30 results about "Aminolevulinic Acid Synthetase" patented technology

The invention discloses a method to prepare engineering bacteria of 5-amino levulinic acid, which comprises the following steps: 1) extracting total genom DNA from bacterial liquid of red ball bacterium; 2)augmenting 5- glycyl ethylformic acid synthase gene with polyose chain reaction; 3) connecting with colonic carrier pMD-18T simple; proceeding DNA testing sequence; 4) connecting the goal gene segment with expressing carrier pET28a; constructing recon pET28a-R. S. hemA; 5) transforming the recon pET28a-R. S. hemA to host bacteria; getting the product. This invention possesses higher enzyme activity, which output of external cell ALA can reach 6. 6g/L.

The invention provides a 5-aminolevulinic acid (ALA) synthetase. The amino acid sequence of the ALA synthetase has mutation at the amino acid residues of the 40th site, the 365th site, the 75th site,the 29th site and the 44th site corresponding to the amino acid sequence as shown in SEQ ID NO:1. The ALA synthetase provided by the invention obviously improves the activity and also partially relieves the feedback inhibition of hemachrome. Furthermore, the invention also provides methods for preparing and modifying the ALA synthetase, an expression vector containing the ALA synthetase, host cells containing the ALA synthetase, and application of the ALA synthetase in ALA production.

The invention discloses a method to prepare engineering bacteria of 5-glycyl propionic acid and methods for using them, which comprises the following steps: incorporating radial agrobacteriocin; setting the preserved number at CGMCC No. 1938; activating engineering bacteria with LB medium flat; getting monoclonal of the engineering bacteria; seeding to shake; culturing; getting first-grade seed; seeding the first-grade in shake; culturing; getting second-grade seed; seeding the second-grade seed in fermenter; culturing; cooling and evoking with isopropyl-beta-D-sulfo-galactose-glycoside; culturing continually; proceeding supplementing material culture. This engineering bacteria possesses higher ALA synthase expression, simple craft, low cost and good industrial prospect.

An engineered bacterium able to generate 5-aminolevulic acid is disclosed, which contains the 5-aminolevulic acid, synthetase gene of radial Agrobacterium. Its configuring process includes such steps as extracting genom DNA from Agrobacterium liquid, PCR amplification of 5-aminoevulic acid synthetase gene, linking it with carrier pGEM-T, sequencing linking sequenced target gene fragment with expression carrier pET28 a to configue recombination pET28a-A.R-hemA, and transfering it in host.

The invention discloses a recombinant escherichia coli strain for producing 5-aminolevulinic acid (5-ALA). The invention also discloses a way for efficiently synthesizing the 5-aminolevulinic acid, wherein the 5-aminolevulinic acid synthetase (HemL and HemA) of the escherichia coli is enhanced, and the strain has the capability of synthesizing the 5-aminolevulinic acid preliminarily; the expression of the 5-aminolevulinic acid efflux protein eamA gene is enhanced, and the 5-aminolevulinic acid efflux capability of the strain is improved; an exogenous 5-aminolevulinic acid synthetase hemA gene is introduced, so that the 5-aminolevulinic acid synthesis capability of the strain is enhanced; galR, glk and ppc genes of a glucose utilization way are modified, and the utilization efficiency of glucose is improved; and genes (hemF, poxB and aceB) of metabolic bypasses are knocked out. The recombinant Escherichia coli strain constructed by the invention has the capability of efficiently synthesizing 5-aminolevulinic acid by using glucose and glycine, so that the recombinant Escherichia coli strain has the application of industrially producing 5-aminolevulinic acid.

The invention discloses a method for increasing the yield of 5-aminolevulinic acid synthesized by corynebacterium glutamicum, and belongs to the technical field of bioengineering. According to the method, the construction of the recombinant corynebacterium glutamicum for knocking out alpha-ketoglutaric acid inhibitory protein and succinate dehydrogenase and overexpressing 5-aminolevulinic acid synthetase is successfully realized. A 5-L fermentation tank batch fermentation strategy is adopted, fermentation conditions are optimized, finally, recombinant corynebacterium glutamicum (C. glutamicum) 13032/delta odhI delta sdhA/pXMJ19-hemA is fermented for 84 h, 25.05 g/L of 5-ALA is synthesized within 48 h under the condition that glycine with the final concentration of 4 g/L is added, the productivity is 0.52 g/L/h, and at the moment, the sugar-acid conversion rate reaches 16.79%. The high yield of the 5-aminolevulinic acid is realized, and the highest yield of one-step fermentation starting from glucose as a carbon source to the current is obtained.

The invention provides a rhodobacter capsulatus 5-aminolevulinic acid synthetase mutant and application thereof. The amino acid sequence of the rhodobacter capsulatus 5-aminolevulinic acid synthetase mutant is as shown in SEQ ID NO. 1. Compared with wild type rhodobacter capsulatus 5-aminolevulinic acid synthetase, the rhodobacter capsulatus 5-aminolevulinic acid synthetase mutant has the advantages that the yield of 5-aminolevulinic acid in host cells is increased by about 22%; in the presence of 20 [mu] M heme, the mutant 5-aminolevulinic acid synthetase C201A can maintain relatively high relative enzyme activity.

The invention discloses a method for improving insect resistance of a plant. The method is characterized in that a 5-aminolevulinic acid synthetase gene is introduced and expressed into the plant to obtain the insect resistance of a transgenic plant. The invention also provides a method for preparing the transgenic plant with improved insect resistance, wherein the method comprises the following steps of transforming the plant by using the plant expression vector containing the 5-aminolevulinic acid synthetase gene, and carrying out screening to obtain the transgenic plant with improved insectresistance. Experimental results show that each strain of the 5-ALAS transgenic rice shows enhanced resistance response to the rice planthopper, so that the method has relatively high popularizationand application values.

The invention discloses a method for improving the low-temperature stress resistance of rice and a plant expression vector used in the method. The method is characterized in that a 5-aminolevulinic acid synthetase gene is introduced and expressed in a plant, so that the low-temperature stress resistance of the transgenic plant is improved. The invention also provides a method for preparing transgenic rice with the improved low-temperature stress resistance, which comprises the following steps of transforming the plant by using the plant expression vector containing the 5-aminolevulinic acid synthetase gene, and performing screening to obtain the transgenic plant with the improved low-temperature stress resistance. Experimental results show that the low-temperature stress resistance of eachstrain of the 5-ALAS transgenic rice is greatly enhanced, so that the method has the popularization and application value in the cultivation of low-temperature stress-resistant varieties such as cold-resistant varieties.

The invention provides a 5-aminolevulinic acid synthetase (ALA synthetase). Amino acid residues of the ALA synthetase at the 11 position of the amino acid sequence shown in SEQ ID NO:1 are Ile, Ser, Met, Cys and Val. The invention also provides a method for preparing the ALA synthetase, an expression vector and host cell which contain the ALA synthetase, the application of the ALA synthetase in ALA production and a method for modifying the ALA synthetase to improve the activity of the ALA synthetase.

Systems and methods for increasing protoporphyrin IX accumulation in a target cell population using aminolevulinate synthase variants. Aminolevulinic acid-mediated photodynamic therapy is a promising approach to treating dysplasic disorders such as cancer and atherosclerosis, but is limited by the lack of a means to deliver optimal quantities of aminolevulinic acid selectively to the target cells, and thereby ensure the best therapeutic response. The disclosed invention provides a means for enhancing the natural production of aminolevulinic acid selectively within target cells to levels predetermined to give an optimal therapeutic response, and is expected to lead to increased efficacy of treatment, possibly broadening the scope of diseases treatable by photodynamic therapy considerably. The disclosed invention is also amenable to patient specific therapy, meaning that a patient's target cells could be used to screen for the aminolevulinic acid delivery system most appropriate for the patient's needs.

The invention provides a 5-aminolevulinic acid synthetase (ALA synthetase). The C-terminal tail amino acid residue of the ALA synthetase is deleted or is non-alanine; and/or, the amino acid residue atthe 403rd position corresponding to SEQ ID NO:1 of the amino acid sequence of the ALA synthetase is deleted or is non-alanine; and/or, 1-N arbitrary amino acid residues are added to the tail positionof the C terminal of the ALA synthetase, and have the ALA synthetase activity. The invention also provides a method for preparing the enzyme, an expression vector containing the enzyme, a host cell,application of the enzyme in ALA production and a method for modifying the ALA synthetase to improve the activity of the ALA synthetase.

The invention discloses a pig 5-aminolevulinic acid synthetase 1 gene as a genetic marker of pig litter size traits and an application thereof. The nucleotide sequence of the genetic marker is shown as SEQ ID NO.1, and R at a 26-site base in the sequence is T or C. The mutation causes polymorphism of a 5'flanking region of the porcine 5-aminolevulinic acid synthase 1 gene. According to the invention, the SNP locus of the 5'flanking region of the pig ALAS1 gene is discovered for the first time; the relationship between the SNP locus and the pig litter size traits is analyzed; the molecular marker related to the pig litter size is found; and a rapid detection method is established.