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Han Min Woo
Photoautotrophic cyanobacteria have been developed to convert CO2 to valuable chemicals and fuels as solar-to-chemical (S2C) and solar-to-fuel (S2F) platforms. Here, I describe the rewiring of the metabolic pathways in cyanobacteria to better understand the endogenous carbon flux and to enhance the yield of heterologous products. The plasticity of the cyanobacterial metabolism has been proposed to be advantageous for the development of S2C and S2F processes. The rewiring of the sugar catabolism and of the phosphoketolase pathway in the central cyanobacterial metabolism allowed for an enhancement in the level of target products by redirecting the carbon fluxes...
April 21, 2017: Bioengineered
Chenglin Zhang, Jie Ma, Zhixiang Li, Yunlong Liang, Qingyang Xu, Xixian Xie, Ning Chen
L-Isoleucine dioxygenase (IDO) specifically converts L-isoleucine(L-Ile) to 4-hydroxyisoleucine(4-HIL). To obtain IDO with improved activity, a strategy was developed that is dependent on the restoration of succinate-minus E. coli cell growth by the coupling of L-Ile hydroxylation and the oxidation of α-ketoglutarate(α-KGA) to succinate. Five mutants were obtained with this strategy, and the characteristics of IDO(M3), which exhibited the highest activity, were studied. The catalytic efficiency, thermal stability and catalytic rate of IDO(M3) were significantly improved compared with those of wild-type IDO...
April 21, 2017: Bioengineered
Nan Zhao, Jialiang Cai, Chuchu Zhang, Zhuang Guo, Wenwei Lu, Bo Yang, Feng-Wei Tian, Xiao-Ming Liu, Hao Zhang, Wei Chen
Traditional paocai brine (PB), which is continuously propagated by back-slopping and contains various species of lactic acid bacteria (LAB), is critical for the flavor of paocai. Culture-independent approaches are commonly used to investigate the microbial communities of fermented food. To evaluate the influence of different DNA (DNA) extraction methods on estimates of bacterial community profiles from 4 PBs, the lysis efficiency, DNA yield, purity and denaturing gradient gel electrophoresis (DGGE) profiles of V3 region of a 16S ribosomal ribonucleic acid gene were acquired...
April 14, 2017: Bioengineered
Megan S Lord, MoonSun Jung, John M Whitelock
Heparin has been used clinically as an anti-coagulant for more than 100 y and the major source of this therapeutic is still animal tissues. Contamination issues in some batches of heparin over 10 y ago have highlighted the need to develop alternative methods of production of this essential drug. (1) Bioengineering heparin by expressing serglycin in mammalian cells is a promising approach that was recently reported by the authors. (2) This addendum explores the approaches that the authors are taking to increase the yield of recombinantly expressed serglycin decorated with heparin/heparan sulfate focusing on cell culture and bioreactor conditions and proposes that the cell microenvironment is a key modulator of heparin biosynthesis...
April 10, 2017: Bioengineered
Taner Sar, Gamze Seker, Ayse Gokce Erman, Benjamin C Stark, Meltem Yesilcimen Akbas
This study describes an efficient and reusable process for ethanol production from medium containing whey powder, using alginate immobilized ethanologenic E. coli strains either expressing (TS3) or not expressing (FBR5) Vitreoscilla hemoglobin. Reuseabilities of the FBR5 and TS3 strains were investigated regarding their ethanol production capacities over the course of 15 successive 96-h batch fermentations. The ethanol production was fairly stable over the entire duration of the experiment, with strain TS3 maintaining a substantial advantage over strain FBR5...
April 10, 2017: Bioengineered
Wenwen Ding, Huanjiao Weng, Peng Jin, Guocheng Du, Jian Chen, Zhen Kang
Efficient assembly of multiple DNA fragments is a pivotal technology for synthetic biology. A scarless and sequence-independent DNA assembly method (DATEL) using thermal exonucleases has been developed recently. Here, we present a simplified DATEL (sDATEL) for efficient assembly of unphosphorylated DNA fragments with low cost. The sDATEL method is only dependent on Taq DNA polymerase and Taq DNA ligase. After optimizing the committed parameters of the reaction system such as pH and the concentration of Mg(2+) and NAD+, the assembly efficiency was increased by 32-fold...
April 6, 2017: Bioengineered
Yushan Wang, Yang Shao, Xinyue Zou, Mandi Yang, Lin Guo
With a set of perfect extracellular lignocellulolytic enzymes, white-rot fungus has been recognized as playing an important role in the degradation of lignocellulose materials, which leads to the possibility of creating a composite enzymatic system with high hydrolysis efficiency in vitro. Echinodontium taxodii is a promising white-rot fungus for biological pretreatment. In this study, we extracted the extracellular products of E. taxodii under solid-state fermentation conditions, mixed the extracellular products with cellulase to build a composite enzymatic system, and systematically evaluated the effect of this system on the hydrolysis of acid-pretreated and raw corn stovers...
April 6, 2017: Bioengineered
Roy D Sleator, Niall Smith
No abstract text is available yet for this article.
March 21, 2017: Bioengineered
Leonhard Kick, Marion Kirchner, Sabine Schneider
The adaptive bacterial immune system CRISPR-Cas is revolutionizing all fields of life science and has opened up new frontiers toward personalised medicine. Since the elucidation of the molecular mechanism of Cas9 from Streptococcus pyogenes in 2012 and its development as a genomic engineering tool, genetic modifications in more than 40 species have been performed, over 290 patents have been filed worldwide and the first clinical trials using CRISPR-Cas-modified T-cells have recently been started in China and in the US...
March 13, 2017: Bioengineered
Patrícia Pereira, Augusto Q Pedro, João A Queiroz, Ana R Figueiras, Fani Sousa
RNA interference-based technologies have emerged as an attractive and effective therapeutic option with potential application in diverse human diseases. These tools rely on the development of efficient strategies to obtain homogeneous non-coding RNA samples with adequate integrity and purity, thus avoiding non-targeted gene-silencing and related side-effects that impair their application onto pre-clinical practice. These RNAs have been preferentially obtained by in vitro transcription using DNA templates or via chemical synthesis...
March 10, 2017: Bioengineered
Xiao Li, Yu-Di Luo, Dong-Rui Pan, Xiao-Dan Shi, Ya-Li Tan, Zhi-Hong Li
Using 5 Zn(2+) supplementation strategies in a 50 L batch bioreactor named FUS-50L(A), possible correlations among Zn(2+) content and addition timing, physiologic activity (PA), halohydrin dehalogenase (HheC) accumulation of Escherichia coli P84A/MC1061 were systematically investigated. First, Zn(2+) was confirmed as the significant factor, and its optimal concentration for HheC expression was 3.87 mg/L through fermentation experiments in shaking flasks. Second, based on experimental results from the different strategies, it was found that PA, nutrient consumption rate (NCR) and specific growth rate (μ) for E...
March 10, 2017: Bioengineered
Hitoshi Morita, Kiyohito Taimatsu, Kanoko Yanagi, Atsuo Kawahara
Genome editing technologies, such as transcription activator-like effector nuclease (TALEN) and the clustered regularly interspaced short palindromic repeat (CRISPR)/ CRISPR-associated protein (Cas) systems, can induce DNA double-strand breaks (DSBs) at the targeted genomic locus, leading to frameshift-mediated gene disruption in the process of DSB repair. Recently, the technology-induced DSBs followed by DSB repairs are applied to integrate exogenous genes into the targeted genomic locus in various model organisms...
March 8, 2017: Bioengineered
Huiran Pan, Xiaolin Xu, Zhu Wen, Yanshun Kang, Xinhao Wang, Youshan Ren, Danqi Huang
Anthraquinone dye represents an important group of recalcitrant pollutants in dye wastewater. Aspergillus sp XJ-2 CGMCC12963 showed broad-spectrum decolorization ability, which could efficiently decolorize and degrade various anthraquinone dyes (50 mg L(-1)) under microaerophilic condition. And the decolorization rate of 93.3% was achieved at 120 h with Disperse Blue 2BLN (the target dye). Intermediates of degradation were detected by FTIR and GC-MS, which revealed the cleavage of anthraquinone chromophoric group and partial mineralization of target dye...
March 8, 2017: Bioengineered
Sharmila Narayanan, Pallab Sanpui, Lingaraj Sahoo, Siddhartha Sankar Ghosh
Phytaspase, a plant serine protease, has been demonstrated to play an important role in the programmed cell death of various plants. Phytaspase is synthesized as an inactive proenzyme containing an N-terminal signal peptide followed by a pro-domain and a mature protease catalytic domain. Pre-prophytaspase autocatalytically processes itself into a pro-domain and an active mature phytaspase enzyme. We have recently demonstrated the successful expression of mature phytaspase from tobacco in a bacterial system...
February 28, 2017: Bioengineered
Stephen Tamekou Lacmata, Lan Yao, Mo Xian, Hui Liu, Jules-Roger Kuiate, Huizhou Liu, Xinjun Feng, Guang Zhao
The release of intracellular products, especially polyhydroxyalkanoates, is still a great challenge in industry. To solve this bottleneck, a novel autolysis system strictly controlled with magnesium was constructed and applied to poly(3-hydroxypropionate) production in engineered Escherichia coli. The autolysis system was constructed by inserting the 5'untranslated region (5'UTR) behind promoter PmgtA with lysis genes (S, R, and Rz, from E. coli) overexpressed. The autolysis system functioned well (lysis efficiency of more than 90%) in the P3HP producer with double plasmids containing lysis genes and P3HP biosynthesis genes, whereas the P3HP production was reduced due to plasmid losses...
February 26, 2017: Bioengineered
(no author information available yet)
No abstract text is available yet for this article.
February 25, 2017: Bioengineered
Kamilla Swiech, Virgínia Picanço-Castro, Dimas Tadeu Covas
The main treatment option for Hemophilia A/B patients involves the administration of recombinant coagulation factors on-demand or in a prophylactic approach. Despite the safety and efficacy of this replacement therapy, the development of antibodies against the coagulation factor infused, which neutralize the procoagulant activity, is a severe complication. The production of recombinant coagulation factors in human cell lines is an efficient approach to avoid such complication. Human cell lines can produce recombinant proteins with post translation modifications more similar to their natural counterpart, reducing potential immunogenic reactions...
February 23, 2017: Bioengineered
Luiza Cesca Piva, Maritza Ocampo Bentacur, Viviane Castelo Branco Reis, Janice Lisboa De Marco, Lidia Maria Pepe de Moraes, Fernando Araripe Gonçalves Torres
Komagataella phaffii (formerly Pichia pastoris) is a well-known fungal system for heterologous protein production in the context of modern biotechnology. To obtain higher protein titers in this system many researchers have sought to optimize gene expression by increasing the levels of transcription of the heterologous gene. This has been typically achieved by manipulating promoter sequences or by generating clones bearing multiple copies of the desired gene. The aim of this work is to describe how these different molecular strategies have been applied in K...
February 22, 2017: Bioengineered
Xianghui He, Jinhua Dai, Youfen Fan, Chun Zhang, Xihong Zhao
Cutaneous wound healing is a complex physiological process that requires the efforts of various cell types and signaling pathways and often results in thickened collagen-enriched healed tissue called a scar. Therefore, the identification of the mechanism of cutaneous wound healing is necessary and has great value in providing better treatment. Here, we demonstrated that MMP-1 inhibition could promote cell proliferation in dermal fibroblasts via the MTT assay. Meanwhile, we investigated cell migration by flow cytometry and tested type I collagenase activity...
February 22, 2017: Bioengineered
Hui-Ting Song, Yi-Min Yang, Ding-Kang Liu, Xiao-Qing Xu, Wen-Jing Xiao, Zi-Lu Liu, Wu-Cheng Xia, Chao-Ying Wang, Xiao Yu, Zheng-Bing Jiang
Lignocellulose is a polysaccharide and an abundant biomass resource that widely exists in grains, beans, rice, and their by-products. Over 10 million tons of lignocellulose resources and processing products are produced every year in China. Three recombinant Y. lipolytica strains with cellulase (β-glucosidase, endoglucanase and cellobiohydrolase) were constructed. The enzymatic activities of these enzymes were 14.181 U/mL, 16.307 U/mL, and 17.391 U/mL, respectively. The whole cell cellulases were used for a stover bio-transformation...
February 17, 2017: Bioengineered
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