sialyltransferase, marine bacterium, Genus Photobacterium, Genus Vibrio

Bacterial Sialyltransferases


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Background

Sialic acids are present in a variety of glycoproteins and glycolipids, often at the non-reducing termini of carbohydrate chains. It has been demonstrated that sialic acids play very important roles in various biological and physiological events. Ample supply of sialosides and sialyl-glycoconjugates is indispensable to the study of their biological functions in detail. Transfer of sialic acids by sialyltransferases to appropriate substrates in the final step under a mild reaction condition can prepare these materials in quantity. Therefore, one of the most important tasks in the study of glycobiology is to provide a large amount of bacterial sialyltransferases with diverse characteristics at low prices.

Standard Assay Method

Sialyltransferase activity was assayed by measuring [4,5,6,7,8,9-14C]-NeuAc transferred from CMP-[4,5,6,7,8,9-14C]-NeuAc to lactose as an acceptor substrate. Add 0.5M of NaCl to the reaction mixture. The radioactivity of CMP-[4,5,6,7,8,9-14C]-NeuAc that had transferred to the acceptor was measured with a liquid scintillation counter, and the amount of NeuAc transferred was calculated.

CMP-[4,5,6,7,8,9-14C]-NeuAc + lactose → [4,5,6,7,8,9-14C]-NeuAc-lactose + CMP

Feature and Advantages

Table 1. Product comparison

 

Table 2. Acceptor substrate specificity of a2,6-sialyltransferases from Photobacterium damselae JT0160.

 

Table 3. Comparison of acceptor substrate specificities of a2,3-sialyltransferases from marine bacteria.

 

Figure 1. Enzymatic synthesis of several sialyloligosaccharides employing marine bacterial sialyltransferases.

 

Figure 2. Enzymatic synthesis of several sialyloligosaccharides employing marine bacterial sialyltransferases.

Product List

Recombinant beta-galactoside-alpha-2,6-sialyltransferase (EC No. : 2. 4. 99. 1)

Origin : Photobacterium damselae JT0160


Product Name Cat# Quantity Price

(β)-Galactoside-(α)-2,6-Sialyltransferase DataSheet ,

CSR-GEJ-001

0.5 UNIT N/A

Recombinant beta-galactoside-alpha-2,3-sialyltransferase (EC No. : 2. 4. 99. 4)

Origin : Photobacterium sp.JT-ISH-224

Product Name Cat# Quantity Price

(β)-Galactoside-(α)-2,3-Sialyltransferase DataSheet ,

CSR-GEJ-002

0.5 UNIT N/A

Recombinant beta-galactoside-alpha-2,3-sialyltransferase (EC No. : 2. 4. 99. 4)

Origin : Photobacterium phosphoreum JT-ISH-467

Product Name Cat# Quantity Price

(β)-Galactoside-(α)-2,3-Sialyltransferase DataSheet ,

CSR-GEJ-003

0.5 UNIT N/A

Recombinant beta-galactoside-alpha-2,3-sialyltransferase (EC No. : 2. 4. 99. 4)

Origin: Photobacterium phosphoreum JT-ISH-467

Product Name Cat# Quantity Price

beta-galactoside-alpha-2,3-sialyltransferase from Photobacterium phosphoreum JT-ISH-467 DataSheet ,

CSR-GEJ-004

0.5 UNIT N/A

¡  Reference

[20] Mine, T., Kajiwara, H., Murase, T., Kajihara, Y. and Yamamoto, T. An α2,3-sialyltransferase cloned from Photobacterium sp. JT-ISH-224 transfers N-acetylneuraminic acid to both O-2 and O-3’ hydroxyl groups of lactose. J. Carbohydr. Chem. 29 (2010) 51-60.

[19] Takakura, Y., Oka, N., Kajiwara, H., Tsunashima, M., Usami, S., Tsukamoto, H., Ishida, Y. and Yamamoto, T. Tamavidin, a versatile affinity tag for protein purification and immobilization. J. Biotechnol. 145 (2010) 317-322.

[18] Kushi, Y., Kamimiya, H., Hiratsuka, H., Nozaki, H., Fukui, H., Yanagida, M., Hashimoto, M., Nakamura, K., Watarai, S., Kasama, T., Kajiwara, H. and Yamamoto, T. Sialyltransferases of marine bacteria efficiently utilize glycosphingolipid substrates. Glycobiology 20 (2010) 187-198.

[17] Mine, T., Katayama, S., Kajiwara, H., Tsunashima, M., Tsukamoto, T., Takakura, Y. and Yamamoto, T. An α2,6-sialyltransferase cloned from Photobacterium leiognathi strain JT-SHIZ-119 shows both sialyltransferase and neuraminidase activity. Glycobiology 20 (2010) 158-165.

[16] Yamamoto, T. CMP-N-acetylneuraminic Acid Synthetases and Sialyltransferases from Bacterial Sources in Glycobiology Research Trends, Nova Science Publisher, N.Y., 2009, pp. 85-107.

[15]Kajiwara, H., Mine, T. and Yamamoto, T. Sialyltransferases obtained from marine bacteria. J. Appl. Glycosci. 56 (2009) 77-82.

[14]Iwatani, T., Okino, N., Sakakura, M., Kajiwara, H., Takakura, Y., Kimura, M., Ito, M., Yamamoto, T. and Kakuta, Y. Crystal structure of α/β-galactoside α-2,3-sialyltransferase from a luminous marine bacterium, Photobacterium phosphoreum. FEBS lett. (2009) 2083-2087.

[13]Tsukamoto, H., Takakura, Y., Mine, T., Yamamoto, T. Photobacterium sp. JT-ISH-224 Produces Two Sialyltransferases, α/β-Galactoside α2,3-Sialyltransferase and β-Galactoside α2,6-Sialyltransferase. J. Biochem. (Tokyo) 143 (2008) 187-197.

[12]Yamamoto, T., Ichikawa, M., Takakura, Y. Conserved amino acid sequences in the bacterial sialyltransferases belonging to Glycosyltransferase family 80. Biochem. Biophys. Res. Commun. 365 (2008) 340-343.

[11]Kakuta, Y., Okino, N., Kajiwara, H., Ichikawa, M., Takakura, Y., Ito, M., Yamamoto, T. Crystal structure of Vibrionaceae Photobacterium sp. JT-ISH-224 α2,6-sialyltransferase in a ternary complex with donor product CMP and acceptor substrate lactose: catalytic mechanism and substrate recognition. Glycobiology 18 (2008) 66-73.

[10] Yamamoto, T., Hamada, Y., Ichikawa, M., Kajiwara, H., Mine, T., Tsukamoto, H., Takakura, Y. A β-galactoside α2,6-sialyltransferase produced by a marine bacterium, Photobacterium leiognathi JT-SHIZ-145, is active at pH 8. Glycobiology 17 (2007) 1167-1174.

[9] Tsukamoto, H., Takakura, Y., Yamamoto, T. Purification, cloning and expression of an α-/β-galactoside α2,3-sialyltransferase from a luminous marine bacterium, Photobacterium phosphoreum. J. Biol. Chem. 282 (2007) 29794-29802.

[8] Kajihara, Y., Kamitani, T., Sato, R., Kamei, N., Miyazaki, T., Okamoto, R., Sakakibara, T., Tsuji, T. and Yamamoto, T. Synthesis of CMP-9’’-modified-sialic acids as donor substrate analogues for mammalian and bacterial sialyltransferases. Carbohydr. Res. 342 (2007) 1680-1688.

[7] Takakura, Y., Tsukamoto, H., Yamamoto, T. Molecular Cloning, Expression and Properties of an α/β-Galactoside α2,3-Sialyltransferase from Vibrio sp. JT-FAJ-16. J. Biochem (Tokyo) 142 (2007) 403-412.

[6] Okino, N., Kakuta, Y., Kajiwara, H., Ichikawa, M., Takakura, Y., Ito, M., Yamamoto, T. Purification, crystallization and preliminary crystallographic characterization of the α2,6-sialyltransferase from Vibrionaceae Photobacterium sp. JT-ISH-224. Acta. Crystallogr. Sect. F Struct. Biol. Cryst. Commun. Aug 1 (2007) 662-664.

[5] Yamamato, T., Takakura, Y., Tsukamoto, H. Bacterial sialyltransferases. Trends@Glycosci. Glycotech. 18 (2006) 253-265.

[4] Yamamoto, T., Nagae, H., Kajihara, Y., Terada, I. Mass production of bacterial α2,6-sialyltransferase and enzymatic syntheses of sialyloligosaccharides. Biosci. Biotechnol. Biochem, 62 (1998) 210-214.

[3] Yamamoto, T., Nakashizuka, M., Terada, I. Cloning and expression of a marine bacterial β-galactoside α2,6-sialyltransferase gene from Photobacterium damsela JT0160. J. Biochem. (Tokyo) 123 (1998) 94-100.

[2] Kajihara, Y., Yamamoto, T., Nagae, H., Nakashizuka, M., Sakakibara, T., and Terada, I. A Novel α2,6-Sialyltransferase: Transfer of Sialic Acid to Fucosyl and Sialyl Trisaccharides. J. Org. Chem. 61 (1996) 8632-8635.

[1] Yamamoto, T, Nakashizuka, M, Kodama, H, Kajihara, Y, Terada, I. Purification and characterization of a marine bacterial β-galactoside α2,6-sialyltransferase from Photobacterium damsela JT0160. J. Biochem. (Tokyo) 120 (1996) 104-110.