Efficient activity screening of new glucuronoyl esterases using a pNP-based assay.

Glucuronoyl esterases (CE15, EC 3.1.1.117) catalyze the hydrolysis of ester bonds between lignin and carbohydrates in lignocellulose. They are widespread within fungi and bacteria, and are subjects to research interest due to their potential applicability in lignocellulose processing. Identifying new and relevant glucuronoyl esterase candidates is challenging because available model substrates poorly represent the natural substrate, which leads to inefficient screening for the activity. In this study, we demonstrate how fifteen novel, fungal, putative glucuronoyl esterases from family CE15 were expressed and screened for activity towards a commercially available, colorimetric assay based on the methyl-ester of 4-O-methyl-aldotriuronic acid linked to para-nitrophenol (methyl ester-UX-ß-pNP) and coupled with the activity of GH67 (α-glucuronidase) and GH43 (ß-xylosidase) activity. The assay provides easy means for accurately establishing activity and determining specific activity of glucuronoyl esterases. Out of the fifteen expressed CE15 proteins, seven are active and were purified to determine their specific activity. The seven active enzymes originate from Auricularia subglabra (3 proteins), Ganoderma sinensis (2 proteins) and Neocallimastix californiae (2 proteins). Among the CE15 proteins not active towards the screening substrate (methyl ester-UX-ß-pNP) were proteins originating from Schizophyllum commune, Podospora anserina, Trametes versicolor, and Coprinopsis cinerea. It is unexpected that CE15 proteins from such canonical lignocellulose degraders do not have the anticipated activity, and these observations call for deeper investigations.

The effect of total hip arthroplasty surgical approach on gait.

This study examined the effect of the surgical approach used in total hip arthroplasty (THA) on gait mechanics six months following surgery. Quantitative gait analysis was performed on 29 subjects: 10 anterolateral (A-L) and 10 posterolateral (P-L) THA patients and nine able-bodied, velocity-matched subjects. Discriminant function analysis was used to determine the distinction of the groups with respect to sagittal plane hip range of motion, index of symmetry, trunk inclination, pelvic drop, hip abduction, and foot progression angles. The A-L group had the largest trunk inclination (3.0+/-2.4 degrees) and the smallest hip range of motion (34.0+/-7.4 degrees). Both THA groups demonstrated greater asymmetry as expressed by the smaller symmetry index (0.97+/-0.04 for A-L and 0.98+/-0.05 for the P-L) than the able-bodied group (0.99+/-0.01). The classification procedure correctly classified 89% of the control group cases, 90% of the A-L cases, and 50% of the P-L cases. These results support the conclusion that six months following surgery, the gait of the majority (85%) of THA patients has not returned to normal. The A-L patients displayed distinct gait patterns, while a small percentage (30%) of the P-L patients demonstrated normal gait. While these differences are statistically significant, the clinical significance is unknown and linked to the duration that they persist.