Abstract: A SpyCatcher peptide-modified carrier, a protein immobilizing carrier, and a protein immobilizing method. The method comprises: linking a SpyCatcher peptide to a carrier so as to obtain a SpyCatcher peptide-modified carrier; and then based on a SpyCatcher-SpyTag reaction, immobilizing a target protein containing SpyTag to the SpyCatcher-modified carrier. A protein immobilization process in which the method is used is fast, a condition is mild, no extra chemical reagent is needed, the immobilization efficiency and an activity recovery rate are high, and the homogeneity is good.

Abstract: The present invention provides a fusion polypeptide comprising a target polypeptide moiety and a self-aggregating peptide moiety, and a method of producing and purifying a target polypeptide by expressing the fusion polypeptide.

Abstract: The present disclosure describes a Mtu ?I-CM intein variant containing one or more mutations or a biologically active fragment thereof, and a method for producing and purifying a molecule of interest using the intein variant. Further described are isolated fusion proteins comprising the intein variant and a tag and a molecule of interest. Also described are expression systems for expressing the intein variant as well as polypeptide screening methods employing the intein variant.

Abstract: The present disclosure describes a Mtu ?I-CM intein variant containing one or more mutations or a biologically active fragment thereof, and a method for producing and purifying a molecule of interest using the intein variant. Further described are isolated fusion proteins comprising the intein variant and a tag and a molecule of interest. Also described are expression systems for expressing the intein variant as well as polypeptide screening methods employing the intein variant.

Abstract: The present invention relates to a method for production and purification of polypeptides. In particular, the present invention relates to a fusion protein comprising a solubility-enhancing peptide tag moiety, a self-aggregating peptide moiety and a moiety of target peptide and to a method for production and purification of target peptides through expressing said fusion protein.

Abstract: The present invention relates to a method for production and purification of polypeptides. In particular, the present invention relates to a fusion protein comprising a solubility-enhancing peptide tag moiety, a self-aggregating peptide moiety and a moiety of target peptide and to a method for production and purification of target peptides through expressing said fusion protein.

Abstract: A method for detecting the presence of an oxygenated compound which is produced when a substrate is reacted with an oxygenase for the substrate. The method involves reacting a coupling enzyme with the oxygenated compound to form a polymeric oxygenated compound which is fluorescent or luminescent. Measurement of the fluorescence or luminescence of the polymeric oxygenated compound provides indirect detection of the oxygenated compound produced by reaction of the oxygenase with the substrate. The method is carried out in a whole cell environment wherein the cell is transformed to express both the oxygen a set being screened and the coupling enzyme. The method can be used to measure the activity of monooxygenases and dioxygenases on aromatic substrates. The method is amenable to large scale screening of enzyme mutants to isolate those with maximum oxygenase activity.

Abstract: This invention relates to the improved expression of evolved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly peroxidase enzymes, in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this maruer include improved folding, without formation of inclusion bodies, and retained functional activity. In a particular embodiment, the invention relates to improved expression of the horseradish peroxidase (HRP) gene and HRP enzymes. HRP mutants that are highly expressed, highly active, and/or thermostable, are disclosed.