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Preparation of extracts from mature spruce needles for enzymatic analyses

Preparation of extracts from mature spruce needles for enzymatic analyses It was possible to extract simultaneously several active enzymes involved in the carbohydrate or the amino acid metabolism from spruce needles [Picea abies (L.) Karst.] when a) a 100 mM Na‐Pi buffer of pH 7.5 containing 5% PVPP and 0.5% Triton X‐100 was used and when b) the resulting crude extracts were freed from lowmolecular‐weight compounds by gel‐chromatography using the separation medium Fractogel TSK HW‐40. Besides Triton X‐100, Triton X‐305, Myrij‐52 and Brij‐35 were tested, but 0.5% Triton X‐100 brought about the most active enzyme extracts. In crude extracts prepared from spruce needles during the early summer a high increase in absorbance at 334 nm was observed when the co‐substrate NADP+ was added, thus making reliable spectrophotometric assays impossible. The interfering low‐molecular‐weight substances could be eliminated by gel chromatography. As separation media Bio‐Gel P‐6 DG, Sephadex G‐25 m, Trisacryl GF 05 and Fractogel TSK HW‐40 (F) were tested, with Fractogel yielding the highest activities. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Physiologia Plantarum Wiley

Preparation of extracts from mature spruce needles for enzymatic analyses

Weimar, Maria; Rothe, Gunter M.
Physiologia Plantarum , Volume 69 (4) – Jan 1, 1987
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References (21)

Publisher
Wiley
Copyright
Copyright © 1987 Wiley Subscription Services
ISSN
0031-9317
eISSN
1399-3054
DOI
10.1111/j.1399-3054.1987.tb01986.x
Publisher site
See Article on Publisher Site

Abstract

It was possible to extract simultaneously several active enzymes involved in the carbohydrate or the amino acid metabolism from spruce needles [Picea abies (L.) Karst.] when a) a 100 mM Na‐Pi buffer of pH 7.5 containing 5% PVPP and 0.5% Triton X‐100 was used and when b) the resulting crude extracts were freed from lowmolecular‐weight compounds by gel‐chromatography using the separation medium Fractogel TSK HW‐40. Besides Triton X‐100, Triton X‐305, Myrij‐52 and Brij‐35 were tested, but 0.5% Triton X‐100 brought about the most active enzyme extracts. In crude extracts prepared from spruce needles during the early summer a high increase in absorbance at 334 nm was observed when the co‐substrate NADP+ was added, thus making reliable spectrophotometric assays impossible. The interfering low‐molecular‐weight substances could be eliminated by gel chromatography. As separation media Bio‐Gel P‐6 DG, Sephadex G‐25 m, Trisacryl GF 05 and Fractogel TSK HW‐40 (F) were tested, with Fractogel yielding the highest activities.

Journal

Physiologia Plantarum – Wiley

Published: Jan 1, 1987

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