Abstract: Provided are immunoglobulin-binding protein variants with increased alkali tolerance, and uses thereof, and more particularly to immunoglobulin-binding protein variants with increased alkali tolerance relative to the wild type due to mutations in amino acids at specific positions in the B3 domain of the immunoglobulin-binding proteins

Abstract: The present invention relates to a mutated immunoglobulin-binding protein having increased alkaline tolerance and, more specifically, to an immunoglobulin-binding protein in which, with respect to the A-domain of Staphylococcal protein A, or a functional variant thereof, an amino acid at a specific site is mutated and thereby exhibits an increased chemical stability at an alkaline pH value in comparison to a parental molecule. The present invention can provide an antibody-purifying immunoglobulin-binding protein ligand and matrix which have enhanced alkaline tolerance and accordingly enhanced stability in multiple times of alkaline cleaning.

Abstract: The present invention relates to a mutated immunoglobulin-binding protein having increased alkaline tolerance and, more specifically, to an immunoglobulin-binding protein in which, with respect to the A-domain of Staphylococcal protein A, or a functional variant thereof, an amino acid at a specific site is mutated and thereby exhibits an increased chemical stability at an alkaline pH value in comparison to a parental molecule. The present invention can provide an antibody-purifying immunoglobulin-binding protein ligand and matrix which have enhanced alkaline tolerance and accordingly enhanced stability in multiple times of alkaline cleaning.

Abstract: A method of detecting and quantifying various enzymatic activities using a constructed artificial genetic circuit GESS (genetic enzyme screening system) for sensing phenolic compounds and a method of screening a trace of activities of target enzymes from a metagenome using the artificial genetic circuit, thereby securing target enzyme genes. When the method for screening and quantifying target enzymatic activity is used, useful genes can be screened from various genetic communities, including environmental or metagenomic libraries, at a single cell level in high throughput (million/day). Further, the sensitivity of the genetic circuit to phenol derivatives and the expression thereof can be controlled, and thus the genetic circuit can rapidly sense and quantify various enzymatic activities. Thus, the method can be advantageously used in the protein engineering technology for enzyme modification.

Abstract: A method of performing high-throughput screening of various enzymatic activities with high sensitivity using artificial genetic circuits is provided. Particularly, the invention is screening and quantifying the activity of isoprene biosynthesis enzymes using an artificial genetic circuit capable of sensing isoprene. The artificial genetic circuit comprises an isoprene-sensing transcriptional regulator which recognizes isoprene, at least one reporter gene, a isoprene-sensing transcriptional regulator binding region and promoters for genes encoding isoprene-sensing transcriptional regulators and reporter proteins. The artificial genetic circuit detects isoprene liberated from many enzymatic reactions and measures the activity of reporter genes. This system is widely applicable for high throughput and quantitative screening of isoprene biosynthesis enzymes and MEP/MVA pathway enzymes. Therefore, the invention can be advantageously used in the protein engineering technology for enzyme modification.

Abstract: The present invention relates to a method for detecting protein-protein interactions in living cells, and more particularly, to a method for providing cells comprising a first construct and a second construct, wherein the first construct comprises a polynucleotide encoding a first fusion protein which comprises a bait protein, a first fluorescent protein and a CBD (cellulose-binding domain) protein, and wherein the second construct comprises a polynucleotide encoding a second fusion protein which comprises a prey protein and a second fluorescent protein so as to allow formation of inclusion bodies, and detecting interactions between the bait protein and the prey protein that are displayed by inclusion bodies, a method for isolating the prey protein bound to the bait protein using the cells comprising the constructs, the cells, and a kit for detecting protein-protein interactions, comprising the constructs.

Abstract: A method of performing high-throughput screening of various enzymatic activities with high sensitivity using artificial genetic circuits is provided. Particularly, the invention is screening and quantifying the activity of isoprene biosynthesis enzymes using an artificial genetic circuit capable of sensing isoprene. The artificial genetic circuit comprises an isoprene-sensing transcriptional regulator which recognizes isoprene, at least one reporter gene, a isoprene-sensing transcriptional regulator binding region and promoters for genes encoding isoprene-sensing transcriptional regulators and reporter proteins. The artificial genetic circuit detects isoprene liberated from many enzymatic reactions and measures the activity of reporter genes. This system is widely applicable for high throughput and quantitative screening of isoprene biosynthesis enzymes and MEP/MVA pathway enzymes. Therefore, the invention can be advantageously used in the protein engineering technology for enzyme modification.

Abstract: The present invention relates to a metagenome-derived alkaline phosphatase, and more particularly to a novel, metagenome-derived alkaline phosphatase screened using a artificial genetic circuit that detects phenolic compounds, and a preparation method thereof. A novel alkaline phosphatase according to the present invention has high activity of dephosphorylating DNA and can be inactivated rapidly by simple heat treatment. Thus, it can be used for a dephosphorylation reaction so that genetic manipulations, including genetic cloning, become efficient. In addition, it can be actively expressed in recombinant microorganisms, and thus can be used in various assays, including enzyme immunoassay.

Abstract: The present invention relates to a method for detecting protein-protein interactions in living cells, and more particularly, to a method for providing cells comprising a first construct and a second construct, wherein the first construct comprises a polynucleotide encoding a first fusion protein which comprises a bait protein, a first fluorescent protein and a CBD (cellulose-binding domain) protein, and wherein the second construct comprises a polynucleotide encoding a second fusion protein which comprises a prey protein and a second fluorescent protein so as to allow formation of inclusion bodies, and detecting interactions between the bait protein and the prey protein that are displayed by inclusion bodies, a method for isolating the prey protein bound to the bait protein using the cells comprising the constructs, the cells, and a kit for detecting protein-protein interactions, comprising the constructs.

Abstract: The present invention relates to a metagenome-derived alkaline phosphatase, and more particularly to a novel, metagenome-derived alkaline phosphatase screened using a artificial genetic circuit that detects phenolic compounds, and a preparation method thereof. A novel alkaline phosphatase according to the present invention has high activity of dephosphorylating DNA and can be inactivated rapidly by simple heat treatment. Thus, it can be used for a dephosphorylation reaction so that genetic manipulations, including genetic cloning, become efficient. In addition, it can be actively expressed in recombinant microorganisms, and thus can be used in various assays, including enzyme immunoassay.

Abstract: A method of detecting and quantifying various enzymatic activities using a constructed artificial genetic circuit GESS (genetic enzyme screening system) for sensing phenolic compounds and a method of screening a trace of activities of target enzymes from a metagenome using the artificial genetic circuit, thereby securing target enzyme genes. When the method for screening and quantifying target enzymatic activity is used, useful genes can be screened from various genetic communities, including environmental or metagenomic libraries, at a single cell level in high throughput (million/day). Further, the sensitivity of the genetic circuit to phenol derivatives and the expression thereof can be controlled, and thus the genetic circuit can rapidly sense and quantify various enzymatic activities. Thus, the method can be advantageously used in the protein engineering technology for enzyme modification.