Identification of galacto-N-biose phosphorylase from Clostridium perfringens ATCC13124

Applied Microbiology and Biotechnology
Masahiro NakajimaMotomitsu Kitaoka

Abstract

Lacto-N-biose phosphorylase (LNBP) from bifidobacteria is involved in the metabolism of lacto-N-biose I (Galbeta1-->3GlcNAc, LNB) and galacto-N-biose (Galbeta1-->3GalNAc, GNB). A homologous gene of LNBP (CPF0553 protein) was identified in the genome of Clostridium perfringens ATCC13124, which is a gram-positive anaerobic intestinal bacterium. In the present study, we cloned the gene and compared the substrate specificity of the CPF0553 protein with LNBP from Bifidobacterium longum JCM1217 (LNBPBl). In the presence of alpha-galactose 1-phosphate (Gal 1-P) as a donor, the CPF0553 protein acted only on GlcNAc and GalNAc, and GalNAc was a more effective acceptor than GlcNAc. The reaction product from GlcNAc/GalNAc and Gal 1-P was identified as LNB or GNB. The CPF0553 protein also phosphorolyzed GNB much faster than LNB, which suggests that the protein should be named galacto-N-biose phosphorylase (GNBP). GNBP showed a kcat/Km value for GNB that was approximately 50 times higher than that for LNB, whereas LNBPBl showed similar kcat/Km values for both GNB and LNB. Because C. perfringens possesses a gene coding endo-alpha-N-acetylgalactosaminidase, GNBP may play a role in the intestinal residence by metabolizing GNB that is available ...Continue Reading

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Citations

Jan 10, 2009·Applied Microbiology and Biotechnology·Masahiro NakajimaMotomitsu Kitaoka
Aug 3, 2011·Applied Microbiology and Biotechnology·Takanori NihiraMotomitsu Kitaoka
Jan 7, 2009·The Journal of Biological Chemistry·Masafumi HidakaShinya Fushinobu
Sep 26, 2013·Protein Engineering, Design & Selection : PEDS·Yoshiyuki KoyamaMotomitsu Kitaoka
Aug 30, 2008·Applied and Environmental Microbiology·Masahiro Nakajima, Motomitsu Kitaoka
Dec 15, 2010·Bioscience, Biotechnology, and Biochemistry·Shinya Fushinobu
Mar 22, 2014·PloS One·Masahiro NakajimaMotomitsu Kitaoka
Aug 12, 2010·Bioscience, Biotechnology, and Biochemistry·Masahiro NakajimaMotomitsu Kitaoka
Nov 25, 2011·Enzyme and Microbial Technology·Chao ChenDesmet Tom
Aug 22, 2015·Applied Microbiology and Biotechnology·Motomitsu Kitaoka
Aug 15, 2013·The Journal of Biological Chemistry·Takanori NihiraHiroyuki Nakai
Oct 8, 2014·Acta Crystallographica. Section F, Structural Biology Communications·Masahiro NakajimaHayao Taguchi
Jul 18, 2020·Scientific Reports·Antonio Rubio-Del-CampoMaría J Yebra

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