Patent classifications
C12Y101/01037
Method for promoting acetylglucosamine synthesis of bacillus subtilis
The present invention relates to a method for promoting acetylglucosamine synthesis of Bacillus subtilis, which belongs to the field of genetic engineering. The present invention adopts the recombinant Bacillus subtilis BSGNKAP2 as a starting strain, exogenously introducing pyruvate carboxylase BalpycA derived from Bacillus cereus, eliminating the central carbon metabolism overflow of the Bacillus subtilis and avoiding the synthesis of the by-product acetoin; further, five exogenous reducing force metabolic reactions are introduced to replace the reaction of generating NADH in glycolysis pathway and tricarboxylic acid cycle to reconstruct intracellular reducing force metabolism, which specifically comprise glyceraldehyde-3-phosphate ferredoxin dehydrogenase, isocitrate NAD.sup.+ dehydrogenase, a malate quinone dehydrogenase, a ketoacid ferredoxin oxidoreductase and a nitrogenase ferritin. In a shake-flask fermentation process using a complex medium, acetylglucosamine yield of the recombinant strain BSGNKAP8 is 24.50 g/L, acetylglucosamine/glucose yield is 0.469 g/g, respectively 1.97 times and 2.13 times of those of the starting strain BSGNKAP2.
Malate dehyrogenases
The present invention relates to a recombinant host cell which is capable of producing a dicarboxylic acid and which comprises a mutant malate dehydrogenase resulting in an increased production of the dicarboxylic acid. The invention also relates to a process for producing a dicarboxylic acid, which method comprises fermenting said recombinant host cell in a suitable fermentation medium and producing the dicarboxylic acid.
GENETICALLY ENGINEERED YEAST YARROWIA LIPOLYTICA AND METHODS FOR PRODUCING BIO-BASED GLYCOLIC ACID
The present disclosure provides a method for genetically engineering Yarrowia lipolytica host cell for producing glycolic acid from organic wastes. A subject genetically engineered Y. lipolytica cell comprises the disrupted native genes encoding malate synthase, heterologous enzyme of glyoxylate reductase targeted in the different cellular compartments including mitochondria, peroxisome and cytosol, and a mutant NADP.sup.+-dependent malate dehydrogenase. The pathway with a theoretical yield as high as that 1 g of acetic acid can be converted to 1.27 g of glycolic acid without carbon loss was engineered for glycolic acid production. The methods particularly include process for production of volatile fatty acids (VFAs) mainly comprised of acetic acid from organic waste, and then use of resultant VFAs for biosynthesis of glycolic acid by recombinant Y. lipolytica.
Microorganisms and methods for production of specific length fatty alcohols and related compounds
The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.
SUCCINIC ACID PRODUCTION USING ISSATCHENKIA
Provided herein are genetically modified Issatchenkia yeast and fermentation methods for producing succinic acid.
A Recombinant of Hydrogenophilus Bacterium Producing Lactic Acid
When lactate dehydrogenase gene and/or malate/lactate dehydrogenase gene is/are introduced into a Hydrogenophilus bacterium as well as one or more of the three lactic acid-utilizing enzyme genes on the genome of the Hydrogenophilus bacterium is/are disrupted, lactic acid-producing ability is remarkably increased. The inventors of the present invention have identified the three lactic acid-utilizing enzyme genes of the Hydrogenophilus bacterium. When lactate permease gene is further introduced into the recombinant, lactic acid-producing ability is further increased. The recombinant of the present invention effectively produces lactic acid using carbon dioxide as a sole carbon source, and therefore, it is able to efficiently produce the material of biodegradable plastics, while solving global warming caused by increased emissions of carbon dioxide.
Compositions and methods for succinate production
The present application provides genetically modified yeast cell comprising an active succinate fermentation pathway, as well as methods of using these cells to produce succinate.
Dibasic organic acid producing strain and preparation and application of same
Provided are an engineered strain for synthesizing a dibasic organic acid and preparation and application of same. The engineered strain introduces or up-regulates expression of a positive regulator gene for synthesis of a dibasic organic acid, and/or down-regulates expression of a negative regulator gene for synthesis of a dibasic organic acid, as compared with the origin strain of the engineered strain, the producing capability for producing the dibasic organic acid is improved. The dibasic organic acid comprises malic acid, succinic acid, fumaric acid, oxaloacetic acid, glutaric acid, and adipic acid; the expression product of the positive regulator gene comprises aspartate aminotransferase, glutamic acid-aspartate transporter, C4-dicarboxylic acid transporter, pyruvate carboxylase and malate dehydrogenase, glucose transporter; the expression product of the negative regulatory gene comprises succinyl-CoA synthase, and malic acid-alpha ketoglutarate transporter, and the original strain comprises Myceliophthora thermophila, Thielavia terrestris, Aspergillus, and Rhizopus.
Microorganisms and methods for the production of glycolic acid and glycine via reverse glyoxylate shunt
The present invention provides biochemical pathways, glyoxylate producing recombinant microorganisms, and methods for the production and yield improvement of glycolic acid and/or glycine via a reverse glyoxylate shunt. The reverse glyoxylate shunt comprises an enzyme that catalyzes the carboxylation of phosphoenol pyruvate (PEP) to oxaloacetate (OAA), or an enzyme that catalyzes the carboxylation of pyruvate to oxaloacetate (OAA) or an enzyme that catalyzes the carboxylation of pyruvate to malate or a combination of any of the previous reactions; an enzyme that catalyzes the conversion of malate to malyl-CoA; an enzyme that catalyzes the conversion of malyl-CoA to glyoxylate and acetyl-CoA; and optionally an enzyme that catalyzes the conversion of oxaloacetate (OAA) to malate. Glyoxylate is reduced to produce glycolate. Alternatively, glyoxylate is converted to glycine. The reverse glyoxylate shunt pathway of the present invention can be utilized synergistically with other glycolic acid and/or glycine producing pathways to increase product yield.
CHIMERIC ANTIGEN RECEPTOR POLYPEPTIDES IN COMBINATION WITH TRANS METABOLISM MOLECULES MODULATING KREBS CYCLE AND THERAPEUTIC USES THEREOF
Disclosed herein are genetically engineered hematopoietic cells, which express one or more Krebs cycle modulating polypeptides, and optionally a chimeric receptor polypeptide (e.g., an antibody-coupled T cell receptor (ACTR) polypeptide or a chimeric antigen receptor (CAR) polypeptide) capable of binding to a target antigen of interest. Also disclosed herein are uses of the engineered hematopoietic cells for inhibiting cells expressing a target antigen in a subject in need thereof.