Abstract: The present invention relates to a recombinant OxDC expressed in a filamentous fungal host cell, methods for constructing a recombinant filamentous fungal host cell, methods for producing recombinant OxDC and the application thereof. The recombinant filamentous fungal host cell comprises one or more copies of OxDC expression cassette integrated in its genome; the expression cassette comprises a promoter, a signal peptide coding sequence, an OxDC coding sequence and a transcription terminator. The host cell can be constructed by random integration or site-specific integration. In addition, the present invention also optimizes the medium formulation for different recombinant filamentous fungal host cells. In the production of the recombinant OxDC, the final yield and enzyme activity were greatly improved. The invention effectively solves the problem that the production of OxDC in the prior art cannot be industrialized on a large scale.
Abstract: Described herein are devices and methods for using the same to produce carotenoids. The carotenoids produced by the devices and methods disclosed herein do not require the ultra purification that is common in conventional or commercial methods. The devices and methods disclosed herein also enhance one or more physical properties of plants treated with the devices described herein.
Abstract: This invention relates to the bioproduction of substituted or unsubstituted phenylacetaldehyde, 2-phenylethanol, phenylacetic acid or phenylethylamine by subjecting a starting material comprising glucose, L-phenylalanine, substituted L-phenylalanine, styrene or substituted styrene to a plurality of enzyme catalyzed chemical transformations in a one-pot reaction system, using recombinant microbial cells overexpressing the enzymes. To produce phenylacetaldehyde from styrene, the cells are modified to overexpress styrene monooxygenase (SMO) and styrene oxide isomerase (SOI). To produce phenylacetic acid from styrene, SMO, SOI and aldehyde dehydrogenase are overexpressed. Alternatively, to produce 2-phenylethanol, SMO, SOI and aldehyde reductase or alcohol dehydrogenase are overexpressed, while to produce phenylethylamine, SMO, SOI and transaminase are overexpressed.
Type:
Grant
Filed:
May 23, 2018
Date of Patent:
March 7, 2023
Assignee:
NATIONAL UNIVERSITY OF SINGAPORE
Inventors:
Zhi Li, Shuke Wu, Yi Zhou, Benedict Ryan Lukito
Abstract: A DNA expression construct comprising a polynucleotide encoding an unnatural UstD enzyme, the unnatural enzyme itself, and a method of making gamma-hydroxy amino acids by contacting an aldehyde-containing substrate, an amino acid, and the unnatural, purified UstD enzyme under conditions and for a time sufficient to react at least a portion of the aldehyde-containing substrate with at least a portion of the amino acid, to yield a gamma-hydroxy amino acid product.
Type:
Grant
Filed:
October 16, 2020
Date of Patent:
February 28, 2023
Assignee:
Wisconsin Alumni Research Foundation
Inventors:
Andrew R. Buller, Jonathan M. Ellis, Prasanth Kumar
Abstract: The present invention provides an isolated AZF1 gene sequence, recombinant vectors, and recombinant yeast which are useful in methods of enhanced biofuel production, particularly ethanol production. Methods of bioengineering recombinant yeast with isolated AZF1 gene sequence useful for biofuel production are also provided.
Type:
Grant
Filed:
October 10, 2018
Date of Patent:
February 28, 2023
Assignee:
Wisconsin Alumni Research Foundation (WARF)
Abstract: The present invention provides several non-naturally occurring sulfotransferase enzymes that have been engineered to react with aryl sulfate compounds as sulfo group donors, instead of the natural substrate 3?-phosphoadenosine 5?-phosphosulfate (PAPS), and with heparosan-based polysaccharides, particularly heparan sulfate, as sulfo group acceptors. Each of the engineered sulfotransferase enzymes have a biological activity characterized by the position within the heparosan-based polysaccharide that receives the sulfo group, including glucosaminyl N-sulfotransferase activity, hexuronyl 2-O sulfotransferase activity, glucosaminyl 6-O sulfotransferase activity, or glucosaminyl 3-O sulfotransferase activity. Methods of using the engineered sulfotransferases to produce sulfated heparosan-based polysaccharides, including polysaccharides having anticoagulant activity, are also provided.
Abstract: The present invention provides several non-naturally occurring sulfotransferase enzymes that have been engineered to react with aryl sulfate compounds as sulfo group donors, instead of the natural substrate 3?-phosphoadenosine 5?-phosphosulfate (PAPS), and with heparosan-based polysaccharides, particularly heparan sulfate, as sulfo group acceptors. Each of the engineered sulfotransferase enzymes have a biological activity characterized by the position within the heparosan-based polysaccharide that receives the sulfo group, including glucosaminyl N-sulfotransferase activity, hexuronyl 2-O sulfotransferase activity, glucosaminyl 6-O sulfotransferase activity, or glucosaminyl 3-O sulfotransferase activity. Methods of using the engineered sulfotransferases to produce sulfated heparosan-based polysaccharides, including polysaccharides having anticoagulant activity, are also provided.
Abstract: A DNA fragment to encode a nitrogen fixation enzyme includes a base sequence of SEQ ID NO:1 or a base sequence having not less than 50% identity with the SEQ ID NO:1.
Abstract: According to the present disclosure, on the basis of all existing mutations, the tenth glycine of a BC-PC-PLC is mutated into aspartic acid, a specific enzyme activity thereof is 83% higher than that of a sequence before the mutation, and protein expression and degumming activity of unit enzyme activity do not change, so as to further reduce manufacturing costs.
Type:
Grant
Filed:
December 10, 2018
Date of Patent:
January 31, 2023
Assignee:
Wilmar (Shanghai) Biotechnology Research & Development Center Co., Ltd.
Abstract: Disclosed herein are methods for treating a subject with an alkaline phosphatase deficiency, further comprising monitoring additional analytes, e.g., calcium, parathyroid hormone and/or vitamin D, with treatment modifications as indicated by the levels, e.g., serum levels, of the additional analytes.
Abstract: Disclosed is variants of a cellulase having improved stability in the presence of a protease, and the use of such variants in laundry.
Type:
Grant
Filed:
April 12, 2019
Date of Patent:
January 31, 2023
Assignee:
Novozymes A/S
Inventors:
Svend Gunnar Kaasgaard, Frank Winther Rasmussen, Roland Alexander Pache, Lars Anderson, Mette Louise Dissing Overgaard, Thomas Agersten Poulsen, Dorte Marie Koefoed Klitgaard, Christian Isak Joergensen, Lars Giger
Abstract: Disclosed herein is a rapid genetics-free method for eliciting and detecting cryptic metabolites using an imaging mass spectrometry-based approach. An organism of choice is challenged with elicitors from a small molecule library. The molecules elicited are then imaged by mass spec, which allows for rapid identification of cryptic metabolites. These are then isolated and characterized. Employing the disclosed approach activated production of cryptic glycopeptides from an actinomycete bacterium. The molecules that result, the keratinimicins and keratinicyclins, are metabolites with important structural features. At least two of these, keratinimicins B and C, are highly bioactive against several pathogenic strains. This approach will allow for rapid activation and identification of cryptic metabolites from diverse microorganisms in the future.
Type:
Grant
Filed:
July 30, 2018
Date of Patent:
January 10, 2023
Inventors:
Mohammad R. Seyedsayamdost, Fei Xu, Yihan Wu, Leah Bushin, Katherine Davis
Abstract: The present invention provides compositions and methods for regulated gene expression. In certain aspects, the invention relates to an inducible synthetic promoter that can be used for regulated gene expression or to generate mutations in one or more bacterial cells of the gut microbiota.
Type:
Grant
Filed:
April 18, 2018
Date of Patent:
January 10, 2023
Assignee:
Yale University
Inventors:
Andrew Goodman, Bentley Lim, Natasha Barry
Abstract: A nucleic acid molecule comprising a variant inc coding strand is disclosed as a regulator of plasmid copy number. Also disclosed is a replicon comprising the nucleic acid molecule, a promoter, and an origin of replication. Also disclosed is a vector comprising the replicon. Also disclosed is a recombinant microorganism comprising the vector.
Type:
Grant
Filed:
August 7, 2019
Date of Patent:
January 10, 2023
Assignee:
CJ CHEILJEDANG CORPORATION
Inventors:
Ji Sun Lee, Dong Eun Chang, Sang Jun Kim, So Young Kim
Abstract: Methods of preparing highly purified steviol glycosides, particularly rebaudiosides A, D and M are described. The methods include utilizing recombinant microorganisms for converting various staring compositions to target steviol glycosides. In addition, novel steviol glycosides reb D2 and reb M2 are disclosed, as are methods of preparing the same. The highly purified rebaudiosides are useful as non-caloric sweetener in edible and chewable compositions such as any beverages, confectioneries, bakery products, cookies, and chewing gums.
Type:
Grant
Filed:
June 16, 2020
Date of Patent:
January 3, 2023
Assignee:
PURECIRCLE SDN BHD
Inventors:
Avetik Markosyan, Indra Prakash, Cynthia Bunders, Pankaj Soni, Jarrin Cyrille, Aurélien Badie, Robert ter Halle
Abstract: A fusion tag according to an embodiment of the present invention may increase the water solubility and expression level of a target protein. As the water solubility and expression level of a target protein in host cell can be increased by a recombinant vector including the fusion tag, the fusion tag can be advantageously used in industry.
Type:
Grant
Filed:
December 28, 2018
Date of Patent:
January 3, 2023
Assignee:
INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY
Abstract: Provided herein are genetically modified microbes. In one embodiment, a genetically modified microbe includes an exogenous polynucleotide that includes a pheromone-responsive region. In one embodiment, the pheromone-responsive region is derived from a conjugative plasmid from a member of the genus Enterococcus spp. The pheromone-responsive region includes a pheromone-responsive promoter and an operably linked coding region encoding an antimicrobial peptide. In one embodiment, a genetically modified microbe includes an exogenous polynucleotide that includes a promoter and an operably linked coding sequence encoding an antimicrobial peptide, where expression of the coding region is controlled by a modulator polypeptide and is altered by a modulating agent, and where the coding region encodes an antimicrobial peptide.
Type:
Grant
Filed:
October 9, 2020
Date of Patent:
December 27, 2022
Assignee:
REGENTS OF THE UNIVERSITY OF MINNESOTA
Inventors:
Yiannis J. Kaznessis, Katherine G. Volzing, Juan Borrero Del Pino, Gary Dunny
Abstract: The present disclosure provides engineered ketoreductase enzymes having improved properties as compared to a naturally occurring wild-type ketoreductase enzyme. Also provided are polynucleotides encoding the engineered ketoreductase enzymes, host cells capable of expressing the engineered ketoreductase enzymes, and methods of using the engineered ketoreductase enzymes to synthesize a variety of chiral compounds. The engineered ketoreductase polypeptides are optimized for catalyzing the conversion of N-methyl-3-keto-3-(2-thienyl)-1-propanamine to (S)—N-methyl-3-hydroxy-3-(2-thienyl)-1-propanamine.
Type:
Grant
Filed:
July 17, 2020
Date of Patent:
November 29, 2022
Assignee:
Codexis, Inc.
Inventors:
Christopher Savile, John M. Gruber, Emily Mundorff, Gjalt W. Huisman, Steven J. Collier
Abstract: A method for improving a bio-coupling efficiency between a protein and a nucleic acid based on an ?-helix handle includes the following steps. First, the handle carrying the non-natural amino acid (H-tag) is designed. Then, a recombinant expression plasmid encoding a fusion protein containing the H-tag and the protein to be tested is constructed. Subsequently, the fusion recombinant protein containing the non-natural amino acid in the H-tag is expressed and purified. Finally, the non-natural amino acid in the H-tag-fused protein and the coupling group on the nucleic acid substrate are efficiently connected by click chemistry. Thea-helix handle is used to provide a controllable reaction condition on the protein surface for the non-natural amino acid, avoiding the complex structure, charge and polar nanoenvironment around the surface of the protein to be tested.
Abstract: This document describes biochemical pathways for producing 7-aminoheptanoic acid using a ?-ketoacyl synthase or a ?-ketothiolase to form an N-acetyl-5-amino-3-oxopentanoyl-CoA intermediate. 7-aminoheptanoic acid can be enzymatically converted to pimelic acid, 7-hydroxyheptanoic acid, heptamethylenediamine or 1,7-heptanediol or corresponding salts thereof. This document also describes recombinant microorganisms producing 7-aminoheptanoic acid as well as pimelic acid, 7-hydroxyheptanoic acid, heptamethylenediamine and 1,7-heptanediol or corresponding salts thereof.
Type:
Grant
Filed:
November 14, 2019
Date of Patent:
November 22, 2022
Assignee:
INV Nylon Chemicals Americas, LLC
Inventors:
Alex Van Eck Conradie, Adriana Leonora Botes, Jonathan Kennedy, Nadia Fatma Kadi