Patent classifications
C12Y101/01047
Machine learning gene mining method and phosphinothricin dehydrogenase mutant for amino translocation
Disclosed are a machine learning gene mining method and a phosphinothricin dehydrogenase mutant for amino translocation. The phosphinothricin dehydrogenase mutant for amino translocation is obtained by mutation of a wild-type phosphinothricin dehydrogenase with an amino acid sequence as shown in SEQ ID No.2 at one of the following sites: (1) E263D-K134R-H96A-R290V; (2) E263D-K134R-H96A; (3) E263D-K134R; (4) E263D; (5) E263N; (6) E263C; and (7) E263G. The present invention utilizes the site-saturation mutagenesis technology to mutate a phosphinothricin dehydrogenase gene as shown in SEQ ID No. 1, finds that the 263rd, 134th, 290th and 290th positions are the key sites affecting enzyme activity and stereoselectivity, and obtains a mutant with enzyme activity and ee value much higher than those of the parent phosphinothricin dehydrogenase.
PRODUCTION METHOD OF RECOMBINANT ESCHERICHIA COLI AND HIGH-PURITY URSODEOXYCHOLIC ACID
The present invention belongs to the field of bioengineering technologies, and in particular, to a production method of recombinant Escherichia coli (E. coli) and high-purity ursodeoxycholic acid (UDCA). The present invention constructs novel double-enzyme co-expression gene engineered bacteria, that is, recombinant E. coli. The bacteria simultaneously expresses 7β-hydroxysteroid dehydrogenase (7β-HSDH) and glucose dehydrogenase (GDH). The bacteria is applicable to the production of high-purity UDCA. The yield of a target product is increased through the joint expression and application of 7β-HSDH and GDH. The production method of high-purity UDCA in the present invention is simple, generates a small amount of impurities in a production process, is a green process that satisfies environmental protection requirements, and has significant industrial application value.
CATALYTICALLY ACTIVE PROTEIN FOAMS, AND METHOD FOR PRODUCING SAME
Catalytically active protein foams and methods for producing same by coupling catalytically active fusion proteins with connectors, and the use of catalytically active protein foams in biocatalysis and microfluidics.
MACHINE LEARNING GENE MINING METHOD AND PHOSPHINOTHRICIN DEHYDROGENASE MUTANT FOR AMINO TRANSLOCATION
Disclosed are a machine learning gene mining method and a phosphinothricin dehydrogenase mutant for amino translocation. The phosphinothricin dehydrogenase mutant for amino translocation is obtained by mutation of a wild-type phosphinothricin dehydrogenase with an amino acid sequence as shown in SEQ ID No.2 at one of the following sites: (1) E263D-K134R-H96A-R290V; (2) E263D-K134R-H96A; (3) E263D-K134R; (4) E263D; (5) E263N; (6) E263C; and (7) E263G. The present invention utilizes the site-saturation mutagenesis technology to mutate a phosphinothricin dehydrogenase gene as shown in SEQ ID No. 1, finds that the 263rd, 134th, 290th and 290th positions are the key sites affecting enzyme activity and stereoselectivity, and obtains a mutant with enzyme activity and ee value much higher than those of the parent phosphinothricin dehydrogenase.
ENGINEERED POLYPEPTIDES THAT EXHIBIT INCREASED CATALYTIC EFFICIENCY FOR UNNATURAL COFACTORS AND USES THEREOF
Provided herein are engineered polypeptides that exhibit increased catalytic efficiency for unnatural cofactors and use of said polypeptides in engineered unnatural redox cofactor systems for whole-cell biomanufacturing and in cell-free applications.
Direct electron transfer-type oxidoreductase-modified molecular recognition element
A molecular recognition element comprising a target molecule-recognizing portion, and a direct electron transfer-type oxidoreductase linked to the target molecule-recognizing portion.
ENZYME-CATALYZED METHOD FOR SYNTHESIZING (2S, 3R)-2-SUBSTITUTED AMINOMETHYL-3-HYDROXYBUTYRATE
An enzyme-catalyzed method of synthesizing (2S, 3R)-2-substituted aminomethyl-3-hydroxybutyrate, including: preparing engineered bacteria containing a carbonyl reductase SsCR-encoding gene; preparing a resting cell suspension of the engineered bacteria; preparing a culture containing carbonyl reductase; and mixing the culture containing carbonyl reductase with substrate 2-substituted aminomethyl-3-one butyrate, glucose dehydrogenase, a cosolvent, glucose and a cofactor followed by asymmetric carbonyl reduction to obtain (2S, 3R)-2-substituted aminomethyl-3-hydroxybutyrate. The amino acid sequence of the carbonyl reductase is shown in SEQ ID NO.1.
Recombinant <i>E. coli </i>and method of producing Danshensu by using same
The present disclosure discloses a production method of Danshensu, belonging to the technical field of bioengineering. The present disclosure constructs a novel genetic engineering strain co-expressed by three enzymes, which can be applied to the production of optically pure 3-(3,4-dihydroxyphenyl)-2-hydroxypropionic acid. All of the (D/L)--hydroxycarboxylic acid dehydrogenase selected by the present disclosure have the characteristics of poor substrate specificity and strong optical specificity, and can produce optically pure D-danshensu and L-danshensu. Further, the production efficiency of the recombinant strain is improved by knocking out or enhancing the expression of a related gene on the E. coli genome to promote substrate transport and reduce product decomposition. The method for producing Danshensu and -ketoglutaric acid by using the transformation of the recombinant strain according to the present disclosure is simple, has easily available raw materials, few impurities, and has good industrial application prospects.
SYNTHETIC BIOLOGICAL CIRCUITS FOR THE DETECTION OF TARGET ANALYTES USING A GLUCOSE METER IN A CELL-FREE SYSTEM
Described are methods for generating a reporter molecule in response to a target analyte in a cell-free system. A synthetic biological circuit is used to modify the level of the reporter molecule in response to the presence of the target analyte. The reporter molecule may be glucose or another molecule readily detected using a device such as glucose monitor or other portable sensor. Also provided are kits comprising a cell-free system with a synthetic biological circuit that generates or consumes a reporter molecule in response to a target analyte.
PROCESS FOR ENZYMATIC PRODUCTION OF OXIDATION AND REDUCTION PRODUCTS OF MIXED SUGARS
The present invention provides a process for obtaining n+a oxidation and reduction products from a mixture of n sugars selected from the group consisting of C5 and C6 sugars, wherein n is at least 2 and a is at least 1, wherein at least two of the sugars in the mixture are present at a non-equimolar ratio to each other, wherein, in a first stage, at least one of the sugars which are present at a non-equimolar ratio to each other is oxidized enzymatically and, at the same time, at least one of the other sugars present at a non-equimolar ratio to each other is reduced enzymatically, and wherein, in the first stage, a portion of at least one of the sugars present at a non-equimolar ratio to each other is not converted, and which is characterized in that, in at least a second stage, at least a portion of the sugar not converted in the first stage is oxidized enzymatically by half and, respectively, is reduced enzymatically by the remaining half.