319 results about "Transport protein" patented technology

The invention discloses construction and application of an L-malic acid producing issatchenkia orientalis strain, and belongs to the technical field of microorganisms and the technical field of microbial fermentation and metabolic engineering. Through a metabolic engineering means, a cytoplasm metabolic pathway of an issatchenkia orientalis strain is transformed, and a pyruvate decarboxylase coding gene, glycerol dehydrogenase and lactic dehydrogenase are knocked out; a pyruvate carboxylase gene derived from aspergillus flavus, a malate dehydrogenase gene derived from corynebacterium glutamicum and phosphoenolpyruvate carboxylase derived from escherichia coli are screened and integrated, so that a cytoplasm rTCA cycle synthesis pathway is enhanced; inserting a dicarboxylic acid external transporter gene from schizosaccharomyces cerevisiae at a neutral site GD21, and knocking out endogenous dicarboxylic acid internal transporter; the L-malic acid yield of the modified yeast strain in the fermentation process is remarkably increased and reaches 140.16 g / L. The engineering strain provided by the invention has good industrial application potential in efficient L-malic acid production.

The invention provides an application of a transporter in improving the yield and / or purity of lactose-N-neotetraose (LNnT), and relates to the technical field of genetic engineering. Transporter proteins of different sources are screened, the transporter protein which aims at LNnT synthesis and can achieve efficient transportation is obtained, and a new biological material is provided for LNnT synthesis. Moreover, the efficient transporter obtained by screening is used for transporting the LNnT to the outside of cells, so that the pressure on the cells caused by overhigh concentration of intracellular products is relieved, the conversion of an intermediate product LNT II to the LNnT is improved to the maximum extent, the yield is improved while byproducts are reduced, the yield of a byproduct lactose-N-neohexaose is also reduced, and the separation and purification of later products are facilitated.

The invention belongs to the technical field of gene engineering, and particularly relates to a mutant of polymyxin efflux transporter and application of the mutant. The mutant is a PmxD transporter mutant, the amino acid sequence of the PmxD transporter mutant is shown as SEQ ID NO: 1, and compared with wild type PmxD, the polymyxin transport capacity of the PmxD transporter mutant (T38W) is improved by 450.46%; and the total discharge amount of polymyxin is increased by 85.72%. Meanwhile, the mutant can significantly improve the growth ability of the strain on a plate containing 250 [mu] g / mL of polymyxin B, namely significantly improve the autoresistance of paenibacillus polymyxa to polymyxin.

The invention discloses an L-histidine transporter mutant and a method for increasing the yield of L-histidine, and the mutant has any one mutation of I20A, V28L, I20A-V28L and I20A-V21L-T28S relative to an escherichia coli wild type yeaS. The yield of the L-histidine of the recombinant escherichia coli containing the yeaS mutant protein is remarkably increased, it is indicated that the yeaS mutant protein and the coding gene of the yeaS mutant protein can be used for increasing the yield of the L-histidine, and the mutant can lay a good foundation for industrial efficient production of the L-histidine and downstream metabolites.

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 provides a method for synthesizing D-psicose through fermentation of escherichia coli by using D-glucose and D-xylose. The method comprises the following steps: by taking escherichia coli as a host cell, over-expressing a galactose permease gene galP, a glucokinase gene glk, a 6-phosphate glucose isomerase gene pgi, a D-psicose-6-phosphate epimerase gene alsE with an SUMO tag and a D-psicose-6-phosphate phosphatase gene a6PP, and constructing a path for synthesizing D-psicose by taking D-glucose and D-xylose as substrates; the synthetic route of D-psicose is optimized by knocking out a D-glucose phosphorylation transport pathway gene ptsG, knocking out D-fructose-6-phosphokinase genes pfkA and pfkB, silencing a glucose-6-phosphate dehydrogenase gene zwf, overexpressing glutamate transhydrogenase genes gdh1 and gdh2, knocking out a UDP-glucose-4-epimerase gene galE and knocking out a transport protein key gene fryA in D-psicose; and finally, the optimal proportion of mixed sugar fermentation is explored, so that efficient synthesis of D-psicose from escherichia coli through mixed sugar fermentation of D-glucose and D-xylose is realized.

The invention relates to the technical field of microorganisms, and particularly discloses a recombinant microorganism and application thereof in production of hexamethylenediamine. Compared with an original strain, the recombinant microorganism disclosed by the invention expresses a carboxylic acid reductase CAR variant, a CAR activating enzyme sfp and putrescine transaminase patA; the amino acid sequence of the carboxylic acid reductase CAR variant is as shown in SEQ ID NO. 28; preferably, adipic acid transporters DcaK and DcaP, acyl CoA transferases DcaI and DcaJ, an acylated succinate semialdehyde dehydrogenase sucD, and a 4-aminobutyric acid transaminase gabT are also expressed. By utilizing the recombinant microorganism, 6-aminocaproic acid / adipic acid can be efficiently converted into hexamethylenediamine, and a new method is provided for green biosynthesis of hexamethylenediamine.

The invention belongs to the technical field of biology, and discloses application of a lignin monomer transporter. The amino acid sequence of the lignin monomer transporter is as shown in SEQ ID No: 1. The application is used for constructing a recombinant strain growing in a culture medium of lignin-derived aromatic compounds, or used for constructing a recombinant strain for producing microbial proteins and / or lipids. According to the novel lignin monomer transport protein driven by deep learning, the metabolic bottleneck of lignin utilization is broken through, the key protein transported by the lignin monomer is excavated, the lignin monomer transport capacity can be endowed to a microbial chassis, and a multifunctional microbial platform is provided for sustainable biological refining.

Described herein are compositions and methods for treatment of Age-related Macular Degeneration (AMD). In particular, described herein are polypeptides comprising helical structures having ATP binding cassette transporter membrane stabilization and agonist activity, transporter protein binding activity, and / or that may result in a cholesterol efflux, as well as methods of using these peptides to treat AMD.

The invention discloses a cherry sugar transporter gene CpSWEET10 and application thereof, and belongs to the technical field of cherry gene engineering. A novel sugar transporter gene CpSWEET10 is obtained through cloning, the nucleotide sequence of the novel sugar transporter gene CpSWEET10 is shown as SEQ ID NO.1, and the coded amino acid sequence of the novel sugar transporter gene CpSWEET10 is shown as SEQ ID NO.2. The gene is highly expressed in cherry fruits, and the content of soluble solids and total sugar in mature fruits is remarkably positively correlated with the expression quantity of the soluble solids and the total sugar; through subcellular localization and sugar transport function defect type yeast mutant functional complementation experimental analysis, the protein coded by the CpSWEET10 gene is localized in a cytoplasmic membrane and has a sucrose transport function; the cherry sugar transporter gene CpSWEET10 disclosed by the invention can be applied to the improvement of the sugar degree character of the fruits, and a new gene resource is provided for molecular breeding for improving the high-sugar quality of the cherry fruits.

The invention provides an application of a citrus transcription factor CsAGL9 gene and a protein coded by the citrus transcription factor CsAGL9 gene in regulating and controlling the content of citric acid in citrus fruits, the nucleotide sequence of the CsAGL9 gene is shown as SEQ ID NO.1, and the amino acid sequence of the protein is shown as SEQ ID NO.2. The content of citric acid in the fruits can be reduced through overexpression in the oranges; the content of citric acid in fruits can be increased by silencing the gene. It is further verified that the gene promotes the activity of the citrus citric acid transporter CsTCT1. The discovery provides a new gene resource for citrus quality improvement and molecular design breeding, and provides an efficient tool for fruit quality improvement.

The invention provides a milk protein peptide which takes milk protein as a raw material and has the effects of promoting calcium absorption and improving bone health and application of the milk protein peptide. Specifically, milk protein peptide sequences, including YGGVSL and SQRY, are screened by adopting enzymolysis, sequence identification and molecular docking methods. The milk protein peptide is proved to have high calcium transport promoting capacity in the in-vitro intervention of Caco-2 cells, and the milk protein peptide is proved to have the capacity of promoting osteogenesis and inhibiting bone resorption in the in-vitro intervention of MC3T3-E1 mouse embryo osteoblasts. In animal experiments, by increasing the content of blood calcium and bone calcium, reducing the content of excrement calcium, improving mRNA expression of calcium ion channel protein, calcium binding protein and peptide transporter in intestinal tracts, enhancing the efficiency of calcium entering blood, promoting osteogenesis and inhibiting bone resorption, the milk protein peptide is comprehensively proved to have the capabilities of improving calcium resorption of organisms and improving bone health. The milk protein peptide has both functionality and nutrition, and has a wide application prospect in the aspect of bone health.

The invention provides an application of a peach transport protein PpSWEET11 in regulating and controlling the hexose content of fruits. According to the invention, the function of the PpSWEET11 gene is cloned and verified, and through overexpression of the PpSWEET11 in peach fruits and tomatoes, the glucose and fructose levels in an overexpression group are obviously increased compared with those in a control group, which indicates that the PpSWEET11 has a wide sugar transport function. The invention not only expounds the effect of the PpSWEET11 in the aspect of sugar accumulation and provides a key molecular insight in the aspect of improving sugar accumulation of peaches and other economically important fruit crops, but also can be applied to nutritional quality improvement and germplasm innovation of peach fruits.

The invention belongs to the technical field of genetic engineering, and relates to the field of wheat genetic engineering, in particular to a gene TaABCG42 for regulating and controlling cadmium accumulation of wheat as well as an encoding protein and application of the gene TaABCG42. According to the invention, a TaABCG42 gene is knocked out by using CRISPR / Cas9, a TaABCG42 gene overexpression plant is constructed at the same time, and a homozygous mutant is obtained by using a Hi-Tom sequencing technology and PCR (Polymerase Chain Reaction) successive generation screening. A wild type TaABCG42 homozygous overexpression and knockout plant treated with a 40 [mu] mol / L Cd solution for 5 days is subjected to analysis such as metal content determination, subcellular component metal determination, Cd chemical form determination and Cd ion flow determination, and the Cd absorption and transport approach of TaABCG42 is analyzed, so that the physiological and molecular mechanism of the TaABCG42 metal transporter for absorbing and transporting Cd is explored, and the application prospect is wide. The understanding on the wheat ABC metal transporter is enriched, and a theoretical basis is provided for cultivating high-quality wheat with low Cd accumulation.

The invention discloses a tomato Solyc08g008190 gene and a tomato Solyc08g008200 gene and application of the tomato Solyc08g008190 gene and the tomato Solyc08g008200 gene. A CDS (Coding Sequence) sequence of the Solyc08g008190 gene is shown as SEQ ID NO. 1, and a CDS sequence of the Solyc08g008200 gene is shown as SEQ ID NO. 2. A tomato transporter mutant library is constructed through a CRISPR (clustered regularly interspaced short palindromic repeats) technology, and a mutant 0990-32 which simultaneously covers expression of two homologous genes of an NHE family is screened. Based on observation of the phenotype of the mutant, the inventor preliminarily determines that the two genes are related to tomato fruit quality and fruit volume increase. Compared with a wild type, the tomato fruit of the mutant has the advantages that the brix content is increased, and the fruit becomes larger. It is shown that the Solyc08g008190 gene and the Solyc08g008200 gene play an important role in regulating and controlling the growth, development and quality of the tomato fruits. The invention lays a good gene genetic resource foundation for the cultivation of new varieties of high-quality tomatoes.

The invention belongs to the technical field of biological medicine, and relates to application of a clerodendranthus spicatus tea bag in regulating intestinal excretion and reducing uric acid. One of the purposes of the invention is to provide the application of the clerodendranthus spicatus teabag in regulating intestinal tracts to reduce uric acid According to a preferred embodiment, the clerodendranthus spicatus teabag regulates ATP binding cassette transporter G2 and inhibits uricase activity. The invention provides the application of the clerodendranthus spicatus teabag in regulating the intestinal microbial environment and reducing uric acid, and the clerodendranthus spicatus teabag has remarkable beneficial effects. The clerodendranthus spicatus teabag can effectively regulate the structure and the function of intestinal microflora, especially the enrichment amount of characteristic bacteria Muriaculum in the intestinal tract, further strengthens the probiotic activity of the intestinal flora, and is helpful for maintaining the steady state of uric acid in the body. The multi-way synergistic effect not only provides an innovative strategy for preventing and treating hyperuricemia and related metabolic diseases, but also avoids side effects possibly brought by traditional medicines, and shows good application prospects and clinical values.

Methods of treating age-related macular degeneration (AMD) may include the administration of amphipathic, ATP binding cassette transporter membrane agonists to reduce the amount of drusen in the subject in need thereof. These methods may be particularly useful for treatment of intermediate AMD.

The invention relates to the field of synthetic biology, metabolic engineering and fermentation engineering, and discloses a genetically engineered bacterium and application thereof in production of alpha-ketoglutaric acid.The genetically engineered bacterium comprises the steps that the genetically engineered strain with escherichia coli BW25113 or corynebacterium glutamicum ATCC13032 as a host is constructed; overexpression type isocitrate dehydrogenase genes are integrated through chromosomes, alpha-ketoglutarate dehydrogenase complex genes are knocked out, NADPH invertase is overexpressed through plasmids, and exogenous AKG transporter protein is introduced, so that metabolic flux directional regulation and control and product efflux enhancement are realized; meanwhile, a staged fermentation process is adopted and comprises dynamic gradient adjustment of inorganic phosphorus concentration and two-stage pH control. According to the method, the AKG yield of 78.2 g / L can be obtained under the 5L tank scale, the sugar acid conversion rate reaches 0.68, and the method has the advantages of being high in yield, low in byproduct and controllable in cost. The obtained AKG is suitable for a plurality of fields such as medical intermediates, functional foods and cell culture additives, and has wide industrial application prospects.

The invention discloses a genetically engineered bacterium and an application of the genetically engineered bacterium in production of farnesene. The genetically engineered bacterium takes saccharomyces cerevisiae as a chassis strain; an exogenous beta-farnesene synthase gene is integrated, an ERG9 gene is knocked out, a promoter of a truncated HMG1 gene is replaced with an ANB1 promoter, an exogenous transhydrogenase gene is integrated and driven by a glucose sensitive promoter, beta-farnesene synthase is mutated and fused with a transmembrane structural domain of ABC transporter protein PDR10, and then a chimeric expression unit is constructed. According to the present invention, the chassis bacteria are modified by at least one of the steps of overexpression of phosphatidylcholine synthetase, knockout of sterol ester synthetase and introduction of exogenous alkyl glycerol monooxygenase, and the obtained engineering bacteria are combined with the optimized fermentation process so as to significantly improve the farnesene synthesis efficiency and the production economy; therefore, the genetically engineered bacterium and the fermentation method provided by the invention have good application prospects.

This invention relates to a method for producing chondroitin with improved safety and extracellular secretion levels, belonging to the field of genetic engineering. Using the probiotic *Escherichia coli* Nissle 1917 as the host, this invention designs methods to increase chondroitin production by studying the chondroitin synthase KfoAC and the chondroitin-linking Kdo synthase. The relationship between chondroitin synthesis and secretion in *E. coli* was clarified, and the specificity of the chondroitin-linking Kdo was investigated, resulting in an extracellular chondroitin production of 0.32 g / L. Subsequently, the introduction of the protein KfoB from the chondroitin synthesis gene cluster increased the extracellular chondroitin production to 0.51 g / L. Further screening and optimization of the expression of transport proteins resulted in an extracellular chondroitin production of 0.73 g / L in shake flasks. Further optimization using a 7-L fermenter resulted in an extracellular chondroitin production of 5.9 g / L.

Provided herein is a method of administering levoketoconazole, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, wherein the subject is also being administered a multidrug and toxin extrusion transporter 1 (MATE1) substrate or an organic cation transporter 2 (OCT2) substrate.

The present disclosure provides an aggregate of kidney cells having improved expression of transporter genes. Some aspect of the present invention is an aggregate of kidney cells. The aggregate satisfies at least one of the following conditions. (a) The expression level of the OAT1 gene is 1 * 10 <-3 > or more (GAPDH ratio). (b) The expression level of the OAT3 gene is 1 * 10 <-4 > or more (GAPDH ratio). (c) The expression level of the OCT2 gene is 1 * 10 <-3 > or more (GAPDH ratio). (d) The expression level of MATE1 gene is 1 * 10 <-4 > or more (GAPDH ratio). (e) The expression level of the MATE2 gene is 1 * 10 <-5 > or more (GAPDH ratio). (f) The expression level of the MDR1 gene is 1 * 10 <-3 > or more (GAPDH ratio). (g) The expression level of the UART1 gene is 1 * 10 <-6 > or more (GAPDH ratio).

One aspect of the present invention relates to: a conjugate comprising a peptide of a novel sequence and glycyrrhizin; and a pharmaceutical composition for preventing or treating obesity, the composition comprising the conjugate. The conjugate and composition comprising same according to one aspect were found to inhibit hypertrophy of fat cells, increase the cell membrane expression level of ATP-binding cassette transporter A1 (ABCA1), and reduce the secretion of tumor necrosis factor-α (TNF-α). In addition, the conjugate and composition comprising same were found to remain in a greater amount in the blood and have better pharmacokinetic properties than glycyrrhizin used by itself, and thus can be used in the obesity prevention and / or treatment market / industry.

The invention discloses a method for constructing a high-yield natamycin engineering strain through overexpression of an extracellular transport protein gene pimA and a pathway specific regulation gene pimR, and the high-yield engineering strain is applied to fermentation production of natamycin. The yield of the natamycin can reach 19.49 g / L through fed-batch fermentation in a fermentation tank of 5L of the high-yield strain S.ilvosporeus pimA-pimR, and the yield of the natamycin is improved by 33.12% compared with the yield of a chassis strain S.ilvosporeus TUST01; the production intensity is improved by 33.33% compared with the production intensity of a chassis strain S.ilvosporeus TUST01, and the strain has a relatively high industrial application value.

The invention relates to the technical field of biology, in particular to engineered yeast for high-yield citronellol as well as a preparation method and application of the engineered yeast. According to the invention, genetically engineered bacteria for high yield of citronellol are constructed by means of genetic engineering, SyBESc081300004 is taken as an original strain, Erg20ww, Erg8, tCrGES and CrIS are expressed in a peroxisome region in a room manner, and phosphopentose pathway genes TAL1 and TKL1 and endogenous transporter protein PDR1 are over-expressed, so that the genetically engineered bacteria are obtained; experimental results show that the shake flask fermentation yield of the genetically engineered bacterium reaches 3.38 g / L, the yield of a 100L fermentation tank reaches 10.556 g / L, and the yield of the genetically engineered bacterium is obviously higher than that of other existing citronellol production engineered bacterium.

The invention relates to the technical field of bioengineering, in particular to an AimA mutant as well as a construction method and application thereof. It is found for the first time that the activity of glutamic acid / serine transporter is weakened, and the nucleoside production capacity of microorganisms can be improved. Based on this, the present invention provides a modified microorganism in which the glutamic acid / serine transporter activity is reduced or lost as compared to an unmodified microorganism, and the nucleoside production capacity of the modified microorganism is enhanced as compared to the unmodified microorganism, and more specifically, the modified microorganism has a reduced or lost glutamic acid / serine transporter activity as compared to the unmodified microorganism. The modified bacillus subtilis or bacillus amyloliquefaciens provided by the invention can generate guanine nucleoside or hypoxanthine nucleoside with high efficiency and high speed.

The invention belongs to the field of bioengineering, and particularly relates to an engineering bacterium for expressing transporter TBDT11 by an outer membrane as well as preparation and application of the engineering bacterium. The preparation method of the engineering bacteria comprises the following steps: (1) connecting a TBDT11 gene with an escherichia coli expression vector pET-28a to obtain a recombinant vector pET-TBDT11; (2) transforming the recombinant vector pET-TBDT11 into an escherichia coli competent cell, so as to obtain an engineering bacterium EcN-pET-TBDT11; and (3) carrying out induced expression on the engineering bacterium EcN-pET-TBDT11 by using isopropyl thiogalactoside IPTG, and culturing to obtain the engineering bacterium of the transporter TBDT11 expressed by the outer membrane of the bacterium. The engineering bacteria prepared by the method can be ingested to reduce the benzopyrene content in the intestinal environment, and the preparation method has the advantages of simplicity, high efficiency, safety, no toxicity, no need of protein purification, low cost and the like.

The application discloses a method for promoting transport and improving retinal synthesis, which is obtained by introducing an exogenous ABC transport protein Pdr5 into a host fungus and adding a copy of beta-carotene-15, 15'-monooxygenase BLH; wherein the host fungus is a genetically modified Pichia pastoris strain PP-BCCIV. The application further verifies the retinal production performance of the recombinant strain by screening different surfactants, and further improves the ability of Pichia pastoris to produce retinal. At the same time, all the retinal is extracted into the extracellular, and all the carotenes are left in the intracellular, so as to achieve the co-production of carotenes and retinal. The recombinant Pichia pastoris of the application has a simple construction method, can better promote the synthesis of retinal and other vitamin A, and can make the engineering strain utilize glucose and methanol to efficiently synthesize retinal through a 5 L fermentation tank, which is beneficial to industrial production.

Provided are a recombinant Escherichia coli with improved ability to secrete extracellular endoglucanase (BcsZ) and a construction method therefor. The recombinant Escherichia coli is constructed by using Escherichia coli JBZ-DH5α as a starting strain and inserting a membrane protein coding gene. The membrane protein gene is one of the following coding genes: a coding gene of zinc transporter permease znuB, a coding gene of DUF1449 family inner membrane protein ylaC, and a coding gene of UPF0410 family inner membrane protein ymgE.

The invention discloses a pugionium cornutum cation-Cl-co-transporter protein coding gene PcCCC1 and application thereof. The nucleotide sequence of the cation-Cl-co-transporter protein coding gene PcCCC1 is as shown in SEQ ID No.1, the amino acid sequence of the coding protein of the cation-Cl-co-transporter protein coding gene PcCCC1 is as shown in SEQ ID No.2, and the sequence of a specific promoter of the cation-Cl-co-transporter protein coding gene PcCCC1 is as shown in SEQ ID No.3. The invention further discloses an application of the cation-Cl-co-transporter protein coding gene PcCCC1. Experiments prove that after the gene is driven by a specific promoter of the gene to be overexpressed in arabidopsis thaliana, accumulation of Na < + > and Cl <-> of the overground part under salt stress can be reduced, so that the salt tolerance of the plant is improved, a gene resource is provided for improvement of the salt tolerance of the plant, and the gene has a wide application value.