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Microbial Cell Factories

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https://www.readbyqxmd.com/read/28231787/ethylene-production-with-engineered-synechocystis-sp-pcc-6803-strains
#1
Vinod Puthan Veetil, S Andreas Angermayr, Klaas J Hellingwerf
BACKGROUND: Metabolic engineering and synthetic biology of cyanobacteria offer a promising sustainable alternative approach for fossil-based ethylene production, by using sunlight via oxygenic photosynthesis, to convert carbon dioxide directly into ethylene. Towards this, both well-studied cyanobacteria, i.e., Synechocystis sp PCC 6803 and Synechococcus elongatus PCC 7942, have been engineered to produce ethylene by introducing the ethylene-forming enzyme (Efe) from Pseudomonas syringae pv...
February 23, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28228147/artificially-designed-recombinant-protein-composed-of-multiple-epitopes-of-foot-and-mouth-disease-virus-as-a-vaccine-candidate
#2
Ho-Bin Lee, Da-Chuan Piao, Jun-Yeong Lee, Jae-Yun Choi, Jin-Duck Bok, Chong-Su Cho, Sang-Kee Kang, Yun-Jaie Choi
BACKGROUND: Concerns regarding the safety of inactivated foot-and-mouth disease (FMD) vaccine have been raised since it is produced from cultured live FMD virus (FMDV). To overcome this issue, recombinant protein has been studied as an alternative vaccine. RESULTS AND CONCLUSION: We designed a chimerical multi-epitope recombinant protein (5BT), which is comprised of tandem repeats of five B cell epitopes (residue of VP1 136-162) derived from different FMDV variants and one T-cell epitope (residue of 3A 21-35)...
February 22, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28219382/high-level-extracellular-protein-production-in-bacillus-subtilis-using-an-optimized-dual-promoter-expression-system
#3
Kang Zhang, Lingqia Su, Xuguo Duan, Lina Liu, Jing Wu
BACKGROUND: We recently constructed a Bacillus subtilis strain (CCTCC M 2016536) from which we had deleted the srfC, spoIIAC, nprE, aprE and amyE genes. This strain is capable of robust recombinant protein production and amenable to high-cell-density fermentation. Because the promoter is among the factors that influence the production of target proteins, optimization of the initial promoter, PamyQ from Bacillus amyloliquefaciens, should improve protein expression using this strain. This study was undertaken to develop a new, high-level expression system in B...
February 20, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28212656/using-a-vector-pool-containing-variable-strength-promoters-to-optimize-protein-production-in-yarrowia-lipolytica
#4
Rémi Dulermo, François Brunel, Thierry Dulermo, Rodrigo Ledesma-Amaro, Jérémy Vion, Marion Trassaert, Stéphane Thomas, Jean-Marc Nicaud, Christophe Leplat
BACKGROUND: The yeast Yarrowia lipolytica is an increasingly common biofactory. To enhance protein expression, several promoters have been developed, including the constitutive TEF promoter, the inducible POX2 promotor, and the hybrid hp4d promoter. Recently, new hp4d-inspired promoters have been created that couple various numbers of UAS1 tandem elements with the minimal LEU2 promoter or the TEF promoter. Three different protein-secretion signaling sequences can be used: preLip2, preXpr2, and preSuc2...
February 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28202018/de-novo-biosynthesis-of-pterostilbene-in-an-escherichia-coli-strain-using-a-new-resveratrol-o-methyltransferase-from-arabidopsis
#5
Kyung Taek Heo, Sun-Young Kang, Young-Soo Hong
BACKGROUND: Pterostilbene, a structural analog of resveratrol, has higher oral bioavailability and bioactivity than that of the parent compound; but is far less abundant in natural sources. Thus, to efficiently obtain this bioactive resveratrol analog, it is necessary to develop new bioproduction systems. RESULTS: We identified a resveratrol O-methyltransferase (ROMT) function from a multifunctional caffeic acid O-methyltransferase (COMT) originating from Arabidopsis, which catalyzes the transfer of a methyl group to resveratrol resulting in pterostilbene production...
February 15, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28193276/thermus-thermophilus-as-source-of-thermozymes-for-biotechnological-applications-homologous-expression-and-biochemical-characterization-of-an-%C3%AE-galactosidase
#6
Martina Aulitto, Salvatore Fusco, Gabriella Fiorentino, Danila Limauro, Emilia Pedone, Simonetta Bartolucci, Patrizia Contursi
BACKGROUND: The genus Thermus, which has been considered for a long time as a fruitful source of biotechnological relevant enzymes, has emerged more recently as suitable host to overproduce thermozymes. Among these, α-galactosidases are widely used in several industrial bioprocesses that require high working temperatures and for which thermostable variants offer considerable advantages over their thermolabile counterparts. RESULTS: Thermus thermophilus HB27 strain was used for the homologous expression of the TTP0072 gene encoding for an α-galactosidase (TtGalA)...
February 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28193216/a-high-throughput-expression-screening-platform-to-optimize-the-production-of-antimicrobial-peptides
#7
Christine Schreiber, Hagen Müller, Oliver Birrenbach, Moritz Klein, Doreen Heerd, Tobias Weidner, Denise Salzig, Peter Czermak
BACKGROUND: Antimicrobial peptides (AMPs) are promising candidates for the development of novel antibiotics, but it is difficult to produce sufficient quantities for preclinical and clinical studies due to their toxicity towards microbial expression hosts. To avoid laborious trial-and-error testing for the identification of suitable expression constructs, we have developed a small-scale expression screening platform based on a combinatorial plasmid library. RESULTS: The combinatorial library is based on the Golden Gate cloning system...
February 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28193209/secretion-of-biologically-active-pancreatitis-associated-protein-i-pap-by-genetically-modified-dairy-lactococcus-lactis-nz9000-in-the-prevention-of-intestinal-mucositis
#8
Rodrigo D Carvalho, Natalia Breyner, Zelia Menezes-Garcia, Nubia M Rodrigues, Luisa Lemos, Tatiane U Maioli, Danielle da Gloria Souza, Denise Carmona, Ana M C de Faria, Philippe Langella, Jean-Marc Chatel, Luis G Bermúdez-Humarán, Henrique C P Figueiredo, Vasco Azevedo, Marcela S de Azevedo
BACKGROUND: Mucositis is one of the most relevant gastrointestinal inflammatory conditions in humans, generated by the use of chemotherapy drugs, such as 5-fluoracil (5-FU). 5-FU-induced mucositis affects 80% of patients undergoing oncological treatment causing mucosal gut dysfunctions and great discomfort. As current therapy drugs presents limitations in alleviating mucositis symptoms, alternative strategies are being pursued. Recent studies have shown that the antimicrobial pancreatitis-associated protein (PAP) has a protective role in intestinal inflammatory processes...
February 13, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28187768/current-lipid-extraction-methods-are-significantly-enhanced-adding-a-water-treatment-step-in-chlorella-protothecoides
#9
Xiaojie Ren, Xinhe Zhao, François Turcotte, Jean-Sébastien Deschênes, Réjean Tremblay, Mario Jolicoeur
BACKGROUND: Microalgae have the potential to rapidly accumulate lipids of high interest for the food, cosmetics, pharmaceutical and energy (e.g. biodiesel) industries. However, current lipid extraction methods show efficiency limitation and until now, extraction protocols have not been fully optimized for specific lipid compounds. The present study thus presents a novel lipid extraction method, consisting in the addition of a water treatment of biomass between the two-stage solvent extraction steps of current extraction methods...
February 11, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28179021/switch-a-dynamic-crispr-tool-for-genome-engineering-and-metabolic-pathway-control-for-cell-factory-construction-in-saccharomyces-cerevisiae
#10
Katherina García Vanegas, Beata Joanna Lehka, Uffe Hasbro Mortensen
BACKGROUND: The yeast Saccharomyces cerevisiae is increasingly used as a cell factory. However, cell factory construction time is a major obstacle towards using yeast for bio-production. Hence, tools to speed up cell factory construction are desirable. RESULTS: In this study, we have developed a new Cas9/dCas9 based system, SWITCH, which allows Saccharomyces cerevisiae strains to iteratively alternate between a genetic engineering state and a pathway control state...
February 8, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28179004/understanding-the-interplay-of-carbon-and-nitrogen-supply-for-ectoines-production-and-metabolic-overflow-in-high-density-cultures-of-chromohalobacter-salexigens
#11
María J Salar-García, Vicente Bernal, José M Pastor, Manuel Salvador, Montserrat Argandoña, Joaquín J Nieto, Carmen Vargas, Manuel Cánovas
BACKGROUND: The halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ectoines as compatible solutes. However, metabolic overflow, which is a major drawback for the efficient conversion of biological feedstocks, occurs as a result of metabolic unbalances during growth and ectoines production...
February 8, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28178978/enhanced-extracellular-production-of-recombinant-proteins-in-escherichia-coli-by-co-expression-with-bacillus-cereus-phospholipase-c
#12
Lingqia Su, Qi Jiang, Lingang Yu, Jing Wu
BACKGROUND: Our laboratory has reported a strategy for improving the extracellular production of recombinant proteins through co-expression with Thermobifida fusca cutinase, which increases membrane permeability via its phospholipid hydrolysis activity. However, the foam generated by the lysophospholipid product makes the fermentation process difficult to control in a fermentor. Phospholipase C (PLC) catalyzes the hydrolysis of phospholipids to produce sn1,2-diacylglycerides and organic phosphate, which do not induce foam formation...
February 8, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28178965/effect-of-glucose-on-poly-%C3%AE-glutamic-acid-metabolism-in-bacillus-licheniformis
#13
Wencheng Yu, Zhen Chen, Hong Ye, Peize Liu, Zhipeng Li, Yuanpeng Wang, Qingbiao Li, Shan Yan, Chuan-Jian Zhong, Ning He
BACKGROUND: Poly-gamma-glutamic acid (γ-PGA) is a promising macromolecule with potential as a replacement for chemosynthetic polymers. γ-PGA can be produced by many microorganisms, including Bacillus species. Bacillus licheniformis CGMCC2876 secretes γ-PGA when using glycerol and trisodium citrate as its optimal carbon sources and secretes polysaccharides when using glucose as the sole carbon source. To better understand the metabolic mechanism underlying the secretion of polymeric substances, SWATH was applied to investigate the effect of glucose on the production of polysaccharides and γ-PGA at the proteome level...
February 8, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28153019/the-effect-of-enhanced-acetate-influx-on-synechocystis-sp-pcc-6803-metabolism
#14
Kati Thiel, Eerika Vuorio, Eva-Mari Aro, Pauli Tapio Kallio
BACKGROUND: Acetate is a common microbial fermentative end-product, which can potentially be used as a supplementary carbon source to enhance the output of biotechnological production systems. This study focuses on the acetate metabolism of the photosynthetic cyanobacterium Synechocystis sp. PCC 6803 which is unable to grow on acetate as a sole carbon source but still can assimilate it via acetyl-CoA-derived metabolic intermediates. In order to gain insight into the acetate uptake, associated limitations and metabolic effects, a heterologous acetate transporter ActP from Escherichia coli was introduced into Synechocystis to facilitate the transport of supplemented acetate from the medium into the cell...
February 2, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28143563/adipic-acid-tolerance-screening-for-potential-adipic-acid-production-hosts
#15
Emma Karlsson, Valeria Mapelli, Lisbeth Olsson
BACKGROUND: Biobased processes for the production of adipic acid are of great interest to replace the current environmentally detrimental petrochemical production route. No efficient natural producer of adipic acid has yet been identified, but several approaches for pathway engineering have been established. Research has demonstrated that the microbial production of adipic acid is possible, but the yields and titres achieved so far are inadequate for commercialisation. A plausible explanation may be intolerance to adipic acid...
February 1, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28143533/erratum-to-fermentative-production-and-direct-extraction-of-%C3%AE-bisabolol-in-metabolically-engineered-escherichia-coli
#16
Gui Hwan Han, Seong Keun Kim, Paul Kyung-Seok Yoon, Younghwan Kang, Byoung Su Kim, Yaoyao Fu, Bong Hyun Sung, Heung Chae Jung, Dae-Hee Lee, Seon-Won Kim, Seung-Goo Lee
No abstract text is available yet for this article.
January 31, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28143479/droplet-based-microfluidic-high-throughput-screening-of-heterologous-enzymes-secreted-by-the-yeast-yarrowia-lipolytica
#17
Thomas Beneyton, Stéphane Thomas, Andrew D Griffiths, Jean-Marc Nicaud, Antoine Drevelle, Tristan Rossignol
BACKGROUND: Droplet-based microfluidics is becoming an increasingly attractive alternative to microtiter plate techniques for enzymatic high-throughput screening (HTS), especially for exploring large diversities with lower time and cost footprint. In this case, the assayed enzyme has to be accessible to the substrate within the water-in-oil droplet by being ideally extracellular or displayed at the cell surface. However, most of the enzymes screened to date are expressed within the cytoplasm of Escherichia coli cells, which means that a lysis step must take place inside the droplets for enzyme activity to be assayed...
January 31, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28137282/dynamic-control-of-erg20-expression-combined-with-minimized-endogenous-downstream-metabolism-contributes-to-the-improvement-of-geraniol-production-in-saccharomyces-cerevisiae
#18
Jianzhi Zhao, Chen Li, Yan Zhang, Yu Shen, Jin Hou, Xiaoming Bao
BACKGROUND: Microbial production of monoterpenes provides a promising substitute for traditional chemical-based methods, but their production is lagging compared with sesquiterpenes. Geraniol, a valuable monoterpene alcohol, is widely used in cosmetic, perfume, pharmaceutical and it is also a potential gasoline alternative. Previously, we constructed a geraniol production strain by engineering the mevalonate pathway together with the expression of a high-activity geraniol synthase. RESULTS: In this study, we further improved the geraniol production through reducing the endogenous metabolism of geraniol and controlling the precursor geranyl diphosphate flux distribution...
January 31, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28137256/production-and-characterization-of-a-novel-antifungal-chitinase-identified-by-functional-screening-of-a-suppressive-soil-metagenome
#19
Francesca Berini, Ilaria Presti, Fabrizio Beltrametti, Marco Pedroli, Kjell M Vårum, Loredano Pollegioni, Sara Sjöling, Flavia Marinelli
BACKGROUND: Through functional screening of a fosmid library, generated from a phytopathogen-suppressive soil metagenome, the novel antifungal chitinase-named Chi18H8 and belonging to family 18 glycosyl hydrolases-was previously discovered. The initial extremely low yield of Chi18H8 recombinant production and purification from Escherichia coli cells (21 μg/g cell) limited its characterization, thus preventing further investigation on its biotechnological potential. RESULTS: We report on how we succeeded in producing hundreds of milligrams of pure and biologically active Chi18H8 by developing and scaling up to a high-yielding, 30 L bioreactor process, based on a novel method of mild solubilization of E...
January 31, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28137297/microbial-production-of-vitamin-b12-a-review-and-future-perspectives
#20
REVIEW
Huan Fang, Jie Kang, Dawei Zhang
Vitamin B12 is an essential vitamin that is widely used in medical and food industries. Vitamin B12 biosynthesis is confined to few bacteria and archaea, and as such its production relies on microbial fermentation. Rational strain engineering is dependent on efficient genetic tools and a detailed knowledge of metabolic pathways, regulation of which can be applied to improve product yield. Recent advances in synthetic biology and metabolic engineering have been used to efficiently construct many microbial chemical factories...
January 30, 2017: Microbial Cell Factories
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