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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
Fereshteh Rahimi, Javad Mohammadnejad Arough, Mona Yaghoobi, Hadi Davoodi, Fatemeh Sepehri, Masood Amirabadizadeh
In this study, porous silicon (PSi) was utilized instead of prevalent polystyrene platforms and its capability in biomolecules screening was examined. Here, two types of porous structure, macroporous silicon (Macro-PSi) and mesoporous silicon (Meso-PSi) were produced on silicon wafers by electrochemical etching using different electrolytes. Moreover, both kinds of fresh and Oxidized PSi samples were investigated. Next, osteocalcin as biomarker of bone formation process was used as a model biomarker and the colorimetric detection was performed by competitive Enzyme-linked Immunosorbent Assay (ELISA)...
October 24, 2016: Biotechnology and Applied Biochemistry
Poonam Syal, Rani Gupta
Five lipase genes, ylip4, ylip5, ylip7, ylip13 and ylip15, from Yarrowia lipolytica MSR80 were cloned and expressed in pEZZ18-HB101 system. The lipases shared maximum sequence identity with Candida galli lipase, while they shared structural similarity with YLIP2 of Y. lipolytica CLIB122. The enzymes, purified using IgG Sepharose, had specific activities in the range of 7-25 U/mg. Biochemical characteristics of all the lipases varied with respect to thermostability, substrate specificity and enantioselectivity...
October 24, 2016: Biotechnology and Applied Biochemistry
Fengying Dai, Miao Zhang, Dixin Xu, Yin Yang, Jiaxiao Wang, Mingzhen Li, Meihong Du
Micro- and nano-immunomagnetic beads (MIMBs and NIMBs) used for immunomagnetic separation (IMS) with Polymerase Chain Reaction (PCR) were studied for the rapid detection of Salmonella. The capture efficiency of the two different IMBs was evaluated by a conventional plate counting method, and the binding pattern was studied by scanning electron microscopy. The specificity of the IMBs was tested with Salmonella, Shigella flexneri, Enterhemorrhagic Escherichia coli O157:H7(EHEC O157:H7) and Listeria monocytogenes...
October 12, 2016: Biotechnology and Applied Biochemistry
Robert Millar, Rahman Rahmanpour, Eugenie Wei Jia Yuan, Catharine White, Timothy D H Bugg
An extracellular esterase gene estK was identified in Pseudomonas putida mt-2, and was overexpressed to high levels in Escherichia coli. The recombinant EstK enzyme was purified and characterised kinetically against p-nitrophenyl ester and other aryl-alkyl ester substrates, and was found to be selective for hydrolysis of acetyl ester substrates, with high activity for p-nitrophenyl acetate (kcat 5.5 s(-1) , KM 285 μM). Recombinant EstK was found to catalyse deacetylation of acetylated beech xylan, indicating a possible in vivo function for this enzyme, and partial deacetylation of a synthetic polymer, poly(vinylacetate)...
October 3, 2016: Biotechnology and Applied Biochemistry
Zhe Wu, Wipawee Dejtisakdi, Prasart Kermanee, Chunhong Ma, Wallop Arirob, Ramaraj Sathasivam, Niran Juntawong
To evaluate the potential for algal biotechnology to replace traditional agriculture in northeastern Thailand, an open race-way cultivation system was developed to produce biomass and beta-carotene. Dunaliella salina KU 11 isolated from local saline soil was cultured in open race-way tanks using brine and saline lake water. Grown in modified Johnson's medium (with 2-3.5 M NaCl), the algae reached a maximum cell density on the fourth day (1.8×10(6) cells mL(-1) ). Increasing KNO3 and NaHCO3 from 0.5 and 0.043 g L(-1) to 1 and 2...
October 3, 2016: Biotechnology and Applied Biochemistry
Wei Jiang, Beibei Lv, Xiao Wu, Jinbin Wang, Guogan Wu, Chunhui Shi, Xueming Tang
In this study, trends in synonymous codons usage of V. volvecea have been firstly examined by analysis of complete coding sequences and gene-chip data. The results showed that GC content at three codon positions are obviously different and there were several factors shaping the codon usage of V. volvacea genes, including base composition. The comparison of codon usage among four edible fungi such as V. volvacea, Agaricus bisporus, Coprinopsis cinerea and Pleurotus ostreatus indicated that similarly codon usage pattern was used among V...
October 3, 2016: Biotechnology and Applied Biochemistry
Christos S Karamitros, Nikolaos E Labrou
In the present study, we report on the effect of four different soluble additives (sucrose, lactitol, superfloc c577 and dextran sulphate) on the stability of the enzyme glutathione transferase 1 from Zea mays (ZmGSTF1-1) under free and tethered conditions at 4 and 25 °C. Among all additives, the best stabilizing effects observed in case of superfloc c577 and sucrose in both tested temperatures, yet at distinct concentrations at each condition. Those two stabilizing agents were further combined and potential positive synergistic effects were investigated...
September 28, 2016: Biotechnology and Applied Biochemistry
Jianli Li, Shujie Liu, Juan Wang, Jing Li, Jinxin Li, Wenyuan Gao
This study reports the best culture conditions for roots growth and accumulation of active components by optimizing the parameters. Glycyrrhiza uralensis adventitious roots metabolites were significantly increased after adding Saccharomyces cerevisiae and Meyerozyma guilliermondii. The highest contents of polysaccharide, glycyrrhizic acid, glycyrrhetinic acid and total flavonoids were obtained in Meyerozyma guilliermondii group, the content of glycyrrhizic acid was 5.3-fold higher than the control. In control and treatment groups, twelve compounds were identified by high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS),among which, some new compounds have been detected in elicitor groups including 5,7-Dihydroxyflavanone, glycyrrhisoflavanone, licorice saponin J2, uralsaponin B, (3R)-vestitol, uralenol...
September 20, 2016: Biotechnology and Applied Biochemistry
Soner Donmez, Fatma Arslan, Nurşen Sarı, Elvan Hasanoğlu Özkan, Halit Arslan
In the present study, a novel biosensor that is sensitive to glucose was prepared using the microspheres modified (4-Formyl-3-methoxyphenoxymethyl)polystyrene (FMPS) with L-Glycine (FMPS-Gly). Polymeric microspheres having Schiff bases were prepared from FMPS, Glycine condensation method. Glucose oxidase enzyme (GOx) was immobilized onto modified carbon paste electrode (MCPE) by cross-linking with glutaraldehyde. Oxidation of enzymatically produced H2 O2 (+0.5 V vs.Ag/AgCl) was used for determination of glucose...
September 15, 2016: Biotechnology and Applied Biochemistry
Suzana Jukić, Dijana Bubenik, Nediljko Pavlović, Ana Jurinjak Tušek, Višnja Gaurina Srček
Mammalian cell cultures are the preferred expression systems for the production of biopharmaceuticals requiring posttranslational processing. Usually, cell cultures are cultivated in medium supplemented with serum, which supports cell proliferation, viability, and productivity. However, due to scientific and regulatory concerns, serum-free conditions are required in recombinant protein production. Cell lines that are intended for commercial recombinant protein production have to adapt to serum- or protein-free conditions early in their development...
September 2016: Biotechnology and Applied Biochemistry
Wenhui Liu, Wenbin Zhao, Jun Lai, Qi Shen, Yingchun Xu, Liqiang Pan, Shuqing Chen
Human serum albumin (HSA) and Interleukin-1 receptor antagonist (IL1Ra) fusion protein is a potential long-acting drug in the treatment of type 2 diabetes. Previously, the expression level of HSA/IL1Ra in Pichia pastoris was successfully improved by increasing the gene copy number and co-expression with chaperone (Protein disulfide isomerase, PDI) in our lab. However, the overexpression strain (OE-Strain) resulted in low production in high-cell-density fermentation. In this study, the culture medium had been optimized in both flask and fermenter, and the optimum culture medium notably increased the productivity and stability of HSA/IL1Ra...
August 30, 2016: Biotechnology and Applied Biochemistry
Jia-Wen Chen, Wan-Jun Liu, Dong-Xiong Hu, Xiang Wang, Srinivasan Balamurugan, Adili Alimujiang, Wei-Dong Yang, Jie-Sheng Liu, Hong-Ye Li
Oleaginous microalgae hold great promises for biofuel production. However, commercialization of microalgal biofuels remains impracticable due to lack of suitable industrial strain with high growth rate and lipid productivity. Engineering of metabolic pathways is a potential strategy for the improvement of microalgal strains for the production of lipids and also value-added products in microalgae. Malonyl CoA-acyl carrier protein transacylase (MCAT) has been reported to be involved in fatty acid biosynthesis...
August 30, 2016: Biotechnology and Applied Biochemistry
Ping Yu, Xingge Chen, Peng Li
Fatty acid, isoprenoid and alcohol pathways have been successfully engineered to produce biofuels. By introducing three genes, atfA, adhE and pdc, into Escherichia coli to expand fatty acid pathway, up to 1.28 g/L of fatty acid ethyl esters can be achieved. The isoprenoid pathway can be expanded to produce bisabolene with a high titer of 900 mg/L in Saccharomyces cerevisiae. Short- and long-chain alcohols can also be effectively biosynthesized by extending the carbon chain of ketoacids with an engineered "+1" alcohol pathway...
August 10, 2016: Biotechnology and Applied Biochemistry
Yew Joon Tam, Nazariah Allaudin Zeenathul, Morvarid Akhavan Rezaei, Nor Hidayah Mustafa, Mohd Lila Mohd Azmi, Abdul Rani Bahaman, Sewn Cen Lo, Joo Shun Tan, Homayoun Hani, Abdullah Rasedee
Limit of detection (LOD), limit of quantification (LOQ) and the dynamic range of detection of hepatitis B surface antigen antibody (anti-HBs) using surface plasmon resonance (SPR) chip-based approach with Pichia pastoris derived recombinant hepatitis B surface antigen (HBsAg) as recognition element were established through the scouting for optimal conditions for the improvement of immobilization efficiency and in the use of optimal regeneration buffer. Recombinant HBsAg was immobilized onto the sensor surface of CM5 chip at a concentration of 150 mg/L, in sodium acetate buffer at pH 4 with added 0...
August 10, 2016: Biotechnology and Applied Biochemistry
Anna V Savelyeva, Anna A Nemudraya, Vladimir F Podgornyi, Nadezhda V Laburkina, Yuriy A Ramazanov, Andrey P Repkov, Elena V Kuligina, Vladimir A Richter
The levels of aeration and mass transfer are critical parameters required for an efficient aerobic bioprocess, and directly depend on the design features of exploited bioreactors. A novel apparatus, using gas vortex for aeration and mass-transfer processes was constructed in the Center of Vortex Technologies (Novosibirsk, Russia). In this paper, we compared the efficiency of recombinant E. coli strain cultivation using novel gas-vortex technology with conventional bioprocess technologies such as shake flasks and bioreactors with mechanical stirrers...
August 10, 2016: Biotechnology and Applied Biochemistry
Trevor Alleyne, Diane N Ignacio, Valerie B Sampson, Damian Ashe, Michael Wilson
The mitochondrial enzyme cytochrome c oxidase catalyses the reduction of molecular oxygen in the critical step of oxidative phosphorylation that links the oxidation of food consumed to ATP production in cells. The enzyme catalyses the reduction of oxygen at two vastly different rates that are thought to be linked to two different conformations but the conformation of the 'fast enzyme' remains obscure. In this study we demonstrated how oxygen binding at haem a3 could trigger long distance conformational changes and then simulated a conformational change in an eight residue loop near to the enzyme's substrate (cytochrome c) binding site...
August 4, 2016: Biotechnology and Applied Biochemistry
Juan Dong, Mohammed A A Gasmalla, Wei Zhao, Jingtao Sun, Wenyu Liu, Mingming Wang, Liang Han, Ruijin Yang
A cold-adapted esterase-producing strain named T1-39 was isolated from Glacier No.1, Tianshan, China, and identified as Pseudomonas sp. from 16S rRNA sequence analysis. The esterase (EstT1-39) secreted by this strain preferentially hydrolyzed esters of glycerol with short- and medium-chain fatty acids. Mutants of T1-39 were generated by the atmospheric and room-temperature plasma (ARTP) method and screened for enhanced esterase activity. Among all the mutants, strain TB11 had 4.45-fold higher esterase productivity than T1-39, with high genetic stability over 10 generations of continuous cultivation...
July 13, 2016: Biotechnology and Applied Biochemistry
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