Immobilization of formate dehydrogenase from Candida boidinii through cross-linked enzyme aggregates

We employed a cross-linked enzyme aggregate (CLEA) method to immobilize formate dehydrogenase (FDH) from Candida boidinii. The optimal conditions for the preparation of CLEAs were determined by examining effects of various parameters: the nature and amount of cross-linking reagent, additive concentration, cross-linking time, and pH during CLEA preparation. The recovered activities of CLEAs were significantly dependent on the concentration of glutaraldehyde; however, the recovered activity was not severely influenced by the content of dextran polyaldehyde as a mild cross-linker. Bovine serum albumin (BSA) was also used as a proteic feeder and enhanced the activity recovery by 130%. The highest recovered activity of CLEA was 18% for formate oxidation reaction and 25% for CO₂ reduction reaction. The residual activity of CLEA prepared with dextran polyaldehyde (Dex-CLEA) was over 95% after 10 cycles of reuse. The thermal stability of Dex-CLEA was increased by a factor of 3.6 more than that of the free enzyme. CLEAs of FDH could be utilized efficiently for both NADH regeneration and CO₂ reduction.     

Extended standard curve stability on the CCD magic lite immunoassay system using a two-point adjustment

Ciba Corning Diagnostics has developed a two-point adjustment algorithm for use with the Magic Lite System which allows for extended stability of a full 7–10 point calibration curve over the life of a kit. This adjustment is accomplished by using a master calibration curve established during manufacturing along with two calibrators for each assay. Most conventional non-automated immunoassays contain anywhere from 6 to 10 calibrators which are included with each run of an assay. Eliminating the need to run full standard curves and using the two-point adjustment algorithm results in significant savings in both labour and reagent usage.     

Global stability of a predator-prey system

In this paper we derive a result to ensure the global stability of a predator-prey system. The method used is quite general and may have applications to other situations.Works were partially supported by the National Science Council of the Republic of China     

Protein turnover in a psychrotrophic bacterium

Protein breakdown in pulse-labelled and longlabelled cells of Arthrobacter S ₁-55, a psychrotrophic bacterium, has been assessed at different temperatures. The temperature at which the cells were grown and labelled affected the breakdown of pulsed-labelled but not long-labelled proteins. Inhibitors of ATP synthesis inhibited proteolysis. Miscoding antibiotics stimulated the production of rapidly degradable proteins.     

Long-term studies at a benthic station off the coast of Northumberland

Biannual sampling (March and September) has been carried out over the period 1972 to 1985 at a muddy sand station, 55 m depth, with fauna belonging to the deeper offshore edge of Petersen's Amphiura filiformis community.During the period 1974 to 1980 the community exhibited a high degree of persistence stability. This stability was lost between 1980 and 1983, with rising total numbers and biomass and changes in species ranking. There is some evidence of a downward reversal between 1984–85.Evidence suggests that the principal stabilising process is density dependent mortality mediated by competition in a food limited environment. The principal destabilising process appears to be periodic fluctuations in the organic flux to the bottom. A secondary destabilising process is clearly concerned with fluctuating winter temperature. In competitive terms, cold winters favour increased survival in the dominant species at the expense of the lesser ranked species. This process is, however, more ephemeral and subject to adjustment within the time scale of a year.     

Within-individual changes in developmental stability affect flight performance

Developmental stability, as measured by fluctuating asymmetry,has been purported to be an indicator of individual quality,and low asymmetry can be selected for by sexual selection processes.However, low asymmetry can also arise due to biomechanical constraintsoperating on trait design, as it is predicted that asymmetrywill decrease mechanical efficiency. Specifically, it has beenpredicted that wing length asymmetry will be negatively relatedto avian flight performance. To date, empirical investigationshave only studied the influence of increasing asymmetry beyondnaturally occurring average values. I examined the influenceof within-individual changes in primary feather developmentalstability on flight performance in European starlings by studyingasymmetry and flight before and after wing molt. Individualsthat exhibited a decrease in wing asymmetry through molt experiencedincreased aerodynamic performance in terms of both angle oftakeoff and level flapping-flight speed. Birds that increasedwing asymmetry suffered a decrease in flight performance. Takeoffspeed and the ability to negotiate an aerial obstacle coursewere unaffected by asymmetry. My data provide empirical supportfor the predicted influence of wing asymmetry on flight, eventhough the changes in asymmetry were very small (mean = 0.47%of trait size) and further indicate the importance of biomechanicalconsiderations in any study of developmental stability     

Albumin stabilizes 14,15-leukotriene A4

14,15-Leukotriene A4 is a pivotal biosynthetic intermediate in 15-lipoxygenase initiated leukotriene biosynthesis. This compound hydrolyzes instantaneously in phosphate buffer at pH 7.4. However, addition of human or bovine albumin to otherwise identical buffer solutions increases its stability. Intact 14,15-leukotriene A4 then decomposes by first-order kinetics with rate constants inversely proportional to the albumin concentration. Stabilization of 14,15-leukotriene A4 under certain conditions may influence its proportionate transformation by enzymatic vs non-enzymatic processes.     

Thermal unfolding studies show the disease causing F508del mutation in CFTR thermodynamically destabilizes nucleotide-binding domain 1

Misfolding and degradation of CFTR is the cause of disease in patients with the most prevalent CFTR mutation, an in-frame deletion of phenylalanine (F508del), located in the first nucleotide-binding domain of human CFTR (hNBD1). Studies of (F508del)CFTR cellular folding suggest that both intra- and inter-domain folding is impaired. (F508del)CFTR is a temperature-sensitive mutant, that is, lowering growth temperature, improves both export, and plasma membrane residence times. Yet, paradoxically, F508del does not alter the fold of isolated hNBD1 nor did it seem to perturb its unfolding transition in previous isothermal chemical denaturation studies. We therefore studied the in vitro thermal unfolding of matched hNBD1 constructs ±F508del to shed light on the defective folding mechanism and the basis for the thermal instability of (F508del)CFTR. Using primarily differential scanning calorimetry (DSC) and circular dichroism, we show for all hNBD1 pairs studied, that F508del lowers the unfolding transition temperature (T_m) by 6–7°C and that unfolding occurs via a kinetically-controlled, irreversible transition in isolated monomers. A thermal unfolding mechanism is derived from nonlinear least squares fitting of comprehensive DSC data sets. All data are consistent with a simple three-state thermal unfolding mechanism for hNBD1 ± F508del: N(±MgATP) ⇄ I_T(±MgATP) → A_T → (A_T)_n. The equilibrium unfolding to intermediate, I_T, is followed by the rate-determining, irreversible formation of a partially folded, aggregation-prone, monomeric state, A_T, for which aggregation to (A_T)_n and further unfolding occur with no detectable heat change. Fitted parameters indicate that F508del thermodynamically destabilizes the native state, N, and accelerates the formation of A_T.     

Molecular phylogeny,pigment composition,toxicology and life history of Pseudochattonella cf. verruculosa (Class Dictyochophyceae) from Wellington Harbour,New Zealand

Pseudochattonella verruculosa is a heterokont flagellate and has frequently been found associated with multi-species harmful algal blooms in Wellington Harbour. In this study the partial sequences of the nuclear encoded LSU rDNA and the large subunit of ribulose bisphosphate carboxylase (rbcL) of Pseudochattonella isolated from Wellington Harbour indicate that it is similar to P. verruculosa, while sequences of mitochondrial encoded COI, are similar to those of Pseudochattonella farcimen. As with P. farcimen, the Wellington Pseudochattonella lacked violaxanthin, lutein and anteroxanthin, three pigments detected only in P. verruculosa. The Wellington isolate also contains zeaxanthin which is absent in P. farcimen. Among all Pseudochattonella, cells of the Wellington isolate are the most variable in terms of both size and shape. Mucocysts of the Wellington Pseudochattonella also have the greatest degree of variation – from small, ‘bullet’-shape to large oval, oblong or ‘sausage’-like. In the sexual reproduction phase two gametes of the Wellington isolate fuse to form a zygote which gives rise to a large multi-nucleate cell. At times two or more of these large multi-nucleate cells fuse further to form a ‘massive’, plasmodium-like aggregate (up to 200 μm long). Positive feeding and toxicity tests on rotifers confirmed that the Wellington Pseudochattonella is cytotoxic and probably also contributed to the May 2010 fish kills. As molecular phylogenies do not conclusively support the separation of the Wellington Harbour Pseudochattonella from P. verruculosa or P. farcimen, it is tentatively named as Pseudochattonella cf. verruculosa.     

Aromatic-aromatic interactions: analysis of π-π interactions in interleukins and TNF proteins

Aromatic-aromatic hydrogen bonds are important in many areas of chemistry, biology and materials science. In this study we have analyzed the roles played by the π-π interactions in interleukins (ILs) and tumor necrosis factor (TNF) proteins. Majority of π-π interacting residues are conserved in ILs and TNF proteins. The accessible surface area calculations in these proteins reveal that these interactions might be important in stabilizing the inner core regions of these proteins. In addition to π-π interactions, the aromatic residues also form π-networks in ILs and TNF proteins. The results obtained in the present study indicate that π-π interactions and π-π networks play important roles in the structural stability of ILs and TNF proteins.