Biochemical Engineering Journal (v.6, #2)

Efficient production of protopectinases by Bacillus subtilis using medium based on soybean flour by Takeshi Matsumoto; Yoshitomo Sugiura; Akihiko Kondo; Hideki Fukuda (81-86).
We have developed a culture system for efficient production of protopectinases (PPases) by Bacillus subtilis. PPase shows the pectin-releasing activity and is expected to be utilized in the enzymatic cotton scouring. B. subtilis IFO3134 was cultivated using defatted soybean flour as a main component of culture media. This strain produced three different types of PPases, namely, PPase-C, -N and -R performing endo-arabinase activity, pectate-lyase activity and pectin-lyase activity, respectively. The effects of alkaline solubilization and autoclave treatments to extract nutrients from soybean flour and initial soybean flour concentration (20–80 g/l) on production of PPases in batch fermentation were investigated. Alkaline solubilization of soybean flour with NaOH remarkably reduced enzyme productivity. In addition, a higher initial concentration of soybean flour reduced the enzyme productivity of cells. The pectin-releasing activity was the largest and reached up to 2200–2400 U/ml, when the culture medium containing an initial soybean flour concentration of 40 g/l was autoclaved for 45–60 min without alkaline solubilization treatment.
Keywords: Protopectinase; Pectin; Bacillus subtilis; Soybean flour; Enzyme production;

Chlorella pyrenoidosa was cultivated under photoautotrophic, mixotrophic and cyclic light-autotrophic/dark-heterotrophic conditions. The influence of light on the carbon and energy metabolism of microalgae was investigated by the use of metabolic flux analysis. The respiratory activity of microalgae in the light was assessed from the autotrophic flux distribution. Results showed that the glycolytic pathway, tricarboxylic acid cycle and mitochondrial oxidative phosphorylation maintained high activities during illumination, indicating little effect of light on these pathways, while the flux through the pentose phosphate pathway during illumination was very small due to the light-mediated regulation. The theoretical yields of biomass on ATP decreased in the following order: heterotrophic culture>mixotrophic culture>autotrophic culture, and a significant amount of the available ATP was required for maintenance processes in microalgal cells. The energy conversion efficiency between the supplied energy to culture, the absorbed energy by cells and the free energy conserved in ATP were analyzed for the different cultures. Analysis showed that the heterotrophic culture generated more ATP from the supplied energy than the autotrophic and mixotrophic cultures. The maximum thermodynamic efficiency of ATP production from the absorbed energy, which was calculated from the metabolic fluxes at zero growth rate, was the highest in the heterotrophic culture and as low as 16% in the autotrophic culture. By evaluating the energy economy through the energy utilization efficiency, it was found that the biomass yield on the supplied energy was the lowest in the autotrophic cultivation, and the cyclic culture gave the most efficient utilization of energy for biomass production.
Keywords: Microalgal cells; Metabolic flux; Central metabolism; Energetics;

Surfactant–lactoperoxidase complex catalytically active in organic media by Shin-ya Okazaki; Youichi Uchimura; Masahiro Goto; Shintaro Furusaki (103-107).
A surfactant–lactoperoxidase (LPO) complex catalytically active in organic solvents was developed by the emulsion coating method. The oxidation of 2,6-dimethoxyphenol (2,6-DMP) was conducted by the surfactant–LPO complex in organic media. The LPO complex efficiently catalyzed the oxidation of 2,6-DMP in various organic solvents, although lyophilized LPO did not display the catalytic activity at all. To optimize the preparation and reaction conditions for the surfactant–LPO complex, we examined the effects of pH value in the water pools of W/O emulsions, kinds of oxidants, and the nature of organic solvents on the oxidation reaction. Its optimum activity was obtained when the pH value of the aqueous enzyme solution was adjusted to ca. 8 at the preparation stage. The LPO complex exhibited the highest catalytic activity in chloroform when H2O2 was employed as the oxidant. Furthermore, the storage stability of the surfactant–LPO complex was far better than that of the surfactant–horseradish peroxidase complex. This high storage stability of the LPO complex will be a benefit for industrial usage of peroxidases.
Keywords: Biocatalysis; Enzyme activity; Enzyme technology; Bioreactions; Organic solvents; Surfactant; Peroxidase;

Multiple-impeller systems with a special emphasis on bioreactors: a critical review by Parag R Gogate; Anthony A.C.M Beenackers; Aniruddha B Pandit (109-144).
The multiple-impeller agitated systems are compared with single-impeller agitated systems with a special focus on its applications for bioreactors. Correlations reported in the literature for gas phase hold-up, mass transfer coefficient and power consumption under gassed and ungassed conditions are compared and recommendations have been made regarding their suitability for design and scale-up of bioreactors. The multiple-impeller systems are found to be superior as compared to single-impeller systems in all the above mentioned aspects, except liquid mixing. For all kinds of reactors where the sole purpose is mass transfer, multiple-impeller systems are advantageous and there would be large savings on an industrial scale, especially for the bioreactors where the reaction periods are long and the power consumption cost could be a significant component to the overall production costs.
Keywords: Multiple-impeller systems; Flow patterns and regimes; Power consumption; Mixing; Suspension of solids; Gas hold-up; Mass transfer; Heat transfer;

The biosorption of Fe(III), Cr(VI), Pb(II), Cu(II) and Ni(II) ions on Zoogloea ramigera (activated sludge bacterium) and Rhizopus arrhizus (filamentous fungus) has been studied as a function of initial metal ion concentration and temperature. The applicability of the Langmuir model for each metal–microorganism system has been tested at different temperatures. The enthalpy change for the biosorption process has been evaluated by using the Langmuir constant b, related to the energy of adsorption. Thermodynamic parameters indicate the exothermic nature of Cu(II) and Ni(II) biosorption on both microorganisms. Fe(III), Cr(VI) and Pb(II) biosorption is determined to be an endothermic process since increased binding occurs as the temperature is increased in the range 15–45°C.
Keywords: Waste water; Heavy metal; Zoogloea ramigera; Rhizopus arrhizus; The heat of biosorption; Langmuir model;

Biotechnological potential of coffee pulp and coffee husk for bioprocesses by Ashok Pandey; Carlos R. Soccol; Poonam Nigam; Debora Brand; Radjiskumar Mohan; Sevastianos Roussos (153-162).
Advances in industrial biotechnology offer potential opportunities for economic utilization of agro-industrial residues such as coffee pulp and coffee husk. Coffee pulp or husk is a fibrous mucilagenous material (sub-product) obtained during the processing of coffee cherries by wet or dry process, respectively. Coffee pulp/husk contains some amount of caffeine and tannins, which makes it toxic in nature, resulting the disposal problem. However, it is rich in organic nature, which makes it an ideal substrate for microbial processes for the production of value-added products. Several solutions and alternative uses of the coffee pulp and husk have been attempted. These include as fertilizers, livestock feed, compost, etc. However, these applications utilize only a fraction of available quantity and are not technically very efficient. Attempts have been made to detoxify it for improved application as feed, and to produce several products such as enzymes, organic acids, flavour and aroma compounds, and mushrooms, etc. from coffee pulp/husk. Solid state fermentation has been mostly employed for bioconversion processes. Factorial design experiments offer useful information for the process optimization. This paper reviews the developments on processes and products developed for the value-addition of coffee pulp/husk through the biotechnological means.
Keywords: Coffee pulp; Coffee husk; Submerged fermentation; Solid state fermentation; Biotechnological applications;