Polymorphic crystals and complex multiple melting behavior in an aliphatic biodegradable polyester, poly(butylene adipate) (PBA), were thoroughly examined by wide‐angle X‐ray diffraction (WAXD) and differential scanning calorimetry (DSC). Further clarification on mechanisms of multiple melting peaks related to polymorphic crystal forms in PBA was attempted. More stable α‐form crystal is normally favored for crystallization from melt at higher temperatures (31–35 °C), or upon slow cooling from the melt; while the β‐form is the favored species for crystallization at low temperatures (25–28 °C). We further proved that PBA crystallization could also result in all α‐form even at low temperatures (25–28 °C) if it crystallized with the presence of prior α‐form nuclei. PBA packed with both crystal forms could display as many as four melting peaks (P1 ˜ P4, in ascending temperature order). However, PBA initially containing only the α‐crystal exhibited dual melting peaks of P1 and P3, which are attributed to dual lamellar distributions of the α‐crystal. By contrast, PBA initially containing only the β‐crystal could also exhibit dual melting peaks (P2 and P4) upon scanning. While P2 is clearly associated with melting of the initial β‐crystal, the fourth melting peak (P4), appearing rather broad, was determined to be associated with superimposed thermal events of crystal transformation from β‐ to α‐crystal and final re‐melting of the new re‐organized α‐crystal. Crystal transformation from one to the other or vice versa, lamellae thickening, annealing at molten state, and influence on crystal polymorphism in PBA were analyzed. Relationships and mechanisms of dual peaks for isolate α‐ or β‐crystals in PBA are discussed. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 1662–1672, 2005

Enhancement of the production of soluble recombinant penicillin acylase in Escherichia coli via coexpression of a periplasmic protease/chaperone, DegP, was demonstrated. Coexpression of DegP resulted in a shift of in vivo penicillin acylase (PAC) synthesis flux from the nonproductive pathway to the productive one when pac was overexpressed. The number of inclusion bodies, which consist primarily of protein aggregates of PAC precursors in the periplasm, was highly reduced, and the specific PAC activity was highly increased. DegP was a heat shock protein induced in response to pac overexpression, suggesting that the protein could possibly suppress the physiological toxicity caused by pac overexpression. Coexpression of DegP S210A , a DegP mutant without protease activity but retaining chaperone activity, could not suppress the physiological toxicity, suggesting that DegP protease activity was primarily responsible for the suppression, possibly by degradation of abnormal proteins when pac was overexpressed. However, a shortage of periplasmic protease activity was not the only reason for the deterioration in culture performance upon pac overexpression because coexpression of a DegP-homologous periplasmic protease, DegQ or DegS, could not suppress the physiological toxicity. The chaperone activity of DegP is proposed to be another possible factor contributing to the suppression.The well-known genetic information of Escherichia coli and successful applications of recombinant DNA technology make it possible for a variety of attempts with genetic engineering techniques to overproduce recombinant proteins. Upon performing the cultivation for recombinant protein production, there are two primary goals: high-cell-density cultivation and high-level gene expression. Culture performance can be optimized when the two goals are achieved simultaneously. Highcell-density culture can be obtained by fed-batch cultivation (48), in which concentrated medium is fed gradually into the bioreactor. The primary concern of this operation is developing an optimum feeding strategy, based on which cells can be maintained in a high-energy state for enhancing gene expression while cells are growing.

We study dynamic liquid bridge formation, which is relevant for wet granular flows involving highly viscous liquids and short collisions. Specifically, the drainage process of liquid adhering to two identical, non-porous wet particles with different initial film heights is simulated using Direct Numerical Simulations (DNS). We extract the position of the interface, and define the liquid bridge and its volume by detecting a characteristic neck position. This allows us building a dynamic model for predicting bridge volume, and the liquid remaining on the particle surface. Our model is based on two dimensionless mobility parameters, as well as a dimensionless time scale to describe the filling process. In the present work model parameters were calibrated with DNS data. We find that the proposed model structure is sufficient to collapse all our simulation data, indicating that our model is general enough to describe liquid bridge formation between equally sized particles.

The effects of α‐form and β‐form nuclei on polymorphic morphology of poly(butylene adipate) (PBA) upon recrystallization from the molten state up to various Tmax values were examined by differential scanning calorimetry (DSC), wide‐angle X‐ray diffraction (WAXD) and polarized light microscopy (PLM). In this study, PBA with complex melting and polymorphism behaviour was used as a model for examining different types and extents of residual nuclei. As the PBA initially containing the sole α‐crystal was brought to a molten state of various Tmax, the extents of trace α‐form crystal nuclei varied and were dependent on Tmax. Furthermore, it did not matter whether, initially, the PBA contained α‐ or β‐form crystals (or both) because only a single type of α‐nuclei could be left upon treatment to the molten liquid state at Tmax. Therefore, only the α‐crystal in PBA had ‘memory capacity’ in the molten liquid state while the β‐crystal did not. This was so because the latter had been completely transformed into the solid state prior to being heated into a liquid. PBA crystallized before α‐nuclei could be packed into α‐crystal, regardless of the crystallization temperature (Tc). For recrystallization from molten PBA without any nuclei, the crystalline polymorphism was correspondingly influenced by Tc. Copyright © 2005 Society of Chemical Industry

Cellulases from Bacillus and Geobacillus bacteria are potentially useful in the biofuel and animal feed industries. One of the unique characteristics of these enzymes is that they are usually quite thermostable. We previously identified a cellulase, GsCelA, from thermophilic Geobacillus sp. 70PC53, which is much more thermostable than its Bacillus homolog, BsCel5A. Thus, these two cellulases provide a pair of structures ideal for investigating the mechanism regarding how these cellulases can retain activity at high temperature. In the present study, we applied the SCHEMA non-contiguous recombination algorithm as a novel tool, which assigns protein sequences into blocks for domain swapping in a way that lessens structural disruption, to generate a set of chimeric proteins derived from the recombination of GsCelA and BsCel5A. Analyzing the activity and thermostability of this designed library set, which requires only a limited number of chimeras by SCHEMA calculations, revealed that one of the blocks may contribute to the higher thermostability of GsCelA. When tested against swollen Avicel, the highly thermostable chimeric cellulase C10 containing this block showed significantly higher activity (22%-43%) and higher thermostability compared to the parental enzymes. With further structural determinations and mutagenesis analyses, a 310 helix was identified as being responsible for the improved thermostability of this block. Furthermore, in the presence of ionic calcium and crown ether (CR), the chimeric C10 was found to retain 40% residual activity even after heat treatment at 90°C. Combining crystal structure determinations and structure-guided SCHEMA recombination, we have determined the mechanism responsible for the high thermostability of GsCelA, and generated a novel recombinant enzyme with significantly higher activity.