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Journal of Biotechnology

Joao Diogo Gouveia, Jesus Ruiz, Lambertus A M van den Broek, Thamara Hesselink, Sander Peters, Dorinde M M Kleinegris, Alison G Smith, Douwe van der Veen, Maria J Barbosa, Rene H Wijffels
Botryococcus braunii can produce both long-chain hydrocarbons as well as carbohydrates in large quantities, and is therefore a promising industrial organism for the production of biopolymer building blocks. Many studies describe the use of different strains of Botryococcus braunii but differences in handling and cultivation conditions make the comparison between strains difficult. In this study, 16 B. braunii strains obtained from six culture collections were compared for their biomass productivity and hydrocarbon and carbohydrate content...
March 20, 2017: Journal of Biotechnology
Guoqiang Liu, Yingying Kong, Yajing Fan, Ce Geng, Donghai Peng, Ming Sun
Bacillus velezensis LS69 was found to exhibit antagonistic activity against a diverse spectrum of pathogenic bacteria. It has one circular chromosome of 3,917,761bp with 3,643 open reading frames. Genome analysis identified ten gene clusters involved in nonribosomal synthesis of polyketides (macrolactin, bacillaene and difficidin), lipopeptides (surfactin, fengycin, bacilysin and iturin A) and bacteriocins (amylolysin and amylocyclicin). In addition, B. velezensis LS69 was found to contain a series of genes involved in enhancing plant growth and triggering plant immunity...
March 17, 2017: Journal of Biotechnology
Gongxian Xu, Caixia Li
This paper addresses the problem of identifying the shared metabolic objectives of glycerol bioconversion in Klebsiella pneumoniae for production of 1,3-propanediol (1,3-PD) under different culture conditions. To achieve this goal, we propose a multi-level programming model. This model includes three optimization problems, where the constraint region of the first level problem is implicitly determined by the other two optimization problems. The optimized objectives of the first and second level problems are to minimize the set of fluxes that are of major importance to glycerol metabolism and the difference between the observed fluxes and those computed by the model, respectively...
March 17, 2017: Journal of Biotechnology
Xiaoxue Zhou, Yu Han, Zheng Lv, Xuemei Tian, Han Li, Panpan Xie, Liangyu Zheng
Polyamine tags fused in Candida antarctica lipase B (CalB) can help achieve high soluble expression of CalB in E. coli and can directly mediate silicification, which leads to rapid formation of a CalB-silica particle complex through a one-step approach. After optimization experiments, the fused lipase CalB tagged with 6-histidine at the N terminal and 10-lysine at the C terminal (6His-CalB-10Lys) is effectively expressed with high solubility (0.1mg/mL) and specific activity (10.1U/mg), and easily cross-linked in silica particles with a high immobilization efficiency of 96...
March 16, 2017: Journal of Biotechnology
Xiaolin Pei, Zhengfei Yang, Anming Wang, Lirong Yang, Jianping Wu
Nitrile hydratase (NHase) has attracted considerable attention to synthesize valuable amides. Here, the gene cluster of Co-type NHase from Aurantimonas manganoxydans ATCC BAA-1229 (NHase1229) was identified through Edman degradation and recombinant expression, including α-subunit, β-subunit, and activator. The activator gene contained 408 bases, which was essential for the biosynthesis of NHase1229 in Escherichia coli. The activator gene can be substituted by Co-type activator from Pseudonocardia thermophila JCM3095 and Pseudomonas putida 5B to assist in the functional expression of NHase1229, not by that of Fe-type NHase from Pseudomonas putida F1...
March 16, 2017: Journal of Biotechnology
Longhai Dai, Jiao Li, Peiyuan Yao, Yueming Zhu, Yan Men, Yan Zeng, Jiangang Yang, Yuanxia Sun
Glycosylation is a prominent biological mechanism for structural and functional diversity of natural products. Uridine diphosphate-dependent glycosyltransferases with aglycon promiscuity are generally recognised as effective biocatalysts for glycodiversification of natural products for practical applications. In this study, the aglycon promiscuity of glycosyltransferase Bs-YjiC from Bacillus subtilis 168 was explored. Bs-YjiC, with uridine diphosphate glucose (UDPG) as sugar donor, exhibited robust capabilities to glycosylate 19 structurally diverse types of drug-like scaffolds with regio- and stereospecificities and form O-, N- and S-linkage glycosides...
March 15, 2017: Journal of Biotechnology
Qian Li, Abdullah Al Loman, Anthony M Coffman, Lu-Kwang Ju
Soybean hull consists mainly of three major plant carbohydrates, i.e., cellulose, hemicellulose and pectin. It is inexpensive and a good potential substrate for carbohydrase production because it is capable of inducing a complete spectrum of activities to hydrolyze complex biomass. Aspergillus is known for carbohydrase production but no studies have evaluated and compared, among Aspergillus species and strains, the soybean hull induced production of various carbohydrases. In this study, A. aculeatus, A. cinnamomeus, A...
March 14, 2017: Journal of Biotechnology
Xinyue Zhao, Fang Ma, Cuijie Feng, Shunwen Bai, Jixian Yang, Li Wang
An atrazine-degrading strain Arthrobacter sp. ZXY-2 was originally isolated from Jilin Pesticide Plant (China). Strain ZXY-2 demonstrated excellent atrazine degradation performance and saline tolerance. Here we report the complete genome sequence of strain ZXY-2 contained a circular chromosome and five circular plasmids encoding for the mechanism of salt adaptation and pollutant degradation.
March 14, 2017: Journal of Biotechnology
Daniel Jaeger, Wolfgang Hübner, Thomas Huser, Jan H Mussgnug, Olaf Kruse
Photosynthetic microalgae hold great promise as non-food feedstocks for the sustainable production of a range of bio-products and genetic engineering is an increasingly important strategy to improve the natural cellular features. For the vast majority of microalgal strains, however, genetic engineering is not yet possible and the establishment of efficient genetic tools for a broad range of microalgal species is an important task to reach biotechnological success. Stable DNA transformation is one of the crucial steps for most genetic engineering approaches...
March 13, 2017: Journal of Biotechnology
Smriti Shridhar, Gerald Klanert, Norbert Auer, Inmaculada Hernandez-Lopez, Maciej M Kańduła, Matthias Hackl, Johannes Grillari, Nancy Stralis-Pavese, David P Kreil, Nicole Borth
Chinese Hamster Ovary (CHO) cells are the preferred cell line for production of biopharmaceuticals. These cells are capable to grow without serum supplementation, but drastic changes in their phenotype occur during adaptation to protein-free growth, which typically include the change to a suspension phenotype with reduced growth rate. A possible approach to understand this transformation, with the intention to counteract the reduction in growth by targeted supplementation of protein-free media, is gene expression profiling...
March 13, 2017: Journal of Biotechnology
Koichi Tamano, Ai Miura, Hideaki Koike, Yasushi Kamisaka, Myco Umemura, Masayuki Machida
Free fatty acids (FFAs) are useful for generating biofuel compounds and functional lipids. Microbes are increasingly exploited to produce FFAs via metabolic engineering. However, in many microorganisms, FFAs accumulate in the cytosol, and disrupting cells to extract them is energy intensive. Thus, a simple cost-effective extraction technique must be developed to remove this drawback. We found that FFAs were released from cells of the filamentous fungus Aspergillus oryzae with high efficiency when they were cultured or incubated with non-ionic surfactants such as Triton X-100...
March 12, 2017: Journal of Biotechnology
Xiaotian Zhong, Tao He, Amar S Prashad, Wenge Wang, Justin Cohen, Darren Ferguson, Amy S Tam, Eric Sousa, Laura Lin, Lioudmila Tchistiakova, Scott Gatto, Aaron D'Antona, Yen-Tung Luan, Weijun Ma, Richard Zollner, Jing Zhou, Bo Arve, Will Somers, Ronald Kriz
Protein modifications by intricate cellular machineries often redesign the structure and function of existing proteins to impact biological networks. Disulfide bond formation between cysteine (Cys) pairs is one of the most common modifications found in extracellularly-destined proteins, key to maintaining protein structure. Unpaired surface cysteines on secreted mammalian proteins are also frequently found disulfide-bonded with free Cys or glutathione (GSH) in circulation or culture, the mechanism for which remains unknown...
March 11, 2017: Journal of Biotechnology
Thomas Gundinger, Oliver Spadiut
Horseradish peroxidase (HRP) is used in various biotechnological and medical applications. Since its isolation from plant provides several disadvantages, the bacterium Escherichia coli was tested as recombinant expression host in former studies. However, neither production from refolded inclusion bodies nor active enzyme expression in the periplasm exceeded final titres of 10mg per litre cultivation broth. Thus, the traditional way of production of HRP from plant still prevails. In this study, we revisited the recombinant production of HRP in E...
March 11, 2017: Journal of Biotechnology
Camron S Dawes, Heiko Konig, Chien-Chi Lin
Hypoxia is a critical condition governing many aspects of cellular fate processes. The most common practice in hypoxic cell culture is to maintain cells in an incubator with controlled gas inlet (i.e., hypoxic chamber). Here, we describe the design and characterization of enzyme-immobilized hydrogels to create solution hypoxia under ambient conditions for in vitro cancer cell culture. Specifically, glucose oxidase (GOX) was acrylated and co-polymerized with poly(ethylene glycol)-diacrylate (PEGDA) through photopolymerization to form GOX-immobilized PEG-based hydrogels...
March 8, 2017: Journal of Biotechnology
Jia Zhou, Liyang Yang, Chonglong Wang, Eui-Sung Choi, Seon-Won Kim
The 2C-methyl-D-erythritol 4-phosphate (MEP) pathway is a carbon-efficient route for synthesis of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), the building blocks of isoprenoids. However, practical application of a native or recombinant MEP pathway for the mass production of isoprenoids in Escherichia coli has been unsatisfactory. In this study, the entire recombinant MEP pathway was established with plasmids and used for the production of an isoprenoid, protoilludene. E. coli harboring the recombinant MEP pathway plasmid (ME) and a protoilludene synthesis pathway plasmid (AO) produced 10...
March 7, 2017: Journal of Biotechnology
Kenneth Wei Min Tan, Yuan Kun Lee
Biofuel production from genetically-engineered microalgae is currently among the most widely studied strategies in generating renewable energy. However, microalgae currently suffer from low oil yields which limit the commercial feasibility of industrial-scale production. A major bottleneck in cost-efficient biofuel production from microalgae is the dilemma between biomass productivity and lipid accumulation. When grown under stressful culture conditions such as nitrogen depletion, microalgae accumulate large amounts of neutral lipids, but it comes at the expense of growth which negatively impacts overall lipid productivity...
March 7, 2017: Journal of Biotechnology
Piriya Luangwattananun, Warawan Eiamphungporn, Napat Songtawee, Leif Bülow, Chartchalerm Isarankura Na Ayudhya, Virapong Prachayasittikul, Sakda Yainoy
Synergistic action of major antioxidant enzymes, e.g., superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) is known to be more effective than the action of any single enzyme. Recently, we have engineered a tri-functional enzyme, 6His-MnSOD-TAT/CAT-MnSOD (M-TAT/CM), with SOD, CAT and cell-permeable activities. The protein actively internalized into the cells and showed superior protection against oxidative stress-induced cell death over native enzymes fused with TAT. To improve its molecular size, enzymatic activity and stability, in this study, MnSOD portions of the engineered protein were replaced by CuZnSOD, which is the smallest and the most heat resistant SOD isoform...
March 6, 2017: Journal of Biotechnology
Matthias Steglich, Ulrich Nübel
We tested the capabilities of four different software tools to detect insertions and deletions (indels) in a bacterial genome on the basis of short sequencing reads. We included tools applying the gapped-alignment (VarScan, FreeBayes) or split-read (Pindel) methods, respectively, and a combinatorial approach with local de-novo assembly (ScanIndel). Tests were performed with 151-basepair, paired-end sequencing reads simulated from a bacterial (Clostridioides difficile R20291) genome sequence with predefined indels (indel length, 1-2321bp)...
March 4, 2017: Journal of Biotechnology
Hendrik Pollmann, Jürgen Breitenbach, Gerhard Sandmann
Phytoene is a colorless carotenoid with increasing economic potential for skin care but with limited availability. The red yeast Xanthophyllomyces dendrorhous which has previously been used as a production platform for carotenoids was engineered as a prototype for the yield of this carotene. Phytoene was accumulated by prevention of its metabolization by desaturation in the carotenoid pathway. In a first step, the phytoene desaturase gene crtI was disrupted by insertion of a hygromycin-resistance gene. Most of the resulting transformants were heterozygote for intact and inactivated crtI...
March 3, 2017: Journal of Biotechnology
Lin Lin, Trine Skov Petersen, Kristopher Torp Jensen, Lars Bolund, Ralf Kühn, Yonglun Luo
Mammalian cells repair double-strand DNA breaks (DSB) by a range of different pathways following DSB induction by the engineered clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein Cas9. While CRISPR-Cas9 thus enables predesigned modifications of the genome, applications of CRISPR-Cas9-mediated genome-editing are frequently hampered by the unpredictable and varying pathways for DSB repair in mammalian cells. Here we present a strategy of fusing Cas9 to recombinant proteins for fine-tuning of the DSB repair preferences in mammalian cells...
March 1, 2017: Journal of Biotechnology
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