Our earlier studies with outer membrane permeability in E. coli showed that an insertion mutation in lpp gene (encoding Braun's lipoprotein) drastically changed the outer membrane permeability, resulting in significant acceleration of whole-cell catalyzed reactions. In order to gain a mechanistic understanding of the nature of permeability change, the lpp region was sequenced. The results revealed that Lpp was not expressed in the insertion mutant, suggesting that the absence, rather than the alteration, of Lpp is responsible for the observed permeability change. This surprising result prompts us to investigate the possibility of establishing lpp deletion as a general permeabilization method. Two lpp deletion mutants were generated from strains with different genetic background and the effect of lpp deletion on cell physiology was investigated. While lpp deletion had no significant effect on cell growth, carbon metabolism, and fatty acid compositions, it enhanced permeability of various small molecules, consistent with the results with the insertion mutant. This phenotype is useful in a wide range of biotechnological applications. We illustrate here the use of the mutant with organophosphate hydrolysis and L-carnitine synthesis, where permeability is known to be a limiting factor. Both processes were significantly improved with the mutant because of enhanced permeability through the outer membrane. Therefore, this study has established an easy yet generally applicable method for permeabilizing E. coli cells without significant adverse effects. Further, as lpp homolog is known to exist in gram-negative bacteria, we expect that this method will be applicable to other gram-negative bacteria.
Pb-2.2 and 5.8 wt pct Sb alloys were directionally solidified with a positive thermal gradient of 140 K cm Ϫ1 at growth speeds ranging from 0.8 to 30 m s Ϫ1, and then quenched to retain the mushyzone morphology. Chemical analysis along the length of the directionally solidified portion and in the quenched melt ahead of the dendritic array showed extensive longitudinal macrosegregation. Cellular morphologies growing at smaller growth speeds are associated with larger amounts of macrosegregation as compared with the dendrites growing at higher growth speeds. Convection is caused, mainly, by the density inversion in the overlying melt ahead of the cellular/dendritic array because of the antimony enrichment at the array tip. Mixing of the interdendritic and bulk melt during directional solidification is responsible for the observed longitudinal macrosegregation.
Cellular/dendritic array tip morphology has been examined in directionally solidified and quenched Pb-5.8 wt pct Sb alloy by a serial sectioning and three-dimensional image reconstruction technique. There is a large scatter in the tip radius, the nearest neighbor spacing, and the mushy zone length, even among the immediately neighboring cells and dendrites. This scatter may be caused by the natural convection (in the mushy zone and in the bulk melt at the array tip), which also produces macrosegregation along the length of the directionally solidified samples. Even in the presence of convection, however, the tip radii are observed to be approximately proportional to the square of the primary spacings, and the radii are in a good quantitative agreement with the predictions from the model due to Hunt-Lu.
Adding a cationic polyacrylamide (c-PAM) to either the amylase mediated hydrolysis of corn starch or the hydrolysis of wood fiber by cellulase can enhance the initial hydrolysis rates, although a rate decrease can occur under some conditions. Several c-PAMs can serve as catalysts and the same c-PAM can improve the efficiency of both amylase and cellulase. The initial amylase rate approximately doubles; the analogous cellulase hydrolysis rate increases by about 40%. c-PAMs increase the binding of enzyme to substrate.
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