Novel protecting group strategies in the synthesis of oligosaccharides

The thesis focuses on synthesis strategies in oligosaccharide campaigns, and the influence of protecting groups. New protecting group deprotection methods and protecting groups are developed. These novelties are applied in the synthesis of oligosaccharides which represent interesting biological targets. A solid phase approach is used to construct a library of oligorhamnans, employing a protecting group developed in this thesis. A fast deprotection method is employed to selectively remove fucntionalized benzyl ethers, which allows for the introduction of functional groups on a mannuronic acid molecule. Future synthesis campaigns are drafted combining the methods used in the previous chapters. Bio-organic Synthesi

How the oxygen tolerance of a [NiFe]-hydrogenase depends on quaternary structure

‘Oxygen-tolerant’ [NiFe]-hydrogenases can catalyze H(2) oxidation under aerobic conditions, avoiding oxygenation and destruction of the active site. In one mechanism accounting for this special property, membrane-bound [NiFe]-hydrogenases accommodate a pool of electrons that allows an O(2) molecule attacking the active site to be converted rapidly to harmless water. An important advantage may stem from having a dimeric or higher-order quaternary structure in which the electron-transfer relay chain of one partner is electronically coupled to that in the other. Hydrogenase-1 from E. coli has a dimeric structure in which the distal [4Fe-4S] clusters in each monomer are located approximately 12 Å apart, a distance conducive to fast electron tunneling. Such an arrangement can ensure that electrons from H(2) oxidation released at the active site of one partner are immediately transferred to its counterpart when an O(2) molecule attacks. This paper addresses the role of long-range, inter-domain electron transfer in the mechanism of O(2)-tolerance by comparing the properties of monomeric and dimeric forms of Hydrogenase-1. The results reveal a further interesting advantage that quaternary structure affords to proteins

Time courses of urinary creatinine excretion, measured creatinine clearance and estimated glomerular filtration rate over 30 days of ICU admission

Purpose: Baseline urinary creatinine excretion (UCE) is associated with ICU outcome, but its time course is not known. Materials and methods: We determined changes in UCE, plasma creatinine, measured creatinine clearance (mCC) and estimated glomerular filtration (eGFR) in patients with an ICU-stay 30d without acute kidney injury stage 3. The Cockcroft-Gault, MDRD (modification of diet in renal disease) and CKD-EPI (chronic kidney disease epidemiology collaboration) equations were used. Results: In 248 patients with 5143 UCEs hospital mortality was 24%. Over 30d, UCE absolutely decreased in male survivors and non-survivors and female survivors and nonsurvivors by 0.19, 0.16, 0.10 and 0.05 mmol/d/d (all P < 0.001). Relative decreases in UCE were similar in all four groups: 1.3, 1.4, 1.2 and 0.9%/d respectively. Over 30d, mCC remained unchanged, but eGFR rose by 31% (CKD-EPI) and 73% (MDRD) and creatinine clearance estimated by Cockcroft-Gault by 59% (all P < 0.001). Conclusions: Over 1 month of ICU stay, UCE declined by 1%/d which may correspond to an equivalent decline in muscle mass. These rates of UCE decrease were similar in survivors, non-survivors, males and females underscoring the intransigent nature of this process. In contrast to measured creatinine clearance, estimates of eGFR progressively rose during ICU stay. (c) 2020 Published by Elsevier Inc

Hydrogen activation by [NiFe]-hydrogenases

Hydrogenase-1 (Hyd-1) from Escherichia coli is a membrane-bound enzyme that catalyses the reversible oxidation of molecular H2 The active site contains one Fe and one Ni atom and several conserved amino acids including an arginine (Arg(509)), which interacts with two conserved aspartate residues (Asp(118) and Asp(574)) forming an outer shell canopy over the metals. There is also a highly conserved glutamate (Glu(28)) positioned on the opposite side of the active site to the canopy. The mechanism of hydrogen activation has been dissected by site-directed mutagenesis to identify the catalytic base responsible for splitting molecular hydrogen and possible proton transfer pathways to/from the active site. Previous reported attempts to mutate residues in the canopy were unsuccessful, leading to an assumption of a purely structural role. Recent discoveries, however, suggest a catalytic requirement, for example replacing the arginine with lysine (R509K) leaves the structure virtually unchanged, but catalytic activity falls by more than 100-fold. Variants containing amino acid substitutions at either or both, aspartates retain significant activity. We now propose a new mechanism: heterolytic H2 cleavage is via a mechanism akin to that of a frustrated Lewis pair (FLP), where H2 is polarized by simultaneous binding to the metal(s) (the acid) and a nitrogen from Arg(509) (the base)