Shuzo Tanaka

Ethanol fermentation from biomass resources: current state and prospects

In recent years, growing attention has been devoted to the conversion of biomass into fuel ethanol, considered the cleanest liquid fuel alternative to fossil fuels. Significant advances have been made towards the technology of ethanol fermentation. This review provides practical examples and gives a broad overview of the current status of ethanol fermentation including biomass resources, microorganisms, and technology. Also, the promising prospects of ethanol fermentation are especially introduced. The prospects included are fermentation technology converting xylose to ethanol, cellulase enzyme utilized in the hydrolysis of lignocellulosic materials, immobilization of the microorganism in large systems, simultaneous saccharification and fermentation, and sugar conversion into ethanol.

Biological Removal and Recovery of Toxic Heavy Metals in Water Environment

The authors review presently available or potential biological methods to remove and detoxify toxic heavy metals and metalloids from polluted water and sediments (i.e., biosorption, bioaccumulation, oxidation/reduction, leaching, precipitation, volatilization, degradation, and phytoremediation). In addition, they describe the options for the recovery of metals sequestered by biosorbents (use of appropriate desorbing agents) and microbial and plant biomass (leaching by means of chemical reagents or biological processes, and thermal treatment in controlled systems).

Enzymatic digestion of corncobs pretreated with low strength of sulfuric acid for bioethanol production.

In this study, the effect and the optimum pretreatment condition of corncobs using low strength of H2SO4 were investigated, in which H2SO4 was used to improve the enzymatic digestibility of corncobs for saccharification without degradation of sugars released. The optimum pretreatment condition was found to be the addition of 0.5% (vol./vol.) H2SO4 and autoclaving at 122°C for 20 min. Under this condition, the structural integrity of corncob was altered to make cellulose microfibrils more accessible for cellulase enzymes, and the enzymatic digestion of corncobs could be significantly enhanced. A high yield of sugar, 80% (wt./wt.), could be obtained at a low enzyme dosage of 0.024 g enzymes/g cobs, when pretreated. As a result, the ethanol production was obviously improved by the pretreatment, i.e., the ethanol yield of 77% (wt./wt.) was obtained within 36 h in the SSF fermentation using Saccharomyces cerevisiae NBRC2114.

Enhancement of xylose uptake in 2-deoxyglucose tolerant mutant of Saccharomyces cerevisiae

Chemical mutation of Saccharomyces cerevisiae using ethyl methane sulfonate was performed to enhance its ability of xylose uptake for ethanol production from lignocellulose under microaerobic condition. Among the appeared mutants, the mutant no. 2 (M2) strain screened using inhibitory effects of 2-deoxyglucose (DOG) showed more than 4-fold high ability in xylose uptake compared with the wild type strain, under the presence of glucose. The catabolite repression by glucose was sufficiently reduced in M2 strain due to its tolerance to the high concentration of DOG (0.5%, wt./vol.). Metabolomic analyses of various sugars in the cell revealed that some of xylose was reduced to xylitol in M2 cell, providing the concentration gradient of xylose and more uptake of xylose. Xylulose-5-phosphate was significantly detected in the crude cell extract from M2 strain, indicating higher metabolic activity in pentose phosphate pathway. This was also confirmed by in vitro analyses of key enzymes involved in glucose and xylose metabolism, such as hexokinase, glucose-6-phosphate dehydrogenase and xylose reductase. Glucose uptake was moderately suppressed in the presence of trehalose-6-phosphate inhibiting the activation of hexokinase, resulting in more uptake of xylose through hexose transport system. To our knowledge, this study is the first report verifying that the mutation technique successfully enhances the xylose uptake by S. cerevisiae, particularly under the presence of glucose.

Multiple effects of swelling by sodium bicarbonate after delignification on enzymatic saccharification of rice straw

The multiple effects of pretreatments by chemical delignification using acidified sodium chlorite (ASC) and swelling using sodium bicarbonate (SB) for enzymatic saccharification of rice straw in bioethanol production have been investigated in this study. The treatment with the combination of ASC three times (3× ASC) first and SB later resulted in the significant reduction in Klason lignin content up to 90% (wt./wt.). By the saccharification of the pretreated rice straw with cellulase enzymes, it was confirmed that SB treatment was an important step in the pretreatment process not only to disintegrate the cellulose structure but also to facilitate the amorphization of the crystalline cellulose as well as the extended removal of integrated lignin. Furthermore, FTIR analyses revealed that the crystal type of cellulose appeared to be changed from type I to type II by SB treatment, thereby increasing the cellulose surface area and making it more accessible to the cellulase enzyme. Conversion rate to sugar was remarkably increased when 3× ASC + SB treatments were applied to untreated rice straw, even though the saccharification of the treated rice straw was performed at a low enzyme loading (1/100, wt.-enzymes/wt.-substrate). Conclusively, rice straw could be saccharified at high yield in short time at low cellulase loading, enables the enzymatic saccharification to be more feasible for practical bioethanol production using rice straw as a substrate.