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Qianqian Zhang, Ringo van Wijk, Xavier Zarza, Muhammad Shahbaz, Max van Hooren, Aisha Guardia, Denise Scuffi, Carlos García-Mata, Wim Van den Ende, Susanne Hoffmann-Benning, Michel A Haring, Ana M Laxalt, Teun Munnik
Phospholipase C (PLC) is a well-known signaling enzyme in metazoans that hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2) to produce inositol 1,4,5-trisphosphate and diacylglycerol as second messengers involved in mutiple processes. Plants contain PLC too, but relatively little is known about its function there. The model system Arabidopsis thaliana contains nine PLC genes. Reversed genetics have implicated several roles for PLCs in plant development and stress signaling. Here, PLC5 is functionally addressed...
August 9, 2018: Plant & Cell Physiology
Zhe Kong, Lu Li, Yu-You Li
A lab-scale UASB was operated successfully to anaerobically treat wastewater containing approximately 2000 mg L-1 N, N-dimethylformamide (DMF) by artificially mixing anaerobic granular sludge with DMF-degrading activated sludge. DMF was effectively degraded by the UASB under a low OLR of 1.63-4.22 g COD L-1  d-1 , with over 96% DMF removal efficiency and a high methane production rate. However, the DMF-degrading ability gradually weakened along with increases in the OLR. The analysis of the microbial community structure by high-throughput sequencing revealed a decline in the abundance of the facultatively anaerobic DMF-hydrolyzing bacteria originating from activated sludge with increasing OLR, further deteriorating the methanogenic degradation of DMF...
August 7, 2018: Bioresource Technology
Zheng-Gang Han, Ji-Wen Zhang, Xiao-Fang Jiang, Jiang-Ke Yang
The α-galactosidases, which can catalyze the removal of α-1,6-linked terminal galactose residues from galactooligosaccharide materials, have good potential for industrial applications. The high-level and efficient secretion of the α-galactosidases into the extracellular space has greatly simplified the downstream bioengineering process, facilitating their bioapplications. In this study, the effects of gene dosage and endoplasmic reticulum secretion-associated factors (ERSAs) on the secretory expression of an α-galactosidase gene derived from a Aspergillus oryzae strain were investigated by constructing multicopy expression cassettes and coexpressing the α-galactosidase gene with ERSAs...
August 11, 2018: Protein Expression and Purification
Ren-Kuan Li, Zeng Chen, Xi-Juan Ying, Tzi Bun Ng, Xiu-Yun Ye
An agar-degrading bacterium, strain BN3, was isolated from a coastal soil sample collected in Taiwan Strait, China and identified to be a novel species of the genus Microbulbifer. The gene (N3-1) encoding for a novel β-agarase from the isolate was cloned and sequenced. It encoded a mature protein with 274 amino acids and a calculated molecular mass of 34.3 kDa. The deduced amino acid sequence manifested sequence similarity (61-84% identity) to characterized β-agarases in the glycoside hydrolase family 16...
August 11, 2018: International Journal of Biological Macromolecules
Zishuo Cheng, Jamie VanPelt, Alexander Bergstrom, Christopher Bethel, Andrew Katko, Callie Miller, Kelly Mason, Erin Cumming, Huan Zhang, Robert Kimble, Sarah Fullington, Stacey Lowery Bretz, Jay C Nix, Robert A Bonomo, David Tierney, Richard C Page, Michael W Crowder
In an effort to evaluate whether a recently reported putative metallo-β-lactamase (MβL) contains a novel MβL active site, SPS-1 from Sediminispirochaeta smaragdinae was over-expressed, purified, and characterized using spectroscopic and crystallographic studies. Metal analyses demonstrate that recombinant SPS-1 binds nearly 2 equivalents of Zn(II), and steady-state kinetic studies show that the enzyme hydrolyzes carbapenems and certain cephalosporins but not β-lactam substrates with bulky substituents in the 6/7 position...
August 14, 2018: Biochemistry
Mengke Wang, Dandan Su, Guannan Wang, Xingguang Su
In this work, a facile, label-free, and sensitive fluorometric strategy for detection of trypsin and its inhibitor was established on the basis of the fluorescence resonance energy transfer (FRET) between mercaptoundecanoic acid functionalized gold nanoclusters (AuNCs) and gold nanoparticles (AuNPs) via protamine as a bridge. Protamine can trigger the aggregation of AuNPs and link AuNCs with aggregated AuNPs through electrostatic interaction. Compared with monodisperse AuNPs, the UV-vis absorption band of aggregated AuNPs overlapped considerably with the emission spectrum of AuNCs...
August 13, 2018: Analytical and Bioanalytical Chemistry
Pengbo Cui, Songyi Lin, Ziqi Jin, Beiwei Zhu, Liang Song, Na Sun
In this study, a novel calcium-binding heptapeptide (NDEELNK) that is released during the trypsin hydrolysis of sea cucumber ovum was identified by peptidomics. The calcium binding mode, in vitro digestion profile and calcium absorption of the NDEELNK-calcium complex were investigated. The NDEELNK peptide could spontaneously bind calcium with a 1 : 1 stoichiometry, and the calcium-binding site might involve the carboxyl oxygen and amino nitrogen atoms of two glutamic acid and one aspartic acid residues in the NDEELNK peptide...
August 14, 2018: Food & Function
Thaís Barboni Alves, Pedro Henrique de Oliveira Ornela, Arthur Henrique Cavalcanti de Oliveira, João Atílio Jorge, Luis Henrique Souza Guimarães
The filamentous fungus Aspergillus niveus produced extracellular antifungal chitinase when cultured under submerged fermentation (SbmF) using crab shells as the carbon source. Maximal chitinase production was achieved at 192 h of cultivation using minimal medium containing 1% chitin. The enzyme was purified 1.97-fold with 40% recovery by ammonium sulfate precipitation and Sephadex G-100 gel filtration. The molecular mass was estimated to be 44 kDa by both 12% SDS-PAGE and Sepharose CL-6B gel filtration. Maximal A...
August 2018: 3 Biotech
Manal Elzoheiry, Akram A Da'dara, Rita Bhardwaj, Qiang Wang, Manar S Azab, El-Saeed I El-Kholy, Samar N El-Beshbishi, Patrick J Skelly
Schistosomes are parasitic flatworms that infect the vasculature of >200 million people around the world. These long-lived parasites do not appear to provoke blood clot formation or obvious inflammation around them in vivo . Proteins expressed at the host-parasite interface (such as Schistosoma mansoni alkaline phosphatase, SmAP) are likely key to these abilities. SmAP is a glycoprotein that hydrolyses the artificial substrate p -nitrophenyl phosphate in a reaction that requires Mg2+ and at an optimal pH of 9...
2018: Frontiers in Immunology
Shimin Kang, Jiaming Pan, Guoting Gu, Chong Wang, Zepan Wang, Jionghao Tan, Guiheng Liu
A sequential production of levulinic acid (LA) and porous carbon material (CM) from cellulose was conducted by a two-step process. The cellulose was first acid hydrolyzed, and the preferred reaction conditions required a severity factor of 4.0⁻4.5, in which the yields of LA, formic acid, and solid residue were 38 ± 3 wt%, 17 ± 3 wt%, and 15 ± 3 wt%, respectively. The solid residue was further used for CM preparation through pyrolysis, with or without ZnCl₂ activation. The ZnCl₂ activation promoted the formation of CMs with improved thermal stability, high surface area (1184⁻2510 m²/g), and excellent phenol adsorption capacity (136⁻172 mg/g)...
August 11, 2018: Materials
Peng-Fei Qi, Ya-Zhou Zhang, Cai-Hong Liu, Jing Zhu, Qing Chen, Zhen-Ru Guo, Yan Wang, Bin-Jie Xu, Ting Zheng, Yun-Feng Jiang, Jiang-Ping Wang, Cai-Yi Zhou, Xiang Feng, Li Kong, Xiu-Jin Lan, Qian-Tao Jiang, Yu-Ming Wei, You-Liang Zheng
ATP-binding cassette (ABC) transporters hydrolyze ATP to transport a wide range of substrates. Fusarium graminearum is a major causal agent of Fusarium head blight, which is a severe disease in wheat worldwide. FgABCC9 ( FG05_07325 ) encodes an ABC-C (ABC transporter family C) transporter in F. graminearum , which was highly expressed during the infection in wheat and was up-regulated by the plant defense hormone salicylic acid (SA) and the fungicide tebuconazole. The predicted tertiary structure of the FgABCC9 protein was consistent with the schematic of the ABC exporter...
August 10, 2018: International Journal of Molecular Sciences
Yukihiro Esaka, Kenji Hisato, Takuhei Yamamoto, Hiroya Murakami, Bunji Uno
The substrate selectivities of three endonucleases were studied quantitatively using capillary zone electrophoresis to find one giving N2 -ethyl(Et)-2'-deoxyguanosine-5'-monophosphate (5'-dGMP) and cyclic 1,N2 -propano(CPr)-5'-dGMP from DNAs damaged by acetaldehyde (AA). Six 2'-deoxyribonucleoside-5'-monophosphates to be quantified in the hydrolysis solutions of DNAs, namely, Et-5'-dGMP, CPr-5'-dGMP, and four authentic ones, were completely separated using a 100 mM borate running buffer solution having an optimized pH of 9...
2018: Analytical Sciences: the International Journal of the Japan Society for Analytical Chemistry
Li-Wei Chiu, Matthew J Heckert, You You, Nicholas Albanese, Tamara Fenwick, Daniel L Siehl, Linda A Castle, Yumin Tao
Auxin homeostasis is a highly-regulated process that must be maintained to allow auxin to exert critical growth and developmental controls. Auxin conjugase and hydrolase family proteins play important roles in auxin homeostasis through means of storage, activation, inactivation, response inhibition, and degradation of auxins in plants. We systematically evaluated 60 GH3 proteins from diverse plant species for amino acid conjugation activity with the known substrates jasmonic acid (JA), indole-3-acetic acid (IAA), and 4-hydroxybenzoate (4-HBA)...
August 8, 2018: Plant & Cell Physiology
Snezana Agatonovic-Kustrin, David W Morton
A high-performance thin-layer chromatography (HPTLC) method was developed for quantification of α-amylase inhibitory activity and stigmasterol content in ant plant extracts. An improved HPTLC method for the determination of total free radical scavenging activity in samples using DPPH• is also reported. For quantification of α -amylase inhibitory activity, the developed HPTLC plate is dipped into an α -amylase solution, and the bioautogram is then incubated at 25 °C for 30 min under humid conditions...
2018: MethodsX
Hiromi T Wilson, Masoume Amirkhani, Alan G Taylor
The effect of gelatin, used as a biostimulant, was investigated on plant growth in greenhouse studies. Biostimulants are materials that stimulate plant growth, and gelatin, an animal protein hydrolysate, is classified as one type of biostimulant. Gelatin has a unique amino acid composition with a high percentage of proline and hydroxyproline. In a series of experiments gelatin capsules (#3 hard gelatin) containing 7.1 mg nitrogen each, were placed adjacent to seeds of different crop species, at sowing time in individual growing containers and several growth parameters were measured...
2018: Frontiers in Plant Science
Tao Zhang, Wenbo Yuan, Mengli Li, Ming Miao, Wanmeng Mu
A strain, Alternaria alternata SK37.001, which produces an intracellular α-l-rhamnosidase, was newly isolated from citrus orchard soil. The molecular mass of the enzyme was 66 kDa, as evaluated by SDS-PAGE and 135 kDa, as determined by gel filtration, which indicated that the enzyme is a dimer. The enzyme had a specific activity of 21.7 U mg-1 after step-by-step purification. The optimal pH and temperature were 5.5 and 60 °C, respectively. The enzyme was relatively stable at a pH of 4.0-8.0 and a temperature between 30 and 50 °C compared with other pH levels and temperatures investigated...
December 15, 2018: Food Chemistry
Renate Jonker, Nicolaas E P Deutz, Gerdien C Ligthart-Melis, Anthony J Zachria, Eugene A Veley, Rajesh Harrykissoon, Mariëlle P K J Engelen
BACKGROUND & AIMS: Assessing the ability to respond anabolic to dietary protein intake during illness provides important insight in the capacity of lean body mass maintenance. We applied a newly developed stable tracer approach to assess in one session in patients with chronic obstructive pulmonary disease (COPD) and healthy older adults both the minimal amount of protein intake to obtain protein anabolism (anabolic threshold) and the efficiency of dietary protein to promote protein anabolism (anabolic capacity)...
July 31, 2018: Clinical Nutrition: Official Journal of the European Society of Parenteral and Enteral Nutrition
Sawanan Thongyoo, Tanapati Phakham, Nanthika Khongchareonporn, Onrapak Reamtong, Aphichart Karnchanatat, Songchan Phutong, Thanaporn Wichai, Sajee Noitang, Sarintip Sooksai
The methylotrophic yeasts Pichia pastoris and Hansenula polymorpha have been used for the production of recombinant monomeric insulin precursor (MIP). Recombinant plasmids with one, two and four cassettes of the MIP gene have been successfully constructed in the pPICZαA expression vector to study the effects of gene copy number on MIP production. The MIP protein can be detected by dot-blot analysis from the culture broth of P. pastoris KM71H 24 h after placement in MMH induction medium. The secretion levels of MIP protein in culture broth at 72 h after induction indicated that P...
August 8, 2018: Protein Expression and Purification
Melanija Posavec Marjanovic, Gytis Jankevicius, Ivan Ahel
ADP-ribosylation is the process of transferring the ADP-ribose moiety from NAD+ to a substrate. While a number of proteins represent well described substrates accepting ADP-ribose modification, a recent report demonstrated biological role for DNA ADP-ribosylation as well. The conserved macrodomain fold of several known hydrolyses was found to possess de-ADP-ribosylating activity and the ability to hydrolyze (reverse) ADP-ribosylation. Here we summarize the methods that can be employed to study mono-ADP-ribosylation hydrolysis by macrodomains...
2018: Methods in Molecular Biology
Masato Mashimo, Joel Moss
The ARH family of ADP-ribosyl-acceptor hydrolases is composed of three 39-kDa proteins (ARH1, 2, and 3), which hydrolyze specific ADP-ribosylated substrates. ARH1 hydrolyzes mono(ADP-ribosyl)ated arginine, which results from actions of cholera toxin and other nicotinamide adenine dinucleotide (NAD+ ):arginine ADP-ribosyl-transferases, while ARH3 hydrolyzes poly(ADP-ribose) and O-acetyl-ADP-ribose, resulting from the action of poly(ADP-ribose) polymerases and sirtuins, respectively. ARH2 has not been reported to have enzymatic activity, because of differences in the catalytic domain...
2018: Methods in Molecular Biology
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