Journal of Food and Nutrition Research
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Journal of Food and Nutrition Research. 2014, 2(4), 193-197
DOI: 10.12691/jfnr-2-4-10
Open AccessArticle

Biosynthesis of 2-O-α-D-glucopyranosyl-L-Ascorbic Acid from Maltose by Cyclodextrin Glucanotransferase from Bacillus sp. SK 13.002

Ahmed Eibaid1, Mohanad Bashari1, Ming Miao1, Abubakr Musa1, Tao Zhang1 and Bo Jiang1,

1State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, China

Pub. Date: May 14, 2014

Cite this paper:
Ahmed Eibaid, Mohanad Bashari, Ming Miao, Abubakr Musa, Tao Zhang and Bo Jiang. Biosynthesis of 2-O-α-D-glucopyranosyl-L-Ascorbic Acid from Maltose by Cyclodextrin Glucanotransferase from Bacillus sp. SK 13.002. Journal of Food and Nutrition Research. 2014; 2(4):193-197. doi: 10.12691/jfnr-2-4-10


In this work 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G) was synthesized by Cyclodextrin glucanotransferase (CGTase) from Bacillus sp. SK 13.002 with L-ascorbic acid (AA) as an acceptor and maltose as a glycosyl donor. AA-2G production was analyzed by HPLC and was confirmed by LC/MS results. The reaction parameters, such as pH (4.0-9.0), temperature (25-50C), time (0-30 h), substrate ratios and enzyme concentration were optimized. The results showed that the optimum condition was pH 8.0 at 37C for 24 h, 1:1 maltose to AA substrates mass ratio, and 200 U/mL of CGTase. Under these conditions, the production of AA-2G was 5.5 g/L, this result indicate that CGTase from Bacillus sp. SK 13.002 can effectively uses maltose as a glycosyl donor to produce AA-2G in high yield.

Cyclodextrin glucanotransferase 2-0-α-D-glucopyransyl-L-ascorbic acid maltose

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[1]  Cruz-Rus, E., Amaya, I. and Valpuesta, V. “The challenge of increasing vitamin C content in plant foods”, Biotechnol J, 7 (9). 1110-1121. 2012.
[2]  Phillips, K.M., Tarrago-Trani, M.T., Gebhardt, S.E., Exler, J., Patterson, K.Y., Haytowitz, D.B. and Pehrsson, P.R. “Holden, Joanne M “Stability of vitamin C in frozen raw fruit and vegetable homogenates”, J Food Comp Anal, 23 (3). 253-259. 2010.
[3]  Yamamoto, I., Muto, N., Murakami, K. and Akiyama, J. “Collagen synthesis in human skin fibroblasts is stimulated by a stable form of ascorbate, 2-O-α-D-glucopyranosyl-L-ascorbic acid”, J Nutr, 122. 871-877. 1992.
[4]  Yamamoto, I., Muto, N. Murakami, K., Suga, S. and Yamaguchi, H. “L-Ascorbic acid α-glucoside formed by regioselective transglucosylation with rat intestinal and rice seed α-glucosidases: Its improved stability and structure determination”, Chem Pharm Bull, 38. 3020-3023. 1990.
[5]  Yamamoto, I. and Tai, A. “The current state on development of novel vitamin derivatives”, Nippon Rinsho, 57. 2332-2338. 1997.
[6]  Mima, H., Nomura, H., Imai, Y. and Takashima, H. “Chemistry and application of ascorbic acid phosphate”, Vitamin, 41. 387 1970.
[7]  Mead, C.G. and Finamore, F.J. “Occurrence of ascorbic acid sulfate in the brine shrimp, Artemia salina”, Biochem, 8 (6). 2652-2655. 1969
[8]  Lu, P.W., Lillard, D.W., Seib, P.A., Kramer, K.J. and Liang, Y.T. “Synthesis of the 2-methyl ether of L-ascorbic-acid - stability, vitamin activity, and C-13 nuclear magnetic-resonance spectrum compared to those of the 1-methyl and 3-methyl ethers”, J Agric Food Chem, 32. 21-28. 1984.
[9]  Gregory III, J.F. “Nutritional Properties and Significance of Vitamin Glycosides”, Annu Rev Nutr, 18. 277-96. 1998.
[10]  Han, R., Liu, L., Li, J., Du, G. and Chen, J. “Functions, applications and production of 2-O-D-glucopyranosyl-l-ascorbic acid”, Appl Microbiol Biotechnol, 95 (2). 313-320. 2012.
[11]  Yamamoto, I., Muto, N., Nagata, E., Nakamura, T. and Suzuki, Y. “Formation of a stable L-ascorbic acid α-glucoside by mammalian α-glucosidase-catalyzed transglucosylation”, Biochim Biophys Acta, 1035. 44-50. 1990.
[12]  Nakamura, S. and Oku, T. “Bioavailability of 2-O-α-D-glucopyranosyl-L-ascorbic acid as ascorbic acid in healthy human”, Nutr,. 25. 686-691. 2009
[13]  Markosyan, A.A., Abelyan, L.A., Adamyan, M.O., Akopyan, Z.I. and Abelyan, V.A. “Transglycosylation of L-Ascorbic Acid”, Appl Biochem Biotechnol, 43 (1). 36-40. 2007.
[14]  Lee, S.B., Nam, K., Lee, S.J., Lee, J.H., Inouye, K. and Park, K.H. “Antioxidative effects of glycosyl-ascorbic acids synthesized by maltogenic amylase to reduce lipid oxidation and volatiles production in cooked chicken meat”, Bioscsi Biotechnol Biochem, 68. 36-43. 2004.
[15]  Jun, HK., Bae, KM. and Kim, SK. “Production of 2-O-α-D-glucopyranosyl L-ascorbic acid using cyclodextrin glucanotransferase from Paenibacillus sp.”, Biotechnol Lett, 23. 1793-1797. 2001.
[16]  Prousoontorn, M.H. and Pantatan, S. “Production of 2-O-α-glucopyranosyl L-ascorbic acid from ascorbic acid and b-cyclodextrin using immobilized cyclodextrin glycosyltransferase”, J Incl Phenom Macrocycl Chem, 57. 39-46. 2007.
[17]  Zhang, Z., Li, J., Liu, L., Sun, J., Hua, Z., Du, G. and Chen, J. “Enzymatic Transformation of 2-O-α-D-glucopyranosyl-L-ascorbic Acid by α-cyclodextrin Glucanotransferase from Recombinant Escherichia coli”, Biotechnol Bioprocess Eng, 16. 107-113. 2011.
[18]  Liu, L., Xu, Q., Han, R., Shin, H., Chen, R., Li, J., Du, G. and Chen, J. “Improving maltodextrin specificity for enzymatic synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid by site-saturation engineering of subsite-3 in cyclodextrin glycosyltransferase from Paenibacillus macerans”, J Biotechnol, 166. 198-205. 2013.
[19]  Liu, L., Han, R., Shin, H., Li, J., Du, G. and Chen, J. “Biosynthesis of 2-O-D-glucopyranosyl-L-ascorbic acid from maltose by an engineered cyclodextrin glycosyltransferase from Paenibacillus macerans”, Carbohydr Res, 382. 101-107. 2013.
[20]  Sun, T., Jiang, B. and Pan, B.L. “Purification and Characterization of Novel Cyclodextrin Glucanotransferase from Bacillus sp SK 13.002”, Asian J Chem, 23 (11). 4977-4982. 2011.
[21]  Kwon T., Kim, C.T. and Lee J-H. “Transglucosylation of ascorbic acid to ascorbic acid 2-glucoside by a recombinant sucrose phosphorylase from Bifidobacterium longum”, Biotechnol Lett, 29. 611-5. 2007.
[22]  Zhang, Z., Li, J., Liu, L., Sun, J., Hua, Z., Du, G. and Chen, J. “Enzymatic transformation of 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G) by immobilized α-cyclodextrin glucanotransferase from recombinant Escherichia coli”, J Mol Catal B: Enzym, 68. 223-229. 2011.
[23]  Tanaka, M., Muto, N. and Yamamoto, I. “Characterization of Bacillus stearothermophilus cyclodextrin glucanotransferase in ascorbic acid 2-O-α-glucoside formation”, Biochim Biophys Acta, 1078. 127-132. 1991.