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Bioengineered

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https://www.readbyqxmd.com/read/28644714/genome-mining-in-silico-validation-and-phase-selection-of-a-novel-aldo-keto-reductase-from-candida-glabratea-for-biotransformation
#1
Souvik Basak, Nanda Gopal Sahoo, Angayar K Pavanasam
Previously, we published cloning, over-expression, characterization and subsequent exploitation of a carbonyl reductase (cr) gene, belonging to general family aldo-keto reductase from Candida glabrata CBS138 to convert keto ester (COBE) to a chiral alcohol (ethyl-4-chloro-3-hydroxybutanoate or CHBE). Exploiting global transcription factor CRP, rDNA and transporter engineering, we have improved batch production of CHBE by trinomial bioengineering. Herein, we present the exploration of cr gene in Candida glabrata CBS138 through genome miningapproach, in silico validation of its activity and selection of its biocatalytic phase...
June 23, 2017: Bioengineered
https://www.readbyqxmd.com/read/28632991/uncovering-the-repertoire-of-fungal-secondary-metabolites-from-fleming-s-lab-to-the-international-space-station
#2
Tomasz Boruta
Fungi produce a variety of secondary metabolites (SMs), low-molecular weight compounds associated with many potentially useful biological activities. The examples of biotechnologically relevant fungal metabolites include penicillin, a beta-lactam antibiotic, and lovastatin, a cholesterol-lowering drug. The discovery of pharmaceutical lead compounds within the microbial metabolic pools relies on the selection and biochemical characterization of promising strains. Not all SMs are produced under standard cultivation conditions, hence the uncovering of chemical potential of investigated strains often requires the use of induction strategies to awake the associated biosynthetic genes...
June 20, 2017: Bioengineered
https://www.readbyqxmd.com/read/28594266/simple-fluorescence-based-detection-of-protein-kinase-a-activity-using-a-molecular-beacon-probe
#3
Changbei Ma, Xiaoyuan Lv, Kemin Wang, Shunxin Jin, Haisheng Liu, Kefeng Wu, Weimin Zeng
Protein kinase A was detected by quantifying the amount of ATP utilized after a protein kinase reaction. The ATP assay was performed using the T4 DNA ligase and a molecular beacon (MB). In the presence of ATP, DNA ligase catalyzed the ligation of short DNA. The ligation product then hybridized to MB, resulting in a fluorescence enhancement of the MB. This assay was capable of determining protein kinase A in the range of 12.5∼150 nM, with a detection limit of 1.25 nM. Furthermore, this assay could also be used to investigate the effect of genistein on protein kinase A...
June 8, 2017: Bioengineered
https://www.readbyqxmd.com/read/28574746/microbial-production-of-mevalonate-by-recombinant-escherichia-coli-using-acetic-acid-as-a-carbon-source
#4
Xin Xu, Meng Xie, Qian Zhao, Mo Xian, Huizhou Liu
We sought to produce mevalonate, an important organic acid, by recombinant Escherichia coli, using acetic acid, a less costly alternative feedstock, as a carbon source. In this study, the mevalonate biosynthesis pathway originating with acetate was constructed in recombinant E. coli, resulting in the production of 1.06 g L(-1) mevalonate with a productivity of 0.03 g L(-1) h(-1) in a 5-L bioreactor. The mevalonate concentration and productivity were significantly enhanced with increased cell density during 2-stage aerobic fermentation, reaching 7...
June 2, 2017: Bioengineered
https://www.readbyqxmd.com/read/28509615/mining-of-a-phospholipase-d-and-its-application-in-enzymatic-preparation-of-phosphatidylserine
#5
Wen-Bin Zhou, Jin-Song Gong, Hai-Juan Hou, Heng Li, Zhen-Ming Lu, Hong-Yu Xu, Zheng-Hong Xu, Jin-Song Shi
Phosphatidylserine (PS) is useful as the additive in industries for memory improvement, mood enhancement and drug delivery. Conventionally, PS was extracted from soybeans, vegetable oils, egg yolk, and biomass; however, their low availability and high extraction cost were limiting factors. Phospholipase D (PLD) is a promising tool for enzymatic synthesis of PS due to its transphosphatidylation activity. In this contribution, a new and uncharacterized PLD was first obtained from GenBank database via genome mining strategy...
May 16, 2017: Bioengineered
https://www.readbyqxmd.com/read/28494182/fabg-can-function-as-phab-for-poly-3-hydroxybutyrate-biosynthesis-in-photosynthetic-cyanobacteria-synechocystis-sp-pcc-6803
#6
Haowei Zhang, Yinghui Liu, Changhong Yao, Xupeng Cao, Jing Tian, Song Xue
The production of poly-3-hydroxybutyrate (PHB) by photosynthetic cyanobacteria is a potentially sustainable production method for the biodegradable plastics industry. β-Ketoacyl-ACP reductase (FabG), from the photosynthetic cyanobacterium Synechocystis sp. PCC 6803 (SpFabG), is the first NADPH-dependent reductase in the fatty acid biosynthesis pathway. Its structure is similar to that of acetoacetyl-CoA reductase (SpPhaB), which is critical for PHB synthesis and can replace SpPhaB for acetoacetyl-CoA reduction in vitro...
May 11, 2017: Bioengineered
https://www.readbyqxmd.com/read/28471319/enhanced-microalgal-lipid-production-with-media-engineering-of-potassium-nitrate-as-a-nitrogen-source
#7
Rakesh Singh Gour, Madhusudan Bairagi, Vijay Kumar Garlapati, Anil Kant
Algal biofuels are far from a commercial reality due to the technical challenges associated with their growth and lipid extraction procedures. In this study, we investigated the effect of 4 different media and 5 different nitrogen sources at 5 levels on the growth, biomass and lipid productivity of Scenedesmus sp and Chlorella sp The hypothesis was that a nitrogen source can be identified that provides enough stress to accumulate lipids without compromising significantly on biomass and lipid productivity. A maximum specific growth rate and doubling per day have been observed with algal species using modified BG-11 medium...
May 4, 2017: Bioengineered
https://www.readbyqxmd.com/read/28471311/a-synthetic-hybrid-promoter-for-d-xylonate-production-at-low-ph-in-the-tolerant-yeast-candida-glycerinogenes
#8
Hao Ji, Xinyao Lu, Hong Zong, Bin Zhuge
The tolerant yeast Candida glycerinogenes, with high D-xylonate and low-pH tolerances, was used as the host for D-xylonate production at low pH in this study. A low-pH inducible promoter, pGUKd, was engineered using the core promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (pGAP) combined with the upstream activating sequence of the promoter of the guanylate kinase gene (pGUK1) that had substituted pH-responsive TF binding sites. The recombinant cells that expressed GFP from the hybrid promoter pGUKd displayed dramatically increased fluorescence intensity at pH 2...
May 4, 2017: Bioengineered
https://www.readbyqxmd.com/read/28471292/statistical-optimization-for-the-production-of-recombinant-cold-adapted-superoxide-dismutase-in-e-coli-using-response-surface-methodology
#9
Yatong Wang, Quanfu Wang, Yifan Wang, Han Han, Yanhua Hou, Yonglei Shi
Cold-adapted superoxide dismutase (SOD) with higher catalytic activity at lower temperature has great amount of applications in many aspects as an industrial enzyme. The application of recombinant enzyme in gene engineering and microbial fermentation technology is an effective way to obtain high-yield product. In this study, to obtain the recombinant SOD in E. coli (rPsSOD) with the highest activity, the Box-Behnken design was first applied to optimize the important parameters (lactose, tryptone and Tween-80) affecting the activity of rPsSOD...
May 4, 2017: Bioengineered
https://www.readbyqxmd.com/read/28463573/bioengineering-toward-direct-production-of-immobilized-enzymes-a-paradigm-shift-in-biocatalyst-design
#10
Fabian B H Rehm, Shuxiong Chen, Bernd H A Rehm
The need for cost-effectively produced and improved biocatalysts for industrial, pharmaceutical and environmental processes is steadily increasing. While enzyme properties themselves can be improved via protein engineering, immobilization by attachment to carrier materials remains a critical step for stabilization and process implementation. A new emerging immobilization approach, the in situ immobilization, enables simultaneous production of highly active enzymes and carrier materials using bioengineering/synthetic biology of microbial cells...
May 2, 2017: Bioengineered
https://www.readbyqxmd.com/read/28453429/anti-microbial-coating-innovations-to-prevent-infectious-diseases-amici-cost-action-ca15114
#11
Colum P Dunne, Minna M Keinänen-Toivola, Anne Kahru, Birgit Teunissen, Hulya Olmez, Isabel Gouveia, Luis Melo, Kazimierz Murzyn, Martina Modic, Merja Ahonen, Pete Askew, Theofilos Papadopoulos, Christian Adlhart, Francy R L Crijns
Worldwide, millions of patients are affected annually by healthcare-associated infection (HCAI), impacting up to 80,000 patients in European Hospitals on any given day. This represents not only public health risk, but also an economic burden. Complementing routine hand hygiene practices, cleaning and disinfection, antimicrobial coatings hold promise based, in essence, on the application of materials and chemicals with persistent bactericidal or -static properties onto surfaces or in textiles used in healthcare environments...
April 28, 2017: Bioengineered
https://www.readbyqxmd.com/read/28453385/performance-improvement-of-microbial-fuel-cell-mfc-using-suitable-electrode-and-bioengineered-organisms-a-review
#12
Payel Choudhury, Uma Shankar Prasad Uday, Tarun Kanti Bandyopadhyay, Rup Narayan Ray, Biswanath Bhunia
There is an urgent need to find an environment friendly and sustainable technology for alternative energy due to rapid depletion of fossil fuel and industrialization. Microbial Fuel Cells (MFCs) have operational and functional advantages over the current technologies for energy generation from organic matter as it directly converts electricity from substrate at ambient temperature. However, MFCs are still unsuitable for high energy demands due to practical limitations. The overall performance of an MFC depends on microorganism, appropriate electrode materials, suitable MFC designs, and optimizing process parameters which would accelerate commercialization of this technology in near future...
April 28, 2017: Bioengineered
https://www.readbyqxmd.com/read/28453384/isolation-and-characterization-of-ulva-prolifera-actin1-gene-and-function-verification-of-the-5-flanking-region-as-a-strong-promoter
#13
Chunhui Wu, Peng Jiang, Yang Guo, Jianguo Liu, Jin Zhao, Huihui Fu
Ulva prolifera is a green macroalgae with an extremely high growth rate that can accumulate biomass with considerable protein content. To set up an available seaweed expression system, a prior step is to isolate endogenous and strong constitutive promoters. For this reason, the full-length genomic actin1 gene from U. prolifera (Upactin1) was cloned and its 5' flanking sequence was obtained by genome walking. The Upactin1 open reading frame consisted of 1134 nucleotides encoding 377 amino acid residues. Besides 4 exons and 3 introns in the coding region, an extra leader intron was identified in the 5' untranslated region...
April 28, 2017: Bioengineered
https://www.readbyqxmd.com/read/28448741/chromophore-attachment-to-fusion-protein-of-streptavidin-and-recombinant-allophycocyanin-%C3%AE-subunit
#14
Jing Wu, Huaxin Chen, Peng Jiang
The fusion protein (SLA) of streptavidin and allophycocyanin α subunit (holo-ApcA) was biosynthesized in Escherichia coli by a dual plasmid system. The recombinant SLA, purified by affinity chromatography, showed spectral properties similar to natural allophycocyanin α subunit (ApcA). Spectral and Zinc staining analysis indicated that the recombinant SLA covalently bound phycocyanobilin (PCB). To improve chromophorylation rate of recombinant SLA, an in vitro chromophore attachment reaction system was established, which contained partially chromophylated SLA, PCB and lyase CpcS...
April 27, 2017: Bioengineered
https://www.readbyqxmd.com/read/28430539/metabolic-pathway-rewiring-in-engineered-cyanobacteria-for-solar-to-chemical-and-solar-to-fuel-production-from-co2
#15
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
https://www.readbyqxmd.com/read/28430004/a-strategy-for-l-isoleucine-dioxygenase-screening-and-4-hydroxyisoleucine-production-by-resting-cells
#16
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
https://www.readbyqxmd.com/read/28409998/suitability-of-various-dna-extraction-methods-for-a-traditional-chinese-paocai-system
#17
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
https://www.readbyqxmd.com/read/28394734/optimization-of-bioengineered-heparin-heparan-sulfate-production-for-therapeutic-applications
#18
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
https://www.readbyqxmd.com/read/28394725/repeated-batch-fermentation-of-immobilized-e-coli-expressing-vitreoscilla-hemoglobin-for-long-term-use
#19
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
https://www.readbyqxmd.com/read/28384075/synergistic-action-between-extracellular-products-from-white-rot-fungus-and-cellulase-significantly-improves-enzymatic-hydrolysis
#20
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 biologic 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 maize stovers...
April 6, 2017: Bioengineered
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