Biophysical Characterization of Type III Pantothenate Kinase (PanK) from Acinetobacter baumannii.

BACKGROUND: Type-III Pantothenate kinase from the multi drug resistant bacteria, Acinetobacter baumannii (AbPanK) catalyzes the first step of the essential Coenzyme A biosynthesis pathway. AbPanK is an attractive drug target against the bacteria since it is an essential enzyme and its structure is significantly different from the human PanK. METHODS: AbPanK was cloned, expressed, purified and crystallized. A good quality single crystal was used for X-ray intensity data collection. Dynamic light scattering was done for calculating the hydrodynamic radii and its oligomeric nature in the solution. Binding studies of this protein with its two substrates, Pantothenate and ATP were done using spectrofluorometer. RESULTS: Our results indicated that AbPanK shows a strong affinity with pantothenate with dissociation constant of 1.2 x 10- 8 M and moderate affinity towards ATP of 3.7x 10-3 M. This fact was further substantiated by the calculations of Km of both substrates using kinase assay kit. Dynamic light scattering studies have shown that it exists as homogenous solution with hydrodynamic radii corresponding to the molecular weight of 29.55 kDa. A low-resolution X-ray intensity data set was collected, which shows that AbPank crystallizes in P2 space group with cell dimensions of a= 165 Å, b= 260 Å, and, c= 197 Å and α= 90.0, ß= 113.60, γ= 90.0. DISCUSSION: Recombinant Pantothenate kinase from Acinetobacter baumannii was purified to homogeneity and crystallized. The enzyme exhibits very low sequence identity (28%) to other corresponding enzymes. CONCLUSION: The recombinant enzyme was active and its binding affinities with its substrates pantothenate and ATP have been studied. This information would be very useful while designing the inhibitors of this enzyme in order to fight bacterial infections associated to this pathogen.

Effect of dietary supplementation of Emblica officinalis fruit pomace on methane emission, ruminal fermentation, nutrient utilization, and milk production performance in buffaloes.

Effects of dietary supplementation of Emblica officinalis fruit (Indian gooseberry) pomace (EFP), a waste from fruit processing plants and rich in polyphenolic compounds, were investigated for ruminal fermentation, nutrient utilization, methane production, and milk production performance in buffaloes. An in vitro experiment was conducted using 0 to 50 g/kg of EFP (six treatments) to select an optimum dose for feeding of buffaloes. Organic matter (OM) degradability, total volatile fatty acid concentration, and acetate proportion decreased, but propionate proportion increased at the higher doses (> 30 g/kg). Methane production also decreased at the higher doses (≥ 20 g/kg). In the in vivo study, ten lactating buffaloes were randomly allotted into control and EFP groups (n = 5/group). The control group was fed a total mixed ration, whereas the EFP group was fed the control ration along with EFP at 20 g/kg of dry matter (DM) intake for 120 days. Feeding of EFP to buffaloes improved milk yield (P < 0.01) and milk production efficiency (P < 0.01). Concentration of milk protein tended (P = 0.071) to increase and that of solid not fat increased (P = 0.032) due to the EFP feeding. Yields (kg/day) of milk fat (P = 0.026), solid not fat (P = 0.011), and protein (P = 0.002) were greater in the EFP group than the control group. Somatic cell count in milk decreased (P = 0.032) due to EFP feeding. Digestibility of ether extract (P < 0.001) increased and OM (P = 0.051) tended to increase by EFP feeding. Methane production (g/d), yield (g/kg DM intake or g/kg digestible organic matter intake), and intensity (g/kg milk, g/kg milk fat, or g/kg milk protein), and methane conversion rate (percentage of gross energy intake) were lower (P < 0.01) in the EFP group than the control group. For milk fatty acid (FA) profiles, total saturated FA proportion tended to be greater (P = 0.057) in the EFP group than the control group, which was due to increased (P = 0.045) proportion of total short- and medium-chain FA (C4 to C14). Feed intake, digestibility of crude protein and fiber, and total n-6, n-3, mono-unsaturated FA, poly-unsaturated FA, and long-chain FA (C18 to C24) proportions were similar between the groups. This study suggests that feeding of EFP at 20 g/kg DM intake increases milk production and decreases methane production and intensity without impacting health of buffaloes and FA profiles of milk. This is a win-win situation for sustainable and cleaner buffalo production by improving milk production and decreasing environmental burdens of greenhouse gas emission and EFP residue disposal problems.

The first darter (Aves: Anhingidae) fossils from India (late Pliocene).

New fossils from the latest Pliocene portion of the Tatrot Formation exposed in the Siwalik Hills of northern India represent the first fossil record of a darter (Anhingidae) from India. The darter fossils possibly represent a new species, but the limited information on the fossil record of this group restricts their taxonomic allocation. The Pliocene darter has a deep pit on the distal face of metatarsal trochlea IV not reported in other anhingids, it has an open groove for the m. flexor perforatus et perforans digiti II tendon on the hypotarsus unlike New World anhingid taxa, and these darter specimens are the youngest of the handful of Neogene records of the group from Asia. These fossil specimens begin to fill in a significant geographic and temporal gap in the fossil record of this group that is largely known from other continents and other time periods. The presence of a darter and pelican (along with crabs, fish, turtles, and crocodilians) in the same fossil-bearing horizon strongly indicates the past presence of a substantial water body (large pond, lake, or river) in the interior of northern India in the foothills of the Himalayan Mountains.