TY - JOUR T1 - Structural and biochemical analysis of the asc operon encoding 6-phospho-beta-glucosidase in Pectobacterium carotovorum subsp. carotovorum LY34. AU - An,Chang Long, AU - Lim,Woo Jin, AU - Hong,Su Young, AU - Shin,Eun Chule, AU - Kim,Min Keun, AU - Lee,Jong Reoul, AU - Park,Sang Ryeol, AU - Woo,Jong Gyu, AU - Lim,Yong Pyo, AU - Yun,Han Dae, Y1 - 2004/12/10/ PY - 2004/06/03/received PY - 2004/09/10/revised PY - 2004/09/13/accepted PY - 2005/3/8/pubmed PY - 2005/7/7/medline PY - 2005/3/8/entrez SP - 145 EP - 53 JF - Research in microbiology JO - Res Microbiol VL - 156 IS - 2 N2 - An asc operon of Pectobacterium carotovorum subsp. carotovorum LY34 (Pcc LY34) was isolated from a genomic library in a screen for beta-glucosidase activities. Sequence analysis of the 5618-bp cloned DNA fragment (accession number AY622309) showed three open reading frames (ascG, ascF, and ascB) that are predicted to encode 375, 486, and 476 amino acid proteins, respectively. The AscG ORF shared a high similarity with the Escherichia coli AscG repressor. The AscF ORF shared 81% identity with the E. coli AscF PTS enzyme II(asc), while the AscB ORF was highly similar to 6-phospho-beta-glucosidases and is a member of the glycosyl hydrolase family 1. The purified AscB enzyme hydrolyzed salicin, arbutin, pNPG, and MUG. It exhibited maximal activity at pH 7.0 and 40 degrees C, and its activity was enhanced in the presence of Mg(2+) and Ca(2+). The molecular weight of the enzyme was estimated to be 53 000 Da by SDS-PAGE. Two conserved glutamate residues (Glu(182) and Glu(374)) were shown to be important for AscB activity. SN - 0923-2508 UR - https://www.unboundmedicine.com/medline/citation/15748978/Structural_and_biochemical_analysis_of_the_asc_operon_encoding_6_phospho_beta_glucosidase_in_Pectobacterium_carotovorum_subsp__carotovorum_LY34_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0923-2508(04)00258-X DB - PRIME DP - Unbound Medicine ER -