Journal of Protein Chemistry

, Volume 15, Issue 3, pp 305–313

Analysis of the active center of branching enzyme II from maize endosperm

  • Takashi Kuriki
  • Hanping Guan
  • Mirta Sivak
  • Jack Preiss


Analysis of the primary structure of mBEII, with those of other branching and amylolytic enzymes as reference, identifies four highly conserved regions which may be involved in substrate binding and in catalysis. When one of the amino acid residues corresponding to the putative catalytic sites of mBEII, i.e., Asp-386, Glu-441, and Asp-509, was replaced, activity disappeared. These putative catalytic residues are located in three different regions (regions 2–4) of the four highly conserved regions (regions 1–4) which exist in the primary structure of most starch hydrolases and related enzymes, including branching enzymes. Region 3, which contains Glu-441 as one of the putative catalytic residues, was located downstream of the carboxyl-terminal position previously reported. The importance of the carboxyl amino acid residues was also demonstrated by chemical modification of the branching enzyme protein using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide.

Key words

Branching enzyme active center site-directed mutagenesis EDAC modification α-amylase family 


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Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • Takashi Kuriki
    • 1
  • Hanping Guan
    • 1
  • Mirta Sivak
    • 1
  • Jack Preiss
    • 1
  1. 1.Department of BiochemistryMichigan State UniversityEast Lansing
  2. 2.Biochemical Research LaboratoriesEzaki Glico Co., Ltd.OsakaJapan
  3. 3.ExSeed Genetics LLCIowa State UniversityAmes

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