Researchers Database

mizushima daiki

    InfectionandImmunityMedicalZoologyandParasitology Research Associate
Last Updated :2021/12/07

Researcher Information

J-Global ID

Academic & Professional Experience

  • 2019/04 - Today  Jichi Medical University医学部 感染・免疫学講座 医動物学部門助教
  • 2017/04 - 2019/03  Obihiro University of Agriculture and Veterinary MedicineNational Research Center for Protozoan DiseasesPostdoctoral fellow
  • 2016/04 - 2017/03  National Agriculture and Food Research OrganizationFood Research InstitutePostdoctoral fellow

Published Papers

  • Ahmed Tabbabi, Shinya Watanabe, Daiki Mizushima, Abraham G. Caceres, Eduardo A Gomez, Daisuke S. Yamamoto, Longzhu Cui, Yoshihisa Hashiguchi, Hirotomo Kato
    Microorganisms 9 (1) 68 - 68 2020/12 
    Differences in the gut microbial content of Lutzomyia (Lu.) ayacuchensis, a primary vector of Andean-type cutaneous leishmaniasis in Ecuador and Peru, may influence the susceptibility of these sand flies to infection by Leishmania. As a first step toward addressing this hypothesis, a comparative analysis of bacterial and fungal compositions from Lu. ayacuchensis populations with differential susceptibilities to Leishmania was performed. Bacterial 16S rRNA gene amplification and Illumina MiSeq sequencing approaches were used to characterize the bacterial composition in wild-caught populations from the Andean areas of Ecuador and southern Peru at which the sand fly species transmit Leishmania (Leishmania) mexicana and Leishmania (Viannia) peruviana, respectively, and a population from the northern Peruvian Andes at which the transmission of Leishmania by Lu. ayacuchensis has not been reported. In the present study, 59 genera were identified, 21 of which were widely identified and comprised more than 95% of all bacteria. Of the 21 dominant bacterial genera identified in the sand flies collected, 10 genera had never been detected in field sand flies. The Ecuador and southern Peru populations each comprised individuals of particular genera, while overlap was clearly observed between microbes isolated from different sites, such as the number of soil organisms. Similarly, Corynebacterium and Micrococcus were slightly more dominant bacterial genera in the southern Peru population, while Ochrobactrum was the most frequently isolated from other populations. On the other hand, fungi were only found in the southern Peru population and dominated by the Papiliotrema genus. These results suggest that variation in the insect gut microbiota may be elucidated by the ecological diversity of sand flies in Peru and Ecuador, which may influence susceptibility to Leishmania infection. The present study provides key insights for understanding the role of the microbiota during the course of L. (L.) mexicana and L. (V.) peruviana infections in this important vector.
  • Ahmed Tabbabi, Abraham G Cáceres, T Pershing Bustamante Chauca, Chisato Seki, Yanisa Choochartpong, Daiki Mizushima, Daisuke S Yamamoto, Yoshihisa Hashiguchi, Hirotomo Kato
    PLoS neglected tropical diseases 14 (10) e0008797  2020/10 
    Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mannose phosphate isomerase (mpi) gene was applied to 134 skin samples collected from patients with cutaneous leishmaniasis (CL) in Peru for identification of the infecting parasite at the species level, and the results were compared with those of cytochrome b (cyt b) gene sequencing obtained in previous studies. Although most results (121/134) including 4 hybrids of Leishmania (Viannia) braziliensis and L. (V.) peruviana corresponded to those obtained in the previous study, PCR-RFLP analyses revealed the distribution of putative hybrid strains between L. (V.) peruviana and L. (V.) lainsoni in two samples, which has never been reported. Moreover, parasite strains showing discordance between kinetoplast and nuclear genes (kDNA and nDNA), so-called mito-nuclear discordance, were identified in 11 samples. Of these, six strains had the kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) guyanensis, and three strains had the kDNAs of L. (V.) shawi and nDNAs of L. (V.) braziliensis. The rest were identified as mito-nuclear discordance strains having kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) lainsoni, and kDNAs of L. (V.) lainsoni and nDNAs of L. (V.) braziliensis. The results demonstrate that Leishmania strains in Peru are genetically more complex than previously considered.
  • Daiki Mizushima, Tovuu Amgalanbaatar, Batdorj Davaasuren, Mitsunori Kayano, Sandagdorj Naransatsral, Punsantsogvoo Myagmarsuren, Davaajav Otgonsuren, Batsaikhan Enkhtaivan, Batbold Davkharbayar, Bayasgalan Mungun-Ochir, Purevdorj Baatarjargal, Uranbileg Nyamdolgor, Gurdorj Soyolmaa, Adilbish Altanchimeg, Myagmar Zoljargal, Thu-Thuy Nguyen, Badgar Battsetseg, Banzragch Battur, Noboru Inoue, Naoaki Yokoyama, Keisuke Suganuma
    Parasite epidemiology and control 10 e00158  2020/08 [Refereed][Not invited]
     
    In Mongolia, horses play important roles, not only in livestock production, but also in terms of culture, tradition, and Mongolian beliefs. Although the presence of non-tsetse-transmitted horse trypanosomoses, which are caused by infections with Trypanosoma evansi (surra) and T. equiperdum (dourine), has been reported in the country, whether there is a nationwide epidemic of these infectious diseases is unknown. In the present study, a nationwide surveillance of horse trypanosomoses was performed. The sample sizes for each province, the whole country, and male and female horses were, respectively, 96, 2,400, and 316 and 306. In total, 3,641 samples of horse sera were collected by simple random sampling. The rTeGM6-4r-based ELISA, which was applied for surra against cattle and water buffalo and dourine against horse, revealed that the overall sero-prevalence of the diseases in Mongolia was 4.8%. Among them, high sero-prevalences were observed in the central provinces (5.2-11.0%, p < 0.05) of the country. The sero-prevalence was significantly higher in females than in males (6.0% and 4.0%, p < 0.05, respectively) and in non-castrated males (8.4%, p < 0.01) compared with castrated males (3.0%). These results suggested that currently, horse trypanosomoses are a nationwide endemic problem in Mongolia. Knowledge of the nationwide endemic status of non-tsetse-transmitted horse trypanosomoses in Mongolia will be useful to prevent these diseases.
  • Batbold Davkharbayar, Batdorj Davaasuren, Sandagdorj Narantsatsral, Banzragch Battur, Myagmarsuren Punsantsogvoo, Badgar Battsetseg, Daiki Mizushima, Noboru Inoue, Keisuke Suganuma
    Journal of equine veterinary science 87 102905 - 102905 2020/04 [Refereed][Not invited]
     
    Dourine is a lethal protozoan disease of equids, and it is caused by Trypanosoma equiperdum infection via coitus. To date, treatment strategies against the dourine are not recommended because of the frequent relapses; therefore, the World Organisation for Animal Health recommends the stamping-out policy for the control of dourine. Our previous studies have revealed a number of horses with dourine in Mongolia that is the fifth largest horse-breeding country. It is difficult to apply the stamping-out policy for cases of dourine in Mongolia because of an inadequate livestock guarantee system. Therefore, the development of effective treatment measures is an urgent need. In this study, an 8-year-old stallion was definitely diagnosed with dourine based on clinical signs, molecular analysis, and microscopic examination of trypanosomes. Combination therapy with diminazene aceturate and quinapyramine sulfate was applied. Before the treatment, the characteristic clinical signs of dourine were observed, and trypanosomes were detected in the urogenital tract mucosal swab samples by microscopic examination and polymerase chain reaction (PCR). Moreover, positive serological results were obtained. After the treatment, we observed an improvement in the health of the treated horse and no trypanosome infection in its urogenital tract by microscopic examination and PCR. Moreover, serological tests showed seronegative results. The horse has showed no relapse for at least 2.5 years after the treatment, and its reproductive ability has improved. Our result suggests that trypanosomes did not invade cerebrospinal fluid when we started the therapy. In conclusion, the combination therapy has therapeutic potential against dourine at an early phase.
  • アジアに分布する吸血性サシガメTriatoma rubrofasciataの唾液腺トランスクリプトーム解析
    加藤 大智, 水島 大貴, Tabbabi Ahmed, 山本 大介, 角田 隆, Le Trung Kien
    衛生動物 日本衛生動物学会 71 (Suppl.) 50 - 50 0424-7086 2020/03
  • Daiki Mizushima, Takatsugu Miyazaki, Yuh Shiwa, Keitarou Kimura, Shiho Suzuki, Nobuyuki Fujita, Hirofumi Yoshikawa, Atsuo Kimura, Shinichi Kitamura, Hiroshi Hara, Kazumi Funane
    Applied Microbiology and Biotechnology 103 (16) 6581 - 6592 1432-0614 2019/08 [Refereed][Not invited]
     
    Paenibacillus sp. 598K produces cycloisomaltooligosaccharides (CIs) in culture from dextran and starch. CIs are cyclic oligosaccharides consisting of seven or more α-(1 → 6)-linked-D-glucose residues. The extracellular enzyme CI glucanotransferase (PsCITase), which is the member of glycoside hydrolase family 66, catalyzes the final stage of CI production and produces mainly cycloisomaltoheptaose. We have discovered a novel intracellular CI-degrading dextranase (PsDEX598) from Paenibacillus sp. 598K. The 69.7-kDa recombinant PsDEX598 does not digest isomaltotetraose or shorter isomaltooligosaccharides, but digests longer ones of at least up to isomaltoheptaose. It also digests oligoCIs of cycloisomaltoheptaose, cycloisomaltooctaose, and cycloisomaltononaose better than it does with megaloCIs of cycloisomaltodecaose, cycloisomaltoundecaose, and cycloisomaltododecaose, as well as an α-(1 → 6)-D-glucan of dextran 40. PsDEX598 is produced intracellularly when culture medium is supplemented with cycloisomaltoheptaose or dextran, but not with isomaltooligosaccharides (a mixture of isomaltose, isomaltotriose, and panose), starch, or glucose. The whole genomic DNA sequence of the strain 598K implies that it harbors two genes for enzymes belonging to glycoside hydrolase family 66 (PsCITase and PsDEX598), and PsDEX598 is the only dextranase in the strain. PsDEX598 does not have any carbohydrate-binding modules (CBMs) and has a low similarity (< 30%) with other family 66 dextranases, and the catalytic amino acids of this enzyme are predicted to be Asp191, Asp303, and Glu368. The strain Paenibacillus sp. 598K appears to take up CI-7, so these findings indicate that this bacterium can degrade CIs using a dextranase within the cells and so utilize them as a carbon source for growth.
  • Keisuke Suganuma, Daisuke Kondoh, Thillaiampalam Sivakumar, Daiki Mizushima, Afra'a Tajelsir Mohamed Elata, Oriel M M Thekisoe, Naoaki Yokoyama, Noboru Inoue
    Parasitology research 118 (6) 1927 - 1935 2019/06 [Refereed][Not invited]
     
    Trypanosoma (Megatrypanum) theileri is a cosmopolitan, usually non-pathogenic, trypanosome of cattle transmitted by blood-sucking arthropods, mainly tabanid flies. Several T. theileri strains isolated from domestic and wild ruminants via co-culturing with mammalian feeder cells or blood cells have been characterized morphologically and genetically. Here, we cultured a new trypanosome isolate from a Holstein cow in Hokkaido, Japan, and performed morphological and molecular characterization studies. The new isolate (Obihiro strain) was co-cultivated with Madin-Darby bovine kidney (MDBK) cells in GIT medium supplemented with 10% fetal bovine serum. Trypomastigotes and epimastigotes, but not intracellular parasites, were identified in the culture. Analysis of the V7-V8 region of 18S rRNA sequences showed that the Obihiro strain is positioned within the subgenus Megatrypanum. A dendrogram based on whole internal transcribed spacer rDNA sequence showed that the Obihiro strain clustered in the lineage TthII together with the Japanese isolates of T. theileri, Esashi 9, and Esashi 12, and isolates from Zambia and the USA. T. theileri of the KM strain and a T. theileri-like trypanosome isolated from deer (TSD1 strain) clustered in the lineage TthI, separate from the Obihiro strain. Based on a partial cathepsin L-like protein gene analysis, the Obihiro strain clustered with isolates of the TthIIF genotype, which includes T. theileri from Vietnam, Sri Lanka, and Brazil. Our analyses of the T. theileri Obihiro strain provide relevant insights into its genetic diversity in Japanese cattle and corroborate the host specificity of cattle and deer trypanosomes of the subgenus Megatrypanum.
  • Bayasgalan Mungun-Ochir, Noriyuki Horiuchi, Adilbish Altanchimeg, Kenji Koyama, Keisuke Suganuma, Uranbileg Nyamdolgor, Ken-Ichi Watanabe, Purevdorj Baatarjargal, Daiki Mizushima, Banzragch Battur, Naoaki Yokoyama, Badgar Battsetseg, Noboru Inoue, Yoshiyasu Kobayashi
    Neuromuscular disorders : NMD 29 (6) 437 - 443 2019/06 [Refereed][Not invited]
     
    Dourine is an equine protozoan disease caused by Trypanosoma equiperdum. Dourine-afflicted animals die after developing neurological clinical signs, such as unilateral paresis. The disease has been a problem for many years; however, the pathogenesis regarding the neurological clinical signs of dourine has been unclear. In the present study, we conducted a histopathological examination in order to investigate the mechanisms by which dourine-afflicted horses develop the accompanying neurological clinical signs. Four dourine-afflicted horses in Mongolia were evaluated. An apparently healthy horse exhibited multifocal neuritis without axonal or myelin degeneration. The other horses, which had obvious neurological clinical signs, also exhibited multifocal neuritis. In particular, the nerves that innervated areas associated with neurological clinical signs exhibited neuritis with demyelination in the latter horses. Inflamed, non-demyelinating nerves were infiltrated with B lymphocytes and T lymphocytes; while inflamed, demyelinating nerves were infiltrated with mononuclear phagocytes. Our observations revealed lesion progression in the nerves, such that polyradiculoneuropathy could explain the accompanying neurological clinical signs of dourine. To our knowledge, this is the first report to describe a pathogenic mechanism for the development of the neurological clinical signs found in dourine-afflicted horses.
  • Batdorj Davaasuren, Junya Yamagishi, Daiki Mizushima, Sandagdorj Narantsatsral, Davaajav Otgonsuren, Punsantsogvoo Myagmarsuren, Badgar Battsetseg, Banzragch Battur, Noboru Inoue, Keisuke Suganuma
    Microbiology resource announcements 8 (9) 2019/02 [Refereed][Not invited]
     
    Trypanosoma equiperdum primarily parasitizes the genital organs and causes dourine in equidae. We isolated a new T. equiperdum strain, T. equiperdum IVM-t1, from the urogenital tract of a horse definitively diagnosed as having dourine in Mongolia. Here, we report the whole-genome sequence, the predicted gene models, and their annotations.
  • Daiki Mizushima, Tovuu Amgalanbaatar, Batdorj Davaasuren, Nthatisi Innocentia Molefe, Banzragch Battur, Badgar Battsetseg, Noboru Inoue, Naoaki Yokoyama, Keisuke Suganuma
    Parasitology research 117 (9) 2913 - 2919 2018/09 [Refereed][Not invited]
     
    Our previous studies report epidemics of non-tsetse-transmitted equine trypanosomosis in Mongolia. However, the current status of non-tsetse-transmitted equine trypanosomosis endemicity remains to be clarified in some parts of Mongolia. We previously reported the potential application of rTeGM6-4r-based diagnostic tools, an rTeGM6-4r-based immunochromatographic test (ICT) and an enzyme-linked immunosorbent assay (ELISA), in the serological surveillance of equine trypanosomosis in Mongolia. In the present study, the utility of the rTeGM6-4r-based ICT was validated. The rTeGM6-4r-based ICT accurately diagnosed positive reference sera that had been prepared from dourine horses in Mongolia, similarly to the rTeGM6-4r-based ELISA. The diagnostic performance of the rTeGM6-4r-based ICT was maintained when the strips were preserved for at least 2 months under dry conditions. The ICT detected 42 positive serum samples from a total of 1701 equine sera that had been collected from all 21 provinces of Mongolia. The κ-value, sensitivity and specificity of rTeGM6-4r-based ICT were 0.58, 50.0% (95% CI, 37.7-62.3%) and 99.3% (95% CI, 98.7-99.6%), respectively, in comparison to the rTeGM6-4r-based ELISA. Our field-friendly rTeGM6-4r-based ICT was found to be useful for the serological diagnosis of non-tsetse-transmitted equine trypanosomosis in rural areas of Mongolia.
  • モンゴルの牛におけるBabesia bovisとBabesia bigeminaの疫学マッピング(Mapping the epidemiology of Babesia bovis and Babesia bigemina in cattle in Mongolia)
    Battsetseg Badgar, Sivakumar Thillaiampalam, Narantsatsral Sandagdorj, Myagmarsuren Punsantsogvoo, 水島 大貴, 五十嵐 郁男, Battur Banzragch, 横山 直明
    日本獣医学会学術集会講演要旨集 161回 319 - 319 1347-8621 2018/08
  • モンゴルにおける馬媾疫診断を促進する迅速診断キットの上市(Transferring a rapid test kit to market to facilitate diagnosis of dourine in Mongolia)
    Narantsatsral Sandagdorj, 菅沼 啓輔, 水島 大貴, 井上 昇, 横山 直明, Battur Banzragch, Battsetseg Badgar
    日本獣医学会学術集会講演要旨集 161回 319 - 319 1347-8621 2018/08
  • Badgar Battsetseg, Thillaiampalam Sivakumar, Khandsuren Naranbaatar, Sandagdorj Narantsatsral, Punsantsogvoo Myagmarsuren, Batsaikhan Enkhtaivan, Batdorj Davaasuren, Daiki Mizushima, Gayani Weerasooriya, Ikuo Igarashi, Banzragch Battur, Naoaki Yokoyama
    Veterinary Parasitology: Regional Studies and Reports 13 85 - 91 2405-9390 2018/08 [Refereed][Not invited]
     
    Mongolia is an agriculturally rich country with large livestock populations that contribute significantly to its national economy. However, the export market for live animals and livestock products is often constrained for various reasons including infectious diseases. Babesia bovis and B. bigemina, which are bovine hemoprotozoan parasites, cause severe forms of clinical babesiosis, in cattle. However, a country-wide survey to determine the exposure rates in various provinces in Mongolia was not conducted to determine the risk for infections with these parasite species. Therefore, we investigated the frequency of antibodies to B. bovis and B. bigemina in cattle reared throughout Mongolia. B. bovis-and B. bigemina-specific enzyme-linked immunosorbent assays (ELISAs) were used to screen the serum samples sourced from 1946 cattle in 19 of 21 provinces and a provincial municipality (Ulaanbaatar) in Mongolia. We found 351 (18.0%) samples positive for B. bovis and 435 (22.4%) samples positive for B. bigemina infections. The B. bovis- and B. bigemina-positive rates ranged from 0.8 to 61.5% and 4.0 to 50.6%, respectively, among the surveyed provinces. The positive rates of B. bovis and B. bigemina infections were relatively higher in the provinces located in northernmost, northern, eastern, southeastern, and southern Mongolia. Additionally, the B. bovis- and B. bigemina-positive rates were not significantly different between females (18.2 and 22.2%, respectively) and males (17.2 and 18.8%, respectively) or between the 1–3-year-old (16.2 and 19.4%, respectively) and > 3-year-old (17.1 and 20.9%, respectively) age groups. The differential seropositivity for B. bovis and B. bigemina infections among the provinces may reflect the variations in the risk of cattle being infected with these parasite species. The findings of the present study highlight the need for country-wide control measures, including tick control programs, to minimize the rates of B. bovis and B. bigemina infections in Mongolian cattle.
  • Zui Fujimoto, Naomi Kishine, Nobuhiro Suzuki, Ryuichiro Suzuki, Daiki Mizushima, Mitsuru Momma, Keitarou Kimura, Kazumi Funane
    BIOSCIENCE REPORTS 37 (2) 0144-8463 2017/04 [Refereed][Not invited]
     
    Paenibacillus sp. 598K cycloisomaltooligosaccharide glucanotransferase (CITase), a member of glycoside hydrolase family 66 (GH66), catalyses the intramolecular transglucosylation of dextran to produce CIs with seven or more degrees of polymerization. To clarify the cyclization reaction and product specificity of the enzyme, we determined the crystal structure of PsCITase. The core structure of PsCITase consists of four structural domains: a catalytic (beta/alpha)(8)-domain and three beta-domains. A family 35 carbohydrate-binding module (first CBM35 region of Paenibacillus sp. 598K CITase, (PsCBM35-1)) is inserted into and protrudes from the catalytic domain. The ligand complex structure of PsCITase prepared by soaking the crystal with cycloisomaltoheptaose yielded bound sugars at three sites: in the catalytic cleft, at the joint of the PsCBM35-1 domain and at the loop region of PsCBM35-1. In the catalytic site, soaked cycloisomaltoheptaose was observed as a linear isomaltoheptaose, presumably a hydrolysed product from cycloisomaltoheptaose by the enzyme and occupied subsites -7 to -1. Beyond subsite -7, three glucose moieties of another isomaltooiligosaccharide were observed, and these positions are considered to be distal subsites -13 to -11. The third binding site is the canonical sugar-binding site at the loop region of PsCBM35-1, where the soaked cycloisomaltoheptaose is bound. The structure indicated that the concave surface between the catalytic domain and PsCBM35-1 plays a guiding route for the long-chained substrate at the cyclization reaction.
  • Hisashi Iwata, Yosuke Kobayashi, Daiki Mizushima, Taisuke Watanabe, Jun Ogihara, Takafumi Kasumi
    AMB EXPRESS 7 (1) 45  2191-0855 2017/02 [Refereed][Not invited]
     
    Two transketolase isogenes, MmTKL1 and MmTKL2, isolated from Moniliella megachiliensis were investigated for their roles in stress response and erythritol biosynthesis. The encoded proteins were highly homologous in amino acid sequence and domain structure. Two stress response elements (STREs) were found upstream of MmTKL1, while no STRE was found upstream of MmTKL2. In contrast, two Ap-1 elements were present upstream of MmTKL2, but none were detected upstream of MmTKL1. MmTKL2 partially complemented the aromatic amino acid auxotrophy of a Saccharomyces cerevisiae tkl1 deletion mutant, suggesting that at least one of the MmTKLs functioned as a transketolase in vivo. In response to short-term osmotic stress (20% glucose or 1.2 M NaCl) in Moniliella cells, MmTKL1 expression increased rapidly through the first 40 min before subsequently decreasing gradually, while MmTKL2 expression showed no significant change. In contrast, short-term oxidative stress (0.15 mM menadione) induced considerable increases in MmTKL2, while MmTKL1 expression remained low under the same conditions. Long-term osmotic stress (20% glucose) yielded increased expression of both genes starting at 12 h and continuing through 72 h. During either osmotic or oxidative stress, intracellular erythritol accumulation could clearly be correlated with the pattern of expression of either MmTKL1 or MmTKL2. These results strongly suggested that MmTKL1 is responsible primarily for the response to osmotic stress, while MmTKL2 is responsible primarily for the response to oxidative stress. Thus, we postulate that the two transketolase isoforms of M. megachiliensis play distinct and complementary roles in coordinating erythritol production in response to distinct environmental stresses.
  • Daiki Mizushima, Hisashi Iwata, Yuki Ishimaki, Jun Ogihara, Jun Kato, Takafumi Kasumi
    JOURNAL OF BIOSCIENCE AND BIOENGINEERING 121 (5) 523 - 529 1389-1723 2016/05 [Refereed][Not invited]
     
    Two isogenes of glycerol 3-phosphate dehydrogenase (GPD) from Candida versatilis SN-18 were cloned and sequenced. These intronless genes (Cagpd1 and Cagpd2) were both predicted to encode a 378 amino acid polypeptide, and the deduced amino acid sequences mutually showed 76% identity. Interestingly, Cagpd1 and Cagpd2 were located tandemly in a locus of genomic DNA within a 262 bp interval. To our knowledge, this represents a novel instance of isogenic genes relating to glucose metabolism. The stress response element (STRE) was found respectively at 93 to 89 bp upstream of the 5'end of Cagpd1 and 707 to 703 bp upstream of Cagpd2, indicating that these genes are involved in osmotic stress response. In heterologous expression using a gpd1 Delta gpd2 Delta double deletion mutant of Saccharomyces cerevisiae, Cagpd1 and Cagpd2 transformants complemented the function of GPD, with Cagpd2 being much more effective than Cagpd1 in promoting growth and glycerol synthesis. Phylogenetic analysis of the amino acid sequences suggested that Cagpd1p and Cagpd2p are NADP(+)-dependent GPD5 (EC 1.1.1.94). However, crude enzyme extract from Cagpd1 and Cagpd2 transformants showed GPD activity with only NAD(+) as cofactor. Hence, both Cagpd1p and Cagpd2p are likely NAD(+)-dependent GPD5 (EC 1.1.1.8), similar to GPDs from S. cerevisiae and Candida magnoliae. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.
  • Y. Kobayashi, H. Iwata, D. Mizushima, J. Ogihara, T. Kasumi
    LETTERS IN APPLIED MICROBIOLOGY 60 (5) 475 - 480 0266-8254 2015/05 [Refereed][Not invited]
     
    The number of naphtha plants is being reduced due to a worldwide shift in energy sources. Consequently, a shortage of chemical materials heavily dependent on naphtha-oil, especially C4 compounds such as butene and butane-diol, is an urgent issue in chemical manufacturing. Erythritol is a rare C4 compound produced by fermentation processes using glucose as the carbon source. Since erythritol is considerably more expensive than hydrocarbons derived from naphtha-oil, a reduction in its cost is critical. We found that Moniliella megachiliensis, a highly osmotolerant yeast strain, can utilize nonrefined glycerol waste derived from palm oil or beef tallow and convert it to erythritol. Cell growth on glycerol was almost the same as on glucose, and the cells could grow in up to 300mgml(-1) glycerol. When 200mgml(-1) nonrefined glycerol was supplied, the yield of erythritol from the glycerol was approx. 60%, which is slightly higher than that obtained using glucose. The cost of glycerol waste is considerably lower than that of glucose. Thus, the conversion of glycerol waste into valuable erythritol, proposed here, is attractive and promising from the viewpoint of ensuring a supply of C4 hydrocarbons and utilizing a waste natural resource. Significance and Impact of the StudyA shortage of C4 hydrocarbon depending much on naptha-oil has become urgent problem due to rapid reduction of naphtha plants together with global energy revolution. Erythritol, obtained by fermentation, is a rare C4 polyol that can be converted to C4 hydrocarbons. Erythritol is considerably expensive than hydrocarbons, a reduction in cost is critical issue. To meet this, we proposed to utilize low-cost glycerol waste from bio-diesel fuel as a carbon source. Moniliella megachiliensis successfully converted glycerol waste to erythritol. This proposal is promising to obtain C4 hydrocarbon substitute, and concomitantly to dispose a large amount of glycerol waste discharged.
  • Hisashi Iwata, Daiki Mizushima, Yosuke Kobayashi, Tetsuya Ookura, Jun Ogihara, Jun Kato, Takafumi Kasumi
    JOURNAL OF BIOSCIENCE AND BIOENGINEERING 119 (2) 148 - 152 1389-1723 2015/02 [Refereed][Not invited]
     
    We cloned and sequenced two transaldolase genes from Moniliella megachiliensis, a microorganism known to produce a significant amount of erythritol under hyper-osmotic stress. The amino acid sequences encoded by these two genes (MmTAL1, MmTAL2) showed 72% homology to each other. An AP-1 (ap response element) associated with oxidative stress was found in the promoter region of MmTAL1, while four STREs (stress response element) associated with osmotic stress were found in the promoter region of MmTAL2. In early-stage cultivation (up to 2 h), MmTAL1 was specifically expressed in response to oxidative stress generated by the presence of 0.15 mM menadione; expression level 3-fold higher than before stress loading. MmTAL2 was expressed in response to osmotic stress caused by 1.2 M NaCl; expression level was 21-fold higher than stress-free control. Erythritol accumulated intracellularly under osmotic and oxidative stress, approximately 30-fold and 35-fold, respectively. We therefore concluded that M. megachiliensis selectively uses two isogenes and produces erythritol during early-stage response to stress, depending on the type of environmental stress. (C) 2014, The Society for Biotechnology, Japan. All rights reserved.

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