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Marc Goetz, Anne Guivarc'h, Jörg Hirsche, Martin Andreas Bauerfeind, María-Cruz González, Tae Kyung Hyun, Seung Hee Eom, Dominique Chriqui, Thomas Engelke, Dominik K Großkinsky, Thomas Roitsch
Pollination in flowering plants is initiated by germination of pollen grains on stigmas followed by fast growth of pollen tubes representing highly energy-consuming processes. The symplastic isolation of pollen grains and tubes requires import of sucrose available in the apoplast. We show that the functional coupling of sucrose cleavage by invertases and uptake of the released hexoses by monosaccharide transporters are critical for pollination in tobacco. Transcript profiling, in situ hybridization and immunolocalization of extracellular invertases and two monosaccharide transporters in vitro and in vivo support the functional coupling in supplying carbohydrates for pollen germination and tube growth evidenced by spatiotemporally coordinated expression...
December 6, 2016: Plant Physiology
Lai-Sheng Meng, Ying-Qiu Li, Meng-Qian Liu, Ji-Hong Jiang
Anthocyanin accumulation specifically depends on sucrose (Suc) signaling/levels. However, the gene cascades specifically involved in the Suc signaling/level-mediated anthocyanin biosynthetic pathway are still unknown. Arabidopsis ANGUSTIFOLIA3 (AN3), a transcription coactivator, is involved in the regulation of leaf shape and drought tolerance. Recently, an AN3-CONSTITUTIVE PHOTOMORPHOGENIC 1 gene cascade has been reported to regulate the light signaling-mediated anthocyanin accumulation. Target gene analysis indicates that AN3 is associated with the YODA (YDA) promoter, a mitogen-activated protein kinase kinase kinase, in vivo for inducing anthocyanin accumulation...
2016: Frontiers in Plant Science
Paula E Jameson, Pragatheswari Dhandapani, Ondrej Novak, Jiancheng Song
Transporter genes and cytokinins are key targets for crop improvement. These genes are active during the development of the seed and its establishment as a strong sink. However, during germination, the seed transitions to being a source for the developing root and shoot. To determine if the sucrose transporter (SUT), amino acid permease (AAP), Sugar Will Eventually be Exported Transporter (SWEET), cell wall invertase (CWINV), cytokinin biosynthesis (IPT), activation (LOG) and degradation (CKX) gene family members are involved in both the sink and source activities of seeds, we used RT-qPCR to determine the expression of multiple gene family members, and LC-MS/MS to ascertain endogenous cytokinin levels in germinating Pisum sativum L...
December 1, 2016: International Journal of Molecular Sciences
Xun Liu, Weiling Shi, Wang Yin, Jichun Wang
Potato (Solanum tuberosum L.) vacuolar invertase (β-fructofuranosidase; EC inhibitor 2 (StInvInh2) plays an important role in cold-induced sweetening (CIS) of potato tubers. The transcript levels of StInvInh2 were increased by prolonged cold in potato tubers with CIS-resistance but decreased in potato tubers with CIS-sensitivity. However, the transcript regulation mechanisms of StInvInh2 responding to prolonged cold are largely unclear in CIS-resistant and CIS-sensitive genotypes. In the present study, the 5'-flanking sequence of the StInvInh2 was cloned, and cis-acting elements were predicted...
November 29, 2016: Plant Physiology and Biochemistry: PPB
Justine Ferey, David Da Silva, Sophie Bravo-Veyrat, Pierre Lafite, Richard Daniellou, Benoît Maunit
This paper presents a kinetic study of invertase, a specific fructofuranosidase cloned from the Leishmania major genome. The kinetic parameters of the β-d-fructofuranosidase from Leishmania major (BfrA) were determined using Thin-Layer Chromatography (TLC) and UV-densitometry (TLC@UV) specifically developed for the separation and detection of three carbohydrates namely sucrose, glucose and fructose. Separation was performed on TLC silica gel 60 F254 plates impregnated with sodium bisulphate and citrate and heated prior to development...
November 25, 2016: Journal of Chromatography. A
Derek J Herman, Lisa O Knowles, N Richard Knowles
Tolerance to heat stress for retention of low-temperature sweetening-resistant phenotype in potato is conferred by insensitivity of acid invertase activity to cold induction. Heat stress exacerbated cold sweetening (buildup of reducing sugars) of the LTS (low-temperature sweetening)-susceptible potato (Solanum tuberosum L.) cultivars, Ranger Russet and Russet Burbank, and completely abolished the resistance to cold sweetening in the LTS-resistant cultivars/clones, Sage Russet, GemStar Russet, POR06V12-3 and A02138-2...
November 30, 2016: Planta
Caixia Wang, Qingming Zhang, Feifei Wang, Wenxing Liang
The objective of this study was to evaluate the toxicity of the fungicide dimethomorph to soil microbial activity and the earthworm Eisenia fetida. Multiple biomarkers, namely, four soil enzymes (urease, dehydrogenase, invertase, and acid phosphatase), four earthworm biochemical indices (dismutase, catalase, cellulase, and malondialdehyde), and the transcriptional levels of both target genes (dismutase and catalase) were measured at 1, 10, and 100 mg kg(-1) after 1, 7, 21, and 28 days. The degradation rate of dimethomorph in soil was also determined, and the results indicated that most parameters did not differ from the controls at 1 and 10 mg kg(-1) dimethomorph by the last exposure time (28 d)...
November 22, 2016: Chemosphere
Nooshin Mohandesi, Kamahldin Haghbeen, Omid Ranaei, Seyed Shahriar Arab, Sorour Hassani
Engineering of invertases has come to attention because of increasing demand for possible applications of invertases in various industrial processes. Due to the known physicochemical properties, invertases from micro-organisms such as Saccharomyces cerevisiae carrying SUC2 gene are considered as primary models. To improve thermostability and catalytic efficiency of SUC2 invertase (SInv), six influential residues with Relative Solvent Accessibility<5% were selected through multiple-sequence alignments, molecular modelling, structural and computational analyses...
January 2017: Enzyme and Microbial Technology
Heng Yue, Song Fang, Yizhi Zhang, Yang Ning, Weisong Yu, Fanyu Kong, Jun Qiu
The enantioselective effects of the chiral pesticide metalaxyl on soil enzyme activity were investigated. Incubation experiments were conducted to investigate the effects of metalaxyl enantiomers at different concentrations on the activities of urease, invertase, and catalase as well as the type of activity change (activation vs. inhibition) at different times during incubation. The results indicated that the effects of metalaxyl on the activity of soil enzymes were not only related to the concentration of the enantiomers and soil incubation time, but also to the chiral configuration, suggesting the effects were enantioselective...
November 10, 2016: Chirality
Ranfeng Ye, Chengzhou Zhu, Yang Song, Junhua Song, Shaofang Fu, Qian Lu, Xu Yang, Mei-Jun Zhu, Dan Du, He Li, Yuehe Lin
Protein-protein conjugates play a vital role in bioassays with their inherent functions of biological recognition and signal amplification. Herein, a one-pot green method for synthesis of all-inclusive protein-protein nanoflowers has been developed. The protein-protein nanoflowers integrate both essential functions of biological recognition and signal amplification, and they were used as ideal signal labels for the sensitive point-of-care detection of Escherichia coli O157:H7. Especially noteworthy, the prepared Con A-invertase-CaHPO4 hybrid nanoflowers simultaneously loaded sufficient invertase and enhanced the activity of the immobilized invertase, which fits well with the requirements of signal labels for bioassays...
December 7, 2016: Nanoscale
Hongfei Dai, Maorun Fu, Xiaoying Yang, Qingmin Chen
The aim of this study was to investigate the role of ethylene to control sprouting of potatoes by observing the effect of exogenous ethylene on carbohydrate metabolism and key enzymes. The initial time of potato tuber sprouting and sprouting index were recorded, and rate of respiration, total sugar, total reducing sugar, starch, fructose, glucose, sucrose and the activities of acid invertase (AI), neutral invertase (NI), sucrose synthase (SS), sucrose phosphate synthase (SPS), starch phosphorylase and amylase during sprouting were measured...
August 2016: Journal of Food Science and Technology
Jin Xie, Kun Cai, Hai-Xi Hu, Yong-Liang Jiang, Feng Yang, Peng-Fei Hu, Dong-Dong Cao, Wei-Fang Li, Yuxing Chen, Cong-Zhao Zhou
Invertases catalyze the hydrolysis of sucrose to glucose and fructose, thereby playing a key role in primary metabolism and plant development. According to the optimum pH, invertases are classified into acid invertases (Ac-Invs) and alkaline/neutral invertases (A/N-Invs), which share no sequence homology. Compared with Ac-Invs that have been extensively studied, the structure and catalytic mechanism of A/N-Invs remain unknown. Here we report the crystal structures of Anabaena alkaline invertase InvA, which was proposed to be the ancestor of modern plant A/N-Invs...
December 2, 2016: Journal of Biological Chemistry
Kemal Çetin, Işık Perçin, Fatma Denizli, Adil Denizli
The aim of this study is to investigate the usability of cryogel columns for the purification of invertase from Saccharomyces cerevisiae. Poly(2-hydroxyethyl methacrylate) monolithic columns were produced via cryogelation. Ester groups of the poly(2-hydroxyethyl methacrylate) structure were then converted to imine groups by the reaction with poly(ethylene imine) in the presence of NaHCO3. Transition metal ions, Cu(II), Co(II), and Ni(II), were chelated on the PEI-modified cryogel columns. Purification of invertase from natural source namely S...
October 24, 2016: Artificial Cells, Nanomedicine, and Biotechnology
Aleksandra Margetić, Zoran Vujčić
Yeast Saccharomyces cerevisiae is the most significant source of enzyme invertase. It is mainly used in the food industry as a soluble or immobilized enzyme. The greatest amount of invertase is located in the periplasmic space in yeast. In this work it was isolated two forms of enzyme from yeastS. cerevisiae cell, soluble and cell wall invertase (CWI). Both forms of enzyme showed same temperature optimum (60°C), similar pH optimum and kinetic parameters. The significant difference between these biocatalysts was observed in their thermal stability, stability in urea and methanol solution...
October 13, 2016: Preparative Biochemistry & Biotechnology
Chanchal Kumar, Jitendra Wagh, G Archana, G Naresh Kumar
Enterobacter asburiae PSI3 solubilizes mineral phosphates in the presence of glucose by the secretion of gluconic acid generated by the action of a periplasmic pyrroloquinoline quinone dependent glucose dehydrogenase. In order to achieve mineral phosphate solubilization phenotype in the presence of sucrose, plasmids pCNK4 and pCNK5 containing genes encoding the invertase enzyme of Zymomonas mobilis (invB) and of Saccharomyces cerevisiae (suc2) under constitutive promoters were constructed with malE signal sequence (in case of invB alone as the suc2 is secreted natively)...
December 2016: World Journal of Microbiology & Biotechnology
Nore Struyf, Jitka Laurent, Bianca Lefevere, Joran Verspreet, Kevin J Verstrepen, Christophe M Courtin
It is generally believed that maltose drives yeast-mediated bread dough fermentation. The relative importance of fructose and glucose, released from wheat fructan and sucrose by invertase, compared to maltose is, however, not documented. This is surprising given the preference of yeast for glucose and fructose over maltose. This study revealed that, after 2h fermentation of wheat flour dough, about 44% of the sugars consumed were generated by invertase-mediated degradation of fructan, raffinose and sucrose...
March 1, 2017: Food Chemistry
Zahra Gerivani, Elham Vashaee, Hamid Reza Sadeghipour, Mahnaz Aghdasi, Zahra-Sadat Shobbar, Majid Azimmohseni
Tree seed dormancy release by cold stratification accompanies with the embryo increased gluconeogenesis competence. Cyanide also breaks seed dormancy however, integrated information about its effects on carbon metabolism is lacking. Accordingly, the impacts of HCN on germination, lipid gluconeogenesis and sugar transport capacity of walnut (Juglans regia L.) kernels were investigated during 10-days period prior to radicle protrusion. HCN increased walnut kernel germination and within four days of kernel incubation, hastened the decline of starch, reducing and non-reducing sugars and led to greater activities of alkaline invertase and glucose-6-phosphate dehydrogenase...
November 2016: Plant Science: An International Journal of Experimental Plant Biology
Chunsha Zhang, Hongwei Zhang, Zongxiang Zhan, Bingjiang Liu, Zhentai Chen, Yi Liang
Allium cepa L. is a widely cultivated and economically significant vegetable crop worldwide, with beneficial dietary and health-related properties, but its sucrose metabolism is still poorly understood. To analyze sucrose metabolism during bulb swelling, and the development of sweet taste in onion, a global transcriptome profile of onion bulbs was undertaken at three different developmental stages, using RNA-seq. A total of 79,376 unigenes, with a mean length of 678 bp, was obtained. In total, 7% of annotated Clusters of Orthologous Groups (COG) were involved in carbohydrate transport and metabolism...
2016: Frontiers in Plant Science
Guozheng Qin, Zhu Zhu, Weihao Wang, Jianghua Cai, Yong Chen, Li Li, Shiping Tian
Fruit ripening is a complex process that involves a series of physiological and biochemical changes that ultimately influence fruit quality traits, such as color and flavor. Sugar metabolism is an important factor in ripening, and there is evidence that it influences various aspects of ripening, although the associated mechanism is not well understood. In this study, we identified and analyzed the expression of 36 genes involved in Suc metabolism in ripening tomato (Solanum lycopersicum) fruit. Chromatin immunoprecipitation and gel mobility shift assays indicated that SlVIF, which encodes a vacuolar invertase inhibitor, and SlVI, encoding a vacuolar invertase, are directly regulated by the global fruit ripening regulator RIPENING INHIBITOR (RIN)...
November 2016: Plant Physiology
Kai Yu, Kok Siong Ang, Dong-Yup Lee
Codon optimization has been widely used for designing native or synthetic genes to enhance their expression in heterologous host organisms. We recently developed Codon Optimization On-Line (COOL) which is a web-based tool to provide multi-objective codon optimization functionality for synthetic gene design. COOL provides a simple and flexible interface for customizing codon optimization based on several design parameters such as individual codon usage, codon pairing, and codon adaptation index. User-defined sequences can also be compared against the COOL optimized ones to show the extent by which the user's sequences can be evaluated and further improved...
2017: Methods in Molecular Biology
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