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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 enzyme 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, FMN was found to be the preferred flavin cofactor; the binding process was exothermic and primarily enthalpy driven. The enzyme can reduce multiple nitro compounds and flavins, including antibacterial drug nitrofurazone, priority pollutants 2,4-dinitrotoluene and 2,4,6-trinitrotoluene, as well as key chemical intermediates 3-nitrophthalimide, 4-nitrophthalimide and 4-nitro-1,8-naphthalic anhydride...
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'un-translated region (3'UTR) of hSOD2b...
September 7, 2018: Biotechnology and Applied Biochemistry
Sadegh Rezapour, Mustafa Shiravand, Mahnaz Mardani
One of the epigenetic-modifying factors is regular and continuous physical activity. This article attempts to investigate the effects of physical activity and exercise on changes in histone proteins and gene expression, as well as the effect of these exercises on the prevention of certain cancers and the ejection of age-related illnesses and cellular oxidation interactions. All of this is due to epigenetic changes and gene expression. Most studies have reported the positive effects of regular exercises on the expression of histone proteins...
August 25, 2018: Biotechnology and Applied Biochemistry
Ang Kai-Cheen, Gam Lay-Harn
Differentially expressed aqueous soluble proteins between Mycobacterium tuberculosis H37Ra and H37Rv were identified. The protein extracts were separated by two-dimensional gel electrophoresis followed by tandem mass spectrometric analysis. Twelve proteins were detected to be differentially expressed significantly between virulent strain H37Rv and attenuated strain H37Ra. The differentially expression of these proteins was validated by a recently isolated clinical virulent strains of M. tuberculosis, TB138...
August 21, 2018: Biotechnology and Applied Biochemistry
Narjes Nakhaee, Sedigheh Asad, Khosro Khajeh, Seyed Shahriar Arab, Mohammad Ali Amoozegar
Azoreductases mainly reduce azo dyes, the largest class of colorants, to colorless aromatic amines. AzoH, a new azoreductase from the halophilic bacterium, Halomonas elongata, has been recently cloned and expressed in Escherichia coli. The aim of this study was to improve thermal stability of this enzyme by introducing new disulfide bonds. Since X-ray crystallography was not available, homology modeling and molecular dynamics was used to construct the enzyme three-dimensional structure. Potential disulfide bonds for increasing thermal stability were found using DIScover online software...
August 21, 2018: Biotechnology and Applied Biochemistry
Hamidreza Hajfarajollah, Parisa Eslami, Babak Mokhtarani, Kambiz Akbari Noghabi
Among microorganisms, bacteria are the main group of biosurfactant-producing organisms. Different types of bacteria including Pseudomonas sp., Acinetobacter sp., Bacillus sp., and Arthrobacter sp. are among the most commonly studied bacteria in the realm of scientific research. However, due to the pathogenic nature of the producing organisms, the application of these compounds is restricted, therefore, not suitable for use in food-related industries. Given that probiotic bacteria impact human health, applying probiotics as nonpathogenic and safe organisms have gained much attention for the production of biosurfactants in recent years...
August 18, 2018: Biotechnology and Applied Biochemistry
Muniyan Rajiniraja, Akella Sivaramakrishna, Varatharajan Sabareesh, Gurunathan Jayaraman
Fatty acids of specific chain lengths have been shown to inhibit the growth of Mycobacterium tuberculosis. In the present study, specific synthetic aromatic derivatives of n-octyl esters were investigated for their property to inhibit the growth of M. tuberculosis H37Ra. Agar well diffusion assay indicated that the crude synthetic derivatives obtained by the esterification of phthalic acid (PA) and n-octanol exhibited antimycobacterial activity. Further, the activity was authenticated with the Miroplate Alamar Blue Assay (MABA)...
August 7, 2018: Biotechnology and Applied Biochemistry
Haifeng Liu, Zhongmin Zhang, Yuting Li, Xiaoli Wang, Yufei Zhang, Yanhui Chu, Xiaohuan Yuan, Xiaohua Wang
Production of excessive transforming growth factor-beta 1 (TGF-β1) with elevated TGF-β1 activity has been implicated in renal fibrosis via renal epithelial cells activation and collagen deposition. As such, attenuating the binding of TGF-β1 to its receptor TGF-beta receptor type II (TGF-βRII) in TGF-β1-dependent signaling is an attractive target for the control of renal fibrosis. Here, we verified the interaction between novel truncated human TGF-βRII (thTβRII, Thr23-Gln166) and TGF-β1, prepared thTβRII in Escherichia coli, and assessed the effects of thTβRII on TGF-β1-induced human kidney epithelial cells (HK-2) and unilateral ureteral obstruction (UUO) model of renal fibrosis...
August 1, 2018: Biotechnology and Applied Biochemistry
Lijuan Zhang, Wenjing Bao, Renquan Wei, Shuilin Fu, Heng Gong
NADH:quinone oxidoreductases (NQOs) act as the electron entry sites in bacterial respiration and oxidize intracellular NADH that is essential for the synthesis of numerous molecules. Klebsiella pneumoniae contains three NQOs (NDH-1, NDH-2, and NQR). The effects of inactivating these NQOs, separately and together, on cell metabolism were investigated under different culture conditions. Defective growth was evident in NDH-1-NDH-2 double and NDH-1-NDH-2-NQR triple deficient mutants, which was probably due to damage to the respiratory chain...
July 31, 2018: Biotechnology and Applied Biochemistry
Aidong Ruan, Yuan Gao, Chao Fang, Yaofei Xu
Nicotine is a potent parasympathomimetic stimulant and an important natural alkaloid mainly found in the Nicotiana genus of plants. It can directly threaten ecological security and human health in tobacco waste, wastewater, and other forms of tobacco production. Therefore, it is the basis of nicotine pollution prevention and of great application value to explore efficient and a wide range of nicotinophilic bacteria for tobacco industry and environmental protection. In this study, one nicotinophilic bacterium was isolated from the soil, which accumulated tobacco waste over 50 years at a Hefei cigarette factory...
July 4, 2018: Biotechnology and Applied Biochemistry
Meghdad Abdollahpour-Alitappeh, Sepand Razavi-Vakhshourpour, Mohsen Abolhassani
Acute myeloid leukemia (AML) is a clonal hematologic malignancy arising from a small population of leukemic cells initiating the disease. CD123 is differentially expressed in AML blasts compared with normal hematopoietic stem and progenitor cells. The aim of this study was to develop specific monoclonal antibodies (mAbs) directed against AML. Three BALB/c mice were immunized with the human CD123 antigen, and the immune spleen cells were fused with the SP2/0 myeloma cell line. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA), and the positive hybrids were cloned by limiting dilution...
July 4, 2018: Biotechnology and Applied Biochemistry
Anastasija Panic, Julijana Stanimirovic, Milan Obradovic, Emina Sudar-Milovanovic, Milan Perovic, Milena Lackovic, Nina Petrovic, Esma R Isenovic
This study aimed to investigate in vivo effects of estradiol on the regulation of hepatic inducible nitric oxide synthase (iNOS) expression in the high fat (HF) diet-induced obesity. Also, we aimed to investigate whether activation of the extracellular signal-regulated kinase (ERK1/2), adenosine monophosphate-activated protein kinase (AMPK), Src kinase, and miR-221 is involved in estradiol-mediated regulation of iNOS in the liver of obese male Wistar rats. Male Wistar rats were fed a standard laboratory diet or a HF diet for 10 weeks...
June 29, 2018: Biotechnology and Applied Biochemistry
Umesh Kalathiya, Monikaben Padariya, Maciej Baginski
HIV-1 integrase (IN) is crucial for integration of viral DNA into the host genome and a promising target in development of antiretroviral inhibitors. In this work, six new compounds were designed by linking the structures of two different class of HIV-1 IN inhibitors (active site binders and allosteric IN inhibitors (ALLINIs)). Among newly designed compounds, INRAT10b was found most potent HIV-1 IN inhibitor considering different docking results. To further validate protein-ligand interactions obtained from dockings, molecular dynamics simulations were performed for inhibitor raltegravir and INRAT10b placed either at active site or allosteric site of HIV-1 IN (monomer or dimer)...
July 2018: Biotechnology and Applied Biochemistry
Chengtuo Niu, Feiyun Zheng, Yongxian Li, Chunfeng Liu, Qi Li
β-Amylase is of important biotechnological aid in maltose syrup production. In this study, the extraction condition of β-amylase from brewer's malt and the optimal dosage of β-amylase in maltose syrup production were optimized using response surface methodology and uniform design method. The optimal extraction condition of β-amylase from brewer's malt was composed of 1:17 (g/v) material/liquid ratio, 44°C extraction temperature, pH 6.4 buffer pH, 2.3 H extraction time, and 1.64 g L-1 NaSO3 dosage with a predicted β-amylase activity of 1,290...
July 2018: Biotechnology and Applied Biochemistry
Jun Lu, Jinxin Li, Shihui Wang, Lu Yao, Wenxia Liang, Juan Wang, Wenyuan Gao
In this paper, we reviewed the advances in ginsenoside biosynthesis and metabolic regulation. To begin with, the application of elicitors in the ginsenoside biosynthesis was discussed. Methyl jasmonate (MJ) and analogues have the best effect on accumulation of ginsenoside compared with other elicitors, and few biotic elicitors are applied in Panax genus plants tissue culture. In addition, so far, more than 40 genes encoding ginsenoside biosynthesis related enzymes have been cloned and identified from Panax genus, such as UDP-glycosyltransferases (UGT) genes UDPG, UGTAE2, UGT94Q2, UGTPg100, and UGTPg1...
July 2018: Biotechnology and Applied Biochemistry
Svetlana P Ikonomova, Megan T Le, Neha Kalla, Amy J Karlsson
Single-chain variable fragment antibodies (scFvs) are attractive for use in applications that require high specificity and binding to a target, such as biosensors. Previously, we demonstrated that a variety of scFvs can be immobilized onto a streptavidin surface through in vivo biotinylation of the biotin carboxyl carrier protein (BCCP) or smaller AviTag fused to the scFvs. However, the BCCP constructs showed better immobilization than the AviTag constructs. In this work, we investigated whether the discrepancy between the biotinylation tags could be alleviated by incorporating a flexible (G4 S)n linker of varying lengths or a rigid (EA3 K)3 linker between the biotinylation tags and the scFvs scFv13R4 and scFv5...
July 2018: Biotechnology and Applied Biochemistry
Rajender Vadija, Kiran Kumar Mustyala, Vasavi Malkhed, Ramasree Dulapalli, Hymavathi Veeravarapu, Ramesh Malikanti, Uma Vuruputuri
Mycobacterium tuberculosis (Mtb) is the pathogen, which causes tuberculosis. The development of multidrug-resistant and extensively drug-resistant strains in Mtb is due to an efflux mechanism of antibiotics in the bacteria. The efflux pump proteins in the bacteria are implicated in the active efflux of antibiotics. The efflux pump protein, "fluoroquinolones export ATP-binding protein Rv2688c" (FEAB), is considered as a potential therapeutic target to prevent tuberculosis. In the present work, in silico protocols are applied to identify inhibitors for the FEAB protein to arrest the efflux mechanism...
July 2018: Biotechnology and Applied Biochemistry
Jin-Jin Gu, Ying Zhou, Yan-Bin Tong, Jian-Jiang Lu, Bang-Ce Ye
Aliphatic polyester, poly(3-hydroxyvalerate) (PHV), is commonly produced as a granular component in bacterial cells of various species. Based on 16S rDNA gene sequence analysis, strain PJC48 was identified as a Bacillus species. The current study is aimed to screen for a high-yield strain that can produce PHV efficiently and to increase PHV product yield by optimizing the fermentative process. We identified a high-producer strain based on Nile red staining. Characterization of the PHV produced by PJC48 by nuclear magnetic resonance spectroscopy revealed that it consisted of (R)-3-hydroxyvalerate monomers...
July 2018: Biotechnology and Applied Biochemistry
Dengyang Zhang, Tianzhu Sun, Jingyun Yan, Xuanjun Wang, Jun Sheng
Notch signaling is a highly conserved pathway existed in multicellular organisms. It plays roles in normal human body development, human cancer initiation, progression and metastasis. The Notch negative regulatory region (NRR) is critical for Notch signaling, and cleavage at the S2 site in the NRR ultimately leads to the activation of Notch signaling. To study the function of human NRR1, we expressed the recombinant human NRR1 (rhNRR1) domain in Escherichia coli. After purification, rhNRR1 was obtained with approximately 94% purity according to SDS-PAGE analysis...
July 2018: Biotechnology and Applied Biochemistry
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