA biopesticides, such as microbial host organisms engineered to produce interfering RNA (iRNA) molecules. Such iRNA molecules inhibit the expression of a gene in an insect, such as an ant, by RNA interference. Also disclosed herein are polynucleotides, such as expression cassettes encoding iRNA molecules and facilitating integration, such as stable integration into the genome of a host. Further disclosed herein are compositions including the disclosed nucleotide sequences and host organisms, along with methods of using the same to control ant populations.

Provided are: a transformant capable of improving the productivity of 3-HBA; and a method for producing 3-HBA using said transformant. In one aspect, the present disclosure pertains to a transformant for producing 3-hydroxybenzoic acid (3-HBA), the transformant being obtained by expressibly introducing, into a microbial host having chorismic acid-producing ability, a gene (A) encoding an enzyme (3-HBA synthase) having activity for producing 3-HBA from chorismic acid, wherein the microbial host is Corynebacterium glutamicum. In another aspect, the present disclosure pertains to a method for producing 3-HBA, the method comprising a step for culturing this transformant to produce 3-HBA.

The invention discloses application of a cytidine triphosphate synthetase mutant in cytidine production, and belongs to the technical field of recombinant microorganisms. According to the invention, site-specific mutagenesis is carried out on a pyrG gene for coding cytidine triphosphate synthetase, and the 152nd amino acid of a protein coded by the pyrG gene from W3110 is mutated into alanine, so that the mutant pyrG is obtained. Recombinant plasmids carrying the mutant gene are transformed into microbial host bacteria (such as escherichia coli or bacillus subtilis), and the expression level and / or catalytic activity of CTP synthetase can be remarkably improved, so that a speed limiting step in a cytidine synthesis route is eliminated, and more metabolic flows are guided to biosynthesis of cytidine. Compared with an unmodified strain, the cytidine yield of the engineering strain constructed by the method disclosed by the invention can be increased by more than 20%. The method is accurate and efficient, the uncertainty caused by traditional random mutagenesis is avoided, and a technical scheme with important application value is provided for industrial production of cytidine.

This document discloses methods for producing interfering RNA (RNA) biopesticides, such as microbial host organisms engineered to produce interfering RNA molecules that inhibit gene expression in disease-carrying mosquitoes through RNA interference. This document also discloses polynucleotides, such as expression cassettes encoding these interfering RNA molecules and facilitating integration (e.g., stable integration) into the host organism's genome. This document further discloses compositions comprising the disclosed nucleotide sequences and a host organism, and methods for using them to control mosquito populations.

The present invention relates to mutant galactoside permeases, in particular microbial host cells and fermentation processes employing mutant galactoside permeases that display reduced or abolished proton-coupled transport activity while retaining substrate translocation. These permease variants, when expressed at levels exceeding endogenous wild-type expression, enable cultivation and production processes, in particular in the presence of elevated lactose concentrations without the growth limitations typically associated with lactose-induced energetic collapse. The invention further relates to genetic constructs, metabolic pathways, and process strategies suitable for the production of lactose-based products, such as human milk oligosaccharides.

The present invention relates to metabolic engineering of microbial hosts for the synthesis of products from oil palm empty fruit bunches (OPEFB). In one embodiment, the genetically engineered microorganism is Escherichia coli, which contains a metabolic pathway consisting of nine enzymes (11 genes) to utilize depolymerized lignin, namely vanillin, p-coumaric acid, p-hydroxybenzaldehyde, vanillic acid, p-hydroxybenzoic acid, and ferulic acid, to produce β-ketoadipate, which can then be converted into important derivatives such as adipic acid and levulinic acid. The enzymes are feruloyl-CoA synthetase (fcs), enoyl-CoA hydratase (ech), vanillin dehydrogenase (vdh), vanillate O-demethylase (vanAB; vanA and vanB), p-hydroxybenzoate hydroxylase (pobA), protocatechuate 3,4-dioxygenase {pcaGH; pcaG and pcaH), 3-carboxy-cis,cis-muconate cycloisomerase (pcaB), 4-carboxymuconolactone decarboxylase (pcaC), and β-ketoadipate enol-lactone hydrolase (pcaD).

The instant disclosure provides, inter alia, novel nucleases comprising enhanced performance properties. In certain embodiments, the disclosure is therefore related to nucleases having enhanced performance properties (benefits), nuclease compositions and protein preparations thereof, recombinant polynucleotides (expression construct) encoding mature nuclease of the disclosure, recombinant microbial host cells expressing / producing / secreting nucleases of the disclosure, recombinant microbial host cells co-expressing / co-producing nucleases and proteins of interest, and the like.

The invention discloses a standardized environment-tolerant microbial element library and application thereof, and belongs to the technical field of bioengineering. According to the element library, site modification is carried out on an environment-tolerant gene or a regulatory element thereof, and a prefix and a suffix containing specific enzyme cutting sites are added for sequence standardization and cloning construction, so that a plurality of standardized environment-tolerant microbial elements are obtained and form the element library. Wherein the 5'end of the standardized sequence contains EcoRI and XbaI sites, and the 3 'end of the standardized sequence contains SpeI and PstI sites, so that modular assembly among elements can be realized, and no redundant enzyme cutting sites exist after connection. The element library covers various functional elements such as freezing resistance, high salt resistance, high temperature resistance, acid resistance, heavy metal resistance and the like, microbial hosts can be modified in a plug-and-play mode, the microbial hosts are endowed with physical and chemical environmental stress tolerance or specific functions, and the element library has important application value in the fields of industrial fermentation, biomining industry, environmental governance, agriculture, space biology and the like.

Disclosed herein are methods for producing interfering RNA larvicides for Spotted Wing Drosophila, D. suzukii, such as microbial host organisms engineered to produce interfering RNA (iRNA) molecules which selectively undermine the survival of female D. suzukii larva, to produce a predominantly male population of adult SWD flies. Such iRNA molecules selectively inhibit the expression of a gene in female SWD larvae population by RNA interference. Also disclosed herein are polynucleotides, such as expression cassettes encoding iRNA molecules and facilitating integration, such as stable integration into the genome of a host cell. Further disclosed herein are compositions including the disclosed nucleotide sequences and host organisms, along with methods of using the same to generate a predominately male SWD population.

The invention relates to the technical field of genomics and metabonomics, in particular to a substance for relieving intestinal injury of crisp tilapia and an application effect analysis method of the substance. According to the invention, the tauroursodeoxycholic acid (TUDCA) is applied to relieving intestinal inflammation damage caused by feed embrittlement of the crisp tilapia for the first time, and intestinal inflammation is relieved. By establishing a three-dimensional evaluation system of'intestinal histomorphology + metabonomics + metagenomics', the relieving effect of TUDCA on intestinal inflammation is systematically quantified. Through combined analysis of intestinal metabolome and metagenome, a'microorganism-host interaction 'health judgment model is constructed, and the maintenance of the quality of fish meat after inflammation is relieved is verified through muscle texture parameters, so that a'repair-evaluation-quality maintenance' closed-loop technical scheme is formed.

The invention discloses a novel gene expression plasmid pTAEP2 capable of realizing efficient and stable overexpression in brevibacillus laterosporus (Brevibacillus laterosporus), and a preparation method of the novel gene expression plasmid pTAEP2 capable of realizing efficient and stable overexpression in brevibacillus laterosporus. The plasmid contains a toxin-antitoxin system gene cassette, so that the plasmid can stably exist in a specific non-mode host; the P2 promoter can drive high-level expression of a target gene; the RepF replicon is contained, so that the plasmid can be effectively replicated in a non-mode host; the composition contains ampicillin antibiotics and erythromycin which are used as selection markers. The invention also provides a method for target protein expression by transforming specific non-mode brevibacillus laterosporus with the pTAEP2 plasmid, the plasmid contains a TraJ element, so that the plasmid is efficiently introduced into a non-mode microbial host including brevibacillus laterosporus in a conjugational transfer manner, the gene introduction process is greatly simplified, and the production cost is reduced. And stable and high expression of a target gene is realized. The invention provides a powerful molecular biological tool for developing efficient and stable gene overexpression research and industrial application, and is beneficial to efficient gene manipulation in the non-model bacteria.

The present invention relates to arecombinant microbial host cell producing a tetraketide or derivatives thereof from one or more substrates selected from cinnamoyl-CoA, p-Coumaroyl-CoA, Caffeoyl-CoA, Feruloyl-CoA, malonyl-CoA, sinapoyl-CoA and dihydro derivatives thereof, comprising an operative biosynthetic metabolic pathway for the tetraketide or derivatives thereof comprising a chalcone isomerase-like (CHIL) polypeptide heterologous to the host cell and a Type 3 polyketide synthase (PKS).

Embodiments of the present invention provides recombinant systems, methods, and compositions for the biological production of Testosteronan (Testan), a linear α-(1→4)-linked glucuronic acid–N-acetylglucosamine polysaccharide. Microbial hosts engineered to express full-length CtTS, N-terminal truncation variants (including d64-CtTS), or the Pseudomonas CIPTS homolog synthesize high–molecular-weight Testan (≥800 kDa) with low polydispersity. Structural analysis by ¹H NMR and 2D HSQC demonstrates that all recombinant enzymes produce Testan that is chemically indistinguishable from native polymer. Purification using ultrafiltration and strong-anion exchange chromatography yields highly defined molecular-weight fractions, including monodisperse populations (PDI ~1.02). The invention further provides sulfated derivatives of Testan produced via aqueous sulfation methods, as well as Testan and functionalized Testan compositions useful for chromatographic stationary phases, biomaterial scaffolds, and chemically modifiable polymer platforms. These systems provide the first scalable, fermentation-based production methods for Testan and Testan-derived materials.