Toru Onishi

Genetically engineered wine yeast produces a high concentration of L-lactic acid of extremely high optical purity.

ABSTRACT For mass production of lactic acid, we newly constructed a transgenic wine yeast strain that included six copies of the bovine l-lactate dehydrogenase gene on the genome. On fermentation in inexpensive cane juice-based medium, l-lactate production of this recombinant reached 122 g/liter and the optical purity was 99.9% or higher.

The Effect of Pyruvate Decarboxylase Gene Knockout in Saccharomyces cerevisiae on L-Lactic Acid Production

A plant- and crop-based renewable plastic, poly-lactic acid (PLA), is receiving attention as a new material for a sustainable society in place of petroleum-based plastics. We constructed a metabolically engineered Saccharomyces cerevisiae that has both pyruvate decarboxylase genes (PDC1 and PDC5) disrupted in the genetic background to express two copies of the bovine L-lactate dehydrogenase (LDH) gene. With this recombinant, the yield of lactate was 82.3 g/liter, up to 81.5% of the glucose being transformed into lactic acid on neutralizing cultivation, although pdc1 pdc5 double disruption led to ineffective decreases in cell growth and fermentation speed. This strain showed lactate productivity improvement as much as 1.5 times higher than the previous strain. This production yield is the highest value for a lactic acid-producing yeast yet reported.

Impact of Al Doping Concentration at Channel Region on Mobility and Threshold Voltage Instability in 4H-SiC Trench N-MOSFETs

The effect of Al doping concentration (NA) at channel regions ranging from 1.0×1017 to 4.0×1017 cm-3 on the effective channel mobility of electron (μeff) and the threshold voltage (Vth) instability under the positive bias-temperature-stress conditions has been investigated througu the use of trench-gate 4H-SiC MOSFETs with m-face (1-100) channel regions. It was found that μeff degraded with an increase in NA. On the other hand, the increase of NA enlarged the Vth instability. These results indicate that NA has a large impact not only on the Vth value but also on the channel resistance and reliability in 4H-SiC trench MOSFETs.

Screening and evolution of a novel protist xylose isomerase from the termite Reticulitermes speratus for efficient xylose fermentation in Saccharomyces cerevisiae

BackgroundThe yeast Saccharomyces cerevisiae, a promising host for lignocellulosic bioethanol production, is unable to metabolize xylose. In attempts to confer xylose utilization ability in S. cerevisiae, a number of xylose isomerase (XI) genes have been expressed heterologously in this yeast. Although several of these XI encoding genes were functionally expressed in S. cerevisiae, the need still exists for a S. cerevisiae strain with improved xylose utilization ability for use in the commercial production of bioethanol. Although currently much effort has been devoted to achieve the objective, one of the solutions is to search for a new XI gene that would confer superior xylose utilization in S. cerevisiae. Here, we searched for novel XI genes from the protists residing in the hindgut of the termite Reticulitermes speratus.ResultsEight novel XI genes were obtained from a cDNA library, prepared from the protists of the R. speratus hindgut, by PCR amplification using degenerated primers based on highly conserved regions of amino acid sequences of different XIs. Phylogenetic analysis classified these cloned XIs into two groups, one showed relatively high similarities to Bacteroidetes and the other was comparatively similar to Firmicutes. The growth rate and the xylose consumption rate of the S. cerevisiae strain expressing the novel XI, which exhibited highest XI activity among the eight XIs, were superior to those exhibited by the strain expressing the XI gene from Piromyces sp. E2. Substitution of the asparagine residue at position 337 of the novel XI with a cysteine further improved the xylose utilization ability of the yeast strain. Interestingly, introducing point mutations in the corresponding asparagine residues in XIs originated from other organisms, such as Piromyces sp. E2 or Clostridium phytofermentans, similarly improved xylose utilization in S. cerevisiae.ConclusionsA novel XI gene conferring superior xylose utilization in S. cerevisiae was successfully isolated from the protists in the termite hindgut. Isolation of this XI gene and identification of the point mutation described in this study might contribute to improving the productivity of industrial bioethanol.

Impact of optical phonon scattering on inversion channel mobility in 4H-SiC trenched MOSFETs

Temperature characteristics of the channel mobility were investigated for 4H-SiC trenched MOSFETs in the range from 30 to 200 °C. The conventional model of channel mobility limited by carrier scattering is based on Si-MOSFETs and shows a greatly different channel mobility from the experimental value, especially at high temperatures. On the other hand, our improved mobility model taking into account optical phonon scattering yielded results in excellent agreement with experimental results. Moreover, the major factors limiting the channel mobility were found to be Coulomb scattering in a low effective field (<0.7 MV/cm) and optical phonon scattering in a high effective field.