An  2,6-sialyltransferase cloned from Photobacterium leiognathi strain JT-SHIZ-119 shows both sialyltransferase and neuraminidase activity

Abstract

We cloned, expressed, and characterized a novel beta-galactoside alpha2,6-sialyltransferase from Photobacterium leiognathi strain JT-SHIZ-119. The protein showed 56-96% identity to the marine bacterial alpha2,6-sialyltransferases classified into glycosyltransferase family 80. The sialyltransferase activity of the N-terminal truncated form of the recombinant enzyme was 1477 U/L of Escherichia coli culture. The truncated recombinant enzyme was purified as a single band by sodium dodecyl sulfate polyacrylamide gel electrophoresis through 3 column chromatography steps. The enzyme had distinct activity compared with known marine bacterial alpha2,6-sialyltransferases. Although alpha2,6-sialyltransferases cloned from marine bacteria, such as Photobacterium damselae strain JT0160, P. leiognathi strain JT-SHIZ-145, and Photobacterium sp. strain JT-ISH-224, show only alpha2,6-sialyltransferase activity, the recombinant enzyme cloned from P. leiognathi strain JT-SHIZ-119 showed both alpha2,6-sialyltransferase and alpha2,6-linkage-specific neuraminidase activity. Our results provide important information toward a comprehensive understanding of the bacterial sialyltransferases belonging to the group 80 glycosyltransferase family in the CAZy database.

Loss-of-Function Mutation in Bi-Functional Marine Bacterial Sialyltransferase

“…Escherichia coli, 11) Photobacterium damselae, 12) Photobacterium phosphoreum, 13) Photobacterium leiognathi, 14,15) Photobacterium sp., 16) Vibrio sp., 17) Pasteurella multocida, 18) Haemophilus influenza, 19) and Streptococcus agalactiae. 20) Among these, some sialyltransferases are multi-functional enzymes.…”

Marine Bacterial Sialyltransferases

“…Genes encoding sialyltransferases have been cloned from various types of bacteria, including N. gonorrhoeae [29], Neisseria meningitidis [30], C. jejuni [31], E. coli [32], P. damselae [33], Photobacterium phosphoreum [34], Photobacterium leiognathi [35,36], Photobacterium sp. [37], Vibrio sp.…”

“…JT-ISH-224 [37], P. leiognathi JT-SHIZ-145 [35] and P. leiognathi JT-SHIZ-119 [36]. These genes have all been expressed as recombinant active form enzymes in an E. coli protein expression system.…”

Diversity, Abundance and Distribution of O-linked Glycosylation Pathway Enzymes in Prokaryotes-A Comparative Genomics Study

“…In addition, their promiscuous substrate specificity in response to variations in the assay conditions is advantageous for in vitro glycan synthesis [23,24]. Experimental approaches for the identification of new GTs include the use of probes derived from the sequences of hitherto characterized GTs [25] and screening cell lysates for activity [26]. The main disadvantage of such approaches is that they are very time-consuming.…”