9 results about "Glycerol phosphate" patented technology

This invention belongs to the field of bioengineering, specifically disclosing a 3-phosphoglycerate dehydrogenase mutant with improved enzyme activity and thermostability. Compared to the wild-type 3-phosphoglycerate dehydrogenase from *Corynebacterium glutamicum*, the mutant contains one mutation from among P85Q, D365Y, A389T, M196V, A183V, and I231V. Enzyme activity assays after induced expression and purification showed that the mutant's enzyme activity was 1.5–3.6 times higher than the wild type, and its thermostability was significantly improved. Therefore, the beneficial mutant provided by this invention lays a good foundation for the industrial fermentation production of L-serine and L-cysteine.

The present application provides a recombinant E. coli for synthesizing heme and derivatives thereof, wherein the recombinant E. coli is knocked out of genes such as pyruvate oxidase poxB, pyruvate formate-lyase pflB and protoporphyrinogen peroxidase yfeX, and meanwhile, the genes such as phosphopyruvate carboxylase ppc, glycerol transporter glpF and 3-phosphoglycerate dehydrogenase glpD are enhanced, and the genes such as 5-aminovaleryl-CoA synthase hemA and ferrous chelatase hemH are introduced, so that the recombinant E. coli can synthesize heme and derivatives thereof by using glycerol as a raw material, and has application prospects in the field of biological manufacturing.

This invention provides a multi-enzyme cascade catalytic reaction system and its application, belonging to the field of enzyme catalysis production technology. Using glycerol as a raw material, it includes a first module, a second module, and a third module. The first module involves the production of glyceric acid from glycerol under the catalysis of alditol oxidase. The second module contains glyceric acid, ATP, and NAD+. + L-glutamic acid is synthesized into phosphoserine via glycerate kinase, D-3-phosphoglycerate dehydrogenase, and phosphoserine transaminase. The third module involves phosphoserine and nucleophiles being catalyzed by phosphoserine thioylase to synthesize L-cysteine ​​or a non-natural L-α-amino acid. The multi-enzyme cascade catalytic reaction system provided by this invention can catalyze a variety of nucleophiles, yields high products, is low in cost, causes minimal environmental pollution, and is easily industrialized.

This invention provides a recombinant bacterium and its applications. After specific genetic engineering modifications, this recombinant bacterium can synthesize polyhydroxybutyrate (PHB) using one-carbon compounds such as methanol or carbon dioxide as a carbon source. Specifically, the metabolic pathway of the recombinant bacterium, after optimization, enables the efficient synthesis of PHB from methanol as a raw material via metabolic intermediates such as formaldehyde, 3-phosphoglyceric acid, and acetyl-CoA. The recombinant bacterium constructed in this invention can effectively utilize low-cost carbon sources such as methanol or carbon dioxide to produce PHB, and has promising prospects for industrial application.

This invention discloses a magnetic nanoprobe and kit for assessing postherpetic neuralgia. The magnetic nanoprobe is a complex formed by thiolized carbonized polymer dots, a chelating agent, superparamagnetic molecules, and 2-phospho-D-glyceric acid. 2-phospho-D-glyceric acid specifically reacts with NSE, dehydrating to generate phosphoenolpyruvate and releasing water molecules. The magnetic nanoprobe is specific for NSE and can be used for assessing postherpetic neuralgia. The kit includes the magnetic nanoprobe and instructions, and can determine the NSE marker content in a single sample, with a detection limit better than 1 pg / mL, thereby achieving effective assessment of postherpetic neuralgia.

This invention relates to a genetically engineered bacterial strain that synthesizes indigo from glucose, its construction method, and its applications. The genetically engineered strain overexpresses indole-3-glycerol phosphate lyase derived from plants, and the α subunit of mono / dioxygenase and tryptophan synthase derived from bacteria. This application employs a genetic modification strategy, utilizing metabolic pathway enhancement and the integration of exogenous genes, to achieve the complete metabolism of glucose by *E. coli* to produce indigo. After 72 hours of shake-flask culture, the highest indigo yield can reach 3.11 g / L, reducing production costs and further promoting industrial production.

The application discloses a high-yield resveratrol Yarrowia lipolytica engineering strain and construction and application thereof, and belongs to the technical field of synthetic biology and metabolic engineering. The Yarrowia lipolytica engineering strain provided by the application can realize high-yield resveratrol by using glucose as a substrate, and the yield of resveratrol reaches 3.157 g / L in shake flask fermentation, and the yield of resveratrol is increased to 30.7 g / L in 5L scale batch feeding fermentation, which is the highest value reported at present.

The application relates to a mutant enzyme of 3-phosphoglyceroyltransferase, a coding gene and application thereof, and belongs to the fields of biochemistry, molecular biology and metabolism. 58 GPAT9 mutant genes are constructed by using a site-directed mutagenesis technique, and the influence of single and multiple amino acid site changes on GPAT9 enzyme activity is analyzed by combining a GPAT-specific yeast genetic complementation method. It is found that the changes of six amino acid residues (85, 114, 119, 230, 237 and 322) located outside the acyltransferase conserved domain in AtGPAT9 can significantly affect the enzyme activity. There is interaction among the amino acids, for example, the simultaneous mutation of Y85W / N119H / S237N of the three sites can greatly increase the activity of AtGPAT9, accelerate the growth of yeast and promote the synthesis of triacylglycerol, and the triacylglycerol content in the yeast cells expressing the mutant enzyme is increased by 45.7% compared with that of the yeast cells expressing the wild type BnGPAT9.

The application provides a kit for culturing osteosarcoma organoids, the kit comprising a special culture medium, a sample dissociation digestive solution and a digestion termination solution, the special culture medium being based on a basic culture medium and adding dexamethasone, glycerophosphate disodium salt and calcium chloride, the osteosarcoma organoids carrying or not carrying an immune feature. The application also provides a culture method of the osteosarcoma organoids carrying the immune feature. The osteosarcoma organoids are cultured by using the culture method and the kit, the culture period is short, the stability is high, the in situ tumor heterogeneity can be reserved, the tumor microenvironment immune feature is stable, and the kit can be used for high-throughput drug screening, target gene identification, immunotherapy drug screening, oncolytic virus killing experiment, omics research such as single cell sequencing, and animal model construction. The application fills the blank of the interstitial-derived osteosarcoma organoid immune co-culture, and has a wide application prospect in the field of osteosarcoma basic research and clinical diagnosis and treatment.