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Biotechnology and Applied Biochemistry

Carlos A Duarte, Mickel Palomino
Virion infectivity factor (Vif) is a 23 KDa protein which protects HIV-1 from deamination of its proviral DNA by APOBEC3G. The active form of Vif is a multimer that interacts simultaneously with CBF-beta, the Elongin B and C subunits, Cullin 5 and APOBEC3G to form a ubiquitin ligase complex targeting the latter for degradation. Vif clearly represents an attractive target for developing novel antiviral drugs for the therapy of HIV/AIDS, and this goal requires a source of well folded, readily available protein...
February 8, 2017: Biotechnology and Applied Biochemistry
Chang Su, Zhiyang Chen, I-Son Ng
Proteus hauseri ZMd44, a biodecolorizing bacterium, has been known to produce electricity and multi-copper oxidase (Mco-laccase) under copper induction. However, optimization and regulation of production have not been explored. This study is the first attempt to evaluate several parameters on biomass and Mco-laccase production of P. hauseri ZMd44. Through orthogonal experiments with Taguchi's L9 found that P. hauseri ZMd44 was sensitive to pH value. The cells grew relatively quickly at pH 7, thus the biomass and Mco-laccase-production reached 1...
February 1, 2017: Biotechnology and Applied Biochemistry
Jaruwan Siritapetawee, Chutima Talabnin, Jarunee Vanichtanankul, Chomphunuch Songsiriritthigul, Kanjana Thumanu, Chun-Jung Chen, Nantarat Komanasin
In this study, the binding of a glycosylated serine protease (EuP-82) with human fibrinogen was investigated by the isothermal titration calorimetry (ITC). The ITC analysis indicated that the binding of EuP-82 to fibrinogen in the conditions with or without the activator (Ca(2+) ) was an exothermic reaction (dominant negative enthalpy), which tended to be driven by hydrogen bonding and van der Waals interactions. In contrast, the binding of fibrinogen-EuP-82 in the condition with the inhibitor (Zn(2+) ) was an unfavorable endothermic reaction...
February 1, 2017: Biotechnology and Applied Biochemistry
Bin Xia, Yang Zou, Zhiling Xu, Yonggang Lv
Low-intensity pulsed ultrasound (LIPUS) is a non-invasive technique that has been shown to affect the cell proliferation, migration, differentiation, and promote the regeneration of damaged peripheral nerve. Our previous studies had proved that LIPUS can significantly promote the neural differentiation of induced pluripotent stem cells-derived neural crest stem cells (iPSCs-NCSCs) and enhance the repair of rat transected sciatic nerve. To further explore the underlying mechanisms of LIPUS treatment on iPSCs-NCSCs, this study reported the gene expression profiling analysis of iPSCs-NCSCs before and after LIPUS treatment using RNA-sequencing (RNA-Seq) method...
January 26, 2017: Biotechnology and Applied Biochemistry
Meng-Jie Tan, Xue-Wei Cao, Peng-Fei Li, Yi-Zhou Zhai, Yu Zhou, Ye-Jun Liu, Jian Zhao, Fu-Jun Wang
Alpha-momorcharin (α-MMC), a type I ribosome inactivating protein, has attracted a great deal attention because of its antitumor activity. However, the cytotoxicity of α-MMC is limited due to insufficient cellular internalization in cancer cells. To enhance the cytotoxicity of α-MMC, a heparin-binding domain derived from HB-EGF (named HBP) was used as a cell-penetrating peptide and fused to the C-terminus of α-MMC. This novel α-MMC-HBP fusion protein was expressed and purified with a Ni(2+) -resin. The N-glycosidase activity and DNase activity assay indicated that the introduction of HBP did not interfere with the intrinsic bioactivities of α-MMC...
January 16, 2017: Biotechnology and Applied Biochemistry
Maria Lepore, Marianna Portaccio
A novel sol-gel based biosensor exploiting the optical absorption properties of sol-gel immobilized laccase has been constructed in the attempt to increase enzyme specificity towards different phenolic substrates. Laccase from Trametes versicolor has been immobilized in optically transparent sol-gel matrices. Using Fourier transform infrared spectroscopy and data analysis based on a wavelet algorithm the successful enzyme immobilization has been evidenced. The changes in the optical absorption spectra of laccase reaction products at 425 nm, 375 nm and 400 nm have been used for hydroquinone, resorcinol and catechol concentration determination, respectively...
December 16, 2016: Biotechnology and Applied Biochemistry
Jianbin Ye, Zhan Zhang, Ji Yan, Hui Hao, Xiangzhen Liu, Zongcan Yang, Ke Ma, Xuepeng Yang, Duobin Mao, Hao Zhou
Phytosterols have been demonstrated to be precursors of polycyclic aromatic hydrocarbons (PAHs) formed during biomass pyrolysis. Here, a novel Paenibacillus sp. was evaluated for its ability to degrade phytosterols in tobacco waste extract (TWE). The optimal conditions for cell growth and stigmasterol (a representative of phytosterols) degradation were 37°C, pH 7.0, 1.0 g/l yeast extract and 6.0 g/l glucose. Paenibacillus sp. could degrade stigmasterol under high concentrations of glucose (up to 130 g/l) and tolerate wide pHs (5...
December 13, 2016: Biotechnology and Applied Biochemistry
Sena Yaman, Pınar Çalık
The aim of this work was to develop an effective fed-batch feeding strategy to enhance recombinant glucose isomerase (r-GI) production by recombinant Escherichia coli BL21 (DE3) pLysS on an industrially relevant feedstock without the application of an exogenous inducer. Following the batch operation (0 < t < 7 h), the effects of pulse and/or continuous feeding of hydrolysed-beet molasses were investigated under five different feeding strategies. The two most promising strategies with respect to r-GI activity were: (i) PM-0...
December 13, 2016: Biotechnology and Applied Biochemistry
Xinliang Mao, Shengjie He, Ting Zhang, Xiaolei Guo, Yazhong Ge, Chungwah Ma, Xuewu Zhang
In this study, the whole proteins from Chinese three-striped box turtle (Cuora trifasciata) were extracted, and hydrolyzed using three proteases (alcalase, papain and protamex). By orthogonal experiments, the optimal hydrolysis conditions for producing peptides with the highest cancer cells growth inhibition activity were determined. Such as, the maximum inhibition on MCF-7 cancer cells (92.37% at 1 mg/mL) was achieved by papain hydrolysis (pH 8, 37 ℃, enzyme to substrate ratio (E/S) 1.5%), and the maximum inhibition on HepG2 cancer cells (94...
December 13, 2016: Biotechnology and Applied Biochemistry
Timothy Kernan, Alan C West, Scott Banta
A. ferrooxidans is an important iron- and sulfur-oxidizing acidophilic chemolithoautotroph that is used extensively in metal extraction and refining, and more recently in the bioproduction of chemicals. However, a lack of genetic tools has limited the further development of this organism for industrial bioprocesses. Using prior microarray studies that identified genes which may express differentially in response to the availability of iron and sulfur, the cycA1 and tusA promoter sequences have been characterized for their ability to drive green fluorescent protein (GFP) expression...
November 22, 2016: Biotechnology and Applied Biochemistry
Monikaben Padariya, Umesh Kalathiya, Maciej Baginski
Reverse transcriptase (RT) inhibitors are currently used to treat HIV-1 infections. In this work, novel triethylamine derivatives were designed and studied by rigid and flexible docking and molecular dynamics (MD) approaches. An apo form of HIV-1 RT was also studied by MD simulation in order to analyze comparative response of protein in ligand bound and unbound form. Among newly designed HIV-1 RT inhibitors, compound HIV104 was the most potent inhibitor considering different docking results. Molecular docking results were further validated by MD simulations of HIV-1 RT/HIV104 complex using two independent softwares (Discovery Studio Client 3...
November 8, 2016: Biotechnology and Applied Biochemistry
Xiaoming Pan, Jing Wang, Yanfang Zhang, Ping Dong, Chunchuan Li, Xingguo Liang
The sensitivity and specificity of DNA detection may decrease when the target DNA is in very low abundance. To effectively detect trace amounts of target DNA from massive background of nucleic acids, we have developed a powerful multiplex pre-amplification method based on ligation-mediated PCR (LM-PCR) that can greatly enrich multiple target DNAs from massive backgrounds. By employing type IIS restriction endonuclease (REase) and specifically designed oligonucleotide adapters, target DNA can be pre-amplified with high efficiency and sensitivity...
November 8, 2016: Biotechnology and Applied Biochemistry
ZeHong Xu, XiaoDan Cheng, JianHong Tan, Xianxue Gan
A simple and high sensitive cholesterol amperometric biosensor, which is based on in situ electropolymerization of multi-walled carbon nanotube-polyaniline (MWCNT-PANI) nanocomposite and electrodeposition of platinum nanoparticle (nano-Pt) films onto the glassy carbon electrode surface for cholesterol oxidase immobilization, was constructed in this study. The preparation process of the modified electrode was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, scanning electron microscopy, and chronoamperometry...
November 2016: Biotechnology and Applied Biochemistry
Wenjian Ma, Jianli Wang, Ye Li, Xiaoqing Hu, Feng Shi, Xiaoyuan Wang
Three genes, gnd, pgl, and fbp, relevant to the pentose phosphate pathway (PPP) were overexpressed in Corynebacterium glutamicum IWJ001, leading to increase of l-isoleucine production. The transcriptional levels of gnd, pgl, and fbp significantly increased in IWJ001/pDXW-8-gnd-fbp-pgl. Compared with the control strain IWJ001/pDXW-8, intracellular NADPH/NADP(+) ratios in IWJ001/pDXW-8-gnd and IWJ001/pDXW-8-gnd-fbp cells grown for 36 H increased threefold and fourfold, respectively, indicating that overexpression of gnd and fbp redirected the carbon flux to PPP...
November 2016: Biotechnology and Applied Biochemistry
Jeyachandran Sivakamavalli, Chandrabose Selvaraj, Sanjeev Kumar Singh, Baskaralingam Vaseeharan
Prophenoloxidase (ProPO) cascade is a principal defense system in crustaceans, which consists of a variety of pattern recognition proteins (lipopolysaccharide and β-glucan-binding protein [β-GBP], β-GBP, and β-glucan recognition protein), proteases (serine protease), and protease inhibitors (α2-macroglobulin and pacifastin) to regulate the protection mechanism in crustaceans. In the prophneoloxidase pathway, the protein-protein interactions (PPIs) and other immune-related analyses still have not been reported...
November 2016: Biotechnology and Applied Biochemistry
Kouta Sakaguchi, Toshihiko Katoh, Kenji Yamamoto
Glycan conversion of glycoprotein via the transglycosylation activity of endo-β-N-acetylglucosaminidase is a promising chemoenzymatic technology for the production of glycoproteins including bio-medicines with a homogeneous glycoform. Although Endo-M is a key enzyme in this process, its product undergoes rehydrolysis, which leads to a lower yield, and limits the practical application of this enzyme. We developed several Endo-M mutant enzymes including N175Q with glycosynthase-like activity and/or transglycosidase-like activity...
November 2016: Biotechnology and Applied Biochemistry
Xueqing Fu, Pu Shi, Qian Shen, Weimin Jiang, Yueli Tang, Zongyou Lv, Tingxiang Yan, Ling Li, Guofeng Wang, Xiaofen Sun, Kexuan Tang
Artemisinin, a sesquiterpene lactone isolated from Artemisia annua L. (sweet wormwood), is extensively used in the treatment of malaria. In order to better understand the metabolism of terpenes in A. annua and the influence of terpene synthases on artemisinin yield, the expression pattern of a monoterpene alcohol dehydrogenase (ADH2) has been studied using transgenic plants expressing promoter-β-glucuronidase (GUS) fusion. ADH2 played a major role in monoterpenoid biosynthesis including carveol, borneol, and artemisia ketone through in vitro biochemical analysis...
November 2016: Biotechnology and Applied Biochemistry
Qianli Zheng, Shuhong Gao
Soybean oil is an important carbon source in kitasamycin fermentation by Streptomyces kitasatoensis. In this study, three different surfactants, Tween 80, Tween 85, and sodium dodecyl sulfate (SDS), were added in the fermentation medium to improve soybean oil utilization. Results indicated that all of these surfactants promote kitasamycin biosynthesis. When 0.5 g/L SDS was added at the beginning of fermentation, kitasamycin production increased by 55% and A5 content improved by 12%, compared with the control treatment (i...
November 2016: Biotechnology and Applied Biochemistry
Nuobei Zhang, Xin Chen
O-GlcNAcylation is a monosaccharide modification by a residue of N-acetylglucosamine (GlcNAc) attached to serine or threonine moieties on nuclear and cytoplasmic proteins. O-GlcNAcylation is dynamically regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Increasing evidence suggests that O-GlcNAcylation is involved in a variety of human cancers. However, the exact role of O-GlcNAcylation in tumor progression remains unclear. Here, we show that O-GlcNAcylation accelerates oncogenic phenotypes of gastric cancer...
November 2016: Biotechnology and Applied Biochemistry
Soudeh Khanamani Falahati-Pour, Abbas Sahebghadam Lotfi, Gholamreza Ahmadian, Amin Baghizadeh, Reza Behroozi, Fereshteh Haghighi
Organophosphorus (OPs) compounds are widely used in many pesticides, insecticides, and chemical nerve agents. These compounds are hazardous for humans and the environment. There are many reports on detoxification of these compounds, among them enzymatic cleavage of these compounds with organophosphorus hydrolase (OPH) has been taken into more consideration. Several studies have been performed to improve OPH secretion in Escherichia coli by different signal peptides, but have not been successful. In this study, to achieve the extracellular secretion of OPH in E...
November 2016: Biotechnology and Applied Biochemistry
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