Comparison of the hydrolysis of bovine κ-casein by camel and bovine chymosin: a kinetic and specificity study

Bovine chymosin constitutes a traditional ingredient for enzymatic milk coagulation in cheese making, providing a strong clotting capacity and low general proteolytic activity. Recently, these properties were surpassed by camel chymosin, but the mechanistic difference behind their action is not yet clear. We used capillary electrophoresis and reversed-phase liquid chromatography-mass spectrometry to compare the first site of hydrolysis of camel and bovine chymosin on bovine κ-casein (CN) and to determine the kinetic parameters of this reaction (pH 6.5; 32 °C). The enzymes showed identical specificities, cleaving the Phe105-Met106 bond of κ-CN to produce para-κ-CN and caseinomacropeptide. Initial formation rates of both products validated Michaelis-Menten modeling of the kinetic properties of both enzymes. Camel chymosin bound κ-CN with ～30% lower affinity (K(M)) and exhibited a 60% higher turnover rate (k(cat)), resulting in ～15% higher catalytic efficiency (k(cat)/K(M)) as compared to bovine chymosin. A local, less dense negatively charged cluster on the surface of camel chymosin may weaken electrostatic binding to the His-Pro cluster of κ-CN to simultaneously impart reduced substrate affinity and accelerated enzyme-substrate dissociation as compared to bovine chymosin.     

Comment on "Saturation of the southern ocean CO2 sink due to recent climate change"

Unlike Le Quéré et al. (Reports, 22 June 2007, p. 1735), we do not find a saturating Southern Ocean carbon sink due to recent climate change. In our ocean model, observed wind forcing causes reduced carbon uptake, but heat and freshwater flux forcing cause increased uptake. Our inversions of atmospheric carbon dioxide show that the Southern Ocean sink trend is dependent on network choice.     

Gastrointestinal protectants and cathartics.

The purpose of this article is to provide the reader with an overview of gastrointestinal cathartics and protectants and to point out possible applications for use in the horse with gastrointestinal disease. Most of the treatments described in this article have been used by the authors with apparent success; however, controlled studies with subsequent publication in the scientific literature with respect to these treatments in the horse are, for the most part, lacking. The authors view this emerging field of treatment as exciting and look forward to substantiating the efficacy of several of the treatments discussed.     

Percolation and leachate composition in a disturbed soil layer mulched with sewage biosolids

The effect of sludge and sludge compost as a mulch for dissipating raindrop impact and maintaining a relatively high percolation rate was studied using a Paleudalf from South Africa. Three kg soil were packed in perforated boxes in a making a two cm layer, and subjected to five consecutive simulated rainstorms during a period of 26 days. A 4-day incubation was allowed between the first 4 storms and 10 days before the 5th storm. The soil was amended with either a dry waste activated sludge or a sludge compost at a rate equivalent to 45 Mg ha⁻¹. The amendments (<2-mm) were applied either on the surface or mixed with the whole soil. The application of both sludge and sludge compost reduced the final percolation rate (FPR) compared with the unamended control. The effect of the consecutive rainstorms on the average FPR from all the treatments was as follows: FPR after 1st storm 2nd3rd4th>5th. The effect of amendment on average FPR from all storms was: control>sludge mixed with the soil>surface-applied sludge>sludge compost mixed with the soil≥ surface-applied sludge compost. The decrease of FPR was not related to the electrical conductivity or clay concentration of the leachates. The main chemical species that appeared in the leachates from the treated soils, and persisted throughout the five rainstorms, were ammonium and sulfate. Other ions, such as calcium and magnesium, were present in high concentrations in the leachate during the first storm but at low concentrations in subsequent storms. Nitrate concentrations were high during the first and fifth storms. It is suggested that the adverse effect of the amendments resulted from mechanical and microbial clogging of soil pores.     

Population crash in Lesotho's endemic Maloti minnow Pseudobarbus quathlambae following invasion by translocated smallmouth yellowfish Labeobarbus aeneus

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Substantial advancement in aquaria rearing methods to assist recovery of an Endangered soft coral

1. The temperate soft coral Dendronephthya australis has been listed as Endangered in NSW, Australia, owing to its restricted geographic range, rapidly declining populations, and a number of escalating threats. Research to assess practical methods for aquarium cultivation of D. australis for rehabilitation of wild populations is considered a high priority.
2. Previous attempts at restoration by transplanting D. australis fragments have had limited success, but identified key challenges, which included achieving attachment to substrates and identifying appropriate foods to maximize survival.
3. This study presents the results of experiments promoting the attachment of cuttings (nubbins) removed from donor colonies to moveable substrates (small concrete discs). Using a balanced orthogonal experimental design, three feeding regimes were tested: live artemia, live rotifers and dry powdered zooplankton.
4. Donor colony survival and growth were also monitored over a 6 month period in the field.
5. An overall nubbin survival rate of 100% was achieved under all three feeding regimes. Attachment rates were similar across feeding treatments, although nubbin behavioural responses (percentage of open polyps before feeding) suggested some preference for live rotifers.
6. The cutting removal process had no significant effect on donor colonies in the wild, indicating that this is a sustainable approach for restoration.
7. Overall, this study substantially advances the methodology for maintaining an Endangered azooxanthellate soft coral in aquaria and provides valuable input for management programmes aimed at implementing transplantation into the wild to aid species recovery.