Fluorescent microplate assay method for high-throughput detection of lipase transesterification activity

Zheng, Jingwu; Wei, Wei; Lan, Xing; Zhang, Yinjun; Zhao, Wang

doi:10.1016/j.ab.2018.03.010

Cited by 6 publications

(5 citation statements)

References 22 publications

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“…It has been reported that Proteobacteria is involved in the prevention of Rhizoctonia solani in soil ( 23 ). Zheng et al ( 24 ) reported that the abundances of Actinobacteria and Gemmatimonadetes declined when bacterial wilt disease occurred ( 24 ). The 16S rRNA gene of the Gemmatimonadetes was frequently and abundantly detected in various terrestrial environments, which is currently recognized as one of the dominant soil phyla ( 25 ).…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies showed that the abundance of bacteria is influenced by the acidity of the soil ( 9 ). The imbalance of the bacterial community caused by excessively acidic soil is one of the main causes of soil-borne diseases ( 7 , 24 , 28 ). Zhang et al found that the occurrence of bacterial wilt and the abundance of R. solanacearum significantly reduced when regulating soil pH from 5.45 to 6.0; meanwhile, a slightly acidic level could promote the enrichment of beneficial bacteria to suppress bacterial wilt ( 9 ).…”

Section: Discussionmentioning

confidence: 99%

“…Soil enzyme activity was detected by the fluorescence microplate assay method. After hydrolysis of standard substrates (MUB) of different enzymes, 4-methylumbelliferyl or 7-amino-4-methyl coumarin are produced ( 24 ). The activity of soil extracellular enzymes is characterized by measuring the intensity of their fluorescence values.…”

Section: Methodsmentioning

confidence: 99%

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Effects of the invasion of Ralstonia solanacearum on soil microbial community structure in Wuhan, China

Wu,

Ma,

Wang

et al. 2024

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This study investigated the change in the microbiome of tomato rhizosphere soils after the invasion of Ralstonia solanacearum and analyzed the correlation between microbes and soil physicochemical properties. Diversity analyses of the bacteria in healthy and diseased rhizosphere soil samples (HRS and DRS) revealed that HRS had a higher species diversity and were compositionally different from DRS ( P ≤ 0.05). Substantial differences in the relative abundance of Actinobacteria (37.52% vs 28.96%, P ≤ 0.05) and Proteobacteria (29.20% vs 35.59%, P ≤ 0.05) were identified in HRS and DRS, respectively. Taxonomic composition analysis showed ten differentially abundant genera, and seven of them ( Gaiella , Roseisolibacter , Solirubrobacter , Kribbella , Acidibacter , Actinomarinicola , and Marmoricola ) are more abundant in HRS. Soil pH and enzyme activities were negatively correlated with the abundance of R. solanacearum . The contents of total nitrogen (TN), total phosphorus (TP), total potassium (TK), alkaline nitrogen (alkaline N), available phosphorus (AP), available potassium (AK), NO3—N(NN), NH4 + -N (AN), and organic matter (OM) were all significantly increased in DRS. The composition and richness of protozoa in the samples show significant differences. Cephalobus , Acrobeles , Heteromita , norank_Tylenchida, and Rotylenchulus were enriched in DRS. Microbial interaction networks revealed that the HRS networks were more complex than the DRS networks. Overall, the results of this study demonstrate that healthy soil has a more complex microbial community structure and higher enzyme activity, and the invasion of R. solanacearum damages the soil microbial system. IMPORTANCE How does the invasion of Ralstonia solanacearum affect tomato rhizosphere bacteria and protozoa? Which microbial changes can affect the growth of R. solanacearum ? To date, most research studies focus on bacteria, with little research on protozoa, and even less on the synergistic effects between protozoa and bacteria. Here, we analyzed the correlation between tomato rhizosphere bacterial and protozoan communities and soil physicochemical properties during the invasion of R. solanacearum . We found that the diversity and abundance of rhizosphere microorganisms in healthy rhizosphere soil samples (HRS) were significantly higher than those in diseased rhizosphere soil samples (DRS), and there were significant changes in soil pH and enzyme activity. Overall, in this study, the analysis of microbial changes during the invasion of R. solanacearum provides a theoretical basis for the prevention and control of bacterial wilt.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Effects of the invasion of Ralstonia solanacearum on soil microbial community structure in Wuhan, China

Wu,

Ma,

Wang

et al. 2024

mSphere

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“…A fluorometric analytical method placed on microplates was developed by Zheng and coworkers in order to scale lipase transesterification activity [66]. They performed the assay with two substrates: 4-methyl umbelliferone and methanol in tert -butanol.…”

Section: Optical Lipase Biosensors and Bioassaysmentioning

confidence: 99%

Biosensors and Bioassays Based on Lipases, Principles and Applications, a Review

Pohanka

2019

Molecules

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Lipases are enzymes responsible for the conversion of triglycerides and other esterified substrates, they are involved in the basic metabolism of a wide number of organisms, from a simple microorganism and to mammals. They also have broad applicability in many fields from which industrial biotechnology, the production of cleaning agents, and pharmacy are the most important. The use of lipases in analytical chemistry where it can serve as a part of biosensors or bioassays is an application of growing interest and has become another important use. This review is focused on the description of lipases chemistry, their current applications and the methods for their assay measurement. Examples of bioassays and biosensors, including their physical and chemical principles, performance for specific substrates, and discussion of their relevance, are given in this work.

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“…To build on this observation and interrogate DnrK permissivity, herein we describe evaluation of 100 structurally and functionally diverse natural products and natural product mimetics of wide-ranging architectural complexity as potential DnrK substrates. Included in the demonstrated DnrK substrates is 4-methylumbelliferone (4-MeUmb, 75 , a fluorescent hydroxycoumarin drug frequently used as a real-time fluorescent indicator in enzyme assays) , and a range of complex natural products including anthracyclines, angucyclines, anthraquinone-fused enediynes, flavonoids, pyranonaphthoquinones, and polyketides. This study also provides previously undetermined DnrK kinetic parameters and a comparative evaluation of the impact of anthracycline 4- O -methylation on cancer cell line cytotoxicity, 4E-BP1 phosphorylation, and axolotl embryo tail regeneration.…”

mentioning

confidence: 99%

Biochemical and Structural Studies of the Carminomycin 4-O-Methyltransferase DnrK

Jalali,

Wang,

Overbay

et al. 2024

J. Nat. Prod.

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Structural and functional studies of the carminomycin 4-O-methyltransferase DnrK are described, with an emphasis on interrogating the acceptor substrate scope of DnrK. Specifically, the evaluation of 100 structurally and functionally diverse natural products and natural product mimetics revealed an array of pharmacophores as productive DnrK substrates. Representative newly identified DnrK substrates from this study included anthracyclines, angucyclines, anthraquinone-fused enediynes, flavonoids, pyranonaphthoquinones, and polyketides. The ligand-bound structure of DnrK bound to a nonnative fluorescent hydroxycoumarin acceptor, 4-methylumbelliferone, along with corresponding DnrK kinetic parameters for 4-methylumbelliferone and native acceptor carminomycin are also reported for the first time. The demonstrated unique permissivity of DnrK highlights the potential for DnrK as a new tool in future biocatalytic and/or strain engineering applications. In addition, the comparative bioactivity assessment (cancer cell line cytotoxicity, 4E-BP1 phosphorylation, and axolotl embryo tail regeneration) of a select set of DnrK substrates/products highlights the ability of anthracycline 4-O-methylation to dictate diverse functional outcomes.

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Fluorescent microplate assay method for high-throughput detection of lipase transesterification activity

Cited by 6 publications

References 22 publications

Effects of the invasion of Ralstonia solanacearum on soil microbial community structure in Wuhan, China

Effects of the invasion of Ralstonia solanacearum on soil microbial community structure in Wuhan, China

Biosensors and Bioassays Based on Lipases, Principles and Applications, a Review

Biochemical and Structural Studies of the Carminomycin 4-O-Methyltransferase DnrK

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