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Heterologous Expression and Characterization of an N-Acetyl-beta-D-hexosaminidase from Lactococcus lactis ssp. lactis IL1403
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SYSNO ASEP 0377491 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Heterologous Expression and Characterization of an N-Acetyl-beta-D-hexosaminidase from Lactococcus lactis ssp. lactis IL1403 Author(s) Nguyen, A. H. (AT)
Nguyen, T.-H. (AT)
Křen, Vladimír (MBU-M) RID, ORCID
Eijsink, V. G. H. (NO)
Haltrich, D. (AT)
Peterbauer, C. (AT)Source Title Journal of Agricultural and Food Chemistry. - : American Chemical Society - ISSN 0021-8561
Roč. 60, č. 12 (2012), s. 3275-3281Number of pages 7 s. Language eng - English Country US - United States Keywords N-acetyl-beta-D-hexosaminidase ; Lactococcus lactis ssp lactis IL1403 ; pNP-GlcNAc Subject RIV CE - Biochemistry R&D Projects GAP207/11/0629 GA ČR - Czech Science Foundation (CSF) UT WOS 000301969300042 DOI 10.1021/jf204915e Annotation The lnbA gene of Lactococcus lactis ssp. lactis IL1403 encodes a polypeptide with similarity to lacto-N-biosidases and N-acetyl-beta-D-hexosaminidases. The gene was cloned into the expression vector pET-21d and overexpressed in Escherichia coli BL21* (DE3). The recombinant purified enzyme (LnbA) was a monomer with a molecular weight of approximately 37 kDa. Studies with chromogenic substrates including p-nitrophenyl N-acetyl-beta-D-glucosamine (pNP-GlcNAc) and p-nitrophenyl N-acetyl-beta-D-galactosamine (pNP-GalNAc) showed that the enzyme had both N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D-galactosaminidase activity, thus indicating that the enzyme is an N-acetyl-beta-D-hexosaminidase. K-m and k(cat) for pNP-GlcNAc were 2.56 mM and 26.7 s(-1), respectively, whereas kinetic parameters for pNP-GalNAc could not be determined due to the Km being very high (>10 mM) Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2013
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