Abstract: The present invention discloses a method for establishing a colorectal cancer HK2 reporter gene cell line, specifically including: firstly, designing a site-specific sgRNA sequence of an HK2 gene, and cloning same into a PX459 plasmid; integrating a homologous recombination sequence of an HK2 gene and a green fluorescent protein DNA fragment (EGFP), and transforming the plasmid and the integrated fragment together into a colorectal cancer cell line HCT116 by electroporation; and performing signal cell screening through a flow cytometer to obtain EGFP-expressing cells, and amplifying a monoclonal cell line; and identifying a positive HK2 reporter gene cell line through PCR identification and Western blot, among screened EGFP-expressing cell lines. The colorectal cancer cell line HK2 gene and EGFP are co-expressed, and the expression level of the EGFP is highly consistent with that of the HK2 gene.
Abstract: Disclosed are methods and compositions for increasing the triacylglycerol content of a cell by increasing the activity of a type 1 diacylglycerol acyltransferase (i.e., DGA2) and increasing the activity of a type 2 diacylglycerol acyltransferase (i.e., DGA1). In some embodiments, the triacylglycerol content of a cell is also modified my decreasing the activity of a triacylglycerol lipase in the same cell. Also disclosed are methods and compositions for increasing the triacylglycerol content of a cell by increasing the activity of a type 1 diacylglycerol acyltransferase (i.e., DGA2), or by increasing the activity of a type 3 diacylglycerol acyltransferase (i.e., DGA3).
Type:
Grant
Filed:
April 5, 2019
Date of Patent:
November 8, 2022
Assignee:
Ginkgo Bioworks, Inc.
Inventors:
Vasiliki Tsakraklides, Elena E. Brevnova
Abstract: The present invention relates to mannanase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.
Type:
Grant
Filed:
April 26, 2018
Date of Patent:
November 8, 2022
Assignee:
Novozymes A/S
Inventors:
Vivek Srivastava, Markus Klinger, Rakhi Saikia, Vijaya Shankar Nataraj, Sohel Dalal, Jens Erik Nielsen, Rajendra Kulothungan Sainathan
Abstract: Methods of fractionating lignocellulosic biomass using maleic acids are provided. Also provided are methods of forming lignin particles, furans, sugars, and/or lignocellulosic micro- and nanofibrils from the liquid and solid fractions produced by fractionation process. The fractionation can be carried out at low temperatures with short reaction times to carboxylate and dissolve lignin with a low degree of condensation.
Abstract: The present invention relates to a method for chemical production of 3,6-anhydro-L-galactitol (L-AHGoI), which is a novel sugar alcohol, and agarobititol (ABol), which is a disaccharide having the same agarobititol as a reductant end thereof, from sea algae.
Type:
Grant
Filed:
September 5, 2019
Date of Patent:
October 25, 2022
Assignee:
Korea Univercity Research and Business Foundation
Inventors:
Kyoung Heon Kim, Dong Hyun Kim, Eun-Ju Yun
Abstract: The invention is directed to compositions and methods for the manufacture of pigmented solids structures for which can be used for construction and/or decoration. Manufacturing comprises fixing one or more pigments to an aggregate material such as crushed rock, stone or sand. The pigmented aggregate is incubated with urease or urease producing microorganisms, an amount of a nitrogen source such as urea, and an amount of calcium source such as calcium chloride forming calcite bridges between particles of aggregate. The resulting solid has a hardness and colorfastness for most any construction material. Using selected aggregate and pigment, the process also provides for the manufacture of simulated-stone materials such as clay or granite bricks or blocks, marble counter-tops, and more. The invention is also directed to composition containing microorganisms and pigment as kits that can be added to most any aggregate materials.
Type:
Grant
Filed:
July 8, 2020
Date of Patent:
October 18, 2022
Assignee:
BIOMASON INC.
Inventors:
Thomas A. Hill, Steven W. McAllister, J. Michael Dosier, Ginger K. Dosier
Abstract: Strains of Saccharomyces cerevisiae yeast that are genetically modified so as to co-express a gene coding a glucoamylase of fungal origin, a gene coding a glucoamylase of Saccharomyces cerevisiae var. diastaticus, and a gene coding a xylanase of fungal origin. The production yield of bioethanol through these strains is greater than that of strains that are otherwise identical but that do not include the gene coding the xylanase of fungal origin. Also, a method for obtaining these yeasts, as well as the use of these yeasts in the production of bioethanol.
Type:
Grant
Filed:
July 2, 2019
Date of Patent:
October 18, 2022
Assignee:
LESAFFRE ET COMPAGNIE
Inventors:
Georges Pignede, Maud Petit, Benoît Thomas, Sébastien Hulin
Abstract: There is provided SHC/HAC derivatives, amino acid sequences comprising the SHC/HAC derivatives, nucleotide sequences encoding the SHC/HAC derivatives, vectors comprising nucleotide sequences encoding the SHC/HAC derivatives, recombinant host cells comprising nucleotide sequences encoding the SHC/HAC derivatives and applications of the recombinant host cells comprising either SHC/HAC derivatives or WT SHC/HAC enzymes in methods to prepare (?)-Ambrox and SHC/HAC enzymes in methods to prepare (?)-Ambrox.
Type:
Grant
Filed:
April 26, 2021
Date of Patent:
October 11, 2022
Assignee:
GIVAUDAN S.A.
Inventors:
Eric Eichhorn, Boris Schilling, Denis Wahler, Laurent Fourage, Esther Locher
Abstract: Supplemented mixotrophic method. A mixotrophic fermentation method is disclosed including providing a naturally acetogenic organism; providing a fermentation medium comprising a carbon source and a supplemented non-sugar reductant; and culturing the organism in the fermentation medium, where both the carbon source and the non-sugar reductant are metabolized and a fermentation broth is formed, which contains at least one carbon-containing bioproduct.
Type:
Grant
Filed:
May 31, 2017
Date of Patent:
October 11, 2022
Assignee:
SUPERBREWED FOOD INC.
Inventors:
Bryan Patrick Tracy, Shawn William Jones, Aharon M. Eyal
Abstract: Nodulisporic acids (NAs) comprise a group of indole diterpenes known for their potent insecticidal activities; however, biosynthesis of NAs by its natural producer, Hypoxylon pulicicidum (Nodulisporium sp.) is exceptionally difficult to achieve. The identification of genes responsible for NA production could enable biosynthetic pathway optimization to provide access to NAs for commercial applications. Obtaining useful quantities of NAs using published fermentations methods is challenging, making gene knockout studies an undesirable method to confirm gene function. Alternatively, heterologous gene expression of H. pulicicidum genes in a more robust host species like Penicillium paxilli provides a way to rapidly identify the function of genes that play a role in NA biosynthesis. In this work, we identified the function of four secondary-metabolic genes necessary for the biosynthesis of nodulisporic acid F (NAF) and reconstituted these genes in the genome of P.
Type:
Grant
Filed:
September 28, 2018
Date of Patent:
September 27, 2022
Inventors:
Matthew Joseph Nicholson, Sarah Adeline Kessans, Emily Jane Parker, Leyla Yolanda Bustamante Rodriguez, David Barry Scott, Kyle Cornelius Van de Bittner, Craig John Van Dolleweerd
Abstract: Aqueous fermentation feedstock and method of producing same. The feedstock includes glucose and dextrose oligomers, wherein (i) glucose concentration is in a range between 10 gram/Liter (g/L) and 150 g/L; (ii) dextrose oligomers concentration is in a range between 50 g/L and 300 g/L; and optionally (iii) slurried particles of less than 0.5 micron; (iv) slurried particles of more than 0.5 micron, wherein a content of such suspended particles of more than 0.5 micron is less than 30 g/L; (v) ash at a concentration in a range between 20 g/L and 50 g/L; (vi) lactate at a concentration in a range between 0.5 g/L and 10 g/L; (vii) protein at a concentration in a range between 5 g/L and 50 g/L; (viii) corn oil at a concentration of less than 10 g/L; and/or (ix) glycerol at a concentration in a range between 1 g/L and 30 g/L.
Type:
Grant
Filed:
September 10, 2020
Date of Patent:
September 20, 2022
Assignee:
SUPERBREWED FOOD INC.
Inventors:
Bryan P. Tracy, John Randall Phillips, Daniel Knox Mitchell
Abstract: The purpose of the present invention is to provide a protein, said protein having an activity of synthesizing rhamnose from glucose, and a polynucleotide encoding the same. Provided are a rhamnose synthase derived from stevia and a method for producing rhamnose from glucose using a gene. Also provided is a method for producing a steviol glycoside using the rhamnose synthase derived from stevia.
Abstract: The present invention discloses a kind of xylanase mutant and its preparation method and application, which relates to the technical field of genomic engineering and genetic engineering. Such mutant includes one or more mutants obtained by taking xylanase HwXy110A as female parent to conduct saturation mutagenesis to the site of Gly363. Specifically, relates to obtaining 19 mutants through site-directed mutagenesis, and then conducting yeast expression to them, after that, obtaining two mutants with significantly improved specific activity and thermal stability through screening of thermal stability and catalytic activity; the present invention can significantly improve the thermal stability and catalytic efficiency of xylanase through modifying the site of Gly363, and is of important guiding significance for improving the thermal stability and the catalytic efficiency of the 10th family of xylanases and other glycoside hydrolases as well as lays the foundation for its application in industrial production.
Type:
Grant
Filed:
March 31, 2021
Date of Patent:
September 20, 2022
Assignee:
JIANGSU UNIVERSITY OF SCIENCE AND TECHNOLOGY
Inventors:
Shuai You, Wenxin Zhang, Ziqian Cha, Chen Xie, Yan Ge, Yanghao Hu, Jun Wang
Abstract: Yeast strains and fermentation process for producing D-lactic acid and L-lactic acid are disclosed with higher titer, higher yield, shorter time, lower pH, and higher average specific productivity.
Type:
Grant
Filed:
February 15, 2019
Date of Patent:
September 20, 2022
Assignee:
PTT Global Chemical Public Company Limited
Inventors:
Sudhanshu Vijay Dole, Theron Hermann, Sean Joseph Regan, Mark Andrew Sheff, Russell Lizardo Udani, R. Rogers Yocum
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 chiral compounds.
Type:
Grant
Filed:
October 26, 2020
Date of Patent:
August 30, 2022
Assignee:
Codexis, Inc.
Inventors:
Jack S Liang, Stephan Jenne, Emily Mundorff, Rama Voladri, James J. Lalonde, Gjalt W. Huisman
Abstract: The present disclosure relates generally to methods, isolated polypeptides and polynucleotides, expression vectors, and host cells for the production of olivetolic acid and phytocannabinoids. A method of producing olivetolic acid (OVLa) and/or a phytocannabinoid in a heterologous host cell having OVLa-producing or phytocannabinoid-producing capacity comprises transforming the host cell with a nucleotide encoding a variant olivetolic acid cyclase (OAC) protein having at least 6 amino acid mutations relative to the wild type OAC protein, and culturing the transformed host cell to produce OVLa and/or phytocannabinoids therefrom. The variant OAC protein (SEQ ID NO:92) has at least 85% sequence identity with the wild type OAC protein (SEQ ID NO:91). Exemplary variants having improved OVLa or phytocannabinoid production capacity are described.
Type:
Grant
Filed:
November 20, 2020
Date of Patent:
August 30, 2022
Assignee:
Hyasynth Biologicals Inc.
Inventors:
Timothy S. Liao, Letian Song, Louis Hom, Curtis Walton, Daniel Furlong, Mindy Melgar, Devanshi Bhargava
Abstract: The present disclosure provides systems and methods for increasing production of an alkaloid product through the epimerization of a (S)-1-benzylisoquinoline alkaloid to a (R)-1-benyzlisoquinoline alkaloid via an engineered epimerase in an engineered host cell. A (S)-1-benzylisoquinoline alkaloid is contacted with said engineered epimerase. Contacting said (S)-1-benzylisoquinoline alkaloid with said engineered epimerase converts said (S)-1-benzylisoquinoline alkaloid to said (R)-1-benzylisoquinoline alkaloid.
Abstract: A method for oxidizing an alkyl, including a) contacting the alkyl with an aqueous solution comprising a microorganism where the microorganism has a reduced fatty acid degradation capacity compared to its wild type, wherein the fatty acid degradation capacity is reduced by deletion, inhibition or inactivation of a gene encoding an enzyme involved in the ?-oxidation pathway; and the microorganism expresses a recombinant alkane oxidase, and b) contacting the aqueous solution from a) with a water-immiscible organic solvent.
Type:
Grant
Filed:
October 7, 2020
Date of Patent:
August 23, 2022
Assignee:
Evonik Operations GmbH
Inventors:
Thomas Haas, Oliver Thum, Jan Christoph Pfeffer, Philip Engel, Christian Gehring, Markus Poetter
Abstract: The present disclosure relates to the use of beta-glucosidase to enhance the production efficiency of desired steviol glycosides, such as rebaudioside M (reb M).
Type:
Grant
Filed:
December 23, 2019
Date of Patent:
August 16, 2022
Assignee:
Conagen Inc.
Inventors:
Guohong Mao, Jacob Edward Vick, Michael Batten, Oliver Yu