Báo cáo khoa học: Directed evolution of formate dehydrogenase from Candida boidinii for improved stability during entrapment in polyacrylamide ppt

... Directed evolution of formate dehydrogenase from Candida boidinii for improved stability during entrapment in polyacrylamide Marion B. Ansorge-Schumacher 1 , ... instead of the gel itself. As an investigation system, formate dehydrogenase [(FDH) E.C.1.2.1.2] from Candida boidinii was chosen for its outstanding importance as a cofactor regener- ation...

Ngày tải lên: 16/03/2014, 13:20

... breadth of information available on DHFR and the significance of it in the treatment of a range of conditions makes it an ideal subject for examining the ability of directed evolution to aid us in understanding ... but lacking in TMP (as shown in Fig. 1). This allows for the formation of an additional hydrogen bond with DHF, stabilizing its binding, while destabiliz...

Ngày tải lên: 19/02/2014, 02:20

... half of the initial activity is lost in 5 min. The activity half-life t 1 ⁄ 2, 48 °C of the proteins was determined at 48 °C by incu- bating the proteins for 0–1800 min at 48 °C, followed by incubation ... enhance the thermostability of proteins. This approach involves the generation of genetic diversity in the gene encoding the protein of interest, followed by screening for...

Ngày tải lên: 23/03/2014, 06:20

... mutant protein, was incubated in 500 lL reaction mixture for 30 min, after which detection was performed as described for glutaryl-7-ACA. Kinetic parameters were obtained from Eadie–Hofstee plots, ... mature protein. Removal of the spacer peptide of 10 amino acids leaves a catalytically active enzyme consisting of an a-subunit of 161 amino acids weighing 17.7 kDa and a b-subun...

Ngày tải lên: 31/03/2014, 09:20

... is blocked. A conformational transition upon binding to BchH may bring the integrin I domain of BchD into contact with the integrin-binding motif of BchH, simultaneously triggering porphyrin metallation. ... shown binding of Mg 2+ to this subunit [21]. An integrin I domain, suggested to exist in the C-terminus of the 70 kDa subunit, is a metal binding site (MIDAS motif) that can be...

Ngày tải lên: 19/02/2014, 12:20

... molybdopterin guanine dinucletoide cofactors of Fdh H of E. coli (MGD 801 and MGD 802 )that were found to be invariant among molybdopterin-containing FDH [34] are shown in red. Amino acids coordinating MGD801 ... analogue of formate and therefore a general inhibitor of FDHs [39]. A 50% inhibitory effect was observed in the presence of 0.9 m M sodium azide in standard assay c...

Ngày tải lên: 20/02/2014, 23:20

... (25 lL) was injected and eluted with a linear gradient of sodium nitrate (0–10 min, increasing from 0% to 4%; 10–12 min, 4%; 12–32 min, increasing from 4% to 8%; 32–48 min, increasing from 8% to ... of microgram quantities of protein util- izing the principle of protein-dye binding. Anal Biochem 72, 248–254. 27 Habeeb AFSA (1966) Determination of free amino acid groups in pr...

Ngày tải lên: 07/03/2014, 10:20

... the silver-staining method. Activity-staining for DNase I was performed using a DNA casting-PAGE method [25], and conventional methods were used for the assay of b-galactosidase [26]. Purification of shark ... humans during the course of evolution, it could be postulated that the DN ase I protein has acquired incre asing structural stability through the introduction of the two...

Ngày tải lên: 19/02/2014, 16:20

... setting frainework of Chomsky (1981) claims that learning involves fix- ing the wdues of a finite set of finite-valued param- eters to select a single fully-specified grammar from within the ... by one-point crossover of (and, optionally, single point mutation of) their ini- tial p-settings, ensuring neo-Darwinian rather than Lamarckian inheritance. The encoding of p-set...

Ngày tải lên: 31/03/2014, 21:20

... P i containing 0.05% gelatin (DPBSG). Bacteria [10 lL containing 10 6 colony-forming units (CFU)] were inoculated into 190 lL reaction wells containing the diluted normal human serum, 25% heat- inactivated ... dnastar software (DNASTAR Inc., Madison, WI, USA). Cloning of lpxX and construction of the knockout mutant O35ElpxX A DNA sequence containing lpxX was amplified from the chromosoma...

Ngày tải lên: 18/02/2014, 14:20