TABLE 1

Activity data, obtained by HPLC analysis, for combination β-fructofuranosidase variantsa

No. of substitutionsSubstitution presenceConcn (g/liter) of:
F140YA178PA178SG321NQ490SGlucoseFructoseSucrose1-KestoseNystose
4 (V1)xxxx74.6 (± 3.4)14.5 (± 0.9)177.7 (± 1.3)67.2 (± 0.5)53.8 (± 2.0)
4xxxx50.3 (± 19.6)14.6 (± 6.1)186.1 (± 3.0)71.9 (± 6.4)33.3 (± 1.5)
3 (V4)xxx68.1 (± 1.6)11.2 (± 0.6)177.8 (± 0.6)63.1 (± 0.9)50.8 (± 1.5)
3 (V5)xxx69.0 (± 3.9)12.0 (± 1.2)180.2 (± 3.0)61.0 (± 3.7)47.4 (± 2.5)
3 (V3)xxx67.0 (± 0.5)14.1 (± 2.0)176.5 (± 2.2)65.3 (± 1.4)47.0 (± 0.7)
3xxx64.8 (± 0.9)10.2 (± 1.6)173.4 (± 2.9)75.0 (± 1.0)42.4 (± 2.3)
3xxx60.6 (± 0.6)12.0 (± 0.4)168.4 (± 0.8)80.7 (± 0.2)34.2 (± 0.6)
3xxx58.7 (± 0.6)22.3 (± 0.5)167.1 (± 1.5)80.6 (± 0.6)34.0 (± 0.4)
3xxx47.3 (± 1.0)18.4 (± 1.5)119.9 (± 1.1)86.5 (± 1.1)15.6 (± 0.7)
2 (V8)xx67.1 (± 1.4)15.3 (± 0.8)174.1 (± 0.9)69.2 (± 2.5)47.4 (± 0.6)
2xx57.7 (± 7.5)11.5 (± 0.6)176.5 (± 3.2)72.2 (± 1.1)44.7 (± 1.6)
2xx66.3 (± 1.1)12.9 (± 1.9)173.1 (± 0.5)79.3 (± 1.1)44.3 (± 1.2)
2xx67.3 (± 5.8)14.0 (± 0.8)172.2 (± 0.2)81.4 (± 1.3)42.0 (± 0.9)
2xx21.0 (± 9.4)12.2 (± 1.8)178.7 (± 3.5)59.0 (± 8.0)37.6 (± 5.2)
2xx54.2 (± 9.2)16.0 (± 5.9)173.2 (± 3.3)76.8 (± 2.4)34.7 (± 4.1)
2xx59.3 (± 1.3)17.0 (± 1.2)165.0 (± 0.7)85.2 (± 1.8)34.4 (± 0.6)
2xx57.8 (± 1.7)15.0 (± 1.4)170.1 (± 4.4)76.1 (± 7.1)34.2 (± 4.4)
2xx61.1 (± 1.8)13.1 (± 0.3)167.5 (± 2.7)87.5 (± 2.4)33.6 (± 0.9)
1x43.5 (± 21.3)10.8 (± 0.4)180.8 (± 5.7)77.2 (± 2.6)40.1 (± 2.1)
1x49.0 (± 11.5)13.2 (± 0.6)175.3 (± 6.2)79.1 (± 1.8)37.6 (± 1.4)
1x48.5 (± 13.0)16.3 (± 3.1)174.9 (± 3.0)83.7 (± 3.0)33.8 (± 3.0)
1x59.2 (± 1.0)15.3 (± 1.2)165.1 (± 2.9)82.9 (± 1.3)32.9 (± 1.9)
1x46.7 (± 6.4)18.7 (± 4.0)164.2 (± 4.7)68.0 (± 19.7)22.8 (± 2.3)
Parent44.1 (± 14.9)17.7 (± 3.7)166.1 (± 2.2)69.1 (± 12.8)32.6 (± 1.2)
  • a The parent (wild-type) data are given in boldface. Data are arranged in order of decreasing nystose concentration per number of substitutions. Values in parentheses are standard errors (n = 3). The five most improved variants are coded as V1, V3, V4, V5, and V8.