The structural stability of calmodulin (CaM) has been investigated previously by chemical and thermal methods. The calcium-loaded form of CaM has been found to be exceptionally stable, because it can be exposed to temperatures >90°C or to a 9 M urea solution without a marked change in its tertiary structure, and is therefore not experimentally accessible for unfolding studies using conventional analytical methods. In this study, we have developed a system for measuring the force for mechanically unfolding CaM using an atomic force microscope (AFM) by stretching the protein from its N-and C-terminal residues; we have been successful in obtaining force versus extension (F-E) curves for both apo and holo forms of CaM. In our experiment, distinguishable F-E curves have been obtained upon stretching of apoCaM and holoCaM to their full extensions. A very low force observed upon stretching of apoCaM indicated a relatively high flexibility of the apo form. On the contrary, a relatively high unfolding force and the appearance of a characteristic force peak were noted during full stretching of holoCaM. The F-E curve of the latter form of CaM most likely reflects a more rigid and probably more organized conformation of holoCaM than that of apoCaM. These experiments confirmed that the AFM is able to clearly distinguish two functionally distinct forms of CaM in terms of their mechanical properties.

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The process of acid hydrolysis using conventional methods at high concentrations results in products having lower yields, and it needs a longer time of process; therefore, it becomes less effective. In this study, we analyzed the effects of microwave-assisted pretreatment and cofermentation on bioethanol production from elephant grass (Pennisetum purpureum). We used a combination of delignification techniques and acid hydrolysis by employing a microwave-assisted pretreatment method on elephant grass (Pennisetum purpureum) as a lignocellulosic material. This was followed by cofermentation with Saccharomyces cerevisiae ITB-R89 and Pichia stipitis ITB-R58 to produce bioethanol. The optimal sugar mixtures (fructose and xylose) of the hydrolysis product were subsequently converted into bioethanol by cofermentation with S. cerevisiae ITB-R89 and P. stipitis ITB-R58, carried out with varying concentrations of inoculum for 5 days (48 h) at 30°C and pH 4.5. The high-power liquid chromatographic analysis revealed that the optimal inoculum concentration capable of converting 76.15% of the sugar mixture substrate (glucose and xylose) to 10.79 g/L (34.74% yield) of bioethanol was 10% (v/v). The optimal rate of ethanol production was 0.45 g/L/d, corresponding to a fermentation efficiency of 69.48%.

The main objective of this study is to investigate the possibility of powdered eggshell used as an adsorbent material for removal of humic acid from peat water. The influences of contact time, dose of eggshells, pH, and temperature were the factors considered in the removal processes of humic acid from peat water. In addition, adsorption isotherms of humic acid onto the powdered eggshell were also evaluated with the Langmuir and Freundlich approximations. Our results showed that the equilibrium of the process was eventually established after 60 min of the contact time, and also found that using 5 g of the powdered eggshell nearly 95 % of humic acid has been successfully removed from the peat water. The removal of humic acid gave better result when it was conducted at low pH, and it was almost unaffected the temperatures variation. The data was well fitted to Freundlich isotherm with the correlation coefficient of not \0.999, and could uptake the humic acid about 126.58 mg/g at pH 4.01, estimated from the Langmuir model. The kinetic experimental data proportionally correlated with the pseudo-second-order kinetic model with a rate constant in the range of 0.016-0.112 g mg -1 min -1 , while intra-particle-diffusion were the main rate determining step in the humic acid removal process. The powdered eggshell investigated in this study, thus, exhibited as a high potential adsorbent for the removal of humic acid from peat water.