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Journal of Biotechnology

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https://www.readbyqxmd.com/read/30021117/rational-engineering-of-ornithine-decarboxylase-with-greater-selectivity-for-ornithine-over-lysine-through-protein-network-analysis
#1
Eun Young Hong, Jin Young Kim, Roopali Upadhyay, Byung Jun Park, Jong Min Lee, Byung-Gee Kim
Ornithine decarboxylase (ODC) converts C5 ornithine into C4 putrescine, a monomer for polyamide synthesis. However, ODC also has minor activity towards cell metabolite C6 lysine and yields C5 cadaverine. The accumulation of cadaverine in the reaction solution causes increase in the operational cost of subsequent distillation process for putrescine purification. Here, to increase ODC substrate specificity toward ornithine over lysine, molecular modelling and protein network analysis, specifically k-clique community analysis, around the substrate tunnel were performed...
July 15, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30016741/potato-snakin-1-gene-enhances-tolerance-to-rhizoctonia-solani-and-sclerotinia-sclerotiorum-in-transgenic-lettuce-plants
#2
Flavia S Darqui, Laura M Radonic, Paulina M Trotz, Nilda López, Cecilia Vázquez Rovere, H Esteban Hopp, Marisa López Bilbao
Snakin-1 is a cysteine-rich antimicrobial peptide (AMP) isolated from potato tubers, with broad-spectrum activity. It belongs to the Snakin/GASA family, whose members have been studied because of their diverse roles in important plant processes, including defense. To analyze if this defensive function may lead to disease tolerance in lettuce, one of the most worldwide consumed leafy vegetable, we characterized three homozygous transgenic lines overexpressing Snakin-1. They were biologically assessed by the inoculation with the fungal pathogens Rhizoctonia solani and Sclerotinia sclerotiorum both in vitro and in planta at the greenhouse...
July 14, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30016740/biosynthesis-of-4-vinylguaiacol-from-crude-ferulic-acid-by-bacillus-licheniformis-dlf-17056
#3
Li-Hui Sun, Shi-Wen Lv, Feng-Yu, Sheng-Nan Li, Lei-Yu He
4-vinylguaiacol, a kind of volatile phenolic compound with tobacco flavor, is widely used as a component of edible flavor and intermediate of medicine. Ferulic acid is usually used as substrate for the biosynthesis of 4-vinylguaiacol. However, the price of ferulic acid is high, leading to high production cost. In this study, a feasible low-cost process for the production of 4-vinylguaiacol was developed. The ultrasonic assisted weak alkali was used to extract protein from rice bran, and waste liquid and residue were then mixed to extract crude ferulic acid by alkaline hydrolysis...
July 14, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30016739/intracellular-transformation-rates-of-fatty-acids-are-influenced-by-expression-of-the-fatty-acid-transporter-fadl-in-escherichia-coli-cell-membrane
#4
Eun-Yeong Jeon, Ji-Won Song, Hee-Jeong Cha, Sun-Mee Lee, Jinwon Lee, Jin-Byung Park
Fatty acids have a low permeability through the cell membrane. Therefore, the intracellular biotransformation of fatty acids can be slow due to supply limitations. The effects of expression level of the fatty acid transporter FadL in Escherichia coli on the biotransformations were investigated. The enhanced expression of FadL led to 5.5-fold increase of the maximum reaction rate Vmax (i.e., 200 μmol/min per g dry cells (200 U/g dry cells)) of the recombinant E. coli expressing a hydratase of Stenotrophomonas maltophilia in the periplasm with respect to hydration of oleic acid...
July 14, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30012463/differential-expression-of-micrornas-in-recombinant-chinese-hamster-ovary-cells-treated-with-sodium-butyrate-using-digital-rna-counting
#5
Sung-Min Lim, Sun-Hye Park, Joo-Hyoung Lee, Sun Hong Kim, Jee Yon Kim, Jeong-Ki Min, Gyun Min Lee, Yeon-Gu Kim
Sodium butyrate (NaBu) is an efficient supplement for increasing recombinant protein production in Chinese hamster ovary (CHO) cell culture. To elucidate the effects of NaBu on miRNA expression profile in recombinant CHO (rCHO) cells, differentially expressed miRNAs in NaBu-treated rCHO cells were assessed by NanoString nCounter analysis. This result showed that eight mature mouse miRNAs (let-7b, let-7d, miR-15b, miR-25, miR-27a, miR-99a, miR-125a-5p, and miR-125b-5p) were differentially expressed. Furthermore, quantitative real-time RT-PCR analysis of eight mature CHO miRNAs, annotated using a miRBase database, confirmed the transcriptomic findings...
July 13, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30009844/optimisation-of-enzyme-cascades-for-chiral-amino-alcohol-synthesis-in-aid-of-host-cell-integration-using-a-statistical-experimental-design-approach
#6
Maria F Villegas-Torres, John M Ward, Frank Baganz
Chiral amino alcohols are compounds of pharmaceutical interest as they are building blocks of sphingolipids, antibiotics, and antiviral glycosidase inhibitors. Due to the challenges of chemical synthesis we recently developed two TK-TAm reaction cascades using natural and low cost feedstocks as substrates: a recycling cascade comprising of 2 enzymes and a sequential 3-step enzyme cascade yielding 30% and 1% conversion, respectively. In order to improve the conversion yield and aid the future host strain engineering for whole cell biocatalysis, we used a combination of microscale experiments and statistical experimental design...
July 12, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30017654/the-contributions-of-individual-galactosyltransferases-to-protein-specific-n-glycan-processing-in-chinese-hamster-ovary-cells
#7
Nina Bydlinski, Daniel Maresch, Valerie Schmieder, Gerald Klanert, Richard Strasser, Nicole Borth
Galactosylation as part of N-glycan processing is conducted by a set of beta-1,4-galactosyltransferases (B4 GALTs), with B4 GALT1 as the dominant isoenzyme for this reaction. Nevertheless, the exact contributions of this key-player as well as of the other isoenzymes involved in N-glycosylation, B4 GALT2, B4 GALT3 and B4 GALT4, have not been studied in-depth. To increase the understanding of the protein- and site-specific activities of individual galactosyltransferases in Chinese Hamster Ovary cells, a panel of triple deletion cell lines was generated that expressed only one isoform of B4 GALT each...
July 11, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30006300/enhancement-of-the-antibacterial-activity-of-an-e-faecalis-strain-by-the-heterologous-expression-of-enterocin-a
#8
Carlos Eduardo Serrano-Maldonado, Maricarmen Quirasco
The genus Enterococcus occurs as native microbiota of fermented products due to its broad environmental distribution and its resistance to salt concentrations. Enterococcus faecalis F, a non-pathogenic strain isolated from a ripened cheese, has demonstrated useful enzymatic capabilities, a probiotic behavior and antibacterial activity against some food-borne pathogens, mainly due to peptidoglycan hydrolase activity. Its use as a natural pathogen-control agent could be further enhanced through the production of a bacteriocin, e...
July 10, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30003975/extraction-of-astaxanthin-from-microalga-haematococcus-pluvialis-in-red-phase-by-using-generally-recognized-as-safe-solvents-and-accelerated-extraction
#9
Antonio Molino, Juri Rimauro, Patrizia Casella, Antonietta Cerbone, Vincenzo Larocca, Simeone Chianese, Despina Karatza, Sanjeet Mehariya, Angelo Ferraro, Evangelos Hristoforou, Dino Musmarra
Solvent Extraction was tested to extract astaxanthin from Haematococcus pluvialis in red phase (HPR), by investigating effects of solvents, extraction pressure and temperature. Astaxanthin isomers were identified and quantified in the extract. The performances of acetone and ethanol, Generally Recognized As Safe (GRAS) solvents, were explored. Negligible effect of pressure was found, while with increasing extraction temperature astaxanthin recovery increased till a maximum value, beyond which thermal degradation seemed to be greater than the positive effect of temperature on extraction...
July 9, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30003974/the-diploid-genome-of-the-only-sclerotia-forming-wild-type-species-in-the-genus-pleurotus-pleurotus-tuber-regium-provides-insights-into-the-mechanism-of-its-biomass-conversion-from-lignocellulose-substrates
#10
Ka-Lung Lam, Kaiwei Si, Xiyang Wu, Shuze Tang, Xiaohui Sun, Hoi-Shan Kwan, Chi-Keung Peter Cheung
Pleurotus tuber-regium (Fr.) Singer, being a white-rot fungus, is widely used for food and medicine in the Asia-Pacific region. In this study, we sequenced and annotated the genome of a dikaryon P. tuber-regium wild strain to provide a better understanding of the carbohydrate-active enzymes (CAZymes) involved in the bio-conversion of lignocellulose to beta-glucan reserves in this sclerotia-forming Pleurotus mushroom with reference to enzyme participated in cellulosic compound breakdown and glucan reserve biosynthesis...
July 9, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/30003973/over-expression-of-sinal7-increases-biomass-and-drought-tolerance-and-also-delays-senescence-in-arabidopsis
#11
Diego A Peralta, Alejandro Araya, Diego F Gomez-Casati, María V Busi
The seven in absentia like 7 gene (At5g37890, SINAL7) from Arabidopsis thaliana encodes a RING finger protein belonging to the SINA superfamily that possesses E3 ubiquitin-ligase activity. SINAL7 has the ability to self-ubiquitinate and to mono-ubiquitinate glyceraldehyde-3-P dehydrogenase 1 (GAPC1), suggesting a role for both proteins in a hypothetical signaling pathway in Arabidopsis. In this study, the in vivo effects of SINAL7 on plant physiology were examined by over-expressing SINAL7 in transgenic Arabidopsis plants...
July 9, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29990570/deciphering-molecular-consequences-of-the-proprotein-convertase-1-3-inhibition-in-macrophages-for-application-in-anti-tumour-immunotherapy
#12
Franck Rodet, Alice Capuz, Tsukasa Hara, Rinaldo van Meel, Marie Duhamel, Mélanie Rose, Antonella Raffo Romero, Isabelle Fournier, Michel Salzet
During tumour development, macrophages are recruited to the tumour site and orientated towards an anti-inflammatory phenotype. Due to their immunosuppressive function, tumour associated macrophages (TAMs) are recognized as major components in tumour progression. Changing these macrophages to a pro-inflammatory phenotype is thus extensively studied as a potential means for developing novel anti-tumour therapy. In this context, we found that the Proprotein convertase 1/3 (PC1/3) is a relevant target. Proteomic analysis reveals that PC1/3 knockdown (KD) macrophages present all the characteristic of activated pro-inflammatory macrophages...
July 7, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29990569/influence-of-hydraulic-retention-time-and-carbon-loading-rate-on-the-production-of-algae
#13
C M Fernandez-Marchante, Y Asensio, J Lobato, J Villaseñor, P Cañizares, M A Rodrigo
This paper is focused on the assessment of the production of algae in batch bioreactors. Hydraulic retention time, carbon loading rate and light color were the inputs of the study and algae production the main output. Bioreactors were operated in semi-continuous mode and tests lasted two months, more than two times the period required to meet a steady-state response. This steady-state was verified with plateau responses in both, soluble parameters and suspended solids. Results points out the great relevance of temperature...
July 7, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29990568/synthesis-of-butyl-oleate-catalyzed-by-cross-linked-enzyme-aggregates-with-magnetic-nanoparticles-in-rotating-magneto-micro-reactor
#14
Debing Zheng, Shuguang Wang, Shiwei Qiu, Jiang Lin, Xiaojun Diao
In order to increase application of cross-linked enzyme aggregates (CLEAs) in industry production, a novel micro-reactor system that included a rotating magnetic field (RMF), a micro-reactor and CLEAs with magnetic nanoparticles (M-CLEAs) was designed to synthesize butyl oleate. Result showed that the presence of RMF significantly increased the yield of butyl oleate and the maximum increment was 23%. The yield of butyl oleate was impacted by the dosage and distribution of M-CLEAs in micro-reactor. M-CLEAs showed good reusability, since the morphology and the second structure of protein of M-CLEAs did not show evident change after 4 operative cycles...
July 7, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29981450/characterization-of-a-new-microbial-pictet-spenglerase-nscbb-affording-the-%C3%AE-carboline-skeletonsfrom-nocardiopsis-synnemataformans-dsm-44143
#15
Qi Chen, Shaofei Zhang, Yunchang Xie
Theenzymatic Pictet-Spengler (PS) reaction, catalyzed by Pictet-Spenglerase (PSase), is the feature ofβ-carboline (βC) alkaloid biosynthesis. NscbB is a raremicrobial PSase discovered from a cryptic β-carboline alkaloid biosynthetic gene cluster (BGC) in the Nocardiopsis synnemataformans DSM 44143 (kidney transplant patient derived) by homologous alignment with its well-characterized counterpart McbB. Thebiochemical analysis showed that NscbB could catalyze L-tryptophan (KM  = 89.64 ± 8.69 μM) and methylglyoxal (KM  = 147...
July 4, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29981449/enzymatic-characterization-of-a-soluble-aggregate-induced-by-n-terminal-extension-to-a-lipolytic-enzyme
#16
Ji-Min Park, Mi-Hwa Lee, Chul-Hyung Kang, Ki-Hoon Oh, Jung-Sook Lee, Jung-Hoon Yoon
A self-assembling peptide (27PEP) was isolated from an open reading frame (ORF). The ORF consisted of an unknown functional domain and a catalytic (lipolytic and phospholipolytic) domain (MPlaG) on metagenomic fosmid clone. This extension of 27 amino acids prior to the N-terminus of the catalytic domain (27PEP-MPlaG), starting at Met261 , produced an aggregate of high molecular weight (> 700 kDa). Compared with MPlaG, 27PEP-MPlaG showed the same temperature and pH effect for maximum activity but was stable in the presence of inhibitors such as EDTA and PMSF...
July 4, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29981448/a-multifunctional-tag-with-the-ability-to-benefit-the-expression-purification-thermostability-and-activity-of-recombinant-proteins
#17
Weixin Zhao, Liming Liu, Guocheng Du, Song Liu
In this study, a novel multifunctional tag, S1v1 (AEAEAHAH)2 , was generated from a self-assembling amphipathic peptide in the Zuotin protein sequence by replacing lysine residues with histidine residues. After fusing S1v1 at the N-terminus through a PT-linker, the expressions of polygalacturonate lyase (PGL), lipoxygenase (LOX) and green fluorescent protein (GFP) were enhanced by 3.8, 0.2 and 1.52-fold, respectively,compared to the wild-type proteins. However, the frequently used His-tag with a PT-linker had negligible effects on expression...
July 4, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29981447/enhanced-overexpression-purification-of-a-channelrhodopsin-and-a-fluorescent-flux-assay-for-its-functional-characterization
#18
Hiofan Hoi, Zhigang Qi, Hang Zhou, Carlo D Montemagno
Channelrhodopsins (ChRs) are a group of membrane proteins that allow cation flux across the cellular membrane when stimulated by light. They have been emerged as important tools in optogenetics where light is used to trigger a change in the membrane potential of live cells which induces downstream physiological cascades. There is also increased interest in their applications for generating light-responsive biomaterials. Here we have used a two-step screening protocol to develop a Pichia pastoris strain that produces superior yields of an enhance variant of CaChR2 (from Chlamydomonas reinhardtii), called ChIEF...
July 4, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29981446/crosslinked-enzyme-aggregates-clea-of-phytase-with-soymilk-proteins
#19
Tirunagari Harini, Shalini Basetty, Haridas B Rode, Nitin W Fadnavis
Crosslinked Enzyme Aggregates (CLEA) of phytase (E.C. 3.1.3.8) have been successfully prepared using soymilk as protein feeder. Compared to total initial activity of 4428 units used for immobilization, the recovered activity in the CLEA is enhanced to 8000 units. The CLEA retained 100 % phytase activity up to 50 °C and 70 % activity at 70 °C after 6 h incubation while soluble enzyme lost 50 % activity at 50 °C and 80 % at 70 °C. Soluble phytase has an optimum pH of 2.1 while immobilized enzyme maintains high level of activity in a broad pH range of 1-6...
July 4, 2018: Journal of Biotechnology
https://www.readbyqxmd.com/read/29981445/features-of-the-transglutaminase-activating-metalloprotease-from-streptomyces-mobaraensis-dsm-40847-produced-in-escherichia-coli
#20
Norbert E Juettner, Moritz Classen, Felix Colin, Sascha B Hoffmann, Christian Meyners, Felicitas Pfeifer, Hans-Lothar Fuchsbauer
Transglutaminase from Streptomyces mobaraensis (MTG) is an important enzyme for numerous industrial applications. Recombinant production requires proteolytic activation of the zymogen. The study provides a convenient procedure for the preparation of the transglutaminase-activating metalloprotease (TAMP) in Escherichia coli. In contrast to wtTAMP, rTAMP exhibited the P domain of convertases as molecular mass of 55.7 kDa suggested. Protein integrity was beneficially influenced by 2-5 mM CaCl2 . Study of pH and temperature optima assigned rTAMP to the neutral metalloproteases, more heat-resistant than Dispase but not thermolysin...
July 4, 2018: Journal of Biotechnology
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