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.

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.

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.

Abstract: Methods for biosynthesising hydrocarbons from a gaseous substrate in non-naturally occurring acetogens as well as non-naturally occurring acetogens for production of hydrocarbons are provided.

Abstract: Provided are pharmaceutical formulations comprising a recombinant acid ?-glucosidase, wherein the recombinant acid ?-glucosidase is expressed in Chinese hamster ovary (CHO) cells and comprises an increased content of N-glycan units bearing one or two mannose-6-phosphate residues when compared to a content of N-glycan units bearing one or two mannose-6-phosphate residues of alglucosidase alfa; at least one buffer selected from the group consisting of a citrate, a phosphate and combinations thereof; and at least one excipient selected from the group consisting of mannitol, polysorbate 80, and combinations thereof, wherein the formulation has a pH of from about 5.0 to about 7.0. Also provided are methods of treating Pompe disease using these pharmaceutical formulations.

Abstract: This disclosure describes methods for regulating the biosynthesis of pimelic acid, 7-aminoheptanoate, 7-hydroxyheptanoate, heptamethylenediamine, 7-aminoheptanol, or 1,7-heptanediol by channeling increased flux through the biosynthesis pathway to obtain an intermediate required for growth of the host microorganism.

Abstract: Compositions and methods for treating or reducing the severity or likelihood of occurrence of a parasitic worm or helminth infection in a subject are described. The methods include administering to the subject a therapeutically effective amount of a killed or inactivated recombinant bacterium expressing a crystal protein such as a Bacillus thuringiensis crystal protein (Cry) in the cytosol of the bacterium. The crystal proteins may be full length, truncated, variant, or sub-variant Cry proteins. Examples of crystal proteins include Cry5B, Cry21, Cry14A, Cry6A, and Cry13A. The recombinant bacteria may be treated with an anti-microbial agent before or during administration to a subject.

Abstract: The present invention is related to production of a sterol mix in a modified yeast cell, wherein the amount of zymosterol present in said mix is dramatically reduced or abolished via modification of sterol acyltransferase activity within said yeast. The modified yeast cell can be used for production of vitamin D3 or derivatives and/or metabolites thereof.

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.