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https://www.readbyqxmd.com/read/28521794/in-vitro-synthesis-of-9-10-dihydroxyhexadecanoic-acid-using-recombinant-escherichia-coli
#1
Anees Kaprakkaden, Preeti Srivastava, Virendra Swarup Bisaria
BACKGROUND: Hydroxy fatty acids are widely used in food, chemical and cosmetic industries. A variety of dihydroxy fatty acids have been synthesized so far; however, no studies have been done on the synthesis of 9,10-dihydroxyhexadecanoic acid. In the present study recombinant E. coli has been used for the heterologous expression of fatty acid hydroxylating enzymes and the whole cell lysate of the induced culture was used for in vitro production of 9,10-dihydroxyhexadecanoic acid. RESULTS: A first of its kind proof of principle has been successfully demonstrated for the production of 9,10-dihydroxyhexadecanoic acid using three different enzymes viz...
May 18, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28482913/critical-attributes-of-human-early-mesenchymal-stromal-cell-laden-microcarrier-constructs-for-improved-chondrogenic-differentiation
#2
Youshan Melissa Lin, Jialing Lee, Jessica Fang Yan Lim, Mahesh Choolani, Jerry Kok Yen Chan, Shaul Reuveny, Steve Kah Weng Oh
BACKGROUND: Microcarrier cultures which are useful for producing large cell numbers can act as scaffolds to create stem cell-laden microcarrier constructs for cartilage tissue engineering. However, the critical attributes required to achieve efficient chondrogenic differentiation for such constructs are unknown. Therefore, this study aims to elucidate these parameters and determine whether cell attachment to microcarriers throughout differentiation improves chondrogenic outcomes across multiple microcarrier types...
May 8, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28470602/high-yield-of-recombinant-protein-in-shaken-e-coli-cultures-with-enzymatic-glucose-release-medium-enpresso-b
#3
Kaisa Ukkonen, Antje Neubauer, Vinit J Pereira, Antti Vasala
Expression of recombinant proteins in sufficient quantities is essential for protein structure-function studies. The most commonly used method for recombinant protein production is overexpression in E. coli cultures. However, producing high yields of functional proteins in E. coli can be a challenge in conventional shaken cultures. This is often due to nonoptimal growth conditions, which result in low cell yields and insoluble or incorrectly folded target protein. To overcome the shortcomings of shake flask cultivation, we present a cultivation method based on enzymatic glucose delivery...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28465212/cell-culture-media-supplemented-with-raffinose-reproducibly-enhances-high-mannose-glycan-formation
#4
David Brühlmann, Anais Muhr, Rebecca Parker, Thomas Vuillemin, Blanka Bucsella, Franka Kalman, Serena Torre, Fabio La Neve, Antonio Lembo, Tobias Haas, Markus Sauer, Jonathan Souquet, Hervé Broly, Jürgen Hemberger, Martin Jordan
Glycosylation plays a pivotal role in pharmacokinetics and protein physiochemical characteristics. In particular, effector functions including antibody-dependent cell-mediated cytotoxicity (ADCC) can be desired, and it has been described that high-mannose species exhibited enhanced ADCC. In this work we present the trisaccharide raffinose as a novel cell culture medium supplement to promote high mannose N-glycans in fed-batch cultures, which is sought after in the development of biosimilars to match the quality profile of the reference medicinal product (RMP) also...
April 29, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28436005/fast-and-reliable-strain-characterization-of-streptomyces-lividans-through-micro-scale-cultivation
#5
Joachim Koepff, Matthias Keller, Konstantinos C Tsolis, Tobias Busche, Christian Rückert, Mohamed B Hamed, Jozef Anné, Joern Kalinowski, Wolfgang Wiechert, Anastassios Economou, Marco Oldiges
Filamentous organisms of the genus Streptomyces play an important role in industrial production processes, due to their extensive secondary metabolism variability, as well as their ability to secrete efficiently large amounts of (heterologous) proteins. While genetic engineering tools are available to rapidly build up large strain libraries, the subsequent strain screening and bioprocess development still constitutes a bottleneck. This is due to the lack of reliable parallelized and accelerated cultivation techniques for morphologically challenging organisms...
April 24, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28420336/reduced-changes-in-protein-compared-to-mrna-levels-across-non-proliferating-tissues
#6
Kobi Perl, Kathy Ushakov, Yair Pozniak, Ofer Yizhar-Barnea, Yoni Bhonker, Shaked Shivatzki, Tamar Geiger, Karen B Avraham, Ron Shamir
BACKGROUND: The quantitative relations between RNA and protein are fundamental to biology and are still not fully understood. Across taxa, it was demonstrated that the protein-to-mRNA ratio in steady state varies in a direction that lessens the change in protein levels as a result of changes in the transcript abundance. Evidence for this behavior in tissues is sparse. We tested this phenomenon in new data that we produced for the mouse auditory system, and in previously published tissue datasets...
April 18, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28410897/efficient-production-of-antibody-fab-fragment-by-transient-gene-expression-in-insect-cells
#7
Keita Mori, Hirotsugu Hamada, Takafumi Ogawa, Yuki Ohmuro-Matsuyama, Tomohisa Katsuda, Hideki Yamaji
Transient gene expression allows a rapid production of diverse recombinant proteins in early-stage preclinical and clinical developments of biologics. Insect cells have proven to be an excellent platform for the production of functional recombinant proteins. In the present study, the production of an antibody Fab fragment by transient gene expression in lepidopteran insect cells was investigated. The DNA fragments encoding heavy-chain (Hc; Fd fragment) and light-chain (Lc) genes of an Fab fragment were individually cloned into the plasmid vector pIHAneo, which contained the Bombyx mori actin promoter downstream of the B...
April 11, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28402216/a-novel-and-efficient-immunoglobulin-y-extraction-method-using-poloxamer-polyethylene-glycol
#8
Cui Li, Hao Ren, Rüdiger Schade, Xiaoying Zhang
Although many IgY extraction methods (such as polyethylene glycol (PEG) precipitation method, octanoic acid method, water dilution method, etc.) have been established, there is still industrial drive and real need in developing scale-up IgY production methods. Some previous studies have reported that poloxamer degreasing method shows very good result in IgY extraction from egg yolk with high degreasing speed, harmlessness, simpleness in operation and minimal effect on antibody titer. In this study, we developed a new method, poloxamer-polyethylene glycol (PEG) method, to obtain functional IgY with high purity and yield...
April 12, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28389137/sugar-perceived-healthfulness-and-satiety-when-does-a-sugary-preload-lead-people-to-eat-more
#9
Naomi Mandel, Daniel Brannon
In this research, we examine the interplay between physiological and psychological factors that determine whether the sugar level of a preload increases or decreases consumption on a subsequent snack-eating task. In study 1, participants who drank a high-sugar protein shake (which they believed to be healthy) consumed more subsequent snacks than participants who drank a low-sugar protein shake. Study 2 replicated these findings, but only when the shake was labeled as "healthy." When the shake was labeled as "indulgent," the effect was mitigated...
April 4, 2017: Appetite
https://www.readbyqxmd.com/read/28389059/renal-response-to-a-protein-load-in-healthy-young-adults-as-determined-by-iohexol-infusion-clearance-cimetidine-inhibited-creatinine-clearance-and-cystatin-c-estimated-glomerular-filtration-rate
#10
Kyle E Rodenbach, Dana Y Fuhrman, Paula S Maier, George J Schwartz
OBJECTIVE: Renal reserve (RR) measures the increase in glomerular filtration rate (GFR) in response to a protein load; lack of RR could indicate subclinical kidney disease but such a test is not routinely used in clinical practice. The purpose of this study was to compare a meat versus liquid protein load in a cystatin C-based (Cys-C) RR test using cimetidine-inhibited creatinine clearance (Cr Cl) and iohexol infusion clearance (Io Cl) for validation. The design was cross-sectional analysis and the setting was a Clinical Research Center...
April 4, 2017: Journal of Renal Nutrition
https://www.readbyqxmd.com/read/28368321/sport-nutrition-knowledge-behaviors-and-beliefs-of-high-school-soccer-players
#11
Melinda M Manore, Megan M Patton-Lopez, Yu Meng, Siew Sun Wong
For adolescent athletes (14-18 years), data on sport nutrition knowledge, behaviors and beliefs are limited, especially based on sex, race/ethnicity and socioeconomic status. High school soccer players (n = 535; 55% female; 51% White, 41% Latino; 41% National School Lunch Program (NSLP) participants (80% Latino)) completed two questionnaires (demographic/health history and sport nutrition). The sport nutrition knowledge score was 45.6% with higher scores in NSLP-Whites vs. NSLP-Latinos (p < 0.01). Supplement knowledge differed by sex (16% lower in females; p = 0...
April 1, 2017: Nutrients
https://www.readbyqxmd.com/read/28359705/manipulation-of-ges-and-erg20-for-geraniol-overproduction-in-saccharomyces-cerevisiae
#12
Guo-Zhen Jiang, Ming-Dong Yao, Ying Wang, Liang Zhou, Tian-Qing Song, Hong Liu, Wen-Hai Xiao, Ying-Jin Yuan
Manipulation of monoterpene synthases to maximize flux towards targeted products from GPP (geranyl diphosphate) is the main challenge for heterologous monoterpene overproduction, in addition to cell toxicity from compounds themselves. In our study, by manipulation of the key enzymes geraniol synthase (GES) and farnesyl diphosphate synthase (Erg20), geraniol (a valuable acyclic monoterpene alcohol) overproduction was achieved in Saccharomyces cerevisiae with truncated 3-hydroxy-3-methylglutaryl-coenzyme reductase (tHMGR) and isopentenyl diphosphate isomerase (IDI1) overexpressed...
March 27, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/28357545/growth-kinetics-and-scale-up-of-agrobacterium-tumefaciens
#13
Ingrid K Leth, Karen A McDonald
Production of recombinant proteins in plants through Agrobacterium-mediated transient expression is a promising method of producing human therapeutic proteins, vaccines, and commercial enzymes. This process has been shown to be viable at a large scale and involves growing large quantities of wild-type plants and infiltrating the leaf tissue with a suspension of Agrobacterium tumefaciens bearing the genes of interest. This study examined one of the steps in this process that had not yet been optimized: the scale-up of Agrobacterium production to sufficient volumes for large-scale plant infiltration...
March 29, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28356431/acute-peanut-consumption-alters-postprandial-lipids-and-vascular-responses-in-healthy-overweight-or-obese-men
#14
Xiaoran Liu, Alison M Hill, Sheila G West, Rachel M Gabauer, Cindy E McCrea, Jennifer A Fleming, Penny M Kris-Etherton
Background: Postprandial hyperlipidemia is associated with impaired endothelial function. Peanut consumption favorably affects the lipid and lipoprotein profile; however, the effects on endothelial function remain unclear.Objective: The purpose of the study was to evaluate the effects of acute peanut consumption as part of a high-fat meal on postprandial endothelial function.Methods: We conducted a randomized, controlled, crossover postprandial study to evaluate the effect of acute peanut consumption on postprandial lipids and endothelial function as assessed by flow-mediated dilatation (FMD) of the brachial artery in 15 healthy overweight or obese men [mean age: 26...
March 29, 2017: Journal of Nutrition
https://www.readbyqxmd.com/read/28356104/heterologous-biosynthesis-and-manipulation-of-crocetin-in-saccharomyces-cerevisiae
#15
Fenghua Chai, Ying Wang, Xueang Mei, Mingdong Yao, Yan Chen, Hong Liu, Wenhai Xiao, Yingjin Yuan
BACKGROUND: Due to excellent performance in antitumor, antioxidation, antihypertension, antiatherosclerotic and antidepressant activities, crocetin, naturally exists in Crocus sativus L., has great potential applications in medical and food fields. Microbial production of crocetin has received increasing concern in recent years. However, only a patent from EVOVA Inc. and a report from Lou et al. have illustrated the feasibility of microbial biosynthesis of crocetin, but there was no specific titer data reported so far...
March 29, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28343939/a-tale-of-gastric-layering-and-sieving-gastric-emptying-of-a-liquid-meal-with-water-blended-in-or-consumed-separately
#16
Guido Camps, Monica Mars, Cees de Graaf, Paul A M Smeets
BACKGROUND: The process of gastric emptying determines how fast gastric content is delivered to the small intestine. It has been shown that solids empty slower than liquids and that a blended soup empties slower than the same soup as broth and chunks, due to the liquid fraction emptying more quickly. This process of 'gastric sieving' has not been investigated for liquid foods. OBJECTIVE: To determine whether gastric sieving of water can also occur for liquid foods...
March 23, 2017: Physiology & Behavior
https://www.readbyqxmd.com/read/28330255/ascorbyl-palmitate-synthesis-in-an-organic-solvent-system-using-a-celite-immobilized-commercial-lipase-lipolase-100l
#17
Shivika Sharma, Kriti Kanwar, Shamsher S Kanwar
Ascorbyl palmitate was synthesized using a Celite-immobilized commercial lipase (Lipolase 100L) in dimethylsulfoxide (DMSO) as an organic solvent system. Lipase immobilized by surface adsorption onto Celite 545 matrix and subsequently exposed to 1 % glutaraldehyde showed 75 % binding of protein. The Celite-bound lipase was optimally active at 75 °C and pH 8.5 under shaking and showed maximum hydrolytic activity toward p-NPP as a substrate. The bound lipase was found to be stimulated only in the presence of Al(3+) and EDTA...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28323213/oral-delivery-of-bacillus-subtilis-spores-expressing-cysteine-protease-of-clonorchis-sinensis-to-grass-carp-ctenopharyngodon-idellus-induces-immune-responses-and-has-no-damage-on-liver-and-intestine-function
#18
Zeli Tang, Hengchang Sun, TingJin Chen, Zhipeng Lin, Hongye Jiang, Xinyi Zhou, Cunbin Shi, Houjun Pan, Ouqin Chang, Pengli Ren, Jinyun Yu, Xuerong Li, Jin Xu, Yan Huang, Xinbing Yu
Clonorchis sinensis (C. sinensis) is a fish-borne trematode. Human can be infected by ingestion of C. sinensis metacercariae parasitized in grass carp (Ctenopharyngodon idella). For induction of effective oral immune responses, spores of Bacillus subtilis (B. subtilis) WB600 were utilized as vehicle to delivery CsCP (cysteine protease of C. sinensis) cooperated with CotC (B.s-CotC-CP), one of coat proteins, to the gastrointestinal tract. After routine culture of 8-12 h in LB medium, B. subtilis containing CotC-CsCP was transferred into the sporulation culture medium...
May 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28321795/phytase-production-by-aspergillus-niger-ncim-563-for-a-novel-application-to-degrade-organophosphorus-pesticides
#19
Parin C Shah, V Ravi Kumar, Syed G Dastager, Jayant M Khire
The production of phytase using Aspergillus niger NCIM 563 under submerged fermentation conditions was studied using protein rich chickpea flour as substrate. Employing a hybrid statistical media optimization strategy of Plackett-Burman and Box-Behnken experimental designs in shake-flasks gave an increased phytase activity from an initial 66 IU/mL in 216 h to 160 IU/mL in a reduced time of 132 h. Productivity, thus increased by 3.97 times from 7.3 to 29 IU/mL/day. Using the optimized media, the production was successfully scaled-up further and improved up to 164 IU/mL in 96 h by studies carried out employing 2 and 10-L fermenters...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28318397/baiba-does-not-regulate-ucp-3-expression-in-human-skeletal-muscle-as-a-response-to-aerobic-exercise
#20
Flor E Morales, Jeffrey S Forsse, Thomas L Andre, Sarah K McKinley-Barnard, Paul S Hwang, Ian G Anthony, Grant M Tinsley, Mike Spillane, Peter W Grandjean, Alejandro Ramirez, Darryn S Willoughby
OBJECTIVE: β-Aminoisobutyric acid (BAIBA) has shown to modulate uncoupling protein (UCP)-1 expression, which is mainly expressed in white adipose tissue; however, no studies to date have analyzed its potential effect on the main uncoupling protein of skeletal muscle, UCP-3. The main goal of this study was to assess the potential effect of acute aerobic exercise on serum BAIBA and skeletal muscle UCP-3. The secondary goal was to assess the potential involvement of the transcription factors proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and peroxisome proliferator-activated receptor alpha (PPARα), as well as free fatty acids (FFAs) in UCP-3 expression...
March 2017: Journal of the American College of Nutrition
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