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Effective Preparation Method of Phosphopeptides from Phosvitin and the Analysis of Peptide Profiles Using Tandem Mass Spectrometry
journal contribution
posted on 2019-12-10, 18:34 authored by Xi Huang, Sun Hee Moon, Jaehoon Lee, Hyundong Paik, Eun Joo Lee, Byungrok Min, Dong U. AhnThe effect of high-temperature and mild-pressure (HTMP)
pretreatment
on the enzymatic hydrolysis of phosvitin and the structural characteristics
of the phosphopeptides produced were analyzed using tandem mass spectrometry.
The HTMP pretreatment hydrolyzed phosvitin at random sites and helped
the subsequent enzyme hydrolysis of the peptides produced. With the
HTMP pretreatment alone, 154 peptides were produced, while the use
of trypsin, Protex 6L, and Multifect 14L in combination with the pretreatment
produced 252, 280, and 164 peptides, respectively. The use of two
enzyme combinations (trypsin + Protex 6L and trypsin + Multifect 14L)
helped the hydrolysis further. The number of phosphopeptides produced
increased when the modifications within the same amino acid sequences
were considered. This study indicated that HTMP pretreatment was a
breakthrough method to improve
the enzymatic hydrolysis of phosvitin that enabled an easy production
of phosvitin phosphopeptides for their subsequent functional characterizations.