Impact of D2O on the peptidization of L‑alanine

Abstract

This is our fifth consecutive study carried out in an order to collect experimental evidence on the impact of heavy water ( D2O) on the spontaneous peptidization of proteinogenic α-amino acids and this time its subject matter is L-alanine (L-Ala). Our four earlier studies have been focused on the two sulfur-containing α-amino acids (i.e., L-cysteine (L-Cys) and L-methionine (L-Met)), and on two structurally related α-amino acids (i.e., L-proline (L-Pro) and L-hydroxyproline (L-Hyp)). It seemed interesting to assess the effect exerted by D2O on L-Ala, the simplest chiral (endogenous and proteinogenic) α-amino acid with as low molar weight, as 89.09 g mol−1 only. As analytical techniques, we used high-performance liquid chromatography with the diode array detection (HPLC–DAD), mass spectrometry (MS), and scanning electron microscopy (SEM). The obtained results make it clear that the impact of heavy water on the dynamics of the spontaneous peptidization of L-Ala is even stronger than with the four other α-amino acids discussed earlier (although in all five cases, heavy water significantly hampers spontaneous oscillatory peptidization). Unlike in the four previous cases, though, the solubility of L-Ala in pure D2O is quite low and it takes twice as much time to dissolve it in D2O than in MeOH + X, 70:30 (v/v). Consequently, the peptidization of L-Ala in heavy water is even more obstructed than it was the case with the other investigated α-amino acids and it results in considerable yields of the L-Ala crystals (most probably at least partially deuterated) at the expense of the L-Ala-derived peptides. Perhaps it might be interesting to add that out of five α-amino acids investigated so far, which can be divided into two groups of endogenous and exogenous species, two endogenous species (L-Cys and L-Pro) undergo spontaneous oscillatory peptidization in an aqueousorganic solvent (i.e., in the absence of D2O) following the circadian rhythm, whereas two exogenous ones (i.e., L-Met and L-Hyp) do not. The third endogenous species (L-Ala) first undergoes two initials oscillations which are damped (not periodic) and the oscillatory changes are on a scale of ca. 10 h (as estimated with use of the Fourier transform approach) and after that, the system reaches a steady state.