79 results about "Biosynthetic genes" patented technology

The invention relates to the technical field of biology, in particular to glycopeptide antibiotic and application thereof. According to the invention, a biosynthetic gene cluster for synthesizing a brand new glycopeptide antibiotic skeleton is excavated, and brand new glycopeptide antibiotics Varsomycin A, B and C with novel chemical structures are obtained through heterologous expression; the traditional Chinese medicine composition has a remarkable antibacterial effect on clinically drug-resistant strains. Important fermentation strains and precursor compound molecules are provided for research and development of new drugs of glycopeptide antibiotics, and important application prospects and economic values are achieved.

The invention discloses a saccharomyces cerevisiae recombinant strain based on a uracil metabolism gene URA3 as well as a construction method and application of the saccharomyces cerevisiae recombinant strain. The construction method comprises the following steps: taking saccharomyces cerevisiae rich in farnesyl pyrophosphate as an original strain, introducing a uracil metabolism gene URA3, synchronously overexpressing a ribosome biosynthetic gene UTP10 and an iron metabolism gene FIT3, and utilizing the uracil metabolism gene URA3 to express up-regulation correlation with the ribosome biosynthetic gene UTP10 and the iron metabolism gene FIT3, so as to obtain the farnesyl pyrophosphate-rich saccharomyces cerevisiae. The metabolic flux of the terpenoids is enhanced, and the saccharomyces cerevisiae recombinant strain for efficiently producing the terpenoids is constructed. According to the method, a metabolic network of yeast is systematically optimized, and combined regulation and control on precursor flux distribution, protein translation capability and metal ion steady state are realized, so that the microbial synthesis efficiency of sesquiterpenoids such as patchouli alcohol and beta-elemene is remarkably improved.

The invention discloses an application of an overexpressed bellseed AlbHLH175 gene in increasing tropine alkaloid and precursor content thereof, by creating an AlbHLH175 overexpressed transgenic hairy root strain and analyzing expression quantities of AlbHLH175 and TAs biosynthetic genes in the transgenic strain, results show that the AlbHLH175 can positively regulate expression of seven biosynthetic genes, namely AlODC, AlPMT, AlMPO, AlPYKS, AlCYP82M3, AlPPAR and AlHDH, and the expression quantity of the AlbHLH175 and TAs biosynthetic genes in the transgenic strain is analyzed. Therefore, accumulation of putrescine, N-methyl putrescine, tropinone, tropine, conpin, scopolamine, anisodamine and scopolamine is promoted, and an excellent gene resource is provided for creating a stable and high-yield TAs new germplasm of the bellflower through biological breeding and molecular design.

The invention discloses a streptomyces Targetron gene targeting vector as well as a construction method and application of the streptomyces Targetron gene targeting vector. According to the invention, an II-type intron Ll.LtrB from lactococcus lactis is cloned to an escherichia coli-streptomyces shuttle plasmid to construct a streptomyces genetic manipulation tool-vector pSC30 based on targeting of the II-type intron, and gene targeting in streptomyces coelicolor is successfully realized by using the tool. Screening a biosynthetic gene cluster of red pigment in streptomyces roseosporus; and activating a biosynthetic gene cluster of janus doxorubicin in streptomyces spinosus. According to the invention, the conversion rate and the gene targeting efficiency of the Ll.LtrB intron in streptomyces with high GC content are effectively improved, and a new tool is provided for construction of a gene mutation library.

The invention discloses a biosynthetic gene cluster of a novel skeleton sesquiterpenoid (sestercyclic acid) and an application of the biosynthetic gene cluster of the novel skeleton sestercyclic acid (sestercyclic acid). The sesterterpene tercyclic acid biosynthetic gene cluster at least comprises three genes, namely, terA, terB and terC, and the nucleotide sequences of the sesterterpene tercyclic acid biosynthetic gene cluster are as shown in SEQ ID NO.1-3. The gene cluster of the terrecyclic acid compound disclosed by the invention is found for the first time, and each gene function is also confirmed for the first time. Protease TerA catalyzes farnesyl pyrophosphate (FPP) to be cyclized into a new skeleton sesquiterpene 25, then P450 protease TerB is oxidized into a compound 32, and the compound 32 is dehydrogenated through protease TerC to form a final product compound 2. According to the present invention, the effective drug source approach is provided for the development of the terrecyclic acid antitumor compound.

This invention discloses a *Bacillus laterosporus* strain for the stable production of nonribosomal cyclic lipolipeptide and its construction method. The M1A gene in the Bogorol biosynthetic gene cluster is knocked out using CRISPR-Cas9 technology to block the Bogorol pathway, which competes with Brevicidine for substrates. Subsequently, the pMCCas9-S48 plasmid, which integrates S8 and S41 sgRNAs and their homologous arms as repair templates, is used to simultaneously knock out S8 and S41 family peptidase genes, preventing the expression of extracellular degradation enzymes. This invention also provides the application of the above strain in the preparation of nonribosomal cyclic lipolipeptide of Brevicidine and a method for preparing the nonribosomal cyclic lipolipeptide. This strain, through reducing metabolic competition and preventing product degradation, exhibits rapid cell growth, stable production of Brevicidine, and significantly reduced degradation rate, resulting in a 66.13% increase in fermentation yield compared to the unmodified strain. This invention solves the problems of degradation and unstable yield in the production of Brevicidine, provides an effective technical approach for the industrial production of Brevicidine, and provides important technical support for the development and application of novel antimicrobial peptide drugs.

The present invention discloses an anti-inflammatory compound, its biosynthetic gene cluster, synthesis method and application, belonging to the technical field of genetic engineering. An anti-inflammatory compound is provided, and the chemical molecular formula of the anti-inflammatory compound is C<subgt;15< / subgt;H<subgt;22< / subgt;O<subgt;5< / subgt>, and the structural formula is shown in Formula I, named: phaseolinone B. The present invention first discloses the biosynthetic pathway of the anti-inflammatory compound phaseolinone B, laying an important foundation for realizing its green and efficient synthesis; this compound exhibits significant anti-inflammatory activity in the zebrafish Tg 3dpf tail amputation infection model, providing resources for discovering anti-inflammatory drug lead compounds.

The present application belongs to the field of biotechnology, and particularly relates to a biosynthesis gene cluster of fungal polyketide compounds and application thereof. The present application provides a heterologous expression method of an aspertaichunol compound biosynthesis gene combination, and a compound aspertaichunol B is obtained through the method, and a compound aspertaichunol A is prepared through acid hydrolysis of the aspertaichunol B, which has important significance for future research and development of an efficient biosynthesis method for preparing the aspertaichunol compound.

Methods for identifying biosynthetic gene clusters that include genes for producing compounds that interact with specific target proteins are disclosed. Some methods relate to bioinformatics methods for identifying and / or prioritizing biosynthetic gene clusters. Related systems, components, and tools for the identification and expression of such gene clusters are also disclosed.

The application discloses a gene combination for biosynthesis of paclitaxel and application thereof, and relates to the technical field of genetic engineering. The application breaks through the limitation of traditional paclitaxel extraction and synthesis technology, and for the first time adopts a transformation method of Physcomitrella patens to perform multi-gene co-expression on a paclitaxel synthesis pathway. The application has wider gene source compatibility, and opens up a brand-new technical path for paclitaxel synthesis. As a model plant, the Physcomitrella patens has the characteristics of rapid growth and easy cultivation. After a specific gene combination is introduced into the Physcomitrella patens, a metabolic pathway related to paclitaxel synthesis in the Physcomitrella patens cells can be fully activated, and the Physcomitrella patens is prompted to efficiently synthesize paclitaxel.

The invention discloses a 5 / 8 / 5 chitosan clostridium diterpenoid compound, a biosynthetic gene cluster and application of the biosynthetic gene cluster, and relates to the technical field of biological medicines. A 5 / 8 / 5 chitin type diterpenoid biosynthetic gene cluster Thm in trichoderma harzianum is identified through a genome mining system, chitin diterpenoid compounds with diversified structures are obtained by means of heterologous expression, the obtained compounds are subjected to anti-thrombotic activity comprehensive evaluation, and the result shows that the compounds 6 and 31 can obviously inhibit vascular thrombosis, the compound 6 has a remarkable inhibition effect on collagen-induced platelet aggregation, and the compound 31 has a better inhibition capability on thrombin-induced platelet activation. The compounds 6 and 31 show powerful antithrombotic effects in artery, vein and pulmonary artery thrombosis models, so that powerful technical support is provided for development of antithrombotic drugs.

The invention discloses a production strain of demethylated ristomycin aglycone as well as a construction method and application of the production strain. The construction method comprises the following steps: synthesizing a gene cluster lacking a glycosylation modification function by adopting a gene editing mode, and then expressing the edited gene cluster in a heterologous or homologous host; or the glycosyl transferase gene in the strain containing the ristomycin biosynthetic gene cluster is knocked out. At present, no report about biosynthesis of the demethylated ristomycin aglycone exists, and the invention provides a biosynthesis method of the demethylated ristomycin aglycone. Meanwhile, the demethylated ristomycin aglycone production strain provided by the invention has good genetic operability, is fast in growth and short in fermentation time, can reach the peak of producing the ristomycin aglycone after being fermented for 2-3 days, and has excellent industrial development potential.

The present application relates to a kind of cyclic ester peptide compound aglomycin A biosynthesis gene cluster and its application.The nucleotide sequence of the gene cluster is as shown in SEQ ID NO.1, and the use is to produce peptide compound aglomycin A or peptide compound aglomycin D.

NEW SMALL MOLECULES AND METHODS USING THEM FOR SKIN TREATMENT. The disclosure relates to new biosynthetic gene clusters encoding type II polyketides, their products, extracts and compositions comprising the products, and methods for using the products, extracts, and compositions for skin treatment, e.g., to enhance the skin's barrier function and / or immune defenses. Figure for abstract: none

The present disclosure relates to methods and systems for identifying genes associated with biosynthetic gene clusters (BGCs), including embedded target genes (ETaGs) that are homologs of potential therapeutic targets. The methods and systems described herein apply a comparative genomics and manual review or machine-learning model to analyze grid representations such as heat maps, which assess across a plurality of diverse genomes distribution of orthologs (e.g., bidirectional best hits) of a plurality of query genes that are co- localized with an anchor gene (e.g., a core synthase gene) of a BGC in a query genome.

Colistin is an important last-resort antibiotic, but its production involves significant challenges due to the presence of impurities resulting from various analogs and the limited number of native producers. In response, disclosed are mutant Paenibacillus bacteria that produce Colicin A more efficiently and / or Colicin A that is of higher purity and / or has a cleaner metabolite background. The disclosed mutant bacteria are genetically engineered to inactivate expression of one or more biosynthetic gene clusters (BGCs), increase expression of one or more other BCGs, and / or increase expression of one or more L-DABA pathway genes.

The invention relates to the technical field of saccharomyces cerevisiae expression foreign protein, and discloses a saccharomyces cerevisiae strain with ASFV p14.5 displayed on the surface and a construction method and application thereof, and the method comprises the following steps: (1) synthesizing a delta site homologous arm sequence; (2) seamlessly connecting the homologous arm sequence of the delta site to a carrier pET23a, connecting a selective marker tryptophan biosynthetic gene TRP1 to the 5'end of the upstream homologous arm sequence of the delta site, and sequentially connecting an Aga2 gene, an E120R gene, a T2A peptide gene and an mCherry gene to the 5 'end; (3) carrying out enzyme digestion on two sides of the delta site through restriction endonuclease salI, and recovering a linearized fragment through agarose gel; and (4) integrating the recovered fragments into a host saccharomyces cerevisiae genome through a chemical conversion method, and culturing until bacterial colonies grow. The surface display type saccharomyces cerevisiae strain obtained by the invention not only has the stability of integrative saccharomyces cerevisiae for expressing foreign protein, but also can be used for more easily and more intuitively screening a high-expression quantity saccharomyces cerevisiae strain by combining the T2A peptide with mCherry.

Recombinant Sorangium cellulosum for producing de-epoxidized epothilone B by fermentation, insertional inactivation of an epoK gene in an epothilone biosynthetic gene cluster in the recombinant bacteria, and a method for producing de-epoxidized epothilone B using the recombinant bacteria.

The invention discloses streptomyces hygroscopicus ascomycetes subspecies and an obtaining method thereof. Regulating genes fkbN, fkbR1 and fkbR2 on an ascomycin biosynthesis gene cluster of the streptomyces hygroscopicus ascomycetes subspecies ATCC 14891 and a gene fkbS participating in synthesis of an ascomycin precursor ethyl malonyl coenzyme A are subjected to biosynthesis to obtain the streptomyces hygroscopicus ascomycetes subspecies. And carrying out overexpression on a regulatory gene SH03686 of a leucine response regulatory protein family (Lrp) outside the gene cluster, and screening an ascomycin high-yield strain. According to the invention, after combined overexpression of genes fkbS and fkbN and overexpression of a gene fkbR2 are carried out by a strain SN9 and a strain R2-17 in starting bacteria, the transcriptional level of regulatory genes (fkbN, fkbR2) on an ascomycin biosynthesis gene cluster and related genes participating in synthesis of precursors ethyl malonyl coenzyme A (fkbS, fkbE), piperidine acid (fkbL) and DHCHC (fkbO) are improved, and finally, the ascomycin yield is improved.

The application discloses a method for biosynthesis of gene or gene cluster product based on chromatin three-dimensional structure, and belongs to the field of genetic engineering. The method can realize the improvement of the target gene or gene cluster product by the directional integration of the target gene or gene cluster into the strong correlation chromatin region with high frequency interaction on the genome. The method has the characteristics of simple implementation, good repeatability, remarkable effect and the like, and can be widely applied to the improvement of the target gene or gene cluster product.

The application belongs to the field of agricultural biotechnology and microbial engineering technology, and particularly relates to a biosynthesis gene cluster of cystobactamids and application. The nucleotide sequence of the biosynthesis gene cluster is shown as SEQ ID NO:1. The application discloses a novel biological function of cystobactamids except for antibiotics, broadens the application scene, and has a good development prospect in the field of agricultural biological control.

The invention discloses a pear PbrRGL3 gene and application of the pear PbrRGL3 gene. The PbrRGL3 gene which is separated from Dangshan pears and has the effect of inhibiting the formation of pear cell lignin has a nucleotide sequence as shown in SEQ ID No. 1, and an amino acid sequence coded by the PbrRGL3 gene is as shown in SEQ ID No. 2 in a sequence table. The PbrRGL3 gene is subjected to transient overexpression in pear fruits by adopting an agrobacterium transient transformation method, the lignin content of pulp is remarkably reduced, and the expression of a lignin biosynthetic gene is reduced; the effect of silencing the PbrRGL3 gene is opposite to that of silencing the PbrRGL3 gene; an overexpression vector constructed by the gene is introduced into arabidopsis thaliana, the lignin content of inflorescence stems of the obtained PbrRGL3 transgenic arabidopsis thaliana is remarkably reduced, and the lignification degree of conduit cells and vascular intervascular fibrocytes is weakened. Therefore, the PbrRGL3 participates in regulating and controlling the formation of the lignin in the pear fruit stone cells.

The application discloses a biosynthesis gene cluster of a polyene macrolide natural product mandimycin, and a natural product and application of the biosynthesis gene cluster, wherein the nucleotide sequence of the biosynthesis gene cluster is shown as SEQ ID NO. 1, and the compound structural formula of the natural product mandimycin and mandimycin B is shown as formula I and formula II. The natural product mandimycin can target fungal cell membrane phospholipid molecules, especially phosphatidylinositol, cause important ion efflux in fungal cells, and lead to fungal cell death. The natural product mandimycin and mandimycin B have strong in-vivo and in-vitro antibacterial activity and a broad antibacterial spectrum on various WHO-published multiple drug-resistant fungal key pathogens, including candida, aspergillus, cryptococcus, mucor and fusarium.

The invention provides amycolatopsis mediterranei with gene defects as well as a construction method and application of the amycolatopsis mediterranei. The first aspect of the invention provides a construction method of the amycolatopsis mediterranei with the gene defect, and the construction method comprises the step of knocking out a rifamycin biosynthetic gene cluster in a receptor amycolatopsis mediterranei strain U32 (Cas12a) genome and an integrated plasmid pDZLCas12a containing an FnCas12a coding gene to obtain the amycolatopsis mediterranei with the gene defect. According to the invention, a rifamycin biosynthetic gene cluster in a genome of an amycolatopsis mediterranei strain U32 (Cas12a) and an integrated plasmid pDZLCas12a containing an FnCas12a coding gene are knocked out, so that the amycolatopsis mediterranei with gene defects is constructed, and the amycolatopsis mediterranei can be used as a chassis strain for mining identified or unidentified biosynthetic gene clusters in actinomycetes.

The application discloses streptomyces 891-DC with high aurovertin A yield and application thereof. The application uses CRISPR-Cas9 technology to first change an incomplete attB site sequence in the genome of streptomyces 891 into a complete attB site sequence, promote a plasmid containing a target gene chry, an integrase phiC31 gene and an attP site, and improve the integration rate of the target gene through specific combination of the attP site and the attB site on the genome. Secondly, a promoter capable of obviously improving the yield is screened to express the aurovertin biosynthesis gene cluster, so that the more stable and higher-yield streptomyces 891-DC is obtained. The aurovertin A shake flask fermentation yield is 1411.23+ / -29 mg / L, which is 60% higher than that of the original strain, and the genetic stability is good, thereby laying a technical foundation for industrial production of aurovertin.

The present application relates to a kind of antibacterial new skeleton diterpene compound and its synthesis method.The present application obtains a terpene cyclase of unknown function and a P450 oxidase of unknown function from the genomic data disclosed in a strain of Aspergillus fumigatiaffinis CNM-CM8980, by heterologous expression and in vitro enzyme catalytic reaction of Aspergillus oryzae, it is found that the terpene cyclase can be catalyzed to generate a diterpene compound with 6 / 5 / 5 / 5 brand new four-ring skeleton, candidate P450 oxidase can act on the diterpene compound to generate the oxidation product with antibacterial activity.The present application first reports the chemical structure of the diterpene compound, biological activity and biosynthesis gene, on the one hand, provides lead compound resources for discovering new antibiotics, on the other hand, lays a foundation for realizing its green and efficient synthesis.

The invention belongs to the technical field of agricultural biotechnology and microbial engineering, and particularly relates to a biosynthetic gene cluster of cystobactamid compounds and application of the biosynthetic gene cluster. The nucleotide sequence of the biosynthetic gene cluster is as shown in SEQ ID NO: 1. The invention discloses a brand new biological function of the cystobactamid except antibiotics for the first time, broadens the application scene of the cystobactamid, and has a good development prospect in the field of agricultural biological control.

The present disclosure relates to computer-implemented methods and systems for identifying biosynthetic gene clusters (BGCs) that encode pathways for the production of secondary metabolites. Secondary metabolites that target genes or gene products that are homologous to, e.g., human genes or gene products may have utility as potential drug compounds.

Colistin is an important last-resort antibiotic, but its production involves significant challenges due to the presence of impurities resulting from various analogs and the limited number of native producers. In response, disclosed are mutant Paenibacillus bacteria that produce Colicin A more efficiently and / or Colicin A that is of higher purity and / or has a cleaner metabolite background. The disclosed mutant bacteria are genetically engineered to inactivate expression of one or more biosynthetic gene clusters (BGCs), increase expression of one or more other BCGs, and / or increase expression of one or more L-DABA pathway genes.

The invention discloses a method for constructing high-yield engineering bacteria of an oritavancin precursor cloireomycin. A high-yield chassis strain capable of being used for efficiently synthesizing glycopeptide antibiotics is constructed by knocking out an NRPS gene responsible for ristomycin biosynthesis in TNS106, inserting an integration site and then inserting a pJLL10 plasmid with a double-copy bbr gene. A bacterial artificial chromosome (BAC) plasmid containing a glycopeptide antibiotic biosynthetic gene cluster is introduced into the high-yield chassis strain to obtain the glycopeptide antibiotic high-yield engineering strain. The shake flask fermentation yield of the cloirusin high-producing strain obtained by the method disclosed by the invention can reach 1440mg / L, and the cloirusin high-producing strain is the strain with the highest shake flask fermentation yield in a public report.