Fructan chemical structure and sensitivity to an exohydrolase

Bancal, Pierre; Henson, Cynthia A.; Gaudillère, J. P.; Carpita, Nicholas C.

doi:10.1016/0008-6215(91)84124-w

Cited by 49 publications

(72 citation statements)

References 14 publications

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“…4a). Comparable with our 1-FEH (activity), a partially purified FEH from H. vulgare showed a reduced activity when incubated with (l&6)-kestotetraose (Bancal et al, 1991); hence, it is likely that 1-FEH from both monocotyledons and dicotyledons have a reduced affinity for the terminal y5-(2-l)-linkage at a branch-point.…”

Section: Purification Of 1-fehsupporting

confidence: 62%

Seasonal variation of fructan‐β‐fructosidase (FEH) activity and characterization of a β‐(2‐1)‐linkage specific FEH from tubers of Jerusalem artichoke (Helianthus tuberosus)

1997

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SUMMARYThe fructan-y?-fructosidase activity (1-FEH; EC 3.2.1.80) that degrades inulin in tubers oi Helianthus tuberosus L. appears to be developmentally regulated; it was low in growing tubers but increased during dormancy and sprouting. In spite of relatively high 1-FEH activity in vitro, fructose concentration was very low in developing and dormant tubers and increased markedly only during sprouting. A fructan-/S-fructosidase from such sprouting tubers was purified 41-fold to a single protein band on one-dimensional sodium dodecylsulphate-polyacrylamide gels. The purification procedure included ammonium sulphate precipitation, lectin-affinity chromatography on concanavalin A, anion-exchange and cation-exchange chromatography. The enzyme had an apparent molecular mass of 75000 measured by size-exclusion chromatography, and 79000 measured by one-dimensional sodium dodecylsulphate-polyacrylamide gel electrophoresis. It exhibited a high substrate specificity, hydrolysing terminal /?-(2-l)-fructosyl-fructose-linkages in linear and branched fructan oligomers; y^-(2-6)-linkages were hardly hydrolysed. Hydrolysis of inulin oligomers followed normal saturation kinetics: K^ values for 1,1-kestotetraose and 1,1,1-kestopentaose were 8-3 mM and 12 mM, respectively. Fructosyl residues were hydrolysed from inulin oligomers by a multi-chain mechanism. The fructan-y?-fructosidase showed optimal enzyme activity at pH 5-2, and it retained its full activity after pre-incubation for 1 h at up to 40 °C. The release of fructose from 5 mM 1,1-kestotetraose was reduced by 25 % when 1-FEH was assayed in the presence of 10 mM sucrose. It is proposed that the inhibition of 1-FEH activity by sucrose is a mechanism for controlling fructan degradation in planta.

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Section: Purification Of 1-fehsupporting

confidence: 62%

Seasonal variation of fructan‐β‐fructosidase (FEH) activity and characterization of a β‐(2‐1)‐linkage specific FEH from tubers of Jerusalem artichoke (Helianthus tuberosus)

1997

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“…1B). The identity of bifurcose in the extracts was confirmed by Dr. P. Bancal, using cochromatography with an authentic standard on a CI8 reversed-phase column as described previously (Bancal et al, 1991). After 24 h in the light (Fig.…”

Section: Fructan Synthesis In Vivomentioning

confidence: 95%

“…10). This enzymic reaction would explain a key step in the elongation-trimming pathway suggested for fructan synthesis in wheat (Bancal et al, 1991(Bancal et al, , 1992. Although our results are suggestive, further work is needed to conclusively demonstrate that 6-SST is a bifunctional and perhaps multifunctional enzyme in fructan biosynthesis of barley.…”

Section: Literature Cltedmentioning

confidence: 99%

Fructan Synthesis in Excised Barley Leaves (Identification of Two Sucrose-Sucrose Fructosyltransferases Induced by Light and Their Separation from Constitutive Invertases)

Simmen¹,

Obenland²,

Boller³

et al. 1993

Plant Physiol.

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“…Several FEH activities with differing preferences for the /J-{2-6) or /5'-(2-l) linkages were partly purified from L. rigidum by Bonnett & Simpson (1995). Purified 1-FEH from H. tuberosus (Marx et al, 1997) and partly purified FEH from H. vulgare (Bancal et al, 1991) hydrolyse /?-(2-l)-fructose linkages at branch points in (l&6)-kestotetraose at a markedly slower rate when compared to that hydrolyzing linear inulin oligomers. It has been suggested that steric hindrance caused by the (2-6)-linked fructose at the branch point reduced the enzymatic efficiency of hydrolysis (Bancal et al, 1991).…”

Section: Substrates Of 6-fehmentioning

confidence: 99%

“…Purified 1-FEH from H. tuberosus (Marx et al, 1997) and partly purified FEH from H. vulgare (Bancal et al, 1991) hydrolyse /?-(2-l)-fructose linkages at branch points in (l&6)-kestotetraose at a markedly slower rate when compared to that hydrolyzing linear inulin oligomers. It has been suggested that steric hindrance caused by the (2-6)-linked fructose at the branch point reduced the enzymatic efficiency of hydrolysis (Bancal et al, 1991). In our experiments, however, the 6-FEH from L. perenne hydrolysed linear 6,6-kestotetraose and branched (l&6)-kestotetraose at nearly the same rates (Table 2).…”

Section: Substrates Of 6-fehmentioning

confidence: 99%

Hydrolysis of fructan in grasses: a β‐(2‐6)‐linkage specific fructan‐β‐fructosidase from stubble of Lolium perenne

1997

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SUMMARYSeveral different fructan and sucrose hydrolysing enzyme activities were induced in roots and stubble (mainly leaf sheaths) of Lolium perenne L. plants after defoliation. Among those activities, a fructan-/?-fructosidase (EC 3 .2.1.80) that hydrolyses predominantly /?-(2-6)-fructosyl-fructose linkages (6-FEH) was purified from the stubble. The use of the substrate 6,6-kestotetraose and high-performance anion-exchange chromatography with pulsed amperometric detection allowed linkage-specific screening and sensitive analysis of enzyme activity. A 6-FEH was extensively purified to yield one protein band as revealed by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The 6-FEH was separated from the contaminating y9.(2-l)-linkage-specific fructan-/9-fructosidase and invertase activities (EC 3.1.2.26) by ammonium sulphate precipitation, lectin-affinity, anion-exchange and size-exclusion chromatography. The purified 6-FEH was a glycoprotein with an apparent molecular mass of 65000, as determined by size-exclusion chromatography, and of 69000 by SDS-PAGE. The 6-FEH had an activity optimum in the range of pH 5-1 to 5-6. Temperatures above 30 °C affected the stability of the enzyme activity; however, its temperature stability was increased in the presence of 6,6-kestotetraose. The purified 6-FEH activity hydrolysed the/ff-(2-6)-linkages in 6,6-kestotetraose and (1&6)-kestotetraose at rates five times faster than the /?-(2-l)-linkages in 1,1-kestotetraose and (l&6)-kestotetraose. Fructose up to 50 mM did not affect 6-FEH activity; conversely, sucrose substantially inhibited the enzyme activity. Other disaccharides did not affect 6-FEH. It is suggested that sucrose might modulate 6-FEH activity in vivo.

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Fructan chemical structure and sensitivity to an exohydrolase

Cited by 49 publications

References 14 publications

Seasonal variation of fructan‐β‐fructosidase (FEH) activity and characterization of a β‐(2‐1)‐linkage specific FEH from tubers of Jerusalem artichoke (Helianthus tuberosus)

Seasonal variation of fructan‐β‐fructosidase (FEH) activity and characterization of a β‐(2‐1)‐linkage specific FEH from tubers of Jerusalem artichoke (Helianthus tuberosus)

Fructan Synthesis in Excised Barley Leaves (Identification of Two Sucrose-Sucrose Fructosyltransferases Induced by Light and Their Separation from Constitutive Invertases)

Hydrolysis of fructan in grasses: a β‐(2‐6)‐linkage specific fructan‐β‐fructosidase from stubble of Lolium perenne

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