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Journal of Bioscience and Bioengineering

Ei-Tora Yamamura
With the aim of being able to co-express multiple genes, I searched for novel compatible plasmids and isolated two plasmid species, pRET1100 and pRET1200, from Rhodococcus erythropolis IAM1400. Sequencing analysis revealed that the pRET1100 plasmid is a double-stranded DNA molecule of 5444 bp with two possible open reading frames (ORFs), repT and div, and three minor ORFs. The cryptic replication protein, RepT, is not highly homologous to those from other plasmids that have been reported. The Rhodococcus-Escherichia coli shuttle vector pRET1102 was transformed into R...
February 14, 2018: Journal of Bioscience and Bioengineering
Hirotoshi Sushida, Naoki Ishibashi, Takeshi Zendo, Pongtep Wilaipun, Vichien Leelawatcharamas, Jiro Nakayama, Kenji Sonomoto
EnkT is a novel ATP-binding cassette (ABC) transporter responsible for secretion of four bacteriocins, enterocins NKR-5-3A, C, D, and Z (Ent53A, C, D, and Z), produced by Enterococcus faecium NKR-5-3. It is generally recognized that the secretion of a bacteriocin requires a dedicated ABC transporter, although molecular mechanisms of this secretion are yet to be revealed. In order to characterize the unique ability of EnkT to secrete multiple bacteriocins, the role of N-terminal leader peptides of bacteriocin precursors was evaluated using Ent53C precursor as a model...
February 13, 2018: Journal of Bioscience and Bioengineering
Naoki Yamamoto, Rino Isshiki, Yuto Kawai, Daiki Tanaka, Tetsushi Sekiguchi, Shinya Matsumoto, Satoshi Tsuneda
Bacterial persisters are phenotypic variants that survive the treatment of lethal doses of growth-targeting antibiotics without mutations. Although the mechanism underlying persister formation has been studied for decades, how the persister phenotype is switched on and protects itself from antibiotics has been elusive. In this study, we focused on the lactate dehydrogenase gene (ldhA) that was upregulated in an Escherichia coli persister-enriched population. A survival rate assay using an ldhA-overexpressing strain showed that ldhA expression induced persister formation...
February 12, 2018: Journal of Bioscience and Bioengineering
Nguyen Thi Sam, Ryo Misaki, Takao Ohashi, Kazuhito Fujiyama
Sialic acid plays important roles in stabilization and modulation of the interaction of molecules and membranes in organisms. Due to its high electronegativity, sialic acid can promote binding effects of molecules and support the transportation of drugs and ions in cells. This also strengthens cells against degradation from glycosidases and proteases. Hence sialic acid helps glycoproteins extend their half-lives and bioactivity. On the other hand, Chinese hamster ovary (CHO) cells have been widely used as a workhorse in biopharmaceutical fields in part due to the similarity between their glycan properties and those in humans...
February 10, 2018: Journal of Bioscience and Bioengineering
Masatoshi Suganuma, Tsuyoshi Nomura, Yukiko Higa, Yukiko Kataoka, Shunsuke Funaguma, Hironobu Okazaki, Takeo Suzuki, Kazuhito Fujiyama, Hideki Sezutsu, Ken-Ichiro Tatematsu, Toshiki Tamura
A silkworm-baculovirus system is particularly effective for producing recombinant proteins, including glycoproteins. However, N-glycan structures in silkworm differ from those in mammals. Glycoproteins in silkworm are secreted as pauci-mannose type N-glycans without sialic acid or galactose residues. Sialic acid on N-glycans plays important roles in protein functions. Therefore, we developed pathways for galactosylation and sialylation in silkworm. Sialylated N-glycans on proteins were successfully produced in silkworm by co-expressing galactosyltransferase and sialyltransferase and providing an external supply of a sialylation-related substrate...
February 9, 2018: Journal of Bioscience and Bioengineering
Kenta Morita, Takahiro Suzuki, Yuya Nishimura, Kazuhisa Matsumoto, Chiya Numako, Kazuyoshi Sato, Masao Nakayama, Ryohei Sasaki, Chiaki Ogino, Akihiko Kondo
Polyacrylic acid (PAA)-modified titanium peroxide nanoparticles (PAA-TiOx NPs) are promising radiosensitizers. PAA-TiOx NPs were synthesized from commercial TiO2 nanoparticles that were modified with PAA and functionalized by H2O2 treatment. To realize practical clinical uses for PAA-TiOx NPs, their tissue distribution and acute toxicity were evaluated using healthy mice and mice bearing tumors derived from xenografted MIAPaCa-2 human pancreatic cancer cells. Healthy mice were injected with PAA-TiOx NPs at 25 mg/kg body weight via the tail vein, and tumor-bearing mice were injected either into the tumor locally or via the tail vein...
February 7, 2018: Journal of Bioscience and Bioengineering
Chun-Yen Chen, Yu-Han Chang
Microalgae cultivation using wastewater is an approach for simultaneous wastewater treatment and biofuels/chemicals productions. In this study, three microalgae species Chorella vulgaris ESP-31, Chorella sorokiniana CY-1 and Scenedesmus sp. were cultivated using coke-making wastewater generated from a steel-making company. Of these, C.vulgaris ESP-31 had the best tolerance towards wastewater, with maximal biomass concentration of 2.82 g/l and lipid productivity of 32.3 mg/l/d. The highest biomass concentration 3...
February 7, 2018: Journal of Bioscience and Bioengineering
Takatsugu Miyazaki, Tatsuya Kato, Enoch Y Park
β-1,2-N-Acetylglucosaminyltransferase II (GnTII, EC is a Golgi-localized type II transmembrane enzyme that catalyzes the transfer of N-acetylglucosamine to the 6-arm of the trimanosyl core of N-glycans, an essential step in the conversion of oligomannose-type to complex-type N-glycans. Despite its physiological importance, there have been only a few reports on the heterologous expression and structure-function relationship of this enzyme. Here, we constructed a silkworm-based Bombyx mori nucleopolyhedrovirus bacmid expression system and expressed human GnTII (hGnTII) lacking the N-terminal cytosolic tail and transmembrane region...
February 3, 2018: Journal of Bioscience and Bioengineering
Kumiko Kijima, Hajime Mita, Mitsuyasu Kawakami, Kei Amada
In the present study, we confirm that 2,4-dichlorophenoxyacetic acid (2,4-D) oxygenase from Sphingomonas agrestis 58-1 belongs to the family of Rieske non-heme iron aromatic ring-hydroxylating oxygenases, which comprise a core enzyme (oxygenase), ferredoxin, and oxidoreductase. It has previously been shown that cadAB genes are necessary for the conversion of 2,4-D to 2,4-dichlorophenol; however, the respective roles of ferredoxin and oxidoreductase in the 2,4-D oxygenase system from S. agrestis 58-1 remain unknown...
February 2, 2018: Journal of Bioscience and Bioengineering
Yuma Kato, Mee-Hae Kim, Masahiro Kino-Oka
Understanding the fundamental mechanisms that govern the growth kinetics of human induced pluripotent stem cells (hiPSCs) contributes to culture design strategies to improve large-scale production. Two hiPSC lines (Tic and 253G1) were cultured under static and dynamic suspension conditions, and growth kinetics were compared during early (24-48 h), middle (48-72 h), and late (72-96 h) stages. In 2D static culture, similar growth profiles were observed for both hiPSC lines. However, there were significant differences in growth profile patterns and aggregate morphologies between hiPSC lines grown in 3D static and dynamic cultures...
February 2, 2018: Journal of Bioscience and Bioengineering
Akihiro Mori, Teruyo Ojima-Kato, Takaaki Kojima, Hideo Nakano
A new antibody-enzyme fusion protein, named Zipbodyzyme, composed of a Fab antibody (i.e., an antigen-binding fragment of an antibody) and an enzyme, has been successfully produced in the cytoplasm of Escherichia coli. Zipbodyzymes have a leucine zipper (LZ) pair at the C-termini of the heavy chain (Hc) and the light chain (Lc) of Fab, to promote the association of the Hc and the Lc in E. coli cytoplasm, adjoining a fused enzyme. A Zipbodyzyme containing mouse-derived anti-E. coli O157 Fab and a luciferase or a green fluorescent protein retained both the antigen-binding and an enzymatic activity/fluorescence...
February 1, 2018: Journal of Bioscience and Bioengineering
Jingyin Wang, Hongwei Rong, Chaosheng Zhang
The effect of dissolved oxygen concentration (DO) during simultaneous nitrification and denitrification (SND) was investigated in a sequencing batch biofilm reactor (SBBR). In addition, the removal rates of nitrogen and bacterial communities were investigated under different concentrations of DO (1.5, 3.5, and 4.5 mg/L). When the SND rate was 95.22%, the chemical oxygen demand and nitrogen removal was 92.22% and 84.15%, respectively, at 2.5 mg/L DO. The denitrification was inhibited by the increase of oxygen concentration...
February 1, 2018: Journal of Bioscience and Bioengineering
Naoya Shinozaki, Ryuji Hashimoto, Masanori Noda, Susumu Uchiyama
Recently, we showed that immunized rabbit heavy chain variable regions (rVHs) can have strong antigen binding activity comparable to that of the camelid variable domain of the heavy chain of heavy chain antibody (VHH). These rVHs lack the light chain variable regions (rVLs), which exist in the authentic Fab format; thus, molecular surfaces at the interface region of rVHs are exposed to solvent. This physical feature may change physicochemical properties, such as causing reduced stability. By overcoming potential physicochemical issues through engineering the interface region, rVHs could become more useful as single-domain antibodies...
February 1, 2018: Journal of Bioscience and Bioengineering
Dian Yosi Arinawati, Keiko Miyoshi, Ayako Tanimura, Taigo Horiguchi, Hiroko Hagita, Takafumi Noma
The conventional two-dimensional (2D) in vitro culture system is frequently used to analyze the gene expression with or without extracellular signals. However, the cells derived from primary culture and cell lines frequently deviate the gene expression profile compared to the corresponding in vivo samples, which sometimes misleads the actual gene regulation in vivo. To overcome this gap, we developed the comparative 2D and 3D in vitro culture systems and applied them to the genetic study of amelogenesis imperfecta (AI) as a model...
January 31, 2018: Journal of Bioscience and Bioengineering
Xingxuan Chen, Xiahui Wang, Yiyun Xue, Tian-Ao Zhang, Yuhao Li, Jiajun Hu, Yiu Fai Tsang, Hongsheng Zhang, Min-Tian Gao
Rice straw can be used as carbon sources for lactic acid fermentation. However, only a small amount of lactic acid is produced even though Rhizopus oryzae can consume glucose in rice straw-derived hydrolysates. This study correlated the inhibitory effect of rice straw with rice straw-derived dissolved organic matter (DOM). Lactic acid fermentations with and without DOM were conducted to investigate the effect of DOM on lactic acid fermentation by R. oryzae. Fermentation using control medium with DOM showed a similar trend to fermentation with rice straw-derived hydrolysates, showing that DOM contained the major inhibitor of rice straw...
January 31, 2018: Journal of Bioscience and Bioengineering
Keita Fukui, Kei Nanatani, Yoshihiko Hara, Mitsunori Tokura, Keietsu Abe
Enterobacter aerogenes, a gram-negative, rod-shaped bacterium, is an effective producer of succinate from glucose via the reductive tricarboxylic acid cycle under anaerobic conditions. However, to date, succinate-exporter genes have not been identified in E. aerogenes, although succinate exporters have a large impact on fermentative succinate production. Recently, we genetically identified yjjP and yjjB, as genes encoding a succinate transporter in Escherichia coli. Evaluation of the yjjPB homologs in E. aerogenes (EayjjPB genes) showed that succinate accumulation increased from 4...
January 30, 2018: Journal of Bioscience and Bioengineering
April N Htet, Mana Noguchi, Kazuaki Ninomiya, Yota Tsuge, Kosuke Kuroda, Shinya Kajita, Eiji Masai, Yoshihiro Katayama, Kazuhiro Shikinaka, Yuichiro Otsuka, Masaya Nakamura, Ryo Honda, Kenji Takahashi
Actual biomass of microalgae was tested as a fermentation substrate for microbial production of 2-pyrone 4,6-dicarboxylic acid (PDC). Acid-hydrolyzed green microalgae Chlorella emersonii (algae hydrolysate) was diluted to adjust the glucose concentration to 2 g/L and supplemented with the nutrients of Luria-Bertani (LB) medium (tryptone 10 g/L and yeast extract 5 g/L). When the algae hydrolysate was used as a fermentation source for recombinant Escherichia coli producing PDC, 0.43 g/L PDC was produced with a yield of 20...
January 29, 2018: Journal of Bioscience and Bioengineering
Zhi-Peng Zhang, Jun Ma, Yuan-Yuan He, Jun Lu, Di-Feng Ren
Diospyros lotus, a member of the Ebenaceae family, has long been used as a traditional sedative in China. In this study, the antioxidant and hypoglycemic effects of non-fermented and microorganism-fermented D. lotus were explored. The total phenolic and vitamin C contents of microorganism-fermented D. lotus for 24-72 h were less than those of non-fermented. High-performance liquid chromatography showed that the tannic, catechinic, and ellagic acid contents increased significantly upon fermentation for 24 h...
January 27, 2018: Journal of Bioscience and Bioengineering
Elisabeth Bermejo, María C Ruiz-Domínguez, María Cuaresma, Isabel Vaquero, Adrian Ramos-Merchante, José M Vega, Carlos Vílchez, Inés Garbayo
In this study, the effect of abiotic stress on the acidophilic eukaryotic microalga, Coccomyxa onubensis, was analyzed for the production of lutein and PUFAs (polyunsaturated fatty acids). It grows autotrophically at a pH of 2.5. It showed a growth rate of 0.30 d-1, and produced approximately 122.50 mg.L-1.d-1 biomass, containing lipids (300.39 mg g-1dw), lutein (5.30 mg g-1dw), and β-carotene (1.20 mg g-1dw). The fatty acid methyl ester (FAME) fraction was 89.70 mg g-1dw with abundant palmitic acid (28...
January 20, 2018: Journal of Bioscience and Bioengineering
Risa Harada, Masanobu Yuzuki, Kotaro Ito, Kazuki Shiga, Takeshi Bamba, Eiichiro Fukusaki
Soy sauce is a traditional Japanese fermented seasoning that contains various constituents such as amino acids, organic acids, and volatiles that are produced during the long fermentation process. Although studies regarding the correlation between microbes and aroma constituents have been performed, there are no reports about the influences of the microbial products, such as lactic acid, acetic acid, and ethanol, during fermentation. Because it is known that these compounds contribute to microbial growth and to changes in the constituent profile by altering the moromi environment, understanding the influence of these compounds is important...
January 20, 2018: Journal of Bioscience and Bioengineering
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