Journal Article

Purification and cDNA Cloning of UDP-<span class="smallCaps">d</span>-Glucuronate Carboxy-lyase (UDP-<span class="smallCaps">d</span>-xylose Synthase) from Pea Seedlings

Masaru Kobayashi, Hironobu Nakagawa, Izumi Suda, Isao Miyagawa and Toru Matoh

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 11, pages 1259-1265
Published in print November 2002 | ISSN: 0032-0781
Published online November 2002 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcf157
Purification and cDNA Cloning of UDP-d-Glucuronate Carboxy-lyase  (UDP-d-xylose Synthase) from Pea Seedlings

More Like This

Show all results sharing these subjects:

  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

GO

Show Summary Details

Preview

Uridine diphospho-d-glucuronate carboxy-lyase (UDP-d-xylose synthase; EC 4.1.1.35), which catalyzes the conversion of UDP-d-glucuronate to UDP-d-xylose, was purified to apparent homogenity from pea (Pisum sativum L.) seedlings. The pH optimum for enzyme activity was around 5–6, and the activity was not affected by exogeneously supplied NAD+ and NADH. The purified enzyme had a molecular weight of 250 kDa and consisted of 42 kDa polypeptides. Based on the amino acid sequence, a probe (400 bp) was prepared with degenerate primers by a reverse transcriptase-PCR. Using this probe, a clone encoding 346 amino acid residues was screened from a pea cDNA library. The recombinant protein expressed in Escherichia coli catalyzed conversion of UDP-d-glucuronate to UDP-d-xylose, confirming that the isolated clone encoded UDP-d-glucuronate carboxy-lyase.

Keywords: Keywords: Pisum sativum — UDP-d-glucuronate carboxy-lyase (EC 4.1.1.35) — UDP-d-xylose.; Abbreviations: DTT, dithiothreitol; MBP, maltose-binding protein; TFA, trifluoroacetic acid; UXS, UDP-d-xylose synthase.

Journal Article.  4732 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.