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Biotechnology and Applied Biochemistry

Anderson Albino Gomes, Elisa Pazinatto Telli, Luiz Claudio Miletti, Everton Skoronski, Marcos Gomes Ghislandi, Gustavo Felippe da Silva, Maria de Lourdes Borba Magalhães
Optimization of cellulose enzymatic hydrolysis is crucial for cost effective bioethanol production from lignocellulosic biomass. Enzyme immobilization in solid support allows enzyme recycling for reuse, lowering hydrolysis costs. Graphene is a nanomaterial isolated in 2004, which possesses exceptional properties for biomolecule immobilization. This study evaluates the potential for β-glucosidase recycling by immobilization on graphene nanosheets. Data reported here demonstrated that graphene immobilized β-glucosidase can be recycled for at least eight cycles...
June 21, 2017: Biotechnology and Applied Biochemistry
Dong-Fang Xie, Hui Fang, Jia-Qi Mei, Jin-Yan Gong, Hong-Peng Wang, Xiu-Ying Shen, Jun Huang, Le-He Mei
To improve the thermostability of (R)-selective amine transaminase from Aspergillus terreus (AT-ATA), we used computer software Disulfide by Design (DbD) and Modelling of Disulfide Bonds in Proteins (MODIP) to identify mutation sites where the disulfide bonds were most likely to form. We obtained three stabilised mutants (N25C-A28C, R131C-D134C, M150C-M280C) from seven candidates by site-directed mutagenesis. Compared to the wild-type, the best two mutants N25C-A28C and M150C-M280C showed improved thermal stability with a 3...
June 21, 2017: Biotechnology and Applied Biochemistry
Ramesh Prasad Pandey, Prakash Parajuli, Anaya Raj Pokhrel, Jae Kyung Sohng
7,8-Dihydroxyflavone (7,8-DHF) has been conjugated with glucose moiety to produce glucoside derivatives. Three analogues of 7,8-DHF (7-O-β-D-glucosyl-8-hydroxyflavone, 7-hydroxy-8-O-β-D-glucosyl flavone, and 7,8-di-O-β-D-glucosylflavone) have been successfully produced from in vitro reaction using glycosyltransferase of Bacillus licheniformis. Production of these 7,8-DHF derivatives were shifted to cheaper and easier approach in this study by using engineered Escherichia coli BL21 (DE3) ΔpgiΔzwfΔushA cells in which the flow of glucose-6-phospahte toward glycolysis and pentose phosphate pathway and hydrolysis of UDP-α-D-glucose were blocked while directing the carbon flux toward UDP-α-D-glucose by over-expressing UDP-α-D-glucose pathway genes...
June 12, 2017: Biotechnology and Applied Biochemistry
Hui Deng, Mario van der Stelt
2-Arachidonoylglycerol (2-AG) as an important endogenous signaling lipid that activates the cannabinoid receptors (CB1 R and CB2 R), thereby regulating a diverse range of physiological processes including anxiety, appetite, inflammation, memory, pain sensation and nociception. Diacylglycerol lipases (DAGLs) are the principle enzymes responsible for 2-AG biosynthesis. Recently, the (patho)physiological functions of DAGLs have been explored by both genetic methods and chemical tools. This review will focus on the recent efforts to develop highly selective and in vivo active DAGLs inhibitors using activity-based protein profiling...
June 6, 2017: Biotechnology and Applied Biochemistry
Edgar Edurman García-Silvera, Fernando Martínez-Morales, Brandt Bertrand, Daniel Morales-Guzman, Nashbly Sarela Rosas-Galván, Renato León-Rodríguez, María Del Refugio Trejo-Hernández
In this study, extracellular lipase was produced by Serratia marcescens wild type and three mutant strains. The maximum lipase activity (80 U/mL) was obtained with the SMRG4 mutant strain using soybean oil. Using a 2(2) factorial design the lipase production increased 1.55-fold (124 U/mL) with 4% and 0.05 % soybean oil and Triton X-100, respectively. The optimum conditions for maximum lipase activity were 50°C and pH 8. However, the enzyme was active in a broad range of pH (6-10) and temperatures (5-55°C)...
April 26, 2017: Biotechnology and Applied Biochemistry
Veeresh Juturu, Jin Chuan Wu
Thermophilic Bacillus coagulans JI12 was used to ferment hemicellulose hydrolysate obtained by acid hydrolysis of oil palm empty fruit bunch (EFB) at 50°C and pH6, producing 105.4 g/L of L-lactic acid with a productivity of 9.3 g/L/h by fed batch fermentation under unsterilized conditions. Simultaneous saccharification and fermentation (SSF) was performed at pH5.5 and 50°C to convert both hemicellulose hydrolysate and cellulose-lignin complex in the presence of Cellic Ctec2 cellulases using yeast extract (20 g/L) as the nitrogen source, giving 114...
April 24, 2017: Biotechnology and Applied Biochemistry
Rosa Maria Chura-Chambi, Helen Andrade Arcuri, Felipe Lino, Natan Versati, Mario Sergio Palma, Denize C Favaro, Ligia Morganti
Endostatin (ES) is an antiangiogenic protein that exhibits antitumor activity in animal models. However, the activity observed in animals was not observed in human clinical trials. ES-BAX is a fusion protein composed of two functional domains: ES, which presents specificity and is internalized by activated endothelial cells and the proapoptotic BH3 domain of the protein BAX, a peptide inductor of cellular death when internalized. We have previously shown (Chura-Chambi et al., Cell Death Dis, 5, e1371, 2014) that ES-BAX presents improved antitumor activity in relation to wild-type ES...
May 2017: Biotechnology and Applied Biochemistry
Huo-Xi Jin, Xiao-Kun OuYang, Zhong-Ce Hu
An effective epoxide hydrolase (EH) production strain was mutagenized using (60) Co gamma and UV irradiation. Among positive mutant strains, the EH activity of C2-44 reached 33.7 U/g, which was 267% as much as that of the original Aspergillus niger ZJB-09103. Compared with the wild type, there were significant changes in morphology for C2-44, including the color of mycelia on the slants and the shape of conidial head. In addition, glucose and soybean cake were the optimal carbon and nitrogen source in terms of EH activity for the mutant C2-44 instead of soluble starch and peptone for the wild-type strain...
May 2017: Biotechnology and Applied Biochemistry
Sheng Yang, Yan Long, Hong Yan, Huawan Cai, Yadong Li, Xingguo Wang
The cumA, a gene encoding a multicopper oxidase (MCO), was cloned from the soil-dwelling bacterium Pseudomonas sp. 593. Its corresponding product was overexpressed in Escherichia coli BL21 (DE3) pLysS and purified to homogeneity through Ni-affinity chromatography. The amino acid sequence of the CumA of Pseudomonas sp. 593 was strongly homologous to that of CumA as previously reported. The CumA was quite stable in neutral pH and had poor thermostability. Meanwhile, its optimum pH and temperature toward laccase substrates 2,6-dimethoxyphenol (DMP), syringaldazine (SGZ), and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) were 5...
May 2017: Biotechnology and Applied Biochemistry
Massimiliano Bergallo, Elisa Loiacono, Giovanna Rizzo, Marialuisa Caiazzo, Giuseppe Palladino, Alessandro Amore
Bacterial-derived DNA fragments (BDNAs) have been shown to be present in a dialysis fluid, to pass through dialyzer membranes, and to induce interleukin 6 (IL-6) in mononuclear cells. DNA fragments are thought to be derived from microorganisms inhabiting hemodialysis water and fluid. The primary aim of the present study was to develop two degenerated TaqMan real-time quantitative-PCR (Q-PCR) for detection of a broad range of bacterial DNA that specifically detect 16S ribosomal DNA (rDNA) (862 and 241 bp) and evaluate the efficiency of the Bellco Selecta resin to capture the BDNAs in the dialysis fluid...
May 2017: Biotechnology and Applied Biochemistry
Mohammad H Darvishi, Alireza Nomani, Hadi Hashemzadeh, Mohsen Amini, Mohammad A Shokrgozar, Rassoul Dinarvand
A novel biotinylated chitosan-graft-polyethyleneimine (Bio-Chi-g-PEI) copolymer was synthesized and evaluated as a nonviral gene delivery carrier for improvement of the transfection efficiency, endosomal escape, and targeted gene delivery of a plasmid encoding green fluorescent protein N1 (pEGFP-N1) into two different biotin-overexpressing cell lines including HeLa and OVCAR-3 cells. The structure of the obtained copolymers was confirmed by (1) H nuclear magnetic resonance ((1) H NMR) and Fourier transform infrared spectroscopy...
May 2017: Biotechnology and Applied Biochemistry
Tomohide Uno, Takeshi Yanase, Hiromasa Imaishi
Aspergillus oryzae is a fungus widely used in traditional Japanese fermentation industries. Cytochrome P450 (CYP) proteins are ubiquitously distributed in nature and display a broad range of enzymatic activities. A novel CYP52 (CYP52G3) gene was found in A. oryzae. In this study, we report the functional characterization of CYP52G3. The recombinant protein was expressed heterologously in Escherichia coli, and its membrane fraction isolated. CYP52G3 showed activities for 7-ethoxycoumarin and α-naphtoflavone...
May 2017: Biotechnology and Applied Biochemistry
Cinzia Calcabrini, Umberto Mancini, Roberta De Bellis, Anna Rita Diaz, Maddalena Martinelli, Luigi Cucchiarini, Piero Sestili, Vilberto Stocchi, Lucia Potenza
Some epidemiological studies have suggested possible associations between exposure to extremely low-frequency electromagnetic fields (ELF-EMFs) and various diseases. Recently, ELF-EMF has been considered as a therapeutic agent. To support ELF-EMF use in regenerative medicine, in particular in the treatment of skin injuries, we investigated whether significant cell damage occurs after ELF-EMF exposure. Reactive oxygen species (ROS) production was evaluated in the human keratinocyte exposed for 1 H to 50 Hz ELF-EMF in a range of field strengths from 0...
May 2017: Biotechnology and Applied Biochemistry
Wanhong Liu, Huanyan Wang, Yupei Chen, Shunqin Zhu, Min Chen, Xiaozhong Lan, Guoping Chen, Zhihua Liao
Previous publications reported that the artemisinin level was increased in Artemisia annua following a night-frost period. However, the molecular mechanism was not clear. In this study, we found that exogenous jasmonate (JA) effectively enhanced the freezing tolerance of A. annua. The JA biosynthetic genes (LOX1, LOX2, allene oxide cyclase [AOC], and jasmonate resistant 1 [JAR1]) were induced by cold stress, leading to an increase in endogenous JA in cold-treated A. annua. Increased endogenous JA enhanced the expression of three JA-responsive transcription factors, ethylene response factor 1, ethylene response factor 2, and octadecanoid-responsive AP2/ERF, all of which were reported to transcriptionally activate the expression of artemisinin biosynthetic genes, such as amorpha-4,11-diene synthase (ADS), CYP71AV1, DBR2, and aldehyde dehydrogenase 1 (ALDH1)...
May 2017: Biotechnology and Applied Biochemistry
Saranya Paranji, Sekaran Ganesan
The mixed intracellular enzyme (MICE) from Citrobacter freundii, capable of degrading o-phenylene diamine (OPD), was extracted and characterized. Cofactors such as zinc and copper ions enhanced the MICE activity. The functionalized nanoporous-activated carbon (FNAC) matrix, zinc-impregnated FNAC matrix (Zn(2+) -FNAC), copper-impregnated FNAC matrix (Cu(2+) -FNAC), and zinc- and copper-impregnated FNAC matrix (Zn(2+) -Cu(2+) -FNAC) were prepared and characterized to immobilize MICE. The parameters such as time (0-240 Min), pH (1-10), temperature (20-50 ºC), amount of MICE (1-5 mg), particle size of carbon (100-600 μm), and mass of carbon (0...
May 2017: Biotechnology and Applied Biochemistry
Brenda Eugenia Aguilera, Gabriela Chávez-Calvillo, Darwin Elizondo-Quiroga, Mónica Noemí Jimenez-García, Mauricio Carrillo-Tripp, Laura Silva-Rosales, Rodolfo Hernández-Gutiérrez, Abel Gutiérrez-Ortega
Porcine circovirus type 2 (PCV2) still represents a major problem to the swine industry worldwide, causing high mortality rates in infected animals. Virus-like particles (VLPs) have gained attention for vaccine development, serving both as scaffolds for epitope expression and immune response enhancers. The commercial subunit vaccines against PCV2 consist of VLPs formed by the self-assembly of PCV2 capsid protein (CP) expressed in the baculovirus vector system. In this work, a PCV2 protective epitope was inserted into three different regions of papaya ringspot virus (PRSV) CP, namely, the N- and C-termini and a predicted antigenic region located near the N-terminus...
May 2017: Biotechnology and Applied Biochemistry
Małgorzata Pleszczyńska, Adrian Wiater, Teresa Bachanek, Janusz Szczodrak
Biofilm-related infections of the oral cavity, including dental caries and periodontitis, represent the most prevalent health problems. For years, the treatment thereof was largely based on antibacterial chemical agents. Recently, however, there has been growing interest in the application of more preventive and minimally invasive biotechnological methods. This review focuses on the potential applications of enzymes in the treatment and prevention of oral diseases. Dental plaque is a microbial community that develops on the tooth surface, embedded in a matrix of extracellular polymeric substances of bacterial and host origin...
May 2017: Biotechnology and Applied Biochemistry
Na Li, Guangwei Sun, Shujun Wang, Yu Wang, Zhilong Xiu, Dongsheng Sun, Xin Guo, Ying Zhang, Xiaojun Ma
After islet isolation, diffusion has become the main mechanism to transport oxygen and nutrients into the core of islets. However, diffusion has limitations, by which nutrients cannot effectively reach the core of large islets and can eventually cause core cell death and islet loss. This problem can be resolved by dispersing islets into single islet cells, but single islet cells do not exhibit insulin release function in in vitro culture. In this study, we intended to establish a new islet engineering approach by forming islet cell clusters to improve islet survival and function...
May 2017: Biotechnology and Applied Biochemistry
Simone Brixius-Anderko, Frank Hannemann, Michael Ringle, Yogan Khatri, Rita Bernhardt
Escherichia coli has developed into an attractive organism for heterologous cytochrome P450 production, but, in some cases, was restricted as a host in view of a screening of orphan cytochromes P450 or mutant libraries in the context of molecular evolution due to the formation of the cytochrome P450 inhibitor indole by the enzyme tryptophanase (TnaA). To overcome this effect, we disrupted the tnaA gene locus of E. coli C43(DE3) and evaluated the new strain for whole-cell substrate conversions with three indole-sensitive cytochromes P450, myxobacterial CYP264A1, and CYP109D1 as well as bovine steroidogenic CYP21A2...
May 2017: Biotechnology and Applied Biochemistry
Mohadeseh Hashemi, Amir Yadegari, Ghasem Yazdanpanah, Meisam Omidi, Sayena Jabbehdari, Fatemeh Haghiralsadat, Fatemeh Yazdian, Lobat Tayebi
Graphene oxide (GO) has been recently introduced as a suitable anticancer drug carrier, which could be loaded with doxorubicin (DOX) as a general chemotherapy agent. Herein, the attempts were made to optimize the effective parameters on both loading and release of DOX on GO. GO and GO-DOX were characterized using transition electron microscopy , zeta potential, Raman spectroscopy, UV-visible spectroscopy, and Fourier transform infrared spectroscopy. In addition, loading and releasing behaviors of DOX on GO were studied in terms of different temperature and pH values...
May 2017: Biotechnology and Applied Biochemistry
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