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yeast glycosylation

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https://www.readbyqxmd.com/read/28694218/constitutive-expression-of-human-gastric-lipase-in-pichia-pastoris-and-site-directed-mutagenesis-of-key-lid-stabilizing-residues
#1
Laura Sams, Sawsan Amara, Almahdi Chakroun, Sébastien Coudre, Julie Paume, Jacqueline Giallo, Frédéric Carrière
The cDNA encoding human gastric lipase (HGL) was integrated into the genome of Pichia pastoris using the pGAPZα A transfer vector. The HGL signal peptide was replaced by the yeast α-factor to achieve an efficient secretion. Active rHGL was produced by the transformed yeast but its levels and stability were dependent on the pH. The highest activity was obtained upon buffering the culture medium at pH5, a condition that allowed preserving enzyme activity over time. A large fraction (72±2%) of secreted rHGL remained however bound to the yeast cells, and was released by washing the cell pellet with an acid glycine-HCl buffer (pH2...
July 7, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28688204/enhancing-expression-of-the-pseudorabies-virus-glycoprotein-e-in-yeast-and-its-application-in-an-indirect-sandwich-elisa
#2
Ching-Ying Wu, Ching-Wei Wu, Chih-Ming Liao, Maw-Sheng Chien, Chienjin Huang
AIMS: The purpose of this study was to produce a recombinant pseudorabies virus (PRV) glycoprotein E (gE) protein with the correct antigenicity for use as a low-cost diagnostic antigen. METHODS AND RESULTS: The gene fragment encoding the amino-terminal immunodominant region of PRV gE (codons 31-270) (gEN31-270) was codon-optimized and expressed constitutively and secreted using a Pichia pastoris expression system. Yeast-expressed gEN31-270 (ygEN31-270) was harvested from the culture supernatant, and ygEN31-270 was shown to exhibit N-linked glycosylation...
July 8, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/28669180/polysaccharides-and-oligosaccharides-produced-on-malvar-wines-elaborated-with-torulaspora-delbrueckii-cli-918-and-saccharomyces-cerevisiae-cli-889-native-yeasts-from-d-o-vinos-de-madrid
#3
Margarita Garcia, Rafael Apolinar-Valiente, Pascale Williams, Braulio Esteve-Zarzoso, Teresa Arroyo, Julia Crespo Garcia, Thierry Doco
Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. Glycosyl composition and linkages of wine polysaccharides were determined by GC-EI-MS chromatography. Molar-mass distributions were determined by SEC-MALLS and intrinsic viscosity by differential viscometer...
July 1, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28651914/phenylsulfonylfuroxan-no-donor-phenols-synthesis-and-multifunctional-activities-evaluation
#4
Yundong Xie, Yaping Yang, Sen Li, Yanhong Xu, Wenfang Lu, Zizhang Chen, Guangde Yang, Yiping Li, Yongxiao Cao, Xiaoli Bian
Phenylsulfonyfuroxan nitric oxide (NO)-donor phenols were designed, synthesized and evaluated. The compounds were designed through a symbiotic approach using selected phenols and phenylsulfonylfuroxan NO-donor. The antioxidant activities of the hybrid compounds T2-T6 showed to be good in vivo. Compounds T4 and T6 revealed excellent yeast α-glucosidase inhibitory activity and anti-glycosylation activity. All of the compounds exhibited strong NO releasing activity and significant anti-platelet aggregation activity...
June 15, 2017: Bioorganic & Medicinal Chemistry
https://www.readbyqxmd.com/read/28616799/dolichol-phosphate-mannose-synthase-a-glycosyltransferase-with-unity-in-molecular-diversities
#5
REVIEW
Dipak K Banerjee, Zhenbo Zhang, Krishna Baksi, Jesús E Serrano-Negrón
N-glycans provide structural and functional stability to asparagine-linked (N-linked) glycoproteins, and add flexibility. Glycan biosynthesis is elaborative, multi-compartmental and involves many glycosyltransferases. Failure to assemble N-glycans leads to phenotypic changes developing infection, cancer, congenital disorders of glycosylation (CDGs) among others. Biosynthesis of N-glycans begins at the endoplasmic reticulum (ER) with the assembly of dolichol-linked tetra-decasaccharide (Glc3Man9GlcNAc2-PP-Dol) where dolichol phosphate mannose synthase (DPMS) plays a central role...
August 2017: Glycoconjugate Journal
https://www.readbyqxmd.com/read/28608761/biomedical-applications-of-yeast-a-patent-view-part-one-yeasts-as-workhorses-for-the-production-of-therapeutics-and-vaccines
#6
REVIEW
Farzin Roohvand, Mehdi Shokri, Meghdad Abdollahpour-Alitappeh, Parastoo Ehsani
Yeasts, as Eukaryotes, offer unique features for ease of growth and genetic manipulation possibilities, making it an exceptional microbial host. Areas covered: This review provides general and patent-oriented insights into production of biopharmaceuticals by yeasts. Patents, wherever possible, were correlated to the original or review articles. The review describes applications of major GRAS (generally regarded as safe) yeasts for the production of therapeutic proteins and subunit vaccines; additionally, immunomodulatory properties of yeast cell wall components were reviewed for use of whole yeast cells as a new vaccine platform...
August 2017: Expert Opinion on Therapeutic Patents
https://www.readbyqxmd.com/read/28598353/cellular-consequences-of-diminished-protein-o-mannosyltransferase-activity-in-baker-s-yeast
#7
Ewa Zatorska, Lihi Gal, Jaro Schmitt, Daniela Bausewein, Maya Schuldiner, Sabine Strahl
O-Mannosylation is a type of protein glycosylation initiated in the endoplasmic reticulum (ER) by the protein O-mannosyltransferase (PMT) family. Despite the vital role of O-mannosylation, its molecular functions and regulation are not fully characterized. To further explore the cellular impact of protein O-mannosylation, we performed a genome-wide screen to identify Saccharomyces cerevisiae mutants with increased sensitivity towards the PMT-specific inhibitor compound R3A-5a. We identified the cell wall and the ER as the cell compartments affected most upon PMT inhibition...
June 9, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28597972/a-novel-interacting-protein-serp1-regulates-the-n-linked-glycosylation-and-function-of-glp-1-receptor-in-the-liver
#8
Yuanyuan Xiao, Junfeng Han, Qianqian Wang, Yueqin Mao, Meilin Wei, Weiping Jia, Li Wei
Glucagon-like peptide 1 (GLP-1) exerts multiple effects on metabolism through its receptor, GLP-1R, in the liver. Activation and transduction of GLP-1R require complex interactions of largely unknown accessory proteins, and these processes are crucial to the response to endoplasmic reticulum (ER) stress. Using the membrane-based split ubiquitin yeast two-hybrid system (MYTH) and a human liver cDNA library, we obtained the human GLP-1R interactome and identified SERP1 as a potential interacting protein based on its ability to stabilize membrane proteins and facilitate N-linked glycosylation...
June 9, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28585468/sesquiterpene-glucosides-from-shenzhou-honey-peach-fruit-showed-the-anti-aging-activity-in-the-evaluation-system-using-yeasts
#9
Yanhui Wang, Yanfei Lin, Lan Xiang, Hiroyuki Osada, Jianhua Qi
One new (1, SZMT01) and one known (2) anti-aging substances were isolated from Shenzhou honey peach fruit. Their structures were elucidated by spectroscopic methods and chemical derivatization, and the result reveals that these two compounds are sesquiterpene glucosides. SZMT01 possesses a new glycosylation with an ester linkage at one terminal in an acyclic sesquiterpenoid which is the end of a double bond at another terminal. Both compounds extend the replicative lifespan of K6001 yeast strain at doses of 7...
June 6, 2017: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/28512129/mammalian-o-mannosylation-of-cadherins-and-plexins-is-independent-of-protein-o-mannosyltransferases-1-and-2
#10
Ida Signe Bohse Larsen, Yoshiki Narimatsu, Hiren Jitendra Joshi, Zhang Yang, Oliver J Harrison, Julia Brasch, Lawrence Shapiro, Barry Honig, Sergey Y Vakhrushev, Henrik Clausen, Adnan Halim
Protein O-mannosylation is found in yeast and metazoans, and a family of conserved orthologous protein O-mannosyltransferases is believed to initiate this important post-translational modification. We recently discovered that the cadherin superfamily carries O-linked mannose (O-Man) glycans at highly conserved residues in specific extracellular cadherin domains, and it was suggested that the function of E-cadherin was dependent on the O-Man glycans. Deficiencies in enzymes catalyzing O-Man biosynthesis, including the two human protein O-mannosyltransferases, POMT1 and POMT2, underlie a subgroup of congenital muscular dystrophies designated α-dystroglycanopathies, because deficient O-Man glycosylation of α-dystroglycan disrupts laminin interaction with α-dystroglycan and the extracellular matrix...
July 7, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28512128/refined-topology-model-of-the-stt3-stt3-protein-subunit-of-the-oligosaccharyltransferase-complex
#11
Patricia Lara, Karin Öjemalm, Johannes Reithinger, Aurora Holgado, You Maojun, Abdessalem Hammed, Daniel Mattle, Hyun Kim, IngMarie Nilsson
The oligosaccharyltransferase complex, localized in the endoplasmic reticulum (ER) of eukaryotic cells, is responsible for the N-linked glycosylation of numerous protein substrates. The membrane protein STT3 is a highly conserved part of the oligosaccharyltransferase and likely contains the active site of the complex. However, understanding the catalytic determinants of this system has been challenging, in part because of a discrepancy in the structural topology of the bacterial versus eukaryotic proteins and incomplete information about the mechanism of membrane integration...
July 7, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28500401/sources-of-%C3%AE-galactosidase-and-its-applications-in-food-industry
#12
REVIEW
Shaima Saqib, Attiya Akram, Sobia Ahsan Halim, Raazia Tassaduq
The enzyme β-galactosidases have been isolated from various sources such as bacteria, fungi, yeast, vegetables, and recombinant sources. This enzyme holds importance due to its wide applications in food industries to manufacture lactose-hydrolyzed products for lactose-intolerant people and the formation of glycosylated products. Absorption of undigested lactose in small intestine requires the activity of this enzyme; hence, the deficiency of this enzyme leads to lactose intolerance. Lactose intolerance affects around 70% of world's adult population, while the prevalence rate of lactose intolerance is 60% in Pakistan...
May 2017: 3 Biotech
https://www.readbyqxmd.com/read/28499956/group-x-hybrid-histidine-kinase-chk1-is-dispensable-for-stress-adaptation-host-pathogen-interactions-and-virulence-in-the-opportunistic-yeast-candida-guilliermondii
#13
María J Navarro-Arias, Karine Dementhon, Tatiana A Defosse, Emilien Foureau, Vincent Courdavault, Marc Clastre, Solène Le Gal, Gilles Nevez, Yohann Le Govic, Jean-Philippe Bouchara, Nathalie Giglioli-Guivarc'h, Thierry Noël, Hector M Mora-Montes, Nicolas Papon
Hybrid histidine kinases (HHKs) progressively emerge as prominent sensing proteins in the fungal kingdom and as ideal targets for future therapeutics. The group X HHK is of major interest, since it was demonstrated to play an important role in stress adaptation, host-pathogen interactions and virulence in some yeast and mold models, and particularly Chk1, that corresponds to the sole group X HHK in Candida albicans. In the present work, we investigated the role of Chk1 in the low-virulence species Candida guilliermondii, in order to gain insight into putative conservation of the role of group X HHK in opportunistic yeasts...
May 9, 2017: Research in Microbiology
https://www.readbyqxmd.com/read/28470614/not-limited-to-e-coli-versatile-expression-vectors-for-mammalian-protein-expression
#14
Katharina Karste, Maren Bleckmann, Joop van den Heuvel
Recombinant protein expression is not limited to E. coli or other prokaryotic systems. It is inevitable to use eukaryotic systems in order to express challenging mammalian proteins. Eukaryotic systems are able to perform complex posttranslational modifications like protein processing, phosphorylation, glycosylation, which are essential for stability and functionality of many proteins. Different eukaryotic protein expression systems employing yeast, insect, or mammalian cell lines are established with each having its own advantages and disadvantages...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28467350/chemical-profile-and-antioxidant-anti-inflammatory-antimutagenic-and-antimicrobial-activities-of-geopropolis-from-the-stingless-bee-melipona-orbignyi
#15
Helder Freitas Dos Santos, Jaqueline Ferreira Campos, Cintia Miranda Dos Santos, José Benedito Perrella Balestieri, Denise Brentan Silva, Carlos Alexandre Carollo, Kely de Picoli Souza, Leticia Miranda Estevinho, Edson Lucas Dos Santos
Geopropolis is a resin mixed with mud, produced only by stingless bees. Despite being popularly known for its medicinal properties, few scientific studies have proven its biological activities. In this context, the objective of this study was to determine the chemical composition and antioxidant, anti-inflammatory, antimutagenic and antimicrobial activities of the Melipona orbignyi geopropolis. The hydroalcoholic extract of geopropolis (HEGP) was prepared and its chemical composition determined by high performance liquid chromatography coupled to diode array detector and mass spectrometry (HPLC-DAD-MS)...
May 3, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28446609/glycosylation-of-the-core-of-the-hiv-1-envelope-subunit-protein-gp120-is-not-required-for-native-trimer-formation-or-viral-infectivity
#16
Ujjwal Rathore, Piyali Saha, Sannula Kesavardhana, Aditya Arun Kumar, Rohini Datta, Sivasankar Devanarayanan, Raksha Das, John R Mascola, Raghavan Varadarajan
The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines...
June 16, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28433761/a-glycoproteomic-approach-reveals-that-the-s-layer-glycoprotein-of-lactobacillus-kefiri-cidca-83111-is-o-and-n-glycosylated
#17
Gustavo J Cavallero, Mariano Malamud, Adriana C Casabuono, M de Los Ángeles Serradell, Alicia S Couto
In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid...
April 20, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/28411481/a-re-evaluation-of-the-final-step-of-vanillin-biosynthesis-in-the-orchid-vanilla-planifolia
#18
Hailian Yang, Jaime Barros-Rios, Galina Kourteva, Xiaolan Rao, Fang Chen, Hui Shen, Chenggang Liu, Andrzej Podstolski, Faith Belanger, Daphna Havkin-Frenkel, Richard A Dixon
A recent publication describes an enzyme from the vanilla orchid Vanilla planifolia with the ability to convert ferulic acid directly to vanillin. The authors propose that this represents the final step in the biosynthesis of vanillin, which is then converted to its storage form, glucovanillin, by glycosylation. The existence of such a "vanillin synthase" could enable biotechnological production of vanillin from ferulic acid using a "natural" vanilla enzyme. The proposed vanillin synthase exhibits high identity to cysteine proteases, and is identical at the protein sequence level to a protein identified in 2003 as being associated with the conversion of 4-coumaric acid to 4-hydroxybenzaldehyde...
July 2017: Phytochemistry
https://www.readbyqxmd.com/read/28409466/the-use-of-glycosylation-tags-as-reporters-for-protein-entry-into-the-endoplasmic-reticulum-in-yeast-and-mammalian-cells
#19
Judith Buentzel, Sven Thoms
N-glycosylation is a process occurring in the Endoplasmic Reticulum (ER) in nearly every organism. Proteins containing a glycosylation site are quickly glycosylated by oligosaccharyltransferases once the glycosylation site is exposed to the ER lumen. The oligosaccharide tree is then modified and proteins are targeted to specific organelles or subcompartments. For a long time peroxisomal membrane proteins (PMP) were thought to be targeted directly to the peroxisome. However, in the course of recent years, several PMPs were found to be targeted via the ER...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28407119/a-barley-udp-glucosyltransferase-inactivates-nivalenol-and-provides-fusarium-head-blight-resistance-in-transgenic-wheat
#20
Xin Li, Herbert Michlmayr, Wolfgang Schweiger, Alexandra Malachova, Sanghyun Shin, Yadong Huang, Yanhong Dong, Gerlinde Wiesenberger, Susan McCormick, Marc Lemmens, Philipp Fruhmann, Christian Hametner, Franz Berthiller, Gerhard Adam, Gary J Muehlbauer
Fusarium Head Blight is a disease of cereal crops that causes severe yield losses and mycotoxin contamination of grain. The main causal pathogen, Fusarium graminearum, produces the trichothecene toxins deoxynivalenol or nivalenol as virulence factors. Nivalenol-producing isolates are most prevalent in Asia but co-exist with deoxynivalenol producers in lower frequency in North America and Europe. Previous studies identified a barley UDP-glucosyltransferase, HvUGT13248, that efficiently detoxifies deoxynivalenol, and when expressed in transgenic wheat results in high levels of type II resistance against deoxynivalenol-producing F...
April 1, 2017: Journal of Experimental Botany
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