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The protein was hydrolyzed with 6N HCl under vacuum at 110°C for 24 h. Amino acid analysis was performed by pre-column derivatization with phenylisothiocynate. The phenylthiocarbomoylamino acids were analyzed using a Pico Tag column (3.9×150 mm)on a Waters HPLC system, equipped with a 1525 binary pump and Waters 2996-photodiode-array (PDA) detector set at 254 nm. Numbers on the peaks are retention times in minutes.
