The design of enzymes with new functions and properties has long been a goal in protein engineering. Here, we report a strategy to change the catalytic activity of an existing protein scaffold. This was achieved by simultaneous incorporation and adjustment of functional elements through insertion, deletion, and substitution of several active site loops, followed by point mutations to fine-tune the activity. Using this approach, we were able to introduce beta-lactamase activity into the alphabeta/betaalpha metallohydrolase scaffold of glyoxalase II. The resulting enzyme, evMBL8 (evolved metallo beta-lactamase 8), completely lost its original activity and, instead, catalyzed the hydrolysis of cefotaxime with a (kcat/Km)app of 1.8 x 10(2) (mole/liter)(-1) second(-1), thus increasing resistance to Escherichia coli growth on cefotaxime by a factor of about 100. 相似文献
A 5‐year‐old dog presented with pollakiuria and urinary incontinence. Abdominal radiographs revealed an oval, multilayered structure with soft tissue opacity and gas lucency in the urinary bladder. Ultrasonography showed an oval luminal structure with hyper‐ and hypoechoic layers and internal reverberation artifacts. Following cystotomy, the hemisected plane showed a layered, pale whitish matrix with a gas layer around the core. Histopathological examination showed no evidence of cells, bacteria, or fungi. The core and laminae comprised fibrinous and mineral debris. A final diagnosis of a freely floating, gas‐filled laminated debris ball was made. 相似文献
1. To evaluate the performance of visible and near-infrared (Vis/NIR) spectroscopic models for discriminating true pale, soft and exudative (PSE), normal and dark, firm and dry (DFD) broiler breast meat in different conditions of preprocessing methods, spectral ranges, characteristic wavelength selection and water-holding capacity (WHC) indexes were assessed.
2. Quality attributes of 214 intact chicken fillets (pectoralis major), such as lightness (L*), pH and WHC indicators including drip loss (DL), water gain and expressible fluid were measured. Fillets were grouped into PSE, normal and DFD categories based on combination of L*, pH and WHC threshold criteria. Classification models were developed using support vector machine based methods on characteristic wavelengths selected from the unprocessed or 2nd-derivative spectra, respectively, in three spectral subsets of 400–2500, 400–1100 and 1100–2500 nm.
3. Better classification of three meat groups was obtained based on unprocessed spectra (72–94%) than 2nd-derivative spectra (55–72%). The classification based on 400–2500 nm (91% average) and 400–1100 nm (89% average) performed better than that on 1100–2500 nm (78% average). In terms of the three different WHC indicators, the combination of L*, pH and DL produced better results than the other two groups, with recognition accuracy of 94.4% using 400–2500-nm range.
4. These analytical results suggest that for a better classification of true PSE, normal and DFD broiler breast meat with Vis/NIR spectra, unprocessed spectra wavelengths should be used, ranges of 400–1000 nm should be included in the data collection, and DL as an indicator of WHC might provide a better prediction model. 相似文献
Although the influence of mobility on place attachment has received attention in the literature, this relationship varies between groups. Unlike those who move to enjoy their retirement, for example, older migrants arriving in Shenzhen come to the city ‘passively,’ drawn by the needs of their children. This paper advances understanding of the concept of place attachment by illustrating the relationships between its components. Analysis of interview and questionnaire responses revealed the following relationships between three focal components of place attachment. Place dependence first directly influenced affective attachment and then indirectly affected place identity. Leisure involvement had a positive impact on place dependence and affective attachment but no direct effect on place identity. Residential satisfaction was an antecedent of all three focal dimensions of place attachment. No direct relationship was found between leisure involvement and residential satisfaction. 相似文献
The mechanical behavior of cowpea starch gels (10%, w/v) at small and large deformations were investigated in comparison with acorn, corn, and potato starches in storage at 4°C for seven days. The rapid viscograms of starch paste (7%, w/v) revealed that cowpea starch had a larger setback (1,135 cP) than other starches (465–830 cP), although peak viscosity (1,723 cP) and pasting temperature (76°C) were between those of corn and potato starches. Texture profile analysis of cowpea starch gel showed exceptionally higher values for hardness, gumminess, chewiness and initial modulus than other starch gels. Cowpea starch gel also exhibited higher G′ and smaller tan δ compared with other starch gels, regardless of the storage time. A creep test revealed that the cowpea starch gel could remain highly resistant to stress, showing the least deformation among the tested starch gels during storage up to seven days. The overall results disclosed that cowpea starch was capable of forming exceptionally strong and elastic gels with good storage stability. 相似文献