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Journal of Bioscience and Bioengineering

Ryushi Kawakami, Haruhiko Sakuraba, Taketo Ohmori, Toshihisa Ohshima
A novel amino acid racemase with broad substrate specificity (BAR) was recently isolated from the hyperthermophilic archaeon Pyrococcus horikoshii OT-3. Characterization of this enzyme has been difficult, however, because the recombinant enzyme is produced mainly as an inclusion body in Escherichia coli. In this study, expression of the recombinant protein into the soluble fraction was markedly improved by co-expression with chaperone molecules. The purified enzyme retained its full activity after incubation at 80°C for at least 2 h in buffer (pH 7-10), making this enzyme the most thermostable amino acid racemase so far known...
March 24, 2017: Journal of Bioscience and Bioengineering
Makoto Matsui, Tatsuya Yokoyama, Kaoru Nemoto, Toshitaka Kumagai, Koichi Tamano, Masayuki Machida, Takashi Shibata
FR901469 is a secondary metabolite with antifungal activity, produced by fungal sp. No. 11243. In our previous study, we constructed the frbF overexpression mutant (TFH2-2) from the wild-type strain. FR901469 productivity of TFH2-2 was 3.4 times higher than that of the wild-type strain. To further enhance FR901469 productivity in TFH2-2, we attempted to find genes from the genome that limited the productivity as bottlenecks in this study. Based on both correlation analysis of gene expression level against FR901469 productivity and genome annotation information, the cross-pathway control gene A (cpcA) was most predicted as the bottleneck...
March 21, 2017: Journal of Bioscience and Bioengineering
Kouki Matsuo, Takeshi Matsumura
The production of recombinant proteins in plants has many advantages, including safety and reduced costs. However, this technology still faces several issues, including low levels of production. The repression of RNA silencing seems to be particularly important for improving recombinant protein production because RNA silencing effectively degrades transgene-derived mRNAs in plant cells. Therefore, to overcome this, we used RNA interference technology to develop DCL2- and DCL4-repressed transgenic Nicotiana benthamiana plants (ΔD2, ΔD4, and ΔD2ΔD4 plants), which had much lower levels of NbDCL2 and/or NbDCL4 mRNAs than wild-type plants...
March 20, 2017: Journal of Bioscience and Bioengineering
Yu-Kuo Liu, Wei-Chuan Chen, Yu-Ching Huang, Yu-Kaung Chang, I-Ming Chu, Shen-Long Tsai, Yu-Hong Wei
The aim of this study was to use a modified bioreactor system for simultaneous saccharification of cellulose and bioethanol production. We tested Aspergillus niger and Trichoderma reesei for cellulose saccharification and Zymomonas mobilis for bioethanol production simultaneously in this modified bioreactor. The results showed that various carboxymethylcellulose (CMC) concentrations (10, 15, or 20 g/L) as a substrate for A. niger and T. reesei yielded bioethanol production of 0.51, 0.78, and 0.89 g/L and a CMC conversion rate of 10...
March 18, 2017: Journal of Bioscience and Bioengineering
Yasutaka Hirokawa, Ryota Goto, Yoshitaka Umetani, Taizo Hanai
Using engineered cyanobacteria to produce various chemicals from carbon dioxide is a promising technology for a sustainable future. Lactate is a valuable commodity that can be used for the biodegradable plastic, polylactic acid. Typically, lactate production using engineered cyanobacteria was via the conversion of pyruvate in glycolysis by lactate dehydrogenase. In cyanobacteria, the metabolic flux in the Calvin cycle is higher than that in glycolysis under photoautotrophic conditions. The construction of a novel lactate producing pathway that uses metabolites from the Calvin cycle could potentially increase lactate productivity in cyanobacteria...
March 18, 2017: Journal of Bioscience and Bioengineering
Wan Yng Linda Jong, Pau Loke Show, Tau Chuan Ling, Yee Shin Tan
Amauroderma rugosum is a wild mushroom species widely distributed in tropics and is classified under the class of Basidiomycetes. Basidiomycetes are well-known for their abilities of producing lignocellulolytic enzymes such as lignin peroxidase (LiP), laccase (Lac) and manganese peroxidase (MnP). Different factors such as nutrient sources, incubation period and agitation affect the production of lignocellulolytic enzymes. The A. rugosum produced LiP in the medium supplemented with potato dextrose broth (PDB), 0...
March 16, 2017: Journal of Bioscience and Bioengineering
Thuy Duong Nguyen, Hitoshi Takasuka, Yoshihiro Kaku, Satoshi Inoue, Teruyuki Nagamune, Masahiro Kawahara
Intracellular antibodies (intrabodies) are expected to function as therapeutics as well as tools for elucidating in vivo function of proteins. In this study, we propose a novel intrabody selection method in the cytosol of mammalian cells by utilizing a growth signal, induced by the interaction of the target antigen and an scFv-c-kit growth sensor. Here, we challenge this method to select specific intrabodies against rabies virus nucleoprotein (RV-N) for the first time. As a result, we successfully select antigen-specific intrabodies from a naïve synthetic library using phage panning followed by our growth sensor-based intracellular selection method, demonstrating the feasibility of the method...
March 16, 2017: Journal of Bioscience and Bioengineering
Sekar Sudhakar, Rasool Abdul Nazeer
Antioxidant peptides protect biological macromolecules against radical damages. The use of these peptides was evaluated using free radicals scavenging assays [2,2-diphenyl-1 picrylhydrazyl (DPPH) and hydroxyl] with the help of UV-visible and electron spin resonance (ESR) spectroscopy methods. The Octopus aegina mantle protein were tested upon hydrolysis using gastrointestinal enzymes up to 12 h, where pepsin hydrolysate exhibited superior properties (DPPH: 44.39±0.67% and hydroxyl: 38.84±1.07%) compared with trypsin and α-chymotrypsin...
March 16, 2017: Journal of Bioscience and Bioengineering
Ming-Chen Xia, Ya-Ping Wang, Tang-Jian Peng, Li Shen, Run-Lan Yu, Yuan-Dong Liu, Miao Chen, Jiao-Kun Li, Xue-Ling Wu, Wei-Min Zeng
To seek a feasible technique for processing waste printed circuit boards (PCBs), pretreatment of PCBs by table separation and further bioleached by moderate thermophiles in a stirred tank reactor were investigated. The shaking table separation, conducted after grinding and sieving of PCBs, produced two fractions: metal-rich parts (RPCBs), which is more suitable for pyrometallurgy process than untreated PCBs, and metal-poor parts (PPCBs) with only 8.83% metals was then bioleached by a mixed culture of moderate thermophiles effectively...
March 15, 2017: Journal of Bioscience and Bioengineering
Yen-Chun Lai, Chih-Ming Liang, Shu-Chuan Hsu, Ping-Heng Hsieh, Chun-Hsiung Hung
A purple non-sulfur bacterium, Rhodopseudomonas palustris G11, was isolated from an activated sludge plant that treats domestic wastewater. This isolation resulted in the effective accumulation of polyphosphate in cells upon reaching the stationary growth phase. However, when the carbon and/or energy source were/was removed, this bacterium released intracellular polyphosphate or poly-β-hydroxybutyrate to obtain energy to grow or maintain its growth. Furthermore, a novel photo-microbial fuel cell (PMFC) design was proposed...
March 11, 2017: Journal of Bioscience and Bioengineering
Shin Nishiumi, Keisuke Shima, Takeshi Azuma, Masaru Yoshida
Metabolomics has recently been developed, and there have been a considerable number of metabolomics-based biomarker studies in the medical research field. Therefore, as a first step toward the practical use of metabolite biomarkers, a simple and quick sample preparation method involving metabolite extraction, metabolite measurement, and data analysis needs to be developed. In this study, we evaluated whether the use of simpler metabolite extraction methods would facilitate the stable analysis of hydrophilic blood metabolites during liquid chromatography/triple quadrupole mass spectrometry (LC/QqQMS)-based metabolome analysis...
March 10, 2017: Journal of Bioscience and Bioengineering
Menglu Li, Masahiro Kino-Oka
Tumor angiogenesis is an important event in tumor malignancy; and the vasculature formed in tumor region is typically dysfunctional. Multiple factors are associated with tumor vessel abnormalities, but the precise mechanism has not been fully understood. In the present study, a tumor-containing cell sheet was prepared by mixing a small population of human embryonal rhabdomyosarcoma (RMS) cells (RDs) with human skeletal muscle myoblasts (HSMMs) to mimic muscle tissue invaded by RMS cells. Sheet fluidity and the extracellular matrix (ECM) meshwork of the tumor-containing cell sheet were found to be elevated and disordered, demonstrating the disruptive effect of tumor cells on sheet structure...
March 10, 2017: Journal of Bioscience and Bioengineering
Takehiko Shimohira, Katsuhisa Kurogi, Takuyu Hashiguchi, Ming-Cheh Liu, Masahito Suiko, Yoichi Sakakibara
Dietary polyphenols present in fruits and vegetables have been reported to manifest beneficial health effects on humans. Polyphenol metabolites including their sulfated derivatives have been shown to be biologically active. Primarily due to the difficulty in preparing regiospecific sulfated polyphenols for detailed investigations, the exact functions of sulfated polyphenols, however, remain unclear. The current study aimed to develop a procedure for the regioselective production of sulfated polyphenols using Escherichia coli cells expressing human cytosolic sulfotransferases (SULTs)...
March 9, 2017: Journal of Bioscience and Bioengineering
Hsiao-Hsien Lin, Tsung-Yih Lee, Ting-Wei Liu, Ching-Ping Tseng
Glucose is a carbon source for Chinese hamster ovary (CHO) cell growth, while low growth rate is considered to enhance the production of recombinant proteins. The present study reveals that glucose concentrations higher than 1 g/L reduce the growth rate and substantially increase in cAMP (∼300%) at a high glucose concentration (10 g/L). High glucose also enhances the phosphorylation of extracellular signal-regulated kinase (ERK) and p27(kip) by Western blot analysis. To determine whether the phosphorylation of ERK is involved in the mechanism, a cyclic-AMP dependent protein kinase A (PKA) inhibitor (H-8) or MEK (MAPKK) inhibitor (PD98059) was added to block ERK phosphorylation...
March 9, 2017: Journal of Bioscience and Bioengineering
Toshinari Takahashi, Yusuke Ohara, Kazuo Sueno
Sake yeast produces a fruity flavor known as ginjo-ko-which is mainly attributable to ethyl caproate and isoamyl acetate-during fermentation in sake brewing. The production of these flavor components is inhibited by unsaturated fatty acids derived from the outer layer of rice as raw material. We isolated three mutants (hec2, hec3, and hec6) with enhanced ethyl caproate productivity in sake brewing using rice milled at a high polishing ratio from a cerulenin-resistant mutant derived from the hia1 strain, which shows enhanced isoamyl acetate productivity...
March 9, 2017: Journal of Bioscience and Bioengineering
Akihiko Ono, Akira Ito, Tomonaga Sato, Masaki Yamaguchi, Taiga Suzuki, Yoshinori Kawabe, Masamichi Kamihira
Precise control of gene expression using an artificial gene circuit is a major challenge in the application of synthetic biology. Here, we designed a hypoxia-responsive transgene expression system by combining a hypoxia-inducible RTP801 promoter and a tetracycline-responsive transactivator fused with an oxygen-dependent degradation domain (TA-ODD). The reporter gene expression was highly induced by hypoxia when a transactivator-expression plasmid, pRTP801/TA-ODD, harboring a TA-ODD gene driven by the RTP801 promoter, was cotransfected with a reporter plasmid, pTRE/EGFP, harboring an EGFP gene controlled under the transactivator-responsive promoter...
March 9, 2017: Journal of Bioscience and Bioengineering
Yu-Long Li, Jin Wang, Zheng-Bo Yue, Wei Tao, Hai-Bin Yang, Yue-Fei Zhou, Tian-Hu Chen
Biological treatment played an important role in the treatment of landfill leachate. In the current study, acid mine drainage (AMD) was used as a source of sulfate to strengthen the anaerobic treatment of landfill leachate. Effects of chemical oxygen demand (COD) and SO4(2-) mass concentration ratio on the decomposition of organic matter, methane production and sulfate reduction were investigated and the microbial community was analyzed using the high throughout methods. Results showed that high removal efficiency of COD, methane production and heavy metal removal was achieved when the initial COD/SO4(2-) ratio (based on mass) was set at 3...
March 6, 2017: Journal of Bioscience and Bioengineering
Nalok Dutta, Subrata Biswas, Malay Kumar Saha
In the current literature we have devised an immobilization technique for conferring psychrostability to a cold active α-amylase (amy) enzyme by the use of magnesium nanoparticle (MgNP) and graphene oxide (GO). The GO-MgNP-amy nanocomposite showed enhanced enzymatic activity and thermostability at both upper (90°C) and lower (8°C) temperature extremes. The GO-MgNP-amy showed increased affinity towards substrate, reflected in the decrease in its Km by 2.35 and 14.9-fold at 8°C and 90°C, respectively, than the untreated enzyme...
March 3, 2017: Journal of Bioscience and Bioengineering
Takeshi Kawabata, Masayuki Oda, Fusako Kawai
The cutinase-like enzyme, Cut190, from Saccharomonospora viridis AHK190 can degrade the inner block of polyethylene terephthalate (PET) in the presence of Ca(2+), and its mutant, S226P/R228S, exhibited increased activity and higher thermostability. The crystal structures of the Cut190 S226P mutant in the absence and presence of Ca(2+) were determined, and revealed the large conformational change induced upon Ca(2+) binding. However, the substrate-bound 3D structures of Cut190 remained unknown. In this study, to determine the substrate-binding site and improve the enzyme activity, we first built 3D structures of a PET model compound bound to the crystal structures, using the distance restraints between the scissile carbonyl group of the compound and the catalytic site of the enzyme...
March 1, 2017: Journal of Bioscience and Bioengineering
Daisuke Mitsuya, Takuya Hayashi, Yu Wang, Mami Tanaka, Masahiko Okai, Masami Ishida, Naoto Urano
The Yukawa River is an extremely acidic river whose waters on the east foot of the Kusatu-Shirane Volcano (in Gunma Prefecture, Japan) contain sulfate ions. Here we isolated many acid-tolerant yeasts from the Yukawa River, and some of them neutralized an acidic R2A medium containing casamino acid. Candida fluviatilis strain CeA16 had the strongest acid tolerance and neutralizing activity against the acidic medium. To clarify these phenomena, we performed neutralization tests with strain CeA16 using casamino acid, a mixture of amino acids, and 17 single amino acid solutions adjusted to pH 3...
February 28, 2017: Journal of Bioscience and Bioengineering
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