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

Marx, Stefan; Nösberger, J.; Frehner, Monika

doi:10.1046/j.1469-8137.1997.00642.x

Cited by 51 publications

(47 citation statements)

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“…Both 1-FEHs had optimal activities between 30°C and 40°C, but activities remained surprisingly high at the lower temperature range (not shown). Similar properties were observed for a 1-FEH from barley (Henson, 1989) and a 6-FEH from ryegrass (Marx et al, 1997b).…”

Section: Enzymatic Propertiessupporting

confidence: 65%

“…Also, 6-FEH and invertase activities were present in crude extracts, but they eluted in different fractions. As observed for most other plant FEH enzymes Marx et al, 1997aMarx et al, , 1997bDe Roover et al, 1999), the final 1-FEH w1 and w2 preparations showed negligible invertase activity (Table II). In severe contrast to plant FEHs, which are only capable of degrading fructans and not Suc, ␤-fructo(furano)sidases (EC 3.2.1.80) can degrade Suc as well as fructans.…”

Section: Properties Of Plant Fehsmentioning

confidence: 72%

“…From monocots, only partially purified preparations of 1-FEH were obtained from wheat (Jeong, 1991), barley (Henson, 1989), and Lolium rigidum . A fructan 6-exohydrolase (6-FEH) from ryegrass (Marx et al, 1997b) and an FEH that preferentially hydrolyzes ␤-(236) (oat; Henson and Livingston, 1996) or multiple fructofuranosidic linkages (barley; Henson and Livingston, 1998) were purified more recently.…”

mentioning

confidence: 99%

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Fructan 1-Exohydrolases. β-(2,1)-Trimmers during Graminan Biosynthesis in Stems of Wheat? Purification, Characterization, Mass Mapping, and Cloning of Two Fructan 1-Exohydrolase Isoforms,

et al. 2003

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Graminan-type fructans are temporarily stored in wheat (Triticum aestivum) stems. Two phases can be distinguished: a phase of fructan biosynthesis (green stems) followed by a breakdown phase (stems turning yellow). So far, no plant fructan exohydrolase enzymes have been cloned from a monocotyledonous species. Here, we report on the cloning, purification, and characterization of two fructan 1-exohydrolase cDNAs (1-FEH w1 and w2) from winter wheat stems. Similar to dicot plant 1-FEHs, they are derived from a special group within the cell wall-type invertases characterized by their low isoelectric points. The corresponding isoenzymes were purified to electrophoretic homogeneity, and their mass spectra were determined by quadrupole-time-of-flight mass spectrometry. Characterization of the purified enzymes revealed that inulin-type fructans [␤-(2,1)] are much better substrates than levan-type fructans [␤-(2,6)]. Although both enzymes are highly identical (98% identity), they showed different substrate specificity toward branched wheat stem fructans. Although 1-FEH activities were found to be considerably higher during the fructan breakdown phase, it was possible to purify substantial amounts of 1-FEH w2 from young, fructan biosynthesizing wheat stems, suggesting that this isoenzyme might play a role as a ␤-(2,1)-trimmer throughout the period of active graminan biosynthesis. In this way, the species and developmental stage-specific complex fructan patterns found in monocots might be determined by the relative proportions and specificities of both fructan biosynthetic and breakdown enzymes.Starch is the most prominent storage carbohydrate in plants, but about 15% of flowering plant species use fructan (a Fru polymer) as a storage compound (Hendry, 1993). Inulin-type fructan consists of linear ␤-(2,1)-linked fructofuranosyl units and occur mainly in dicotyledonous species. Levan consists of linear ␤-(2,6)-linked fructofuranosyl units, but more complex and branched fructan types (graminan, inulin neoseries, and levan neoseries) are common in monocotyledonous species (Vijn and Smeekens, 1999;Pavis et al., 2001b) Next to their obvious role as reserve compounds, fructan might have other functions in plants like stress protectants (drought and cold) or osmoregulators (Vergauwen et al., 2000; Hincha et al., 2002, and refs. therein). Unlike starch, fructans are water soluble and are believed to be stored in the vacuole , although the exclusive vacuolar localization has been questioned .Although the metabolism of inulin has become clear in dicotyledonous species and the respective biosynthetic and breakdown enzymes have been cloned (Edelman and Jefford, 1968;Van den Ende and Van Laere, 1996a; van der Meer et al., 1998;Hellwege et al., 2000;Van den Ende et al., 2000, fructan metabolism in monocots is not yet completely unraveled. So far, four different fructosyltransferases, each with their own specificity, are believed to be involved in monocot fructan biosynthesis. In addition to inulin biosynthesis by Suc:Suc 1-fructosyl tr...

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Section: Enzymatic Propertiessupporting

confidence: 65%

Section: Properties Of Plant Fehsmentioning

confidence: 72%

mentioning

confidence: 99%

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Fructan 1-Exohydrolases. β-(2,1)-Trimmers during Graminan Biosynthesis in Stems of Wheat? Purification, Characterization, Mass Mapping, and Cloning of Two Fructan 1-Exohydrolase Isoforms,

et al. 2003

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“…In fungi, inulin can be degraded endolytically by inulinases (Uchiyama, 1993) as well; however, in plants, only (exo)-fructan-/5-fructosidases have been identified up to now (Edelman & Jefford, 1964;Rutherford & Deacon, 1972;Yamamoto & Mino, 1985;Wagner & Wiemken, 1986;Henson, 1989;Claessens et al, 1990;Simpson et al, 1991;Bonnett & Simpson, 1995;Henson & Livingston, 1996;Marx et al, 1997). Generally, two different mechanisms of exolytic degradation of sugar oligomers and polymers are kinetically distinguishable.…”

Section: Properties Of 1-fehmentioning

confidence: 99%

“…In the present and the companion paper (Marx, Nosberger & Frehner, 1997), we focus on the processes of fructan degradation in plants. We have identified FEH activities from different plant species and have determined their physiological relevance to fructan degradation in planta.…”

Section: Introductionmentioning

confidence: 99%

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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Carbohydrate Metabolism

Fernie

Willmitzer

2004

Handbook of Plant Biotechnology

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The yield from crop plants has been subject to improvement through conventional breeding and modifications in agricultural practice for many decades. More recently, the use of molecular genetic approaches has allowed more directed approaches towards crop improvement. Specific examples of both successful strategies and of fundamental knowledge gleaned from unsuccessful strategies are presented including manipulations of starch, sucrose and novel sugar content. The utility of metabolic control analysis in the identification of control points in metabolic pathway and its use in the prediction of changes in metabolite accumulation following enzyme modulation is discussed. Further examples of biotechnological manipulation focus on qualitative rather than quantitative aspects of carbohydrate metabolism detailing the introduction of novel pathways for synthesis, or functionalities, of polymers. Particular focus is given to fructan biosynthesis in non‐fructan accumulating species and the broad structural diversity of starch following genetic manipulation of the enzymes involved in its synthesis and degradation. Finally, the chapter concludes with an appraisal of the challenges that remain before the rational manipulation of carbohydrate becomes routine.

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

Cited by 51 publications

References 50 publications

Fructan 1-Exohydrolases. β-(2,1)-Trimmers during Graminan Biosynthesis in Stems of Wheat? Purification, Characterization, Mass Mapping, and Cloning of Two Fructan 1-Exohydrolase Isoforms,

Fructan 1-Exohydrolases. β-(2,1)-Trimmers during Graminan Biosynthesis in Stems of Wheat? Purification, Characterization, Mass Mapping, and Cloning of Two Fructan 1-Exohydrolase Isoforms,

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)

Carbohydrate Metabolism

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