2027 results about "Enzyme Gene" patented technology

The invention provides methods and compositions for engineering microbes to generate Hydrogen. Some methods of the invention involve recoding of hydrogenase genes followed by subjecting the recoded genes to annealing-based recombination methods. The invention further provides methods of mating organisms that are transformed with recoded and recombined hydrogenase genes with other organisms containing different genome sequences.

The present invention relates to fungal Δ-15 fatty acid desaturases that are able to catalyze the conversion of linoleic acid (18:2, LA) to alpha-linolenic acid (18:3, ALA). Nucleic acid sequences encoding the desaturases, nucleic acid sequences which hybridize thereto, DNA constructs comprising the desaturase genes, and recombinant host plants and microorganisms expressing increased levels of the desaturases are described. Methods of increasing production of specific omega-3 and omega-6 fatty acids by over-expression of the Δ-15 fatty acid desaturases are also described herein.

The present invention is a substantially purified sortase-transamidase enzyme from Gram-positive bacteria, such as Staphylococcus aureus. The enzyme having a molecular weight of about 23,539 or about 29,076 daltons and catalyzing a reaction that covalently cross-links the carboxyl terminus of a protein having a sorting signal to the peptidoglycan of a Gram-positive bacterium, the sorting signal having: (1) a motif of LPX3X4G therein; (2) a substantially hydrophobic domain of at least 31 amino acids carboxyl to the motif; and (3) a charged tail region with at least two positively charged residues carboxyl to the substantially hydrophobic domain, at least one of the two positively charged residues being arginine, the two positively charged residues being located at residues 31-33 from the motif, wherein X3 is any of the twenty naturally-occurring L-amino acids and X4 is selected from the group consisting of alanine, serine, and threonine, and wherein sorting occurs by cleavage between the fourth and fifth residues of the LPX3X4G motif. Variants of the enzyme, methods for cloning the gene encoding the enzyme and expressing the cloned gene, and methods of use of the enzyme, including for screening for antibiotics and for display of proteins or peptides on the surfaces of Gram-positive bacteria, are also disclosed.

This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.

Use of polymorphism site gene to predict ACEI medicine, its method and reagent kit are disclosed. It can be used to determine critical enzyme gene polymorphism site gene typing oligonucleotide and critical enzyme gene polymorphism site gene in cysteine metabolic pathway.

The present invention relates to elongase genes, their polypeptides and their control regions, and the use of such genes, polypeptides and control regions in determining compositions for use in the treatment of disease. The identified compositions regulate the expression of the elongase genes or modulate the activity of their protein products. The nucleotide and amino acid sequences are taught for ELG4, ELG6 and ELG7. The control sequences and function are taught for ELG1, ELG2, ELG3, ELG4, ELG5, ELG6 and ELG7.

The invention provides a method for typing three sites of key enzyme genes (MTHFR and MTRR) for folic acid metabolism by utilizing a fluorescence labeling probe and combining with a multiple PCR method. An amplimer and a fluorescent probe are designed according to MTHFR and MTRR genes; three sections of DNA sequences are detected in the amplification of a PCR amplifier; fluorescence signals which are released in amplification and dissolution processes in a reaction system are detected; a result is judged in the manner of (1) judging a typing result according to a Tm value shown by a hybrid peak of wild type and mutant type standard plasmids and (2) simultaneously typing according to the magnitude of a fluorescence value of an amplification curve. The primer and the probe adopted by the invention are high in specificity and sensitivity; the detection for three sites is simultaneously performed; the operation is simple and the result is easily judged; the typing result is more accurate and reliable in the manner of twice calibrating typing. The method provided by the invention can be applied to the aspects of guiding the pregnant woman to orally take and supplement folic acid, prompting the high risk in cerebral apoplexy, coronary heart disease and venous thrombus, assessing the metabolic activity of folic acid, and the like.

The invention provides a method for suppressing the gene expression of insects through feeding dsRNA, which belongs to the growth development field of adjusting and controlling insects through naturally feeding the dsRNA of the silenced and specific gene. The method selects a gene fragment SeA3 on chitin synthetase enzyme gene SeCHS-A, and studies the impact on the insects after the natural feeding. The method mainly comprises the steps: a proper expression vector L4440 is selected to construct a recombinant expression vector L4440-SeA3 with a purpose gene fragment SeA3; the recombinant vector is induced and expressed in specific recipient cell coliform bacteria HT115 (DE3) under vitro adaptive condition; and the recipient bacterium htLSA3 which expresses dsRNA is added into the feedstuff of the insects, the insect larvae are naturally fed with the feedstuff, the situation of the insects suppressing the gene expression is detected after the feeding, and the change of the phenotype of the insects is recorded. By adopting natural feeding to suppress the expression of the insect genes, the method explores new paths for controlling the insects.

The invention discloses an E. coli BL21(pETDuet-ydjLnox) containing a 2R,3R-butanediol dehydrogenase gene ydjL and an NADH oxidase gene nox, wherein the strain is preserved in the 'China Center for Type Culture Collection' on December 23 in 2008, and the preservation number is CCTCC NO: M 208259. The invention also discloses application of a gene recombination bacterium in producing chiral S-AC by catalyzing meso-BD, producing chiral R-AC by catalyzing 2R,3R-BD and producing chiral pure 2S,3S-BD by splitting a 2,3-BD mixture. The concentration of a chiral AC prepared from the recombinant E. coli can reach over 6 grams per liter (the ee value is more than or equal to 96 percent); and the ee value of the chiral pure 2S,3S-BD is more than or equal to 98 percent, and the regeneration of a cofactor is achieved, thus the gene recombination bacterium has great industrial application prospect.

The invention provides chitinase gene sequences and application of dsRNA thereof, which can silence specific chitinase genes and make insects die of retarded development. By cloning, sequencing and analyzing expression of the chitinase genes of a plurality of locusta migratoria manilensis, the chitinase genes with a sequence of SEQ IDNO:1 are obtained; and the chitinase genes with sequences of SEQ ID NO:2 and SEQ ID NO:3 are selected and used for synthesizing the dsRNA. After the dsRNA is injected into a coelom of an insect, the insect shows difficult molting in the development process and dies. The chitinase gene sequences provide a basis for the effective prevention and control of the insects based on RNA interference, and provide a new approach for the development of a safe and pollution-free method for preventing and controlling the insects.

The present invention relates to fungal Δ12 fatty acid desaturases that are able to catalyze the conversion of oleic acid to linoleic acid (LA; 18:2). Nucleic acid sequences encoding the desaturases, nucleic acid sequences which hybridize thereto, DNA constructs comprising the desaturase genes, and recombinant host microorganisms expressing increased levels of the desaturases are described. Methods of increasing production of specific ω-3 and ω-6 fatty acids by over-expression of the Δ12 fatty acid desaturases are also described herein.

A unique carotenogenic biosynthetic gene cluster has been isolated from Panteoa agglomerans strain DC404, wherein the genetic organization of the cluster is crtE-idi-crtY-crtI-crtB-crtZ. The genes contained within this cluster encode geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), isopentenyl pyrophosphate isomerase (Idi), lycopene cyclase (CrtY), phytoene desaturase (CrtI), phytoene synthase (CrtB), and β-carotene hydroxylase (CrtZ). The gene cluster, genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

The invention relates to the technical field of gene engineering, and discloses a recombinant yeast strain, and a building method and application thereof. The recombinant yeast strain includes two specific gene segments recombined and integrated onto a genome in a homologous way through yeast. By selecting the specific group for forming a promoter combination and matching phaffia rhodozyma sourced phytoene synthetase and lycopene cyclase dual-function enzyme gene CrtYB, GGPP synthase CrtE, phytoene desaturase CrtI and specific yeast endogenous gene and the like, the substances are integrated onto the yeast strain genome through modular design; the yp1062w gene is knocked away through homologous recombination; one strain of fire-new recombinant yeast without an inducing agent is obtained; the high yield of the beta-renieratene is ensured.

The invention relates to an improved rhizomucor miehei lipase gene and an application to yeast display. The sequence of the improved rhizomucor miehei lipase gene is SEQ.ID.No2, with respect to a recombinant vector pMD18-T-RML containing the gene, RML means lipase gene; and the collection number of a bacterial strain Escherichia coli TOP10/pMD18-T-RML carrying the plasmid is CCTCC M 208136. In the invention, the gene is transferred into pichia stipitis host strain, so that the rhizomucor miehei lipase is displayed and expressed in the pichia stipitis. The provided pichia stipitis can effectively display the rhizomucor miehei lipase. The lipase can be widely applicable for producing fatty acid methyl ester, ethyl caproate, triglycerides which have different melting points but does not contain various types of fatty acid and a few 'reconstructed esters'.

The invention relates to a bacillus pumilus (Bacillus pumilus) L-9 laccase gene cotA as well as recombinant expression and application thereof, belonging to the field of bioengineering. The cotA is separated from the bacillus pumilus (Bacillus pumilus) L-9, an expression vector pColdII-cotA is constructed, and an escherichia coli expression strain BL21(DE3) is transformed to realize high-efficiency expression. The obtained recombinase laccase has the advantages of high alkaline environment resistance and high high-temperature resistance, has a good effect of discoloring azo dyes and anthraquinone dyes and has high application potential.

The invention relates to lipase mutants with enhanced thermal stability, which belongs to the technical field of enzyme gene engineering. Rhizopus chinensis CCTCC No.M201021 lipase used as the parent is processed by molecular biotechnology to obtain the lipase mutants with enhanced thermal stability. In the amino acid sequences of the mutants, the related amino acid mutation(s) is(are) one or a plurality of Met101Thr, Glu107Gly, Ala129Ser, Ser151Asn, Cys160Leu, Lys161Arg, Pro168Leu, Pro168His, Leu180His, Asp182Tyr, Thr183Ala, Thr218Ser, Lys219Asp, Ala230Phe, Ser234Phe, Val261Gly, His317Pro, Val329Ala, Glu363Arg, Asn366Asp and Ser373Cys. The mutants are expressed by half-life period t50 at 65 DEG C. The thermal stability of the mutants is enhanced as compared with the parent Rhizopus chinensis lipase. The invention also discloses a DNA sequence, an expression carrier and a host cell for coding the lipase mutants.

The invention discloses an expression system and constructing PHB transgene algae method of rhine chlamydomonas exogenesis gene, which comprises the following parts: promotor, exogenesis-goal gene, screening badge and rhine chlamydomonas acceptor algae strain, wherein the method comprises the following: A, selecting and cultivating acceptor algae strain; B, cloning key enzyme gene by PHB biosynthesizing; C, expressing carrier of PHB biosynthesizing key enzyme rhine chlamydomonas; D, using PHB biosynthesizing key enzyme to lead in rhine chlamydomonas from fungus Bacillus alcaligenes with bead-milling method, gene-gun method or electrization; obtaining bivalence or transgene Chlamydomonas of producing PHB by screen selecting and molecule detecting. The poly-hydroxybutyric acid is compounded by acting in conjunction of PHB compounding key enzyme of phbA, phbB and phbC gene coding, which uses acetylcoenzyme A as substrate by photosynthesis. The method simplifies the operating, which reduces cost.

The invention relates to a ZEN (zearalenone) degrading enzyme gene and a high-yield strain. The ZEN degrading enzyme gene comprises DNA molecules of (a), (b) or (c), wherein (a) has DNA molecules of a nucleotide sequence shown in SEQ ID NO.1; (b) hybridizes with the nucleotide sequence in (a) under a strict condition and encodes DNA molecules of protein with ZEN degrading enzyme activity; (c) has DNA molecules of a nucleotide sequence with 90% or higher of homology with the nucleotide sequence in (a) or (b). The invention establishes the high-yield strain for producing ZEN degrading enzymes through the sequences and further provides a high-density fermentation method. ZEN degrading enzymes can be subjected to efficient secretion expression through the high-yield strain with the high-density fermentation method, and ZEN can be quickly and efficiently degraded during fermentation.

The invention discloses escherichia coli genetically engineered bacteria co-expressed by three enzymes. The escherichia coli genetically engineered bacteria are characterized in that an L-amino-acid oxidase gene, a (D/L)-alpha-hydroxyl carboxylic acid dehydrogenase gene and a gene capable of reducing NAD(P) are imported. The invention also discloses a construction method and an application of the recombinant escherichia coli. The method is used to biologically synthesize optically pure alpha-hydroxyl carboxylic acid, has the characteristics of simple operation, low cost, high product synthesizing efficiency and high optical purity, and has a good industrial prospect.

The invention provides recombinant bacillus subtilis for synthesizing 2'-fucosyllactose and an establishment method and application of recombinant bacillus subtilis. Recombinant bacillus subtilis is obtained in the mode that a lactose transport enzyme gene is subjected to reinforcement expression in a gene group of bacillus subtilis 168, and exogenous fucosyltransferase, fucokinase and gtp-mannose-1-phosphate guanylyltransferase genes are expressed. According to recombinant bacillus subtilis, the lactose transport enzyme gene is subjected to reinforcement expression on the gene group, expression of the lactose transport enzyme is effectively reinforced, the efficiency of transferring exogenous lactose into cytoplasm is improved, the concentration of lactose in cytoplasm is increased, and synthesis of 2'-fucosyllactose is promoted. The establishment method of recombinant bacillus subtilis is simple and convenient to use and has good application prospects.

The invention discloses application of panax japonicus transcription factor gene PjWRKY1, namely, application in improving the expression quantity of key enzyme genes in the biological synthesis of panax japonicus saponins, and increasing the content of saponins in panax japonicus callus. The nucleotide sequence of the PjWRKY1 gene is shown as SEQ ID NO: 1, and WRKY transcription factors are coded. Through the adoption of functional genomics and techniques related to metabolic engineering, a panax japonicus PjWRKY1 transcription factor is proved to have the effect of positively regulating the biological synthesis of panax japonicus saponins; when the panax japonicus PjWRKY1 transcription factor gene disclosed by the invention is established on a plant expression vector and transferred into the panax japonicus callus for over expression, the expression quantity of the key enzyme genes in the synthetic route of panax japonicus saponins is increased, and the yield of panax japonicus saponins is increased.