• Mass Spectrometry Letters
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• 제3권2호
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• pp.47-49
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• 2012

Myoglobin was hydrolyzed by microwave-assisted weak acid hydrolysis with 2% formic acid at $37^{\circ}C$, $50^{\circ}C$, and $100^{\circ}C$ for 1 h. The most effective hydrolysis was observed at $100^{\circ}C$. Hydrolysis products were investigated using matrixassisted laser desorption/ionization time-of-flight mass spectrometry. Most cleavages predominantly occurred at the C-termini of aspartyl residues. For comparison, weak acid hydrolysis was also performed in boiling water for 20, 40, 60, and 120 min. A 60- min weak acid hydrolysis in boiling water yielded similar results as a 60-min microwave-assisted weak acid hydrolysis at $100^{\circ}C$. These results strongly suggest that microwave irradiation has no notable enhancement effect on acid hydrolysis of proteins and that temperature is the major factor that determines the effectiveness of weak acid hydrolysis.

• Bulletin of the Korean Chemical Society
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• 제34권1호
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• pp.27-28
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• 2013

• Mass Spectrometry Letters
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• 제9권2호
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• pp.46-50
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• 2018

The effects of temperature and acetonitrile (ACN) concentration on microwave-assisted weak-acid hydrolysis of proteins were investigated. Myoglobin was hydrolyzed for 1 h using 2% formic acid and a microwave with different concentrations of ACN (0, 5, and 10%) at various temperatures (50, 60, 70, 80, 90, and $100^{\circ}C$). The numbers of peptides identified with each concentration of ACN were the same for each temperature. The greatest number of peptides (18 total) was obtained with hydrolysis at $100^{\circ}C$, and 6 of these were a result of additional removal of aspartic acid at the C-terminus. Hydrolysis at $80^{\circ}C$ resulted in 13 peptides, of which only 1 was generated by the additional removal of aspartic acid, and 12 were observed with hydrolysis at $100^{\circ}C$. Our results demonstrate that microwave-assisted weak-acid hydrolysis of proteins can be performed successfully at $80^{\circ}C$, which could be beneficial for limiting side reactions and generating larger peptide sequences.