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Biotechnology and Applied Biochemistry

Eiichiro Takamura, Takuto Nakamura, Hiroaki Sakamoto, Takenori Satomura, Haruhiko Sakuraba, Toshihisa Ohshima, Shin-Ichiro Suye
In this study, multicopper oxidase (MCO) was immobilized on multi-walled carbon nanotubes (MWCNTs) at two different orientations, and the electrochemical properties of the resulting cathodes were investigated. Using N- or C-terminal His-tagged MCO and MWCNTs, we constructed two types of cathodes. We assumed that the distance between the Type 1 (T1)Cu of the C-terminal His-tagged MCO and the MWCNT surface was smaller than that between the T1Cu of the N-terminal His-tagged MCO and the MWCNT surface. In addition, in the C-terminal His-tagged MCO, the T1Cu was expected to be closer to the MWCNT surface than the type 2/type 3 Cu site...
November 14, 2018: Biotechnology and Applied Biochemistry
Anupong Tankrathok, Arpaporn Punpad, Monrudee Kongchaiyapoom, Sirinthip Sosiangdi, Nisachon Jangpromma, Sakda Daduang, Sompong Klaynongsruang
Cathelicidins are effector molecules of vertebrate immunity that play vital roles against microbial invasion. They are widely identified in the mammal, but few have been reported in Crocodilians with considered as powerful immune system species. In the present study, we identified and characterized a novel cathelicidin from the blood of Siamese crocodile, Crocodylus siamensis. A cDNA sequence (501 base pair (bp)) encodes a predicted 166-residue prepropeptide of C. siamensis cathelicidin (Cs-CATH), which comprised of a 21-residue signal peptide, a 109-residue cathelin domain, and a 36-residue mature cathelicidin peptide...
November 10, 2018: Biotechnology and Applied Biochemistry
Monireh ParvizHamidi, Ghazal Haddad, Shiva Ostadrahimi, Nima Ostadrahimi, Sedigheh Sadeghi, Shima Fayaz, Pezhman Fard-Esfahani
Glioblastoma multiform is the most common and lethal primary central nervous system tumor. Circulating microRNAs (miRNAs), present in cell-free bodily fluids, have been gaining importance as cancer biomarkers. The primary aim of this study was to assess whether circulating miRNA-128, -21, -26a in glioblastoma patients can be used as diagnostic biomarkers. Venous blood samples were collected from 11 non-cancerous volunteers and 15 glioblastoma patients pre- and post-operationally. Also, tissue tumor samples were obtained intra-operationally to assay consistency of miRNA levels in serum and tissue samples...
November 8, 2018: Biotechnology and Applied Biochemistry
Masahiro Watanabe, Takao Toyomura, Hidenori Wake, Keyue Liu, Kiyoshi Teshigawara, Hideo Takahashi, Masahiro Nishibori, Shuji Mori
Previously, we found that endogenously produced pro-inflammatory molecules, advanced glycation end products (AGEs), interact with tumor necrosis factor-like weak inducer of apoptosis (TWEAK), and attenuate its immunomodulatory function. In the present study, to elucidate the mechanism by which AGEs attenuate TWEAK function, we searched for regions responsible for TWEAK-AGE interaction using TWEAK deletion mutants. Pull-down assays with the TWEAK mutants and AGEs revealed that the C-terminal half of TWEAK, which is the region essential for receptor stimulation, was required for this interaction...
November 7, 2018: Biotechnology and Applied Biochemistry
Ali Seidkhani-Nahal, Abdolamir Allameh, Masoud Soleimani
This study was carried out to examine the role of intracellular albumin in the modulation of oxidative damage induced by glutathione modifiers in HepG2 cells. Also, the relationship of albumin synthesis with the oxidative stress factors including antioxidants was studied. HepG2 cell culture was supplemented with glutathione modifiers; L-Buthionine-sulfoximine (BSO; 0.1 and 1.0 mM) or N-acetyl cysteine (NAC; 1 and 10 mM) and the cell viability and changes in reduced (GSH), oxidized glutathione (GSSG), reactive oxygen species (ROS), catalase and superoxide dismutase (SOD) were measured...
November 7, 2018: Biotechnology and Applied Biochemistry
Jie Sun, Jie Jiang, Lifeng Liu, Zhao Wang, Chun Wei
The use of lactoferrin antimicrobial peptides and lysozymes as traditional antibiotic alternatives is suitable for solving drug residue and pathogen resistance. In this study bovine lactoferrin (LfcinB) and human lysozyme (hLY) were combined through fusion expressionin Pichia pastoris GS115 driven by constitutive GAP promoter. For neutralizing the toxic property of the antimicrobial peptide, anion antioxidant peptides from porcine myofibrillar protein and enzymatically hydrolyzed chicken egg white were fused to the hybrid antimicrobial peptide LfcinB-hLY...
November 1, 2018: Biotechnology and Applied Biochemistry
Roghayyeh Seyfi, Valiollah Babaeipour, Mohammad Reza Mofid, Fatemeh Abarghooi Kahaki
Scorpine is a cationic protein from the venom of Pandinus imperator, belonging to potassium channel blocker family, which has been shown to have antibacterial, antiviral, and antiplasmodia activities. In the present study, a pET-44a vector containing scorpine synthetic gene with T7 Promoter (pET 44a-His6-Nus-His6-tev-scorpine) was transferred into Escherichia coli Rosetta-gami B (DE3) for soluble expression of the protein in the cytoplasm and its overproduction. After confirming recombinant scorpine peptide expression using SDS-PAGE and Western blot, augmentation of production was performed during two stages...
October 31, 2018: Biotechnology and Applied Biochemistry
Jue Li, Guiguang Chen, Ren Zhang, Hao Wu, Wei Zeng, Zhiqun Liang
In this study, a bacterial cellulose (BC) producing strain was isolated from Kombucha tea and identified as Komagataeibacter hansenii JR-02 by morphological, physiological, biochemical characterization and 16S rRNA sequence. Then, the media components and culture conditions for BC production were optimized. Result showed that the highest BC yield was 3.14 ± 0.22 g/L and 8.36 ± 0.19 g/L after fermentation for 7 days under shaking and static cultivation, respectively. Moreover, it was interesting that JR-02 could produce BC in nitrogen-free medium with the highest yield of 0...
October 25, 2018: Biotechnology and Applied Biochemistry
Yusuf Sürmeli, Hüseyin İlgü, Gülşah Şanlı-Mohamed
α-L-Arabinofuranosidase (Abf) is a potential enzyme because of its synergistic effect with other hemicellulases in agro-industrial field. In this study, directed evolution was applied to Abf from Geobacillus vulcani GS90 (GvAbf) using one round error-prone PCR and constructed a library of 73 enzyme variants of GvAbf. The activity screening of the enzyme variants was performed on soluble protein extracts using p-nitrophenyl α-L-arabinofuranoside as substrate. Two high activity displaying variants (GvAbf L307S and GvAbf Q90H/L307S) were selected, purified, partially characterized, and structurally analyzed...
October 17, 2018: Biotechnology and Applied Biochemistry
Maryam Gazor, Seyed Siamak Ashraf Talesh, Seyed Nezamedin Hosseini, Amin Javidanbardan, Maryam Khatami
In our previous study, we demonstrated that continuous power laser could be a clean, rapid, and convenient alternative to the other conventional disruption techniques for the release of recombinant hepatitis B surface antigen (rHBsAg) from Pichia pastoris. In the current work, we examined the effect of pulsed laser in the continuous laboratory-scale process on cell disruption. Design-of-experiments (DOE) methodology was used for optimization of cell disruption process to obtain the highest protein concentration in the disruption buffer...
October 14, 2018: Biotechnology and Applied Biochemistry
Takami Sho, JianXiong Xu
Inflammation, as a common immune response to various infections or injuries, can cause many dangerous and complicated diseases. Inflammasome is a protein complex playing a vital role in an inflammation process, and the nucleotide-binding oligomerization domain (NOD)-like receptor containing pyrin domain 3 (NLRP3) inflammasome has been the most-widely studied one. Recent evidence suggests the reactive oxygen species (ROS)-NLRP3 signaling pathway to be a possible NLRP3 inflammasome regulation model. Numerous recent preclinical reports indicate that application of antioxidants could scavenge excessive ROS and attenuate inflammatory responses through suppressing NLRP3 inflammasome activation...
October 13, 2018: Biotechnology and Applied Biochemistry
Zhanying Zhang, Yanfei Wang, Yanlei Li, Dongsheng Yu, Haiyan Chen, Yi Cai, Weikai Fang, Zhe Yang, Yufeng Ji, Yifu Guan, Yannan Chu, Chidong Xu
MicroRNAs (miRNAs) diagnostics can be useful for diagnosing or confirming miRNA abundance and are used in screening tests and to assess changes in miRNAs in vivo. At present, the use of traditional nucleic acid amplification assays to detect miRNAs has been limited in laboratory environment because of the time, equipment, and technical expertise required to perform these assays. A specialized, rapid affordable miRNA detection system is necessary when there are limited resources or point-of-care testing needs...
October 12, 2018: Biotechnology and Applied Biochemistry
Wenjian Ma, Jianli Wang, Ye Li, Xiaoyuan Wang
Cysteine synthase A (CysK) catalyzes the last reaction of l-cysteine synthesis in bacteria, but its moonlighting functions have been revealed recently. In this study, CysK was overexpressed in Corynebacterium glutamicum IWJ001, an l-isoleucine producer. Compared with the control IWJ001/pDXW-8, IWJ001/pDXW-8-cysK cells grew fast during log phase, and produced 26.5% more l-isoleucine in flask fermentation and 23.5% more l-isoleucine in fed-batch fermentation. The key genes aspC, lysC, hom, thrB, ilvA, and ilvBN involved in l-isoleucine biosynthesis were all upregulated in IWJ001/pDXW-8-cysK, compared with IWJ001/pDXW-8...
October 12, 2018: Biotechnology and Applied Biochemistry
Tianzhu Guan, Yonghai Sun, Tiezhu Li, Ligang Hou, Jie Zhang, Yongjun Wang
Human exposure to bisphenol compounds (BPs) has been implicated in the development of several chronic diseases. Instead of exploiting the traditional methods for determination of BPs, this work confirms that the human estrogen receptor α ligand binding domain (hERα-LBD) is a powerful recognition element that can be used to monitor multi-residue of BPs in urine samples by fluorescence polarization (FP) assay. Test parameters were optimized for the best performance. Under the optimal conditions, the IC50 values of BPs are in the range of 0...
October 11, 2018: Biotechnology and Applied Biochemistry
Ayodeji O Falade, Leonard V Mabinya, Anthony I Okoh, Uchechukwu U Nwodo
The industrial applications and prospects of microbial peroxidase are on the upwards trend, thus necessitating the search for sources with high turnaround time. Actinobacterial species have been a major source of peroxidase for the obvious reasons of having robust metabolite expression capabilities. However, other bacteria species have been underexplored for peroxidase production, hence the motivation for the investigation into the peroxidase production potential of Raoultella ornithinolytica OKOH-1 (KX640917)...
October 10, 2018: Biotechnology and Applied Biochemistry
Laura S Mazzaferro, Gisela Weiz, Lucas Braun, Michael Kotik, Helena Pelantová, Vladimír Křen, Javier D Breccia
The structure of the carbohydrate moiety of a natural phenolic glycoside can have a significant effect on the molecular interactions and physicochemical and pharmacokinetic properties of the entire compound, which may include anti-inflammatory and anticancer activities. The enzyme 6-O-α-rhamnosyl-β-glucosidase (EC has the capacity to transfer the rutinosyl moiety (6-O-α-l-rhamnopyranosyl-β-d-glucopyranose) from 7-O-rutinosylated flavonoids to hydroxylated organic compounds. This transglycosylation reaction was optimized using hydroquinone (HQ) and hesperidin as rutinose acceptor and donor, respectively...
October 7, 2018: Biotechnology and Applied Biochemistry
Amir Aidun, Ali Zamanian, Farnaz Ghorbani
So far, many studies have focused on biodegradable scaffolds for tissue engineering purposes. Herein, a starch-based biodegradable scaffold was fabricated by the freeze-drying method and cross-linked using a different concentration of 3-glycidoxypropyl-trimethoxysilane (GPTMS). Field emission scanning electron microscopy (FE-SEM) micrographs indicated an interconnected porous microstructure in which the porosity decreased as a function of starch and GPTMS content. Increasing the mechanical stability and decreasing absorption capacity and biodegradation ratio affected by the higher concentration of cross-linker and the changes in structure as a function of cross-linker enhancement...
September 26, 2018: Biotechnology and Applied Biochemistry
Jun Yang, Jing Bai, Mingbo Qu, Bo Xie, Qing Yang
A nitroreductase-encoded gene from an efficient nitro-reducing bacterium Streptomyces mirabilis DUT001, named snr, was cloned and heterogeneously expressed in Escherichia coli. The purified Streptomyces nitroreductase SNR was a homodimer with an apparent subunit molecular weight of 24 kDa and preferred NADH to NADPH as a cofactor. By enzyme incubation and isothermal calorimetry experiments, flavin mononucleotide (FMN) was found to be the preferred flavin cofactor; the binding process was exothermic and primarily enthalpy driven...
September 19, 2018: Biotechnology and Applied Biochemistry
Xiaojun Liu, Zhenming Liu, Can Jin, Haoran Li, Jia-Le Li
Matrix proteins play important roles in molluscan shell biomineralization, which helps in the understanding of mechanisms associated with pearl formation. In this study, we characterized the gene encoding a novel shell-matrix protein, hic24, in Hyriopsis cumingii and investigated its structure and function. The full cDNA sequence of hic24 is 756 bp, with an open reading frame of 654 bp encoding 217 amino acids, including a signal peptide of 18 amino acids. Sequence analysis revealed that the protein is ∼23...
September 8, 2018: Biotechnology and Applied Biochemistry
Lianghua Shen, Shanze Yi, Luyuan Huang, Shuaiguang Li, Fang Bai, Sijia Lei, Mason Breitzig, Alexander Czachor, Hanxiao Sun, Qing Zheng, Feng Wang
Lung cancer is a serious threat to human health. Studies have revealed that human manganese superoxide dismutase (hSOD2) and miRNAs play an essential role in the metastasis process of lung cancer. However, the miRNAs that associated with hSOD2 and involved in metastasis, remain elusive. After databases analysis and dual luciferase reporter validation, we demonstrated that miR-330-3p expression inversely correlated with hSOD2b expression level, and that miR-330-3p directly targeted the 3'untranslated region (3'UTR) of hSOD2b...
September 7, 2018: Biotechnology and Applied Biochemistry
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