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An Approach for In Situ Rapid Detection of Deep-Sea Aromatic Amino Acids Using Laser-Induced Fluorescence.

ABSTRACT: Amino acids are the material basis of almost all life activities. An improved understanding of the source, state, and cycle of amino acids is essential for determining the energy flow and material circulation of marine ecosystems. In the present study, an in situ rapid detection method of ultraviolet (UV; 266 nm) laser-induced fluorescence (LIF) technology was used to detect three natural, aromatic amino acids in the seawater. The laser-induced fluorescence peaks of aromatic amino acids tryptophan, tyrosine, and phenylalanine were located at 350 nm, 300 nm, and 280 nm, respectively. High, linear correlations between the concentrations of the aromatic amino acids and the fluorescence peak heights were observed, and the lowest detectable concentrations of tryptophan, tyrosine, and phenylalanine were 4.70 × 10-9 mol/L, 2.76 × 10-8 mol/L, and 6.05 × 10-7 mol/L, respectively, which allowed us to quantify their concentrations by using laser-induced fluorescence. This paper not only provides a practical method for the detection of aromatic amino acids in seawater, but a new means to further understand the biogeochemical processes of carbon cycles in the deep sea.

SUBMITTER: Du R

PROVIDER: S-EPMC7085558 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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An Approach for In Situ Rapid Detection of Deep-Sea Aromatic Amino Acids Using Laser-Induced Fluorescence.

Du Ranran R Yang Dingtian D Jiang Guangjia G Song Youren Y Yin Xiaoqing X

Sensors (Basel, Switzerland) 20200229 5

Amino acids are the material basis of almost all life activities. An improved understanding of the source, state, and cycle of amino acids is essential for determining the energy flow and material circulation of marine ecosystems. In the present study, an in situ rapid detection method of ultraviolet (UV; 266 nm) laser-induced fluorescence (LIF) technology was used to detect three natural, aromatic amino acids in the seawater. The laser-induced fluorescence peaks of aromatic amino acids tryptoph ...[more]

PMID: 32121409

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Project description:BackgroundAbyssal microorganisms have evolved particular features that enable them to grow in their extreme habitat. Genes belonging to specific functional categories are known to be particularly susceptible to high-pressure; therefore, they should show some evidence of positive selection. To verify this hypothesis we computed the amino acid substitution rates between two deep-sea microorganisms, Photobacterium profundum SS9 and Shewanella benthica KT99, and their respective shallow water relatives.ResultsA statistical analysis of all the orthologs, led to the identification of positive selected (PS) genes, which were then used to evaluate adaptation strategies. We were able to establish "Motility" and "Transport" as two classes significantly enriched with PS genes. The prevalence of transporters led us to analyze variable amino acids (PS sites) by mapping them according to their membrane topology, the results showed a higher frequency of substitutions in the extra-cellular compartment. A similar analysis was performed on soluble proteins, mapping the PS sites on the 3D structure, revealing a prevalence of substitutions on the protein surface. Finally, the presence of some flagellar proteins in the Vibrionaceae PS list confirms the importance of bacterial motility as a SS9 specific adaptation strategy.ConclusionThe approach presented in this paper is suitable for identifying molecular adaptations to particular environmental conditions. The statistical method takes into account differences in the ratio between non-synonymous to synonymous substitutions, thus allowing the detection of the genes that underwent positive selection. We found that positive selection in deep-sea adapted bacteria targets a wide range of functions, for example solute transport, protein translocation, DNA synthesis and motility. From these data clearly emerges an involvement of the transport and metabolism processes in the deep-sea adaptation strategy of both bathytypes considered, whereas the adaptation of other biological processes seems to be specific to either one or the other. An important role is hypothesized for five PS genes belonging to the transport category that had been previously identified as differentially expressed in microarray experiments. Strikingly, structural mapping of PS sites performed independently on membrane and soluble proteins revealed that residues under positive selection tend to occur in specific protein regions.

Project description:Pseudomonas putida is a highly solvent-resistant microorganism and useful chassis for the production of value-added compounds from lignocellulosic residues, in particular aromatic compounds that are made from phenylalanine. The use of these agricultural residues requires a two-step treatment to release the components of the polysaccharides of cellulose and hemicellulose as monomeric sugars, the most abundant monomers being glucose and xylose. Pan-genomic studies have shown that Pseudomonas putida metabolizes glucose through three convergent pathways to yield 6-phosphogluconate and subsequently metabolizes it through the Entner-Doudoroff pathway, but the strains do not degrade xylose. The valorization of both sugars is critical from the point of view of economic viability of the process. For this reason, a P. putida strain was endowed with the ability to metabolize xylose via the xylose isomerase pathway, by incorporating heterologous catabolic genes that convert this C5 sugar into intermediates of the pentose phosphate cycle. In addition, the open reading frame T1E_2822, encoding glucose dehydrogenase, was knocked-out to avoid the production of the dead-end product xylonate. We generated a set of DOT-T1E-derived strains that metabolized glucose and xylose simultaneously in culture medium and that reached high cell density with generation times of around 100 min with glucose and around 300 min with xylose. The strains grew in 2G hydrolysates from diluted acid and steam explosion pretreated corn stover and sugarcane straw. During growth, the strains metabolized > 98% of glucose, > 96% xylose and > 85% acetic acid. In 2G hydrolysates P. putida 5PL, a DOT-T1E derivative strain that carries up to five independent mutations to avoid phenylalanine metabolism, accumulated this amino acid in the medium. We constructed P. putida 5PLΔgcd (xylABE) that produced up to 250 mg l-1 of phenylalanine when grown in 2G pretreated corn stover or sugarcane straw. These results support as a proof of concept the potential of P. putida as a chassis for 2G processes.

Dataset Information

An Approach for In Situ Rapid Detection of Deep-Sea Aromatic Amino Acids Using Laser-Induced Fluorescence.

Publications

An Approach for In Situ Rapid Detection of Deep-Sea Aromatic Amino Acids Using Laser-Induced Fluorescence.

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