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Overproduction of a β-fructofuranosidase1 with a high FOS synthesis activity for efficient biosynthesis of fructooligosaccharides

Aung, Thu, Jiang, Hong, Liu, Guang-Lei, Chi, Zhe, Hu, Zhong, Chi, Zhen-Ming
International journal of biological macromolecules 2019 v.130 pp. 988-996
Aureobasidium, beta-fructofuranosidase, biosynthesis, biotechnology, enzyme activity, fructooligosaccharides, gene overexpression, genes, glycosylation, molecular weight, sucrose, yeasts
Aureobasidium melanogenum 11–1 was found to be able to produce over 281.7 ± 7.1 U/mL of β-fructofuranosidase activity. The protein deduced from the cloned β-fructofuranosidase1 gene had the conserved motif A (IGDP), motif D (RDP) and motif E (ET) and 11 N-glycosylation sites, indicating it was a β-fructofuranosidase with the high-level fructooligosaccharide (FOS) biosynthesis. Overexpression of the β-fructofuranosidase1 gene in the yeast strain 11–1 made a tranformant 33 produce 557.7 U/mL of β-fructofuranosidase activity. The molecular weight of the β-fructofuranosidase1 in which all the carbohydrates were removed by the Endo-H was 82.4 kDa. Within 7 h of the transfructosylation reaction, the yield of FOS was 0.66 g of FOS/g of sucrose and percentages of GF2, GF3 and GF4 were 79.5%, 18.9% and 1.6%. This demonstrated that the β-fructofuranosidase1 and the transformant 33 had highly potential applications in biotechnology for FOS production.