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Protease papain ? enzyme with glycosidic activity?
Authors: Ježová Jana | Bílková Zuzana
Year: 2005
Type of publication: ostatní - přednáška nebo poster
Name of source: 12th Young Investigators´ Seminar on Analytical Chemistry (YISAC 2005), Book of Abstracts
Publisher name: University of Sarajevo
Place:
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Titles:
Language Name Abstract Keywords
cze Proteáza papain - enzym s glykosidázovou aktivitou? určeno pro seznam publikací Papain; imobilizace; magnetické nosiče; fragmentace; chitosan; kyselina hyaluronová
eng Protease papain ? enzyme with glycosidic activity? Papain (EC 3.4.22.2) is a sulfhydryl protease from Carica papaya latex. Its native form is unreactive until activated by a mild disulfide reducing agent such as cysteine. The enzyme catalyzes the hydrolysis of proteins, short-chain peptides, amino acid esters and amide links non-specifically but downstream of basic residues such as Arg and Lys [1]. Papain is a single peptide chain with molecular mass of 23 000 consists of 211 residues folded into two parts that form a cleft [2] with Cys-25, His-159 and Asp-158 in active site [3]. Papain has an isoelectric point of 9.6 and pH optimum range of enzymatic activity of pH 6 ? 7. Enzyme activity may be measured by pH stat using BAEE (N-α-benzoyl-L-arginine ethyl ester) [1]. Papain is especially applied in beer clarification, meat tenderizing and preparation of protein hydrolysis [4]. In order to stabilize proteolytic activity of enzyme we have immobilized papain on the non-magnetic and magnetic macroporous bead cellulose (particle size of 80 ? 250 μm). Prepared enzyme reactor was utilized for the fragmentation of the IgG molecule onto Fab and Fc fragments. Subsequently the nanopeptide with attached carbohydrate moiety presented in the Fc fragment was prepared by the trypsin digestion and the oligosaccharide chain structure was studied using lectin affinity chromatography. Terbojevich et al. (1996) had successfully used immobilized papain for depolymerization of the chitosan; and their data suggest that papain acts on the link between glucosamine and N-acetylglucosamine units [5]. We used papain reactor for the fragmentation of hyaluronic acid (HA), which is composed of disaccharide units consisted of D-glucuronic acid and N-acetyl-D-glucosamine. Immobilized papain has fragmentized HA too, with the most effectiveness in the initial period of the reaction. papain; immobilization; magnetic carriers; fragmentation; chitosan; hyaluronic acid

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