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Wei Zhou, Rui Huang, Zhiguang Zhu, Yi-Heng P Job Zhang
Thermostability and specific activity of enzymes are two of the most important properties for industrial biocatalysts. Here we developed a Petri dish-based double-layer high-throughput screening (HTS) strategy for rapid identification of desired mutants of polyphosphate glucokinase (PPGK) from a thermophilic actinobacterium Thermobifida fusca YX with both enhanced thermostability and activity. Escherichia coli colonies representing a PPGK mutant library were grown on the first-layer Phytagel™-based plates, which can remain solid for 1 h even at heat treatment temperatures of more than 100 °C...
June 8, 2018: Applied and Environmental Microbiology
Maryam Rezaei Shadegan, Mahdi Banaee
Insecticides and fertilizers are the most common agrochemicals that enter aquatic ecosystems from farmlands via different ways and consequently affect fish. In this study, effects of dimethoate, and Bacilar, as a bio-fertilizer, on oxidative stress and biochemical parameters were measured in common carp. For this purpose, fish were exposed to 16 and 32 μg L-1 dimethoate, 0.1 and 0.2 ml L-1 Bacilar and the mixtures of both compounds for 14 days. The results showed that Bacilar increased aspartate aminotransferase (AST) and LDH in gills, AST in liver and the activity of catalase (CAT) (0...
May 29, 2018: Chemosphere
Rodrigo de Cássio da Silva, Sonia Regina Grötzner, Daniele Dietrich Moura Costa, Juan Ramón Esquivel Garcia, Juan Muelbert, Valéria Freitas de Magalhães, Francisco Filipak Neto, Ciro Alberto de Oliveira Ribeiro
Many tropical freshwater ecosystems are impacted by cyanobacteria blooms increasing the risk of cyanotoxins exposure to aquatic organisms while human populations may be exposed by eating fish, drinking water, or dermal swimming. However, few toxicological data are available on the influence of cyanobacteria blooms in particular, cylindrospermopsin (CYN) on Brazilian neotropical fish. A number of studies demonstrated the ability of CYN to bioaccumulate in freshwater organisms and consequently enter the human food chain...
May 15, 2018: Journal of Toxicology and Environmental Health. Part A
Shiwei Wu, Lujun Yu, Xiaozhe Fu, Xi Yan, Qiang Lin, Lihui Liu, Hongru Liang, Ningqiu Li
Infectious spleen and kidney necrosis virus (ISKNV) has caused significant losses in the cultured mandarin fish (Siniperca chuatsi) industry. The molecular mechanisms that underlie interaction between ISKNV and hosts are not fully understood. In this study, the proteomic profile of CPB cells at progressive time points after ISKNV infection was analyzed by isobaric tags for relative and absolute quantitation (iTRAQ). A total of 2731 proteins corresponding to 6363 novel peptides (false discovery rate <0.01) were identified...
May 4, 2018: Fish & Shellfish Immunology
Shuang Zhao, Yang Liu, Han Li, Yi Lan Chen, Bao Guo Li, Guo Hui Qi, Xue Mei Zhang
Two precocious walnut varieties, Lvling and Lvzao were used as materials in this study, the kernel fat contents and 3 related enzyme activities in different development periods after flowe-ring 50 days were analyzed. The key enzymes affecting walnut fat in different periods were illumina-ted. The results showed that the kernel fat accumulation trends of the two varieties were basically the same. The kernel began to solidify 50 days after flowering, and the increase in kernel fat content was rapid 60-90 days after flowering, slowed down 90-120 days after flowering, and stopped 120-130 days after flowering...
November 2017: Ying Yong Sheng Tai Xue Bao, the Journal of Applied Ecology
Junjeong Choi, Eun-Sol Kim, Ja Seung Koo
Purpose: The purpose of this study was to assess the expression of pentose phosphate pathway- (PPP-) related proteins and their significance in clinicopathologic factors of breast cancer. Methods: Immunohistochemical staining for PPP-related proteins (glucose-6-phosphate dehydrogenase [G6PDH], 6-phosphogluconolactonase [6PGL], 6-phosphogluconate dehydrogenase [6PGDH], and nuclear factor-erythroid 2-related factor 2 [NRF2]) was performed using tissue microarray (TMA) of 348 breast cancers...
2018: Disease Markers
Belisa Parmeggiani, Mateus Grings, Nevton Teixeira da Rosa-Junior, Renata Britto, Moacir Wajner, Guilhian Leipnitz
Non-ketotic hyperglycinemia (NKH) is a severe neurological disorder caused by defects in glycine (GLY) catabolism and characterized by a high cerebrospinal fluid/plasma GLY ratio. Treatment is often ineffective and limited to the control of symptoms and detoxification of GLY. In the present work, we investigated the in vivo effects of GLY intracerebroventricular administration on oxidative stress parameters in rat striatum, cerebral cortex, and hippocampus. In vitro effects of GLY were also evaluated in striatum...
April 19, 2018: Molecular Neurobiology
Joe T Sharick, Peter F Favreau, Amani A Gillette, Sophia M Sdao, Matthew J Merrins, Melissa C Skala
While NAD(P)H fluorescence lifetime imaging (FLIM) can detect changes in flux through the TCA cycle and electron transport chain (ETC), it remains unclear whether NAD(P)H FLIM is sensitive to other potential fates of glucose. Glucose carbon can be diverted from mitochondria by the pentose phosphate pathway (via glucose 6-phosphate dehydrogenase, G6PDH), lactate production (via lactate dehydrogenase, LDH), and rejection of carbon from the TCA cycle (via pyruvate dehydrogenase kinase, PDK), all of which can be upregulated in cancer cells...
April 3, 2018: Scientific Reports
Alice Jara De Porcellinis, Hanne Nørgaard, Laura Maria Furelos Brey, Simon Matthé Erstad, Patrik R Jones, Joshua L Heazlewood, Yumiko Sakuragi
Cyanobacteria fix atmospheric CO2 to biomass and through metabolic engineering can also act as photosynthetic factories for sustainable productions of fuels and chemicals. The Calvin Benson cycle is the primary pathway for CO2 fixation in cyanobacteria, algae and C3 plants. Previous studies have overexpressed the Calvin Benson cycle enzymes, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and bifunctional sedoheptulose-1,7-bisphosphatase/fructose-1,6-bisphosphatase (hereafter BiBPase), in both plants and algae, although their impacts on cyanobacteria have not yet been rigorously studied...
March 3, 2018: Metabolic Engineering
Rui Huang, Hui Chen, Wei Zhou, Chunling Ma, Y-H Percival Zhang
Glucose 6-phosphate dehydrogenase (G6PDH) is one of the most important dehydrogenases responsible for generating reduced NADPH for anabolism and is also the rate-limiting enzyme in the Entner-Doudoroff pathway. For in vitro biocatalysis, G6PDH must possess both high activity and good thermostability due to requirements of efficient use and low expense of biocatalyst. Here, we used directed evolution to improve thermostability of the highly active G6PDH from Zymomonas mobilis. Four generations of random mutagenesis and Petri-dish-based double-layer screening evolved the thermolabile wild-type enzyme to the thermostable mutant Mut 4-1, which showed a more than 124-fold increase in half-life time (t1/2 ) at 60 °C, a 3...
April 2018: Applied Microbiology and Biotechnology
Fernanda Dias de Moraes, Francine Perri Venturini, Priscila Adriana Rossi, Ive Marchioni Avilez, Naiara Elisabete da Silva de Souza, Gilberto Moraes
The effects of cypermethrin-based insecticide (CBI), commonly used in aquaculture and agriculture, were evaluated in matrinxa (Brycon amazonicus) exposed to sub-lethal concentration (20% of LC50) for 96 h. Physiological and biochemical effects were studied through biomarkers: lipid peroxidation (LPO), glutathione (GSH), and ascorbic acid concentrations; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glucose-6-phosphate dehydrogenase (G6PDH) assays in the liver and gills. Besides, ions Na+ , Cl- , and K+ ; protein and glucose concentrations were measured in the plasma...
March 2018: Ecotoxicology
Bamisaye Fisayo Abraham, Sulyman Abdulhakeem Olarewaju, Abegunde Ronke, Ajani Emmanuel Oladipo
Background: Diabetes mellitus is considered one of the 5 principal causes of death in the world and is recognized as a global public health issue because of its multifactorial facets affecting essential biochemical processes in the body. This study investigated the antidiabetic and antidyslipidemic activities of the aqueous extract of Cochlospermum planchonii (C. planchonii) leaves in streptozotocin (STZ)-induced diabetic rats. Methods: Thirty adult female rats ( Rattus norvegicus ) weighing 153±3...
November 2017: Iranian Journal of Medical Sciences
Yuhe R Yang, Jinglin Fu, Shaun Wootten, Xiaodong Qi, Minghui Liu, Hao Yan, Yan Liu
In living cells, compartmentalized or membrane-associated enzymes are often assembled into large networks to cooperatively catalyze cascade reaction pathways essential for cellular metabolism. Here, we report the assembly of an artificial 2D enzyme network of two cascade enzymes-glucose-6-phosphate dehydrogenase (G6PDH) and lactate dehydrogenase (LDH)-on a wireframe DNA origami template. Swinging arms were used to facilitate the transport of the redox intermediate of NAD+ /NADH between enzyme pairs on the array...
February 2, 2018: Chembiochem: a European Journal of Chemical Biology
Tao Huang, Hailing Tan, Fangju Lu, Gong Chen, Zhenqiang Wu
BACKGROUND: Monascus pigments are widely used in the food and pharmaceutical industries due to their safety to human health. Our previous study found that glucose concentration induced extracellular oxidoreduction potential (ORP) changes could influence extracellular water-soluble yellow pigment production by Monascus ruber CGMCC 10910 in submerged fermentation. In this study, H2 O2 and dithiothreitol (DTT) were used to change the oxidoreduction potential for investigating the effects of oxidative or reductive substances on Monascus yellow pigment production by Monascus ruber CGMCC 10910...
November 21, 2017: Microbial Cell Factories
Alaa El-Din H Sayed, Hamdy A M Soliman
The antioxidant role of the green tea (Camellia sinensis) extract (GTE) was examined to remedy the toxic effects of (0.2mgl-1 ) 4-nonylphenol(4-NP). Biochemical parameters, antioxidant enzymes, liver lipid peroxidation (LPO), DNA fragmentation, and apoptosis as well as histopathology of liver of African catfish Clarias gariepinus were considered. Catfishes were divided into four groups: first group (control), second group (0.2mgl-1 of 4-NP), third group (0.2mgl-1 of 4-NP +100mg GTE l-1 water), and fourth group (0...
March 2018: Ecotoxicology and Environmental Safety
Maria Rosa Dmengeon Pedreiro de Souza, Tatiana Herrerias, Tania Zaleski, Mariana Forgati, Priscila Krebsbach Kandalski, Cintia Machado, Dilza Trevisan Silva, Cláudio Adriano Piechnik, Maurício Osvaldo Moura, Lucélia Donatti
Carbohydrate metabolism and the antioxidant defence system of heart and muscle of the Antarctic notothenioids Notothenia rossii and Notothenia coriiceps were evaluated in response to heat stress (8 °C) over 144 h. N. rossii heart exhibited decreased glycolysis and aerobic metabolism after up to 12 h of exposure to 8 °C, and anaerobiosis was inhibited within 24 h. However, these pathways were stimulated after 72 h at 8 °C. The consumption of glucose-6-phosphate, derived from hexokinase (HK), by glucose-6-phosphate dehydrogenase (G6PDH) decreased in N...
March 2018: Biochimie
Huan Lu, Huili Zhu
The aim of the present study was to investigate the effects of transketolase (TKT) on cell proliferation, cell migration and interaction with other metabolism-associated genes in A549 lung cancer cells. A549 cells were transfected with three TKT -specific small interfering (si)RNAs, screened for the optimal transfection concentration, and sequenced with flow cytometry and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell viability was evaluated using Cell Counting Kit-8 (CCK-8), cell cycle was assessed by flow cytometric analysis...
November 2017: Oncology Letters
Kishu Kitayama, Masakazu Yashiro, Tamami Morisaki, Yuichiro Miki, Tomohisa Okuno, Haruhito Kinoshita, Tatsunari Fukuoka, Hiroaki Kasashima, Go Masuda, Tsuyoshi Hasegawa, Katsunobu Sakurai, Naoshi Kubo, Kosei Hirakawa, Masaichi Ohira
The aim of this study was to analyze the significance of glucose metabolism-related enzymes in the proliferation of gastric cancer under hypoxia. Four hypoxia-resistant gastric cancer cell lines and four parent cell lines were used. Reverse transcription-PCR was used to evaluate the mRNA expression levels of the following metabolism-related enzymes: pyruvate kinase isozyme M2 (PKM2), glutaminase (GLS), enolase 1 (ENO1), glucose-6-phosphate dehydrogenase (G6PDH), and PKM1. The effects of these enzymes on the proliferation of gastric cancer cells were examined using siRNAs, shikonin as a PKM2 inhibitor, or BPTES as a GLS inhibitor, in vitro and in vivo...
December 2017: Cancer Science
Kevin E Acero-Navarro, Mariella Jiménez-Ramírez, Miguel A Villalobos, Rocío Vargas-Martínez, Hugo V Perales-Vela, Roberto Velasco-García
Glucose-6-phosphate dehydrogenase (G6PDH) (EC plays an important role in the human pathogen Pseudomonas aeruginosa because it generates NADPH, an essential cofactor for several biosynthetic pathways and antioxidant enzymes. P. aeruginosa G6PDH is also a key enzyme in the metabolism of various carbon sources, such as glucose, glycerol, fructose, and mannitol. Understanding the kinetic characteristics and mechanisms that control the activity of this enzyme is crucial for future studies in this context...
February 2018: Protein Expression and Purification
Gong Chen, Qi Bei, Tao Huang, Zhenqiang Wu
BACKGROUND: Monascus pigments are promising sources for food and medicine due to their natural food-coloring functions and pharmaceutical values. The innovative technology of extractive fermentation is used to promote pigment productivity, but reports of pigment trans-membrane secretion mechanism are rare. In this study, tracking of pigment accumulation and secretion in extractive fermentation of Monascus anka GIM 3.592 was investigated. RESULTS: The increased vacuole size in mycelia correlated with fluorescence intensity (r > 0...
October 4, 2017: Microbial Cell Factories
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