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BMC Biotechnology

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https://www.readbyqxmd.com/read/29145860/a-new-single-step-protocol-for-rapid-baculovirus-driven-protein-production-in-insect-cells
#1
Judith Scholz, Sabine Suppmann
BACKGROUND: In the last three decades, the Baculovirus expression vector system (BEV) has evolved to one of the most widely used eukaryotic systems for heterologous protein expression including approved vaccines and therapies. Despite the significant improvements introduced during the past years, the BEV system still has major drawbacks, primarily the time required to generate recombinant virus and virus instability for certain target proteins. In this study we show that the conventional method to generate recombinant Baculovirus using a Tn7 transposition based system can be shortened to a single-step transfection-only procedure without further amplification...
November 16, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29141617/comparison-of-hso4-cl-and-meco2-as-anions-in-pretreatment-of-aspen-and-spruce-with-imidazolium-based-ionic-liquids
#2
Zhao Wang, John Gräsvik, Leif J Jönsson, Sandra Winestrand
BACKGROUND: Ionic liquids (ILs) draw attention as green solvents for pretreatment of lignocellulose before enzymatic saccharification. Imidazolium-based ILs with different anionic constituents ([HSO4](-), [Cl](-), [MeCO2](-)) were compared with regard to pretreatment of wood from aspen and spruce. The objective was to elucidate how the choice of anionic constituent affected the suitability of using the IL for pretreatment of hardwood, such as aspen, and softwood, such as spruce. The investigation covered a thorough analysis of the mass balance of the IL pretreatments, the effects of pretreatment on the cell wall structure as assessed by fluorescence microscopy, and the effects of pretreatment on the susceptibility to enzymatic saccharification...
November 15, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29137618/afeap-cloning-a-precise-and-efficient-method-for-large-dna-sequence-assembly
#3
Fanli Zeng, Jinping Zang, Suhua Zhang, Zhimin Hao, Jingao Dong, Yibin Lin
BACKGROUND: Recent development of DNA assembly technologies has spurred myriad advances in synthetic biology, but new tools are always required for complicated scenarios. Here, we have developed an alternative DNA assembly method named AFEAP cloning (Assembly of Fragment Ends After PCR), which allows scarless, modular, and reliable construction of biological pathways and circuits from basic genetic parts. METHODS: The AFEAP method requires two-round of PCRs followed by ligation of the sticky ends of DNA fragments...
November 14, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29132331/adaptation-of-the-goldenbraid-modular-cloning-system-and-creation-of-a-toolkit-for-the-expression-of-heterologous-proteins-in-yeast-mitochondria
#4
Ana Pérez-González, Ryan Kniewel, Marcel Veldhuizen, Hemant K Verma, Mónica Navarro-Rodríguez, Luis M Rubio, Elena Caro
BACKGROUND: There is a need for the development of synthetic biology methods and tools to facilitate rapid and efficient engineering of yeast that accommodates the needs of specific biotechnology projects. In particular, the manipulation of the mitochondrial proteome has interesting potential applications due to its compartmentalized nature. One of these advantages resides in the fact that metalation occurs after protein import into mitochondria, which contains pools of iron, zinc, copper and manganese ions that can be utilized in recombinant metalloprotein metalation reactions...
November 13, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29126403/optimization-of-physical-conditions-for-the-production-of-thermostable-t1-lipase-in-pichia-guilliermondii-strain-so-using-response-surface-methodology
#5
Mary Ladidi Abu, Hisham Mohd Nooh, Siti Nurbaya Oslan, Abu Bakar Salleh
BACKGROUND: Pichia guilliermondii was found capable of expressing the recombinant thermostable lipase without methanol under the control of methanol dependent alcohol oxidase 1 promoter (AOXp 1). In this study, statistical approaches were employed for the screening and optimisation of physical conditions for T1 lipase production in P. guilliermondii. RESULT: The screening of six physical conditions by Plackett-Burman Design has identified pH, inoculum size and incubation time as exerting significant effects on lipase production...
November 10, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29126399/production-and-purification-of-chimeric-hbc-virus-like-particles-carrying-influenza-virus-lah-domain-as-vaccine-candidates
#6
Andris Kazaks, I-Na Lu, Sophie Farinelle, Alex Ramirez, Vincenzo Crescente, Benjamin Blaha, Olotu Ogonah, Tarit Mukhopadhyay, Mapi Perez de Obanos, Alejandro Krimer, Inara Akopjana, Janis Bogans, Velta Ose, Anna Kirsteina, Tatjana Kazaka, Nicola J Stonehouse, David J Rowlands, Claude P Muller, Kaspars Tars, William M Rosenberg
BACKGROUND: The lack of a universal influenza vaccine is a global health problem. Interest is now focused on structurally conserved protein domains capable of eliciting protection against a broad range of influenza virus strains. The long alpha helix (LAH) is an attractive vaccine component since it is one of the most conserved influenza hemagglutinin (HA) stalk regions. For an improved immune response, the LAH domain from H3N2 strain has been incorporated into virus-like particles (VLPs) derived from hepatitis B virus core protein (HBc) using recently developed tandem core technology...
November 10, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29121909/successful-production-of-the-potato-antimicrobial-peptide-snakin-1-in-baculovirus-infected-insect-cells-and-development-of-specific-antibodies
#7
Natalia Inés Almasia, María Paula Molinari, Guillermo Andrés Maroniche, Vanesa Nahirñak, María Pilar Barrios Barón, Oscar Alberto Taboga, Cecilia Vazquez Rovere
BACKGROUND: Snakin-1 (StSN1) is a broad-spectrum antimicrobial cysteine-rich peptide isolated from Solanum tuberosum. Its biotechnological potential has been already recognized since it exhibits in vivo antifungal and antibacterial activity. Most attempts to produce StSN1, or homologous peptides, in a soluble native state using bacterial, yeast or synthetic expression systems have presented production bottlenecks such as insolubility, misfolding or low yields. RESULTS: In this work, we successfully expressed a recombinant StSN1 (rSN1) in Spodoptera frugiperda (Sf9) insect cells by optimizing several of the parameters for its expression in the baculovirus expression system...
November 9, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29121904/generation-of-murine-macrophage-derived-cell-lines-expressing-porcine-cd163-that-support-porcine-reproductive-and-respiratory-syndrome-virus-infection
#8
Liangliang Li, Chunyan Wu, Gaopeng Hou, Biyun Xue, Sha Xie, Qin Zhao, Yuchen Nan, Gaiping Zhang, En-Min Zhou
BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) exhibits a highly restricted tropism for cells of the monocyte-macrophage lineage, utilizing porcine CD163 (pCD163) as an indispensable cellular receptor for infection. Transfection the gene of pCD163 into several non-permissive cell lines followed by protein expression confers susceptibility to PRRSV. A lack of specialized porcine antibody tools for use with existing porcine-derived primary cells and cell lines has hampered studies of both PRRSV pathogenesis and virus triggering of immune response cascades...
November 9, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29121887/biological-activities-and-chemical-compositions-of-slime-tracks-and-crude-exopolysaccharides-isolated-from-plasmodia-of-physarum-polycephalum-and-physarella-oblonga
#9
Tuyen T M Huynh, Trung V Phung, Steven L Stephenson, Hanh T M Tran
BACKGROUND: The myxomycetes derive their common name (slime molds) from the multinucleate trophic stage (plasmodium) in the life cycle, which typically produces a noticeable amount of slimy materials, some of which is normally left behind as a "slime track" as the plasmodium migrates over the surface of a particular substrate. The study reported herein apparently represents the first attempt to investigate the chemical composition and biological activities of slime tracks and the exopolysaccharides (EPS) which cover the surface of the plasmodia of Physarum polycephalum and Physarella oblonga...
November 9, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29115967/light-induced-expression-of-%C3%AE-glucosidase-in-escherichia-coli-with-autolysis-of-cell
#10
Fei Chang, Xianbing Zhang, Yu Pan, Youxue Lu, Wei Fang, Zemin Fang, Yazhong Xiao
BACKGROUND: β-Glucosidase has attracted substantial attention in the scientific community because of its pivotal role in cellulose degradation, glycoside transformation and many other industrial processes. However, the tedious and costly expression and purification procedures have severely thwarted the industrial applications of β-glucosidase. Thus development of new strategies to express β-glucosidases with cost-effective and simple procedure to meet the increasing demands on enzymes for biocatalysis is of paramount importance...
November 7, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/29115956/de-novo-characterization-of-venom-apparatus-transcriptome-of-pardosa-pseudoannulata-and-analysis-of-its-gene-expression-in-response-to-bt-protein
#11
Rong Li, Zhenzhen Yan, Juan Wang, Qisheng Song, Zhi Wang
BACKGROUND: Pardosa pseudoannulata is a prevailing spider species, and has been regarded as an important bio-control agent of insect pests in farmland of China. However, the available genomic and transcriptomic databases of P. pseudoannulata and their venom are limited, which severely hampers functional genomic analysis of P. pseudoannulata. Recently high-throughput sequencing technology has been proved to be an efficient tool for profiling the transcriptome of relevant non-target organisms exposed to Bacillus thuringiensis (Bt) protein through food webs...
November 7, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28899371/simultaneous-detection-of-%C3%AE-lactoalbumin-%C3%AE-lactoglobulin-and-lactoferrin-in-milk-by-visualized-microarray
#12
Zhoumin Li, Fang Wen, Zhonghui Li, Nan Zheng, Jindou Jiang, Danke Xu
BACKGROUND: α-Lactalbumin (a-LA), β-lactoglobulin (β-LG) and lactoferrin (LF) are of high nutritional value which have made ingredients of choice in the formulation of modern foods and beverages. There remains an urgent need to develop novel biosensing methods for quantification featuring reduced cost, improved sensitivity, selectivity and more rapid response, especially for simultaneous detection of multiple whey proteins. RESULTS: A novel visualized microarray method was developed for the determination of a-LA, β-LG and LF in milk samples without the need for complex or time-consuming pre-treatment steps...
September 12, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28888230/effect-of-supercritical-carbon-dioxide-on-the-enzymatic-production-of-biodiesel-from-waste-animal-fat-using-immobilized-candida-antarctica-lipase-b-variant
#13
Aldricho Alpha Pollardo, Hong-Shik Lee, Dohoon Lee, Sangyong Kim, Jaehoon Kim
BACKGROUND: Waste animal fat is a promising feedstock to replace vegetable oil that widely used in commercial biodiesel process, however the high content of free fatty acid in waste fat makes it unfeasible to be processed with commercial base-catalytic process. Enzymatic process is preferable to convert waste fat into biodiesel since enzyme can catalyze both esterification of free fatty acid and transesterification of triglyceride. However, enzymatic reaction still has some drawbacks such as lower reaction rates than base-catalyzed transesterification and the limitation of reactant concentration due to the enzyme inhibition of methanol...
September 9, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28888227/conversion-of-an-inactive-xylose-isomerase-into-a-functional-enzyme-by-co-expression-of-groel-groes-chaperonins-in-saccharomyces-cerevisiae
#14
Beatriz Temer, Leandro Vieira Dos Santos, Victor Augusti Negri, Juliana Pimentel Galhardo, Pedro Henrique Mello Magalhães, Juliana José, Cidnei Marschalk, Thamy Lívia Ribeiro Corrêa, Marcelo Falsarella Carazzolle, Gonçalo Amarante Guimarães Pereira
BACKGROUND: Second-generation ethanol production is a clean bioenergy source with potential to mitigate fossil fuel emissions. The engineering of Saccharomyces cerevisiae for xylose utilization is an essential step towards the production of this biofuel. Though xylose isomerase (XI) is the key enzyme for xylose conversion, almost half of the XI genes are not functional when expressed in S. cerevisiae. To date, protein misfolding is the most plausible hypothesis to explain this phenomenon...
September 9, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28865427/a-split-ubiquitin-system-to-reveal-topology-and-released-peptides-of-membrane-proteins
#15
Qiu-Ping Li, Shuai Wang, Jin-Ying Gou
BACKGROUND: Membrane proteins define biological functions of membranes in cells. Extracellular peptides of transmembrane proteins receive signals from pathogens or environments, and are the major targets of drug developments. Despite of their essential roles, membrane proteins remain elusive in topological studies due to technique difficulties in their expressions and purifications. METHODS: First, the target gene is cloned into a destination vector to fuse with C terminal ubiquitin at the N or C terminus...
September 2, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28854910/expression-of-vitreoscilla-hemoglobin-enhances-production-of-arachidonic-acid-and-lipids-in-mortierella-alpina
#16
Huidan Zhang, Yingang Feng, Qiu Cui, Xiaojin Song
BACKGROUND: Arachidonic acid (ARA, C20:4, n-6), which belongs to the omega-6 series of polyunsaturated fatty acids and has a variety of biological activities, is commercially produced in Mortierella alpina. Dissolved oxygen or oxygen utilization efficiency is a critical factor for Mortierella alpina growth and arachidonic acid production in large-scale fermentation. Overexpression of the Vitreoscilla hemoglobin gene is thought to significantly increase the oxygen utilization efficiency of the cells...
August 30, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28810845/a-bioinformatics-approach-for-identifying-transgene-insertion-sites-using-whole-genome-sequencing-data
#17
Doori Park, Su-Hyun Park, Yong Wook Ban, Youn Shic Kim, Kyoung-Cheul Park, Nam-Soo Kim, Ju-Kon Kim, Ik-Young Choi
BACKGROUND: Genetically modified crops (GM crops) have been developed to improve the agricultural traits of modern crop cultivars. Safety assessments of GM crops are of paramount importance in research at developmental stages and before releasing transgenic plants into the marketplace. Sequencing technology is developing rapidly, with higher output and labor efficiencies, and will eventually replace existing methods for the molecular characterization of genetically modified organisms...
August 15, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28789688/an-in-vitro-synthetic-biosystem-based-on-acetate-for-production-of-phloroglucinol
#18
Rubing Zhang, Wei Liu, Yujin Cao, Xin Xu, Mo Xian, Huizhou Liu
BACKGROUND: Phloroglucinol is an important chemical, and the biosynthesis processes which can convert glucose to phloroglucinol have been established. However, due to approximate 80% of the glucose being transformed into undesirable by-products and biomass, this biosynthesis process only shows a low yield with the highest value of about 0.20 g/g. The industrial applications are usually hindered by the low current productivity and yield and also by the high costs. Generally, several different aspects limit the development of phloroglucinol biosynthesis...
August 8, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28778182/immobilisation-of-delta-like-1-ligand-for-the-scalable-and-controlled-manufacture-of-hematopoietic-progenitor-cells-in-a-stirred-bioreactor
#19
Rebecca L L Moore, Matthew J Worrallo, Peter D Mitchell, Jon Harriman, Katie E Glen, Robert J Thomas
BACKGROUND: Umbilical cord blood provides a source of hematopoietic stem cells for transplantation with immunological and availability advantages over conventional bone marrow sources. Limited cell numbers and slower engraftment from umbilical cord blood units has led to the clinical development of immobilised Notch ligand Delta-Like 1 to promote ex vivo expansion of a rapidly engrafting cell population. However, current immobilisation methods are not simple to scale in a controlled manner...
August 4, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28768539/comprehensive-in-silico-allergenicity-assessment-of-novel-protein-engineered-chimeric-cry-proteins-for-safe-deployment-in-crops
#20
Maniraj Rathinam, Shweta Singh, Debasis Pattanayak, Rohini Sreevathsa
BACKGROUND: Development of chimeric Cry toxins by protein engineering of known and validated proteins is imperative for enhancing the efficacy and broadening the insecticidal spectrum of these genes. Expression of novel Cry proteins in food crops has however created apprehensions with respect to the safety aspects. To clarify this, premarket evaluation consisting of an array of analyses to evaluate the unintended effects is a prerequisite to provide safety assurance to the consumers. Additionally, series of bioinformatic tools as in silico aids are being used to evaluate the likely allergenic reaction of the proteins based on sequence and epitope similarity with known allergens...
August 2, 2017: BMC Biotechnology
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