A Complete Pathway Model for Lipid A Biosynthesis in Escherichia coli.

Lipid A is a highly conserved component of lipopolysaccharide (LPS), itself a major component of the outer membrane of Gram-negative bacteria. Lipid A is essential to cells and elicits a strong immune response from humans and other animals. We developed a quantitative model of the nine enzyme-catalyzed steps of Escherichia coli lipid A biosynthesis, drawing parameters from the experimental literature. This model accounts for biosynthesis regulation, which occurs through regulated degradation of the LpxC and WaaA (also called KdtA) enzymes. The LpxC degradation signal appears to arise from the lipid A disaccharide concentration, which we deduced from prior results, model results, and new LpxK overexpression results. The model agrees reasonably well with many experimental findings, including the lipid A production rate, the behaviors of mutants with defective LpxA enzymes, correlations between LpxC half-lives and cell generation times, and the effects of LpxK overexpression on LpxC concentrations. Its predictions also differ from some experimental results, which suggest modifications to the current understanding of the lipid A pathway, such as the possibility that LpxD can replace LpxA and that there may be metabolic channeling between LpxH and LpxB. The model shows that WaaA regulation may serve to regulate the lipid A production rate when the 3-deoxy-D-manno-oct-2-ulosonic acid (KDO) concentration is low and/or to control the number of KDO residues that get attached to lipid A. Computation of flux control coefficients showed that LpxC is the rate-limiting enzyme if pathway regulation is ignored, but that LpxK is the rate-limiting enzyme if pathway regulation is present, as it is in real cells. Control also shifts to other enzymes if the pathway substrate concentrations are not in excess. Based on these results, we suggest that LpxK may be a much better drug target than LpxC, which has been pursued most often

Insights into the biology of Escherichia coli through structural proteomics

10.1007/s10969-007-9019-2Journal of Structural and Functional Genomics82-345-55JSFG

Brood‐stock management and early hatchery rearing of Arctic charr (Salvelinus alpinus (Linnaeus))

Arctic charr (Salvelinus alpinus (Linnaeus)) is a stenothermic cold‐water fish, which has been cultured in Northern Europe and North America since the 1980s. The industry has remained relatively small with an annual production between 6000 and 10 000 tonnes, and is still challenged by an unreliable offspring production. This review focuses on offspring production in Arctic charr aquaculture including holding conditions for brood‐stock, fertilisation and egg rearing until hatch. Brood‐stock requires low temperatures during summer (<12°C) with the optimum still unknown. The temperature maximum for egg incubation lies between 6 and 8°C. The composition of an optimal brood‐stock diet is debated regarding fatty acids. A demand for a freshwater‐based diet rich in omega‐6 fatty acids is indicated, but results remain inconclusive. Extensive knowledge has been gained on the timing of spawning and its manipulation through photoperiod, temperature and hormone treatments; spawning can be induced by short‐day photoperiod; and temperature drops to 5°C. Eggs are fertilised dry in ovarian fluid. Egg quality is highly variable and positively related to egg size and energy density. Contrary, little information is available on sperm quality and its impact on egg survival. There may also be profound differences between Arctic charr of stationary or anadromous origin regarding requirements for holding conditions of brood‐stock and their diet. However, these differences have received little attention, and direct comparative studies are in demand