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... resulted in 55% and 51% loss of the initial activities of BHLmL2 and BHAb, respectively. These results strongly suggest that Ser is involved in the catalytic activity of BHLmL2 and BHAb and are consistent ... culture were 0.012, 0.025, 0.050, 0.125 and 0.250 mm) and after fur- ther incubation for 24 h, the mycelia were separated from the culture broth by filtration and used for BH activity assays. GC–MS ... respectively. The influence of pH on the activities of the BH enzymes was investigated in the pH range 6–11. The pH optima were determined to be in the basic range (pH 8.0–10.0) for BHLmL2 and BHAb...

Ngày tải lên: 18/02/2014, 14:20

... evaluated by the statistical potential used in the present study. In the case of the RNA muta- tions to the SRP complex, the mutated RNA residues are located in a double-stranded region of RNA, and do not ... onto the RNA bases nearest the center of the b-sheet structure of the RRM. We then scored these sequence-variant structures with the distance- dependent potential function. Figure 2 shows the ... score; the blue line represents the rank of these sequences using the distance-dependent potential. The points in each colored line are sorted independently by rank; the x-axis is the sort order. The...

Ngày tải lên: 18/02/2014, 16:20

... investigation of the catalytic mechanism and substrate speci city and to understand the differences therein for this class of enzymes. In order to identify the subsites of the ervatamins and to understand ... both the molecules of the asymmetric unit runs along the extended backbone of residues 65–64 and is partly exposed to the solvent (Fig. 2B). On the other hand, the P3 moiety of E-64 in each of the ... using the programs cns [58] and detwin and sfcheck in the CCP4 suite [59] confirmed that the data were twinned, and allowed us to calculate the twinning fraction (0.394) (Fig. 6A,B) and the twinning...

Ngày tải lên: 18/02/2014, 16:20

... binding of the Sfbgly interactions with the aglycone. Thus, the effect of the alteration of the glycone structure in the stabil- ity of the ES à complex (DDG à ) for the wild-type and mutant ... another important issue in understand- ing the aglycone speci city is the contribution of the interactions with the aglycone to the catalysis in b-glycosidases. Mutational studies with ZmGlu1 and SbDhr1 ... is that the interactions with aglycone do not affect the binding of the glycone within the Sfbgly active site. This hypothesis was further investigated by deter- mining the influence on the glycone...

Ngày tải lên: 18/02/2014, 17:20

... oxidized and the negative signals with the reduced form of the enzyme. For the reduced form, the Soret band can be observed at 415 and 442 nm, and for the oxidized form at 403 and 422 nm. The b–band ... be seen at 517 nm and the a–band at 547 nm and 603 nm. The difference signals that can be observed between 400 and 700 nm include the contributions of the hemes c, a and a 3 . The difference signals ... to the different environ- ment of the heme centers. The difference signals observed at 442 nm and 603 nm can be assigned to the contributions of the hemes a and a 3 , with the position of the...

Ngày tải lên: 21/02/2014, 15:20

... internal membrane side is the rate-limiting s tep and not – as proposed for most of the other electrogenic symporters – the return of the unloaded transporter to the outside of the membrane [17]. In ... alanine, serine and proline but also osmolytes such as sarcosine and betaine, and the D -enantiomers of serine and alanine. The apparent affinities of PAT1 substrates are mainly in the range of 2–15 ... The apparent K m values of D -Pro, D -Ala and OH-Pro increased only 1.5- and 2.1-fold by depolarizing the membrane from )120 to )20 mV. On the other hand, the affinity of b-Ala did not change significantly within...

Ngày tải lên: 07/03/2014, 16:20

... 3B). Thus for the SBD tree there is not an obvious border between the hydrolases and transglycosi- dases, but rather there may be one between the compact cluster of Bacillus and Thermoanaerobacter ... to domain A, the catalytic (b/a) 8 -barrel[1].Mostofthem possess a domain B that protrudes from the barrel between the third b-strand and third a-helix and varies greatly in length, sequence and tertiary ... supports the idea that there has been a separate evolution of this domain [30]. This together with the findings of the present study indicates a separate evolution of the domains C and E compared to the...

Ngày tải lên: 08/03/2014, 08:20

... linkage speci city. Within these N-ter- mini, the A1 ⁄ A2 and O1 ⁄ O2 parts of the reuteransucr- ase catalytic domains mainly determin the glucosidic linkages synthesized. Thus, not only the A2 ⁄ ... a) 8 barrel with the three catalytic resi- dues (Fig. 1C), determine the types and ratios of glu- cosidic linkages synthesized (Table 1). The A1 ⁄ O1 and A2 ⁄ O2 regions within the N-termini of the catalytic ... sequencing), both p15-GTFA-dN-RS and p15-GTFO-dN-RS were digested with XbaI and SalI, SalI and SacI, and SacI and KpnI, and corresponding fragments were exchanged, yielding the six constructs p15-GTFA-O1-dN-RS,...

Ngày tải lên: 16/03/2014, 04:20

... speci city is more pronounced and the synthetic capacity is, respectively, increased. In the latter system PA behaves as a typical transferase [14,15]. The nucleophile or S 1 Â [16] specicity of the ... close to the position found in the complex with PG [19]. The bond lengths and valence angles of the nitroaniline moiety were taken from the X-ray data for the system cyclophilin A and the tripeptide ... large when the substrate’s polar group is directed toward it, and small when the polar group points to ArgB263. The interactionenergyofArgB263withtherestofthecomplex, however, remains large and practically...

Ngày tải lên: 16/03/2014, 16:20

... may influence the shape and volume of the acyl binding pocket and thereby alter the binding mode and affinity of the enzyme for PAA and derivatives thereof, while maintaining the hydrophobicity of the binding ... site. To test the effect of the mutations on the speci city for phenylacetylated substrates, the steady-state kinetic param- eters for the hydrolysis of the chromogenic substrate NIPAB and the inhibition ... interactions with PAA and shields the binding site from the solvent. A third phenylalanine, bF57, is located at the bottom of the hydrophobic cleft. The shortest distance between the side chain of bF57 and PAA...

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