Search

A new and unnatural type of phospholipids with the head group attached to the 2-position of the glycerol backbone has been synthesized and shown to be a good substrate for secretory phospholipase A2 (sPLA2). To investigate the unexpected sPLA2 activity, we have compared three different phospholipids by using fluorescence techniques and HPLC, namely: (R)-1,2-dipalmitoyl-glycero-3-phosphocholine (hereafter referred to as 1R), (R)-1-O-hexadecyl-2-palmitoyl-glycero-3-phoshocholine (2R), and (S)-1-O-hexadecyl-3-palmitoyl-glycero-2-phosphocholine (3S). Furthermore, to understand the underlying mechanisms for the observed differences, we have performed molecular dynamics simulations to clarify on a structural level the substrate specificity of sPLA2 toward phospholipid analogues with their head groups in the 2-position of the glycerol backbone. We have studied the lipids above 1R, 2R, and 3S as well as their enantiomers 1S, 2S, and 3R. In the simulations of sPLA2-1S and sPLA2-3R, structural distortion in the binding cleft induced by the phospholipids showed that these are not substrates for sPLA2. In the case of the phospholipids 1R, 2R, and 3S, our simulations revealed that the difference observed experimentally in sPLA2 activity might be caused by reduced access of water molecules to the active site. We have monitored the number of water molecules that enter the active site region for the different sPLA2-phospholipid complexes and found that the probability of a water molecule reaching the correct position such that hydrolysis can occur is reduced for the unnatural lipids. The relative water count follows 1R2R3S. This is in good agreement with experimental data that indicate the same trend for sPLA2 activity: 1R2R3S.

From an environmental point of view, hydrogen peroxide (HP) has beneficial attributes compared with other disinfectants in terms of its ready degradation and neutral by-products. The rapid degradation of HP can, however, cause difficulties with regard to safe and efficient water treatment when applied in different systems. In this study, we investigated the degradation kinetics of HP in biofilters from water recirculating aquaculture systems (RAS). The potential effect of HP on the nitrification process in the biofilters was also examined. Biofilter elements from two different pilot-scale RAS were exposed to various HP treatments in batch experiments, and the HP concentration was found to follow an exponential decay. The biofilter ammonia and nitrite oxidation processes showed quick recuperation after exposure to a single dose of HP up to 30 mg L−1. An average HP concentration of 10–13 mg L−1 maintained over 3 h had a moderate inhibitory effect on the biofilter elements from one of the RAS with relatively high organic loading, while the nitrification was severely inhibited in the pilot-scale biofilters from the other RAS with a relatively low organic loading. A pilot-scale RAS, equipped with two biofilter units, both a moving-bed (Biomedia) and a fixed-bed (BIO-BLOK®) biofilter, was subjected to an average HP concentration of ∼12 mg L−1 for 3 h. The ammonium- and nitrite-degrading efficiencies of both the Biomedia and the BIO-BLOK® filters were drastically reduced. The filters had not reverted to pre-HP exposure efficiency after 24 h, suggesting a possible long-term impact on the biofilters.