Abstract: Some aspects of this disclosure provide a fusion protein comprising a guide nucleotide sequence-programmable DNA binding protein domain (e.g., a nuclease-inactive variant of Cas9 such as dCas9), an optional linker, and a recombinase catalytic domain (e.g., a tyrosine recombinase catalytic domain or a serine recombinase catalytic domain such as a Gin recombinase catalytic domain). This fusion protein can recombine DNA sites containing a minimal recombinase core site flanked by guide RNA-specified sequences. The instant disclosure represents a step toward programmable, scarless genome editing in unmodified cells that is independent of endogenous cellular machinery or cell state.

Abstract: The present invention relates to sensors and methods for detecting carbohydrates, such as lactose, in a sample. The sensors and methods may also be used to determine the amount of carbohydrate in the sample.

Abstract: The invention relates to a process for the production of ethanol comprising fermenting a (optionally liquefied) corn slurry under anaerobic conditions in the presence of a recombinant yeast; and recovering the ethanol, wherein said recombinant yeast functionally expresses a heterologous nucleic acid sequence encoding a glucoamylase having an amino acid sequence according to SEQ ID NO: 1 or which glucoamylase is a functional homologue thereof having a sequence identity of at least 80%, or which glucoamylase is a functional homologue which is derived, by way of one or more amino acid substitutions, deletions or insertions, from the amino acid sequence of SEQ ID NO: 1, and wherein the process comprises dosing a glucoamylase at a concentration of 0.05 g/L or less.

Abstract: The present disclosure provides activated formylglycine-generating enzymes (FGE), methods of producing activated FGE, and their use in methods of producing a protein comprising a formylglycine (FGly) residue. The methods of producing activated FGE, as well as methods of use of activated FGE in producing FGly-containing proteins, include both cell-based and cell-free methods. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Abstract: A modified endoinulinase is provided, comprising modified wild-type T. purpuregenus endoinulinase, or a functional fragment thereof, in which an amino acid residue at each one of one or more positions corresponding to 128, 316, 344, 350 or 504 of wild-type T. purpuregenus endoinulinase is substituted, wherein: (i) a tyrosine residue corresponding to Y128 is substituted with H, K or R; a glutamate residue corresponding to E344 is substituted with K, H or R; and a threonine residue corresponding to T504 is substituted with M, S or Y; and optionally an alanine residue corresponding to A316 is substituted with T, S, C or M; (ii) a tyrosine residue corresponding to Y128 is substituted with H, K or R; a glutamate residue corresponding to E344 is substituted with K, H or R; a threonine residue corresponding to T504 is substituted with M, S or Y; and a glutamine residue corresponding to Q350 is substituted with L, G, A, V or I; or (iii) a tyrosine residue corresponding to Y128 is substituted with H, K or R.

Abstract: The disclosure discloses a genetically engineered strain for producing porcine myoglobin and fermentation and purification thereof, and belongs to the technical field of genetic engineering. The disclosure realizes efficient secretion and expression of porcine myoglobin by integrating the gene of porcine myoglobin in P. pastoris. On this basis, optimization of the medium and culture conditions of recombinant P. pastoris can significantly increase the titer of porcine myoglobin, so that the titer can reach 285.42 mg/L under fermenter conditions. In addition, by creatively adding different concentrations of ammonium sulfate to fermentation broth step by step, the purity of myoglobin obtained by final concentration is up to 88.0%, and the purification rate is up to 66.1%.

Abstract: The present invention is related to a method for adding one or more L-nucleotides to the 3?end of a first L-nucleic acid, wherein the method comprises the step of reacting the one or more L-nucleotides with the first L-nucleic acid in the presence of a protein comprising a mutant enzymatic activity exhibiting moiety, wherein the enzymatic activity is capable of adding one or more L-nucleotides to the 3? end of the first L-nucleic acid, wherein the mutant enzymatic activity exhibiting moiety comprises an amino acid sequence, wherein the amino acids of the amino acid sequence are D-amino acids, wherein the mutant enzymatic activity exhibiting moiety is a variant of an enzymatic activity exhibiting moiety, wherein the enzymatic activity exhibiting moiety consists of an amino acid sequence according to SEQ ID NO: 15 and wherein the amino acids of the amino acid sequence according to SEQ ID NO: 15 are D-amino acids, wherein the amino acid sequence of the mutant enzymatic activity exhibiting moiety differs from the

Abstract: Provided herein are deglycosylating enzymes that remove a broad range of N-glycans from N-glycosylated proteins. Further provided are methods of recombinantly producing and expressing the deglycosylating enzymes. The presently described deglycosylating enzymes can be used to produce free glycans for characterization, and for prebiotic and immunostimulatory uses. In addition, the presently described deglycosylating enzymes can be used to produce deglycosylated proteins for characterization, to improve digestion, and to reduce immunogenicity.

Abstract: The present invention provides: a novel allulose epimerase variant in which an amino acid residue present at a specific position of an amino acid sequence of a wild-type D-allulose 3-epimerase derived from Flavonifractor plautii is substituted with another amino acid residue; and various uses of the novel allulose epimerase variant. The novel allulose epimerase variant according to the present invention has a higher conversion rate of fructose to allulose compared to that of the wild-type D-allulose 3-epimerase derived from Flavonifractor plautii, and has excellent thermal stability especially under high temperature conditions of 60° C. or higher, and thus can prevent contamination during an industrial-scale enzymatic conversion reaction for the mass production of allulose, shorten production time, and reduce production costs.

Abstract: The present invention relates to a method for preparing oligosaccharides which can be used among others as food additives to reduce calorie content, to sweeten food products, to increase the fiber content of food products, to improve the texture of food products and to stimulate the gut microbiome bacteria. Furthermore they can be applied in the fields of animal feed, or other applications. More particularly, this invention is directed to a high temperature hydrolysis of xyloglucan polysaccharide to defined xyloglucan oligosaccharides. The invention further relates to oligosaccharide hydrolysates produced with the method of the invention and to the use of said oligosaccharide hydrolysates in human and/or animal nutrition, as prebiotic or other uses. Further provided are novel endoglucanases for use in the method of the invention as well as in other applications.

Abstract: The present invention provides engineered penicillin G acylase (PGA) enzymes, polynucleotides encoding the enzymes, compositions comprising the enzymes, and methods of using the engineered PGA enzymes.

Abstract: The present invention relates to polypeptides having trehalase activity, particularly derived from Talaromyces. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides for the production of ethanol.

Abstract: Provided herein are systems and methods for characterizing target/ligand engagement. In particular, luciferase-labeled polypeptide targets are used to detect or quantify target/ligand engagement (e.g., within a cell or cell lysate).

Abstract: The present invention describes a process for pretreatment of lignocellulosic biomass that comprises the step of contacting a lignocellulosic biomass with an ionic liquid consisting of a phthalic salt of dicholine in the weight ratio from 1:1 to 1:100 of biomass:ionic liquid, said step taking place for a period of time that varies from 0.4 to 48 hours and in a temperature range that varies from 60 to 200° C. Furthermore, the present invention also relates to the use of the pretreated lignocellulosic biomass in an enzymatic hydrolysis process.

Abstract: A process for producing cellulolytic and/or hemicellulolytic enzymes with a strain of microorganism belonging to the family of filamentous fungi. The process includes growing the fungi in an aqueous culture medium, in the presence of at least one carbon-based growth substrate, in a stirred and aerated bioreactor. It also includes the production of enzymes, starting with the aqueous culture medium in the presence of at least one inductive carbon-based substrate and also inducing the production of hydrophobins. Further, at least a portion of the hydrophobins produced in step (b) are reintroduced into the growth step (a).

Abstract: Provision of a modified promoter derived from a xylanase promoter. A modified promoter comprising a polynucleotide of Xyn3 promoter comprising a polynucleotide that comprises at least one polynucleotide of Xyn1 promoter cis-element or a complementary strand thereof in a region corresponding to the nucleotides at position 374 to 401 of SEQ ID NO:1. The polynucleotide of Xyn3 promoter consist of the following nucleotide sequences: the nucleotide sequence represented by SEQ ID NO:1; the nucleotide sequence represented by the nucleotides at position 350 to 1084 of SEQ ID NO:1; or a nucleotide sequence that has an identity of at least 90% therewith and that comprises the sequence represented by SEQ ID NO:2 in a region corresponding to the nucleotides at position 374 to 401 of SEQ ID NO:1. The polynucleotide of Xyn1 promoter cis-element consists of the nucleotide sequence represented by SEQ ID NO:4.

Abstract: Disclosed are genetically engineered organisms, such as yeast and bacteria, that have the ability to metabolize atypical phosphorus or sulfur sources. Fermentation methods using the genetically engineered organisms are also described. The fermentation methods are robust processes for the industrial bioproduction of a variety of compounds, including commodities, fine chemicals, an pharmaceuticals.

Abstract: Disclosed herein is a functional nucleic acid that includes i) one or more coding nucleotide sequences encoding a genome editing endonuclease; ii) regulatory sequences operably linked to the one or more coding nucleotide sequences; and iii) one or more genome editing endonuclease-recognized sequences, wherein the functional nucleic acid is configured to express the endonuclease in a host cell and thereby provide a cellular endonuclease activity in a sequence-specific manner in the host cell, and wherein cleavage of the functional nucleic acid by the endonuclease inactivates the cellular endonuclease activity. Methods of using the present functional nucleic acid, and systems and kits that find use in performing the same are also provided.

Abstract: A non-naturally occurring eukaryotic or prokaryotic organism includes one or more gene disruptions occurring in genes encoding enzymes imparting increased fumarate, malate or acrylate production in the organism when the gene disruption reduces an activity of the enzyme. The one or more gene disruptions confers increased production of acrylate onto the organism. Organisms that produce acrylate have an acrylate pathway that at least one exogenous nucleic acid encoding an acrylate pathway enzyme expressed in a sufficient amount to produce acrylate, the acrylate pathway comprising a decarboxylase. Methods of producing fumarate, malate or acrylate include culturing these organisms.

Abstract: The present disclosure relates to microbial stem cell technology that enables a growing microbial culture to stably maintain two or more distinct cell types in a ratio that can be genetically programmed and/or dynamically controlled during cultivation. It is contemplated that embodiments described herein can be utilized to increase product yield in microbial fermentations and advanced engineering of biomaterials using genetically engineered microbial cells, among others.