Bulky high-mannose-type N-glycan blocks the taste-modifying activity of miraculin.

• ITO K.

抄録

BACKGROUND: Miraculin (MCL) is a taste-modifying protein that converts sourness into sweetness. The molecular mechanism underlying the taste-modifying action of MCL is unknown. METHODS: Here, a yeast expression system for MCL was constructed to accelerate analysis of its structure-function relationships. The Saccharomyces cerevisiae expression system has advantages as a high-throughput analysis system, but compared to other hosts it is characterized by a relatively low level of recombinant protein expression. To alleviate this weakness, in this study we optimized the codon usage and signal-sequence as the first step. Recombinant MCL (rMCL) was expressed and purified, and the sensory taste was analyzed. RESULTS: As a result, a 2 mg/l yield of rMCL was successfully obtained. Although sensory taste evaluation showed that rMCL was flat in taste under all the pH conditions employed, taste-modifying activity similar to that of native MCL was recovered after deglycosylation. Mutagenetic analysis revealed that the N-glycan attached to Asn42 was bulky in rMCL. CONCLUSIONS: The high-mannose-type N-glycan attached in yeast blocks the taste-modifying activity of rMCL. GENERAL SIGNIFICANCE: The bulky N-glycan attached to Asn42 may cause steric hindrance in the interaction between active residues and the sweet taste receptor hT1R2/hT1R3.

収録刊行物

• Biochimica et biophysica acta

  Biochimica et biophysica acta 1800 (9), 986-992, 2010-09

  Elsevier B.V.

キーワード

• Miraculin
• Sweet protein
• Taste
• Yeast expression system
• N-glycan
• Steric hindrance

詳細情報

• CRID: 1573668926265492096
• NII論文ID: 80021210617
• NII書誌ID: AA00564635
• ISSN: 00063002
• Web Site: http://hdl.handle.net/2433/128855
• 本文言語コード: en
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