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https://www.readbyqxmd.com/read/28653998/cell-free-production-of-pentacyclic-triterpenoid-compound-betulinic-acid-from-betulin-by-the-engineered-saccharomyces-cerevisiae
#1
Jianan Wu, Yongwu Niu, Abdelmoneim Bakur, Hao Li, Qihe Chen
Betulinic acid is a product of plant secondary metabolism which has shown various bioactivities. Several CYP716A subfamily genes were recently characterized encoding multifunctional oxidases capable of C-28 oxidation. CYP716A12 was identified as betulin C-28 oxidase, capable of modifying betulin. This study aimed to induce the transformation of betulin to betulinic acid by co-expressing enzymes CYP716A12 from Medicago truncatula and ATR1 from Arabidopsis thaliana in Saccharomyces cerevisiae. The microsome protein extracted from the transgenic yeast successfully catalyzed the transformation of betulin to betulinic acid...
June 27, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28651874/enhanced-production-of-2-3-butanediol-from-xylose-by-combinatorial-engineering-of-xylose-metabolic-pathway-and-cofactor-regeneration-in-pyruvate-decarboxylase-deficient-saccharomyces-cerevisiae
#2
Soo-Jung Kim, Hee-Jin Sim, Jin-Woo Kim, Ye-Gi Lee, Yong-Cheol Park, Jin-Ho Seo
The aim of this study was to produce 2,3-butanediol (2,3-BDO) from xylose efficiently by modulation of the xylose metabolic pathway in engineered Saccharomyces cerevisiae. Expression of the Scheffersomyces stipitis transaldolase and NADH-preferring xylose reductase in S. cerevisiae improved xylose consumption rate by a 2.1-fold and 2,3-BDO productivity by a 1.8-fold. Expression of the Lactococcus lactis noxE gene encoding NADH oxidase also increased 2,3-BDO yield by decreasing glycerol accumulation. Additionally, the disadvantage of C2-dependent growth of pyruvate decarboxylase-deficient (Pdc(-)) S...
June 9, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28649896/platelet-lysate-as-a-serum-replacement-for-skin-cell-culture-on-biomimetic-pcl-nanofibers
#3
Vera Sovkova, Karolina Vocetkova, Michala Rampichova, Andrea Mickova, Matej Buzgo, Vera Lukasova, Jana Dankova, Eva Filova, Alois Necas, Evzen Amler
Platelets are a popular source of native growth factors for tissue engineering applications. The aim of the study was to verify the use of platelet lysate as a fetal bovine serum (FBS) replacement for skin cell culture. The cytokine content of the platelet lysate was characterized using the Bio-Plex system. The cells (fibroblasts, melanocytes, and keratinocytes) were cultured on PCL nanofibrous scaffolds to mimic their natural microenvironment. The cytokine content of the platelet lysate was determined, and to the cells, a medium containing platelet lysate or platelet lysate in combination with FBS was added...
June 26, 2017: Platelets
https://www.readbyqxmd.com/read/28649761/functional-assessment-of-plant-and-microalgal-lipid-pathway-genes-in-yeast-to-enhance-microbial-industrial-oil-production
#4
Huadong Peng, Lalehvash Moghaddam, Anthony Brinin, Brett Williams, Sagadevan Mundree, Victoria S Haritos
As promising alternatives to fossil-derived oils, microbial lipids are important as industrial feedstocks for biofuels and oleochemicals. Our broad aim is to increase lipid content in oleaginous yeast through expression of lipid accumulation genes and use Saccharomyces cerevisiae to functionally assess genes obtained from oil-producing plants and microalgae. Lipid accumulation genes DGAT (diacylglycerol acyltransferase), PDAT (phospholipid: diacylglycerol acyltransferase) and ROD1 (phosphatidylcholine: diacylglycerol choline-phosphotransferase) were separately expressed in yeast and lipid production measured by fluorescence, solvent extraction, TLC and GC of fatty acid methyl esters...
June 25, 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/28649699/a-wrky-transcription-factor-from-withania-somnifera-regulates-triterpenoid-withanolide-accumulation-and-biotic-stress-tolerance-through-modulation-of-phytosterol-and-defense-pathways
#5
Anup Kumar Singh, Sarma Rajeev Kumar, Varun Dwivedi, Avanish Rai, Shaifali Pal, Ajit K Shasany, Dinesh A Nagegowda
Withania somnifera produces pharmacologically important triterpenoid withanolides that are derived via phytosterol pathway; however, their biosynthesis and regulation remain to be elucidated. A jasmonate- and salicin-inducible WRKY transcription factor from W. somnifera (WsWRKY1) exhibiting correlation with withaferin A accumulation was functionally characterized employing virus-induced gene silencing and overexpression studies combined with transcript and metabolite analyses, and chromatin immunoprecipitation assay...
June 26, 2017: New Phytologist
https://www.readbyqxmd.com/read/28649279/a-simple-method-to-control-glycolytic-flux-for-the-design-of-an-optimal-cell-factory
#6
Jae Hyung Lim, Gyoo Yeol Jung
BACKGROUND: A microbial cell factory with high yield and productivity are prerequisites for an economically feasible bio-based chemical industry. However, cell factories that show a kinetic imbalance between glycolysis and product formation pathways are not optimal. Glycolysis activity is highly robust for survival in nature, but is not optimized for chemical production. RESULTS: Here, we propose a novel approach to balance glycolytic activity with the product formation capacity by precisely controlling expression level of ptsG (encoded glucose transporter) through UTR engineering...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/28649005/crispr-cas9-coupled-recombineering-for-metabolic-engineering-of-corynebacterium-glutamicum
#7
Jae Sung Cho, Kyeong Rok Choi, Cindy Pricilia Surya Prabowo, Jae Ho Shin, Dongsoo Yang, Jaedong Jang, Sang Yup Lee
Genome engineering of Corynebacterium glutamicum, an important industrial microorganism for amino acids production, currently relies on random mutagenesis and inefficient double crossover events. Here we report a rapid genome engineering strategy to scarlessly knock out one or more genes in C. glutamicum in sequential and iterative manner. Recombinase RecT is used to incorporate synthetic single-stranded oligodeoxyribonucleotides into the genome and CRISPR/Cas9 to counter-select negative mutants. We completed the system by engineering the respective plasmids harboring CRISPR/Cas9 and RecT for efficient curing such that multiple gene targets can be done iteratively and final strains will be free of plasmids...
June 22, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28648749/transcriptome-profiling-of-3d-co-cultured-cardiomyocytes-and-endothelial-cells-under-oxidative-stress-using-a-photocrosslinkable-hydrogel-system
#8
Xiaoshan Yue, Aylin Acun, Pinar Zorlutuna
Myocardial infarction (MI) is one of the most common among cardiovascular diseases. Endothelial cells (ECs) are considered to have protective effects on cardiomyocytes (CMs) under stress conditions such as MI; however, the paracrine CM-EC crosstalk and the resulting endogenous cellular responses that could contribute to this protective effect are not thoroughly investigated. Here we created biomimetic synthetic tissues containing CMs and human induced pluripotent stem cell (hiPSC)-derived ECs (iECs), which showed improved cell survival compared to single cultures under conditions mimicking the aftermath of MI, and performed high-throughput RNA-sequencing to identify target pathways that could govern CM-iEC crosstalk and the resulting improvement in cell viability...
June 22, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28648476/constraint-based-metabolic-modelling-of-marine-microbes-and-communities
#9
REVIEW
Marco Fondi, Renato Fani
Constraint-based metabolic modelling (CBMM) consists in the use of computational methods and tools to perform genome-scale simulations and predict metabolic features at the whole cellular level. This approach is rapidly expanding in microbiology, as it combines reliable predictive abilities with conceptually and technically simple frameworks. Among the possible outcomes of CBMM, the capability to i) guide a focused planning of metabolic engineering experiments and ii) provide a system-level understanding of (single or community-level) microbial metabolic circuits also represent primary aims in present-day marine microbiology...
June 22, 2017: Marine Genomics
https://www.readbyqxmd.com/read/28646866/engineering-redox-homeostasis-to-develop-efficient-alcohol-producing-microbial-cell-factories
#10
REVIEW
Chunhua Zhao, Qiuwei Zhao, Yin Li, Yanping Zhang
The biosynthetic pathways of most alcohols are linked to intracellular redox homeostasis, which is crucial for life. This crucial balance is primarily controlled by the generation of reducing equivalents, as well as the (reduction)-oxidation metabolic cycle and the thiol redox homeostasis system. As a main oxidation pathway of reducing equivalents, the biosynthesis of most alcohols includes redox reactions, which are dependent on cofactors such as NADH or NADPH. Thus, when engineering alcohol-producing strains, the availability of cofactors and redox homeostasis must be considered...
June 24, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28646243/provitamin-a-carotenoids-from-an-engineered-high-carotenoid-maize-are-bioavailable-and-zeaxanthin-does-not-compromise-%C3%AE-carotene-absorption-in-poultry
#11
Joana Díaz-Gómez, Jose A Moreno, Eduardo Angulo, Gerhard Sandmann, Changfu Zhu, Teresa Capell, Carmina Nogareda
High-carotenoid (HC) maize, a biofortified staple crop which accumulates β-carotene, β-cryptoxanthin, lutein and zeaxanthin, was used as a feed component in a chicken feeding trial to assess the bioavailability of provitamin A (PVA) carotenoids in the kernel matrix compared to the synthetic and natural color additives routinely used in the poultry industry. We found that the PVA carotenoids in HC maize were not metabolized in the same manner: β-carotene was preferentially converted into retinol in the intestine whereas β-cryptoxanthin accumulated in the liver...
June 23, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28646149/crucial-role-of-4-deoxy-l-erythro-5-hexoseulose-uronate-reductase-for-alginate-utilization-revealed-by-adaptive-evolution-in-engineered-saccharomyces-cerevisiae
#12
Fumiya Matsuoka, Makoto Hirayama, Takayuki Kashihara, Hideki Tanaka, Wataru Hashimoto, Kousaku Murata, Shigeyuki Kawai
In brown macroalgae, alginate and D-mannitol are promising carbohydrates for biorefinery. Saccharomyces cerevisiae is widely used as a microbial cell factory, but this budding yeast is unable to utilize either alginate or D-mannitol. Alginate can be depolymerized by both endo-type and exo-type alginate lyases, yielding a monouronate, 4-deoxy-L-erythro-5-hexoseulose uronate (DEH), a key intermediate in the metabolism of alginate. Here, we constructed engineered two S. cerevisiae strains that are able to utilize both DEH and D-mannitol on two different strain backgrounds, and we also improved their aerobic growth in a DEH liquid medium through adaptive evolution...
June 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28645530/recent-advances-in-microbial-production-of-aromatic-chemicals-and-derivatives
#13
REVIEW
Shuhei Noda, Akihiko Kondo
Along with the development of metabolic engineering and synthetic biology tools, various microbes are being used to produce aromatic chemicals. In microbes, aromatics are mainly produced via a common important precursor, chorismate, in the shikimate pathway. Natural or non-natural aromatics have been produced by engineering metabolic pathways involving chorismate. In the past decade, novel approaches have appeared to produce various aromatics or to increase their productivity, whereas previously, the targets were mainly aromatic amino acids and the strategy was deregulating feedback inhibition...
June 20, 2017: Trends in Biotechnology
https://www.readbyqxmd.com/read/28645173/spatially-resolved-metabolic-analysis-reveals-a-central-role-for-transcriptional-control-in-carbon-allocation-to-wood
#14
Melissa Roach, Stéphanie Arrivault, Amir Mahboubi, Nicole Krohn, Ronan Sulpice, Mark Stitt, Totte Niittylä
The contribution of transcriptional and post-transcriptional regulation to modifying carbon allocation to developing wood of trees is not well defined. To clarify the role of transcriptional regulation, the enzyme activity patterns of eight central primary metabolism enzymes across phloem, cambium, and developing wood of aspen (Populus tremula L.) were compared with transcript levels obtained by RNA sequencing of sequential stem sections from the same trees. Enzymes were selected on the basis of their importance in sugar metabolism and in linking primary metabolism to lignin biosynthesis...
June 22, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28643382/synthesis-and-biological-evaluation-of-arginyl-diosgenin-conjugate-as-a-potential-bone-tissue-engineering-agent
#15
Ai-Mei Liao, Hana Jung, Ji Won Yu, Dong Hee Lee, Sang-Soo Park, Bangrong Cai, ChangJu Chun
Water-soluble arginyl-diosgenin (Arg-DG) conjugate was designed, synthesized, and evaluated for a biological activity. The Arg-DG conjugate was characterized using FT-IR, (1) H NMR, (13) C NMR, and HPLC-MS analyses, followed by a biological activity evaluation. Compared with DG, the Arg-DG conjugate showed a decreased cytotoxicity against L929 cells and an increased anti-proliferative activity against hepatocellular cells. The safety of the Arg-DG conjugate was confirmed using the highly sensitive Alamar Blue assay, which indicated that it increased the cellular metabolic activity at suitable concentrations...
June 23, 2017: Chemical Biology & Drug Design
https://www.readbyqxmd.com/read/28642195/how-lipid-droplets-tag-along-glycerolipid-synthetic-enzymes-and-lipid-storage
#16
REVIEW
Huan Wang, Michael V Airola, Karen Reue
Triacylglycerols (TAG) serve as the predominant form of energy storage in mammalian cells, and TAG synthesis influences conditions such as obesity, fatty liver, and insulin resistance. In most tissues, the glycerol 3-phosphate pathway enzymes are responsible for TAG synthesis, and the regulation and function of these enzymes is therefore important for metabolic homeostasis. Here we review the sites and regulation of glycerol-3-phosphate acyltransferase (GPAT), acylglycerol-3-phosphate acyltransferase (AGPAT), lipin/phosphatidic acid phosphatase (PAP), and diacylglycerol acyltransferase (DGAT) enzyme action...
June 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28642093/functional-characterization-of-3-ketosteroid-9%C3%AE-hydroxylases-in-rhodococcus-ruber-strain-chol-4
#17
Govinda Guevara, Laura Fernández de Las Heras, Julián Perera, Juana María Navarro Llorens
The 3-Ketosteroid-9α-Hydroxylase, also known as KshAB [androsta-1,4-diene-3,17-dione, NADH:oxygen oxidoreductase (9α-hydroxylating); EC 1.14.13.142)], is a key enzyme in the general scheme of the bacterial steroid catabolism in combination with a 3-ketosteroid-Δ(1)-dehydrogenase activity (KstD), being both responsible of the steroid nucleus (rings A/B) breakage. KshAB initiates the opening of the steroid ring by the 9α-hydroxylation of the C9 carbon of 4-ene-3-oxosteroids (e.g. AD) or 1,4-diene-3-oxosteroids (e...
June 19, 2017: Journal of Steroid Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/28640983/multiplexed-crispr-cas9-mediated-metabolic-engineering-of-%C3%AE-aminobutyric-acid-levels-in-solanum-lycopersicum
#18
Rui Li, Ran Li, Xindi Li, Daqi Fu, Benzhong Zhu, Huiqin Tian, Yunbo Luo, Hongliang Zhu
In recent years, the type II CRISPR system has become a widely used and robust technique to implement site-directed mutagenesis in a variety of species including model and crop plants. However, few studies manipulated metabolic pathways in plants using the CRISPR system. Here, we introduced the pYLCRISPR/Cas9 system with one or two single-site guide RNAs to target the tomato phytoene desaturase gene. An obvious albino phenotype was observed in T0 regenerated plants, and more than 61% of the desired target sites were edited...
June 22, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28639622/growth-coupled-overproduction-is-feasible-for-almost-all-metabolites-in-five-major-production-organisms
#19
Axel von Kamp, Steffen Klamt
Computational modelling of metabolic networks has become an established procedure in the metabolic engineering of production strains. One key principle that is frequently used to guide the rational design of microbial cell factories is the stoichiometric coupling of growth and product synthesis, which makes production of the desired compound obligatory for growth. Here we show that the coupling of growth and production is feasible under appropriate conditions for almost all metabolites in genome-scale metabolic models of five major production organisms...
June 22, 2017: Nature Communications
https://www.readbyqxmd.com/read/28639375/challenges-in-regenerating-the-diabetic-heart-a-comprehensive-review
#20
REVIEW
Venkata R Satthenapalli, Regis R Lamberts, Rajesh Katare
Stem cell therapy is one of the promising regenerative strategies developed to improve cardiac function in patients with ischemic heart diseases (IHD). However, this approach is limited in IHD patients with diabetes due to a progressive decline in the regenerative capacity of stem cells. This decline is mainly attributed to the metabolic memory incurred by diabetes on stem cell niche and their systemic cues. Understanding the molecular pathways involved in the diabetes-induced deterioration of stem cell function will be critical for developing new cardiac regeneration therapies...
June 22, 2017: Stem Cells
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