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Mostafa Abdelrahman, Magdi El-Sayed, Shusei Sato, Hideki Hirakawa, Shin-Ichi Ito, Keisuke Tanaka, Yoko Mine, Nobuo Sugiyama, Minoru Suzuki, Naoki Yamauchi, Masayoshi Shigyo
The genus Allium is a rich source of steroidal saponins, and its medicinal properties have been attributed to these bioactive compounds. The saponin compounds with diverse structures play a pivotal role in Allium's defense mechanism. Despite numerous studies on the occurrence and chemical structure of steroidal saponins, their biosynthetic pathway in Allium species is poorly understood. The monosomic addition lines (MALs) of the Japanese bunching onion (A. fistulosum, FF) with an extra chromosome from the shallot (A...
2017: PloS One
Changsheng Li, Yongcai Huang, Ruidong Huang, Yongrui Wu, Wenqin Wang
Starch is the most abundant storage carbohydrate in maize kernel. The content of amylose and amylopectin confers unique properties in food processing and industrial application. Thus, the resurgent interest has been switched to the study of individual amylose or amylopectin rather than total starch, whereas the enzymatic machinery for amylose synthesis remains elusive. We took advantage of the phenotype of amylose content and the genotype of 9,007,194 single nucleotide polymorphisms (SNPs) from 454 inbred maize lines...
August 10, 2017: Plant Biotechnology Journal
Melissa H Mageroy, Sharon Jancsik, Macaire M S Yuen, Michael Fischer, Stephen G Withers, Christian Paetz, Bernd Schneider, John J MacKay, Joerg Bohlmann
Acetophenones are phenolic compounds involved in resistance of white spruce (Picea glauca) against spruce budworm (Choristoneura fumiferiana), a major forest pest in North America. The acetophenones pungenol and piceol commonly accumulate in spruce foliage in the form of the corresponding glycosides, pungenin and picein. These glycosides appear to be inactive against the insect, but can be cleaved by a spruce β-glucosidase, PgβGlu1, which releases the active aglycons. The reverse glycosylation reaction was hypothesized to involve a family-1 UDP-sugar dependent glycosyltransferase (UGT) to facilitate acetophenone accumulation in the plant...
August 9, 2017: Plant Physiology
Ganapati Bhat, Vishwanath-Reddy Hothpet, Ming-Fong Lin, Pi-Wan Cheng
BACKGROUND: There is a pressing need for biomarkers that can distinguish indolent from aggressive prostate cancer to prevent over-treatment of patients with indolent tumor. METHODS: Golgi targeting of glycosyltransferases was characterized by confocal microscopy after knockdown of GM130, giantin, or both. N-glycans on a trans-Golgi enzyme β4galactosyltransferase-1 isolated by immunoprecipitation from androgen-sensitive and independent prostate cancer cells were determined by matrix-assisted laser desorption-time of flight-mass spectrometry...
August 4, 2017: Biochimica et Biophysica Acta
Sonia Covaceuszach, Manuela Bozzi, Maria Giulia Bigotti, Francesca Sciandra, Petr Valeryevich Konarev, Andrea Brancaccio, Alberto Cassetta
Dystroglycan (DG), composed of α and β subunits, belongs to the dystrophin-associated glycoprotein complex. α-DG is an extracellular matrix protein that undergoes a complex post-translational glycosylation process. The bifunctional glycosyltransferase like-acetylglucosaminyltransferase (LARGE) plays a crucial role in the maturation of α-DG, enabling its binding to laminin. We have already structurally analyzed the N-terminal region of murine α-DG (α-DG-Nt) and of a pathological single point mutant that may affect recognition of LARGE, although the structural features of the potential interaction between LARGE and DG remain elusive...
August 2017: FEBS Open Bio
Ali Saleh Hassan, Kelly Houston, Jelle Lahnstein, Neil Shirley, Julian G Schwerdt, Michael J Gidley, Robbie Waugh, Alan Little, Rachel A Burton
In barley endosperm arabinoxylan (AX) is the second most abundant cell wall polysaccharide and in wheat it is the most abundant polysaccharide in the starchy endosperm walls of the grain. AX is one of the main contributors to grain dietary fibre content providing several health benefits including cholesterol and glucose lowering effects, and antioxidant activities. Due to its complex structural features, AX might also affect the downstream applications of barley grain in malting and brewing. Using a high pressure liquid chromatography (HPLC) method we quantified AX amounts in mature grain in 128 spring 2-row barley accessions...
2017: PloS One
Gonzalo H Villarino, Qiwen Hu, Michael J Scanlon, Lukas Mueller, Aureliano Bombarely, Neil S Mattson
One of the primary objectives of plant biotechnology is to increase resistance to abiotic stresses, such as salinity. Salinity is a major abiotic stress and increasing crop resistant to salt continues to the present day as a major challenge. Salt stress disturbs cellular environment leading to protein misfolding, affecting normal plant growth and causing agricultural losses worldwide. The advent of state-of-the-art technologies such as high throughput mRNA sequencing (RNA-seq) has revolutionized whole-transcriptome analysis by allowing, with high precision, to measure changes in gene expression...
August 3, 2017: Genes
Toshihiro Inubushi, Satoshi Nozawa, Kazu Matsumoto, Fumitoshi Irie, Yu Yamaguchi
Multiple hereditary exostoses (MHE) is characterized by the development of numerous benign bony tumors (osteochondromas). Although it has been well established that MHE is caused by mutations in EXT1 and EXT2, which encode glycosyltransferase essential for heparan sulfate (HS) biosynthesis, the cellular origin and molecular mechanisms of MHE remain elusive. Here, we show that in Ext1 mutant mice, osteochondromas develop from mesenchymal stem cell-like progenitor cells residing in the perichondrium, and we show that enhanced BMP signaling in these cells is the primary signaling defect that leads to osteochondromagenesis...
August 3, 2017: JCI Insight
Sung Hyun Cho, Pallinti Purushotham, Chao Fang, Cassandra Maranas, Sara M Díaz-Moreno, Vincent Bulone, Jochen Zimmer, Manish Kumar, B Tracy Nixon
Cellulose, the major component of plant cell walls, can be converted to bioethanol and is thus highly studied. In plants, cellulose is produced by cellulose synthase, a processive family-2 glycosyltransferase. In plant cell walls, individual β-1,4-glucan chains polymerized by CesA are assembled into microfibrils that are frequently bundled into macrofibrils. An in vitro system in which cellulose is synthesized and assembled into fibrils would facilitate detailed study of this process. Here, we report the heterologous expression and partial purification of His-tagged CesA5 from Physcomitrella patens...
August 2, 2017: Plant Physiology
Georg A Sprenger, Florian Baumgärtner, Christoph Albermann
Human milk oligosaccharides (HMO) are almost unique constituents of breast milk and are not found in appreciable amounts in cow milk. Due to several positive aspects of HMO for the development, health, and wellbeing of infants, production of HMO would be desirable. As a result, scientist from different disciplines have developed methods for the preparation of single HMO compounds. Here, we review approaches to HMO preparation by (chemo-)enzymatic syntheses or by whole-cell biotransformation with recombinant bacterial cells...
July 29, 2017: Journal of Biotechnology
Víctor M Castillo-Acosta, Jan Balzarini, Dolores González-Pacanowska
Trypanosomal diseases are in need of innovative therapies that exploit novel mechanisms of action. The cell surface of trypanosomatid parasites is characterized by a dense coat of glycoconjugates with important functions in host cell recognition, immune evasion, infectivity, and cell function. The nature of parasite surface glycans is highly dynamic and changes during differentiation and in response to different stimuli through the action of glycosyltransferases and glycosidases. Here we propose a new approach to antiparasitic drug discovery that involves the use of carbohydrate-binding agents that bind specifically to cell-surface glycans, giving rise to cytotoxic events and parasite death...
July 28, 2017: Trends in Parasitology
Jian-Hua Hao, Li-Ping Huang, Xiao-Tong Chen, Jing-Jing Sun, Jun-Zhong Liu, Wei Wang, Mi Sun
Cyclodextrin glycosyltransferase (CGTase) is an enzyme able to convert starch and other substrates into cyclodextrins (CDs). A marine strain Y112 producing α-CGTase was identified as Bacillus agaradhaerens Y112 by physiological and biochemical characterization, and 16S rDNA analysis. The gene coding for α-CGTase was cloned, sequenced and expressed in Escherichia coli BL21 (DE3) cells. Recombinant α-CGTase was purified in one-step chromatographic separation and its purity evaluated by SDS-PAGE, showing the presence of one band with a molecular mass of about 92 kDa...
July 27, 2017: Protein Expression and Purification
Jianguo Zhang, Yan Zhang, Mengla Li
Cyclodextrin glycosyltransferase (CGTase) catalyzes the conversion of starch into cyclodextrin (CD), which is widely applied in food, pharmaceutical, cosmetic, and agricultural industries. For efficient production of CD, high yield of CGTase with good characteristics is necessary. In this study, the cgt gene from Bacillus pseudalcaliphilus was expressed in Komagataella phaffii after codon optimization and expression vector selection. The β-CGTase activity in the transformant reached 3885.1UmL(-1), which is the highest value reported so far, at 28°C, 6% inoculum ratio, and 1...
July 27, 2017: Journal of Biotechnology
Alejandro H Orrego, Lara Trobo-Maseda, Javier Rocha-Martin, Jose M Guisan
Sucrose synthases (SuSys) can be used to synthesize cost-effective uridine 5'-diphosphate glucose (UDP-glc) or can be coupled to glycosyltransferases (GTs) for the continuous recycling of UDP-glc. In this study, we present the first report of the immobilization-stabilization of a SuSy by multipoint covalent attachment. This stabilization strategy is very complex for multimeric enzymes because a very intense multipoint attachment can promote a dramatic loss of activity and/or stability. The homotetrameric SuSy from Nitrosomonas europaea (SuSyNe) was immobilized on a glyoxyl agarose support through two different orientations...
October 2017: Enzyme and Microbial Technology
Somanath Kallolimath, Clemens Gruber, Herta Steinkellner, Alexandra Castilho
Glyco-modulation of therapeutic proteins produced in plants has shown great success. Plant-based expression platforms for tailored human-like N-glycosylation are based on the overexpression of foreign genes. However, drawbacks such as protein miss targeting, interference with endogenous glycosyltransferases or with plant development hamper the widespread use of the technology. Here we describe a technique that facilitates the generation of recombinant proteins with targeted N-glycosylation at high homogeneity...
July 29, 2017: Biotechnology Journal
Rebecca-Ayme Hughes, Yunlong Zhang, Ran Zhang, Philip G Williams, Jonathan S Lindsey, Eric S Miller
The cyanobacterial culture HT-58-2 was originally described as a strain of Tolypothrix nodosa with the ability to produce tolyporphins, which comprise a family of distinct tetrapyrrole macrocycles with reported efflux pump inhibition properties. Upon reviving the culture from what was thought to be a non-extant collection, studies of culture conditions, strain characterization, phylogeny, and genomics have been undertaken. Herein, HT-58-2 was shown by 16S rRNA analysis to closely align with Brasilonema strains and not with Tolypothrix isolates...
July 28, 2017: Applied and Environmental Microbiology
Zheng Wu, Wenli Gao, Shaotong Zhou, Zhaolin Wen, Xianpu Ni, Huanzhang Xia
Gentamicin B and gentamicin C1a are the direct precursor for Isepamicin and Etimicin synthesis, respectively. Although producing strains have been improved for many years, both gentamicin B titer and gentamicin C1a titer in the fermentation are still low. Because all gentamicin components are biosynthesized using UDP-N-acetyl-d-glucosamine (UDP-GlcNAc) and UDP-xylose as precursors, we tried to explore strategies for development of strains capable of directing greater fluxes of these precursors into production of gentamicins...
October 2017: Microbiological Research
Yangyang Xu, Zhigang Wu, Peiru Zhang, He Zhu, Hailiang Zhu, Qitao Song, Li Wang, Faxing Wang, Peng George Wang, Jiansong Cheng
A novel enzymatic approach was developed for facile production of glycopeptides carrying natural eukaryotic N-glycans. In this approach, peptides can be GlcNAcylated at one or two natural N-glycosylation sites via two-step enzymatic reactions catalyzed by an evolved N-glycosyltransferase (ApNGT(Q469A)) and a glucosamine N-acetyltransferase (GlmA), respectively. The resulting GlcNAc-peptides were further modified by an endo-β-N-acetylglucosaminidase M mutant (EndoM(N175Q)) to generate glycopeptides. In three steps of enzymatic catalysis, glycopeptides carrying complex-type N-glycans can be efficiently synthesized...
July 28, 2017: Chemical Communications: Chem Comm
Xiang-Yi Li, Ya-Qin Wen, Nan Meng, Xu Qian, Qiu-Hong Pan
HIGHLIGHTS A similar trend on accumulation of glycosidically bound monoterpenes was observed in both varietiesTwo VvGT7 alleles mutations occurred at key sites in Muscat blanc à PetitVvGT14 exerted a major role in production of monoterpenyl glycosides in both varieties Terpenoids are the major aroma components and generally exist as both free and glycosidically-bound forms, of which nonvolatile glycosides account for a large fraction in grape berries. Our previous study has indicated that differential accumulation of monoterpenyl glycosides in Vitis vinifera "Muscat blanc à Petit" between two regions is closely correlated to monoterpenyl glucosyltransferase (VvGT14, XM_002285734...
2017: Frontiers in Plant Science
Fan Lin, Brad J Williams, Padmavathi A V Thangella, Adam Ladak, Athena A Schepmoes, Hernando J Olivos, Kangmei Zhao, Stephen J Callister, Laura E Bartley
Internodes of grass stems function in mechanical support, transport, and, in some species, are a major sink organ for carbon in the form of cell wall polymers. This study reports cell wall composition, proteomic, and metabolite analyses of the rice elongating internode. Cellulose, lignin, and xylose increase as a percentage of cell wall material along eight segments of the second rice internode (internode II) at booting stage, from the younger to the older internode segments, indicating active cell wall synthesis...
2017: Frontiers in Plant Science
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