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-Schumacher1, ... 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 system in ... effects of the addi-tional mutations E53V and K56R which wereintroduced during the ﬁrst step of evolution. Stability of mutant FDHs in PAAFinally, the performance of the FDH variants afterentrapment...

Tài liệu Báo cáo khoa học: Directed evolution of trimethoprim resistance inEscherichia coli doc

... breadth of information availableon 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 ... butlacking in TMP (as shown in Fig. 1). This allows for the formation of an additional hydrogen bondwith DHF, stabilizing its binding, while destabilizingthe binding of TMP. The net result of this ... present in the directed evolution process.This allows for a greater understanding of enzymemechanisms than studies of clinical isolates. Directed evolution is gaining popularity as a means of studyingenzymes...

Báo cáo khoa học: Directed evolution of a histone acetyltransferase – enhancing thermostability, whilst maintaining catalytic activity and substrate speciﬁcity doc

... half of the initial activity is lost in 5 min. The activity half-lifet1 ⁄ 2, 48 °C of the proteins was determined at 48 °C by incu-bating the proteins for 0–1800 min at 48 °C, followed byincubation ... 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 mutant proteinswith the ... moreopportunities for salt bridge formation on the surface of the protein.Implications for protein thermostabilityengineeringGenerally, only a few mutations are sufﬁcient toincrease the thermostability of...

Báo cáo khoa học: Directed evolution of a glutaryl acylase into an adipyl acylase potx

... mutant protein, was incubated in 500 lL reaction mixture for 30 min, after which detectionwas performed as described for glutaryl-7-ACA. Kineticparameters were obtained from Eadie–Hofstee plots, ... mature protein.Removal of the spacer peptide of 10 amino acids leaves acatalytically active enzyme consisting of an a-subunit of 161amino acids weighing 17.7 kDa and a b-subunit of 522amino acids ... is included in the top 10 of most sold therapeutics,surpassing the penicillin class of b-lactams [1]. Semi-synthetic cephalosporins are industrially produced from the b-lactam nuclei 7-aminocephalosporanic...

Tài liệu Báo cáo khoa học: ATPase activity of magnesium chelatase subunit I is required to maintain subunit D in vivo ppt

... isblocked. A conformational transition upon binding toBchH may bring the integrin I domain of BchD into contactwith the integrin-binding motif of BchH, simultaneouslytriggering porphyrin metallation. ... shownbinding of Mg2+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 expectedto bind ... arelease of the blockade of the ATP-binding site of BchI bythe integrin I domain, triggering ATP hydrolysis [10]. It isstill an open question which subunit provides the Mg2+tobe inserted into...

Tài liệu Báo cáo khoa học: The tungsten-containing formate dehydrogenase from Methylobacterium extorquens AM1: Puriﬁcation and properties docx

... molybdopterin guanine dinucletoide cofactors of FdhH of E. coli (MGD801and MGD802)thatwere found to be invariant among molybdopterin-containing FDH [34] are shown in red. Amino acids coordinating MGD801 ... analogue of formate and therefore ageneral inhibitor of FDHs [39]. A 50% inhibitory effect wasobserved in the presence of 0.9 mMsodium azide in standard assay conditions. A stabilizing effect of ... participate in the coordination of the centralMo/W atom via two pterin cofactors are also identiﬁed in the sequence of FDH1.The primary sequence of the b-subunit of FDH1 from M. extorquens AM1 indicates...

Báo cáo khoa học: Molecular imprinting of cyclodextrin glycosyltransferases from Paenibacillus sp. A11 and Bacillus macerans with c-cyclodextrin pptx

... (25 lL) was injected and eluted with alinear 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 Biochem72, 248–254.27 Habeeb AFSA (1966) Determination of free amino acidgroups in proteins by trinitrobenzene ... imprinting of the enzymeswith CD8, resulting in high levels of the desired prod-uct being formed, is reported.Results and DiscussionTo provide enough attachment points for crosslinkingof...

Tài liệu Báo cáo khoa học: Molecular evolution of shark and other vertebrate DNases I pptx

... thesilver-staining method. Activity-staining for DNase I wasperformed using a DNA casting-PAGE method [25],and conventional methods were used for the assay of b-galactosidase [26].Purification of shark ... humans during thecourse of evolution, it could be postulated that the DN ase Iprotein has acquired incre asing structural stability throughthe introduction of the two Cys residues and deletion of ... purifiedusing the CONCERT High Purity Plasmid Midi kit(Invitrogen) for transfection.COS-7 cells were maintained in Dulbecco’s modifiedEagle’s medium containing 1 mML-glutamine, 50 UÆmL)1penicillin,...

Báo cáo khoa học: "Co-evolution of Language and of the Language Acquisition Device" docx

... 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 initial p-settings, ensuring neo-Darwinian rather than Lamarckian inheritance. The encoding of p-settings allows ... limitations during learning or during parsing which were affecting the results, another series of runs for "English" was performed with either memory limitations during learning but...

Tài liệu Báo cáo khoa học: Identiﬁcation of two late acyltransferase genes responsible for lipid A biosynthesis in Moraxella catarrhalis doc

... Picontaining 0.05% gelatin(DPBSG). Bacteria [10 lL containing 106colony-formingunits (CFU)] were inoculated into 190 lL reaction wellscontaining the diluted normal human serum, 25% heat-inactivated ... dnastar software (DNASTARInc., Madison, WI, USA).Cloning of lpxX and construction of the knockoutmutant O35ElpxXA DNA sequence containing lpxX was ampliﬁed from thechromosomal DNA of O35E using ... two mutants in bacterial clearance might reﬂectdifferences in the integrity of their outer membrane,binding activity and sensitivity of the murine comple-ment-mediated killing. In conclusion,...

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