Abstract: The present invention relates to a recombinant E. coli cell harbouring a recombinant plasmid wherein said E. coli cell has a thermo sensitive regulation of an essential E. coli gene.

Abstract: The present invention relates to the treatment of tissue adhesions, e.g. tissue adhesions that occur after surgical interventions. More specifically, the invention refers to an enzyme having DNAse activity for use in a method of treating or preventing tissue adhesions. The invention also relates to a pharmaceutical composition that comprises an enzyme having DNAse activity for use in a method of treating or preventing tissue adhesions.

Abstract: Provided is a nucleic acid system introduced into a bacterial strain to generate a genetically engineered bacterial strain that grows in solid tumors but does not grow in non-tumor tissues, the nucleic acid system comprising: a first DNA fragment that encodes a toxin gene that expresses a toxin that kills the genetically engineered bacterial strain; a second DNA fragment that encodes an antidote gene that expresses an antidote that negates the toxin; a first promotor that controls transcription of the antidote gene, such that glucose represses the transcription of the antidote gene; and a first constitutive promoter that causes constitutive expression of the toxin gene; wherein the second DNA fragment is transcribed in the solid tumors but not transcribed in the non-tumor tissues.

Abstract: Bridge helix-modified variant Cas12a and Cas12b proteins having improved DNA cleavage selectivity in comparison to wild type versions of the Cas12a and Cas12b proteins, nucleic acids encoding the variant proteins, host cells containing the nucleic acids, and methods of their use.

Abstract: The present invention belongs to the bioengineering field, and relates to a method for fermentation production of L-theanine by using an Escherichia coli genetically engineered bacterium. The engineered bacterium is obtained by serving a strain as an original strain, wherein the strain is obtained after performing a single copy of T7RNAP, a dual copy of gmas, xylR knockout, and sucCD knockout on an Escherichia coli W3110 genome, and by integrating genes xfp, pta, acs, gltA, and ppc, and knocking out ackA on the genome. The present invention has a high yield, and stable production performance; after 20-25 h, L-theanine has a titer of 75-80 g/L, and the yield is up to 52-55%. The fermentation broth is purified by membrane separation in combination with a cation-anion resin series technique. Moreover, the one-step crystallization yield is 72.3% and the L-theanine final product has a purity of 99%.

Abstract: Provided herein are novel materials and methods for site-specific incorporation of phosphotyrosines into proteins. The novel methods of the invention encompass the use of a novel aminoacyl tRNA synthetase capable of charging compatible tRNAs with a phosphotyrosine precursor. The phosphotyrosine precursor is then incorporated, site-specifically, into a protein at sites where phosphotyrosine residues are desired. The phosphotyrosine precursors are subsequently treated to convert them into phosphotyrosine residues, yielding proteins with phosphotyrosines at selected sites. The scope of the invention encompasses novel aminoacyl tRNA synthetases, novel phosphotyrosine precursors, and methods of using these materials to create site-specific phosphorylated tyrosine residues in a protein.

Abstract: An enzyme-catalyzed synthesis of (1S,5R)-bicyclolactone. A first genetically-engineered bacterium containing Baeyer-Villiger monooxygenase gene and a second genetically-engineered bacterium containing glucose dehydrogenase gene are constructed and then suspended with culture medium to prepare a first suspension and a second suspension, respectively. The first and second suspensions are centrifuged to respectively produce a first supernatant containing Baeyer-Villiger monooxygenase and a second supernatant containing glucose dehydrogenase, which are mixed. The mixed supernatant is then mixed with a raceme of a substituted bicyclo[3.2.0]-hept-2-en-6-one, a solvent, a hydrogen donor and a cofactor to perform an asymmetric Baeyer-Villiger oxidation to produce the (1S,5R)-bicyclolactone, where an amino acid sequence of the Baeyer-Villiger monooxygenase is shown in SEQ ID NO:1.

Abstract: Provided is a method of producing a mixture of pure feedstock-based native polyphenols from a feedstock. Contaminant polyphenols are first removed from an enzyme solution for converting feedstock to a product to produce a polyphenol reduced enzyme solution. The polyphenol reduced enzyme solution is combined with the feedstock and the feedstock is converted to a product and by-product. Heretofore, there has been no process available to reduce or remove the contaminant phenols introduced to the feedstock by commercial enzyme solutions. This method allows for the removal of contaminant phenols prior to introduction to the processing stream and subsequent harvesting of pure feedstock 6 based native polyphenols. The pure feedstock-based polyphenols are removed from the product or by-product to produce a pure mixture of feedstock-based polyphenols.

Abstract: Methods for the production, capturing and purification of recombinant human lysosomal proteins are described. Such recombinant human lysosomal proteins can have high content of mannose-6-phosphate residues. Also described are pharmaceutical compositions comprising such recombinant human lysosomal proteins, as well as methods of treatment and uses of such recombinant human lysosomal proteins.

Abstract: Modified ribosomes that were selected using a dipeptidyl-puromycin aminonucleoside are used to mediate site-specific incorporation of one or more peptides and peptidomimetics into protein in a cell free translation system. In addition, new fluorescent dipeptidomimetics have been synthesized and incorporated into proteins, as well as modified proteins containing one or more non-naturally occurring dipeptides.

Abstract: In various aspects, the present invention is directed to a scalable method of producing rhamnolipids by bacterial fermentation with higher product concentrations, yields and productivities and preventing excessive foaming during the cell growth phase when the cell respiration rate is higher. It has been found that by slowing the growth rate of the bacteria by altering the ratio of the nitrogen source to the non-nitrogen source in the initial fermentation medium and supplementing the nitrogen source, excessive foaming in the growth phase can be prevented. Further, by using the non-nitrogen source as the limiting nutrient that initiates the stationary phase and then supplementing fermentation broth with the nitrogen and carbon sources, the length of the standing phase, and with it the time during which rhamnolipid production occurs can be greatly extended.

Abstract: Provided are various genetically engineered strains of TEKK microorganisms, e.g., yeast in a container of industrial-scale volume containing biomass, wherein the microorganisms are capable of fermenting C5 material to produce desirable organic compounds. Also provided are methods for efficiently producing industrial-scale volumes of the desirable organic compounds, e.g., lactic acid, by TEKK strains of microorganisms such as TEKK-LAC and variants thereof.

Abstract: The instant disclosure is generally related to novel Bacillus sp. mutants capable of producing increased amounts of industrially relevant proteins of interest. Certain embodiments of the disclosure are related to modified Bacillus sp. cells comprising an introduced polynucleotide encoding a variant GlcT protein. Other embodiments are related to methods and compositions for producing endogenous and/or heterologous proteins of interest in the modified Bacillus sp. (daughter) cells, whereas certain other embodiments are directed to nucleic acid sequences, particularly polynucleotide open reading frame (ORF) sequences, vectors thereof and DNA expression constructs thereof, encoding variant GlcT proteins of the disclosure.

Abstract: The present disclosure provides methods and compositions useful for the prophylactic and therapeutic amelioration and treatment of infections caused by Gram-negative bacteria, including Pseudomonas aeruginosa. The disclosure further provides compositions and methods of incorporating and utilizing lysin polypeptides of the present disclosure for augmenting the efficacy of antibiotics generally suitable for the treatment of Gram-negative bacterial infection.

Abstract: Novel hyperthermophilic Dictyoglomus beta-mannanases are provided for use in high temperature industrial applications requiring enzymatic hydrolysis of 1,4-?-D-mannosidic linkages in mannans, galactomannans, and glucomannans. Also provided are methods and compositions for fracturing a subterranean formation in which a gellable fracturing fluid is first formed by blending together a hydratable polymer and a Dictyoglomus beta-mannanase as an enzyme breaker. An optimized and stabilized recombinant Dictyoglomus beta-mannanase is provided that shows superior performance/effectiveness and properties in degrading guar and derivatized guars at pH ranges from 3.0 to 12 and temperatures ranging from 130° F. to in excess of 270° F.

Abstract: The present invention relates to a protease variant which is at least 90% identical to the full length amino acid sequence of a Kumamolisin AS backbone as set forth in any of SEQ ID NOs 1-3, while maintaining proteolytic activity, or a fragment, fraction or shuffled variant thereof maintaining proteolytic activity, which protease variant demonstrates altered or improved stability compared to the Kumamolisin AS wildtype as set forth in SEQ ID NO 4, or the Kumamolisin AS backbone as set forth in any of SEQ ID NOs 1-3.

Abstract: The invention provides computational methods for engineering, selecting, and/or identifying proteins with a desired activity. Further provided are automated computational design and screening methods to engineer proteins with desired functional activities including, but not limited to ligand binding, catalytic activity, substrate specificity, regioselectivity and/or stereoselectivity.

Abstract: The present disclosure relates to a novel protein variant having an activity of exporting 5?-inosine monophosphate, a microorganism comprising the protein variant, and a method for preparing 5?-inosine monophosphate using the microorganism.

Abstract: Methods are provided for enhanced production of one or more microbial biopolymers, the methods comprising co-cultivating Stigmatella aurantiaca and Acinetobacter venetianus. In certain embodiments, the one or more biopolymers are emulsan. In certain embodiments, other microbial growth by-products are produced, such as biosurfactants. Microbe-based products produced according to the subject methods are also provided, as well as their uses in, for example, oil and gas recovery, agriculture, and health care.

Abstract: The invention provides non-naturally occurring microbial organisms containing butadiene or 2,4-pentadienoate pathways comprising at least one exogenous nucleic acid encoding a butadiene or 2,4-pentadienoate pathway enzyme expressed in a sufficient amount to produce butadiene or 2,4-pentadienoate. The organism can further contain a hydrogen synthesis pathway. The invention additionally provides methods of using such microbial organisms to produce butadiene or 2,4-pentadienoate by culturing a non-naturally occurring microbial organism containing butadiene or 2,4-pentadienoate pathways as described herein under conditions and for a sufficient period of time to produce butadiene or 2,4-pentadienoate. Hydrogen can be produced together with the production of butadiene or 2,4-pentadienoate.