Laser Tweezers Raman Spectroscopy (LTRS) to Detect Effects of Chlorine Dioxide on IndividualNosema bombycisSpores

2019 ◽  
Vol 73 (7) ◽  
pp. 774-780
Author(s):  
Yu Zhang ◽  
Zhenbin Miao ◽  
Xuhua Huang ◽  
Xiaochun Wang ◽  
Junxian Liu ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Real Time ◽  
Chlorine Dioxide ◽  
Single Cell Analysis ◽  
Protein Structures ◽  
Protein Secondary Structure ◽  
Spore Wall ◽  
Time Range ◽  
Laser Tweezers ◽  
Α Helix

The microsporidium Nosema bombycis (Nb) causes pebrine, a fatal disease in sericulture. Nb is effectively killed by chlorine dioxide (ClO2); however, the precise killing mechanism remains unclear. We used laser tweezers Raman spectroscopy (LTRS) to monitor the action of ClO2on individual Nb spores in real time. Raman peaks of ClO2appeared in Nb spores, corresponding to decreased peaks of trehalose that gradually disappeared. A peak (1658 cm–1) corresponding to the protein α-helix significantly weakened while that (1668 cm–1) corresponding to irregular protein structures was enhanced; their intensities were negatively correlated in a certain time range and dependent on ClO2concentration. The intensities of peaks at 782 cm–1(nucleic acids) and 1004 cm–1(phenylalanine of protein) did not change evidently even under extremely high ClO2concentrations. Thus, ClO2rapidly permeates the Nb spore wall, changing the protein secondary structure to lose biological function and destroy permeability, causing trehalose to leak out. These effects are ClO2concentration-dependent, but no other obvious changes to biomacromolecules were detected. Single-cell analysis using LTRS is an effective method to monitor the action of chemical sporicides on microbes in real time, providing insight into the heterogeneity of cell stress resistance.

scholarly journals Structural and Technological Approach to Reveal the Role of the Lipid Phase in the Formation of Soy Emulsion Gels with Chia Oil

Gels ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 48
Author(s):  
Ana M. Herrero ◽  
Claudia Ruiz-Capillas
Keyword(s):  
Raman Spectroscopy ◽  
Structural Properties ◽  
Structural Changes ◽  
Protein Secondary Structure ◽  
Lipid Phase ◽  
Α Helix ◽  
Β Sheet ◽  
Shed Light ◽  
Chia Oil

Considerable attention has been paid to emulsion gels (EGs) in recent years due to their interesting applications in food. The aim of this work is to shed light on the role played by chia oil in the technological and structural properties of EGs made from soy protein isolates (SPI) and alginate. Two systems were studied: oil-free SPI gels (SPI/G) and the corresponding SPI EGs (SPI/EG) that contain chia oil. The proximate composition, technological properties (syneresis, pH, color and texture) and structural properties using Raman spectroscopy were determined for SPI/G and SPI/EG. No noticeable (p > 0.05) syneresis was observed in either sample. The pH values were similar (p > 0.05) for SPI/G and SPI/EG, but their texture and color differed significantly depending on the presence of chia oil. SPI/EG featured significantly lower redness and more lightness and yellowness and exhibited greater puncture and gel strengths than SPI/G. Raman spectroscopy revealed significant changes in the protein secondary structure, i.e., higher (p < 0.05) α-helix and lower (p < 0.05) β-sheet, turn and unordered structures, after the incorporation of chia oil to form the corresponding SPI/EG. Apparently, there is a correlation between these structural changes and the textural modifications observed.

scholarly journals Disinfection chemicals mode of action on the bacterial spore structure and their Raman spectra

2020 ◽  
Author(s):  
Dmitry Malyshev ◽  
Tobias Dahlberg ◽  
Krister Wiklund ◽  
Per Ola Andersson ◽  
Sara Henriksson ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Raman Spectra ◽  
Sodium Hypochlorite ◽  
Chlorine Dioxide ◽  
Peracetic Acid ◽  
Structural Changes ◽  
Detection Methods ◽  
Laser Tweezers ◽  
Rapid Diagnostics ◽  
Spore Forming Bacteria

AbstractContamination of toxic spore-forming bacteria is problematic since spores can survive a plethora of disinfection chemicals. It is also problematic to rapidly detect if the disinfection chemical was active, leaving spores dead. Robust decontamination strategies, as well as reliable detection methods to identify dead from viable spores, are thus critical. Vibrational detection methods such as Raman spectroscopy has been suggested for rapid diagnostics and differentiation of live and dead spores. We investigate in this work, using laser tweezers Raman spectroscopy, the changes in Raman spectra of Bacillus thuringiensis spores treated with sporicidal agents such as chlorine dioxide, peracetic acid, and sodium hypochlorite. We also imaged treated spores using SEM and TEM to verify if any changes to the spore structure can be correlated to the Raman spectra. We found that chlorine dioxide did not change the Raman spectrum or the spore structure; peracetic acid shows a time-dependent decrease in the characteristic DNA/DPA peaks and ∼20 % of the spores were degraded and collapsed; spores treated with sodium hypochlorite show an abrupt drop in DNA and DPA peaks within 20 minutes all though the spore structure was overall intact, however, the exosporium layer was reduced. Structural changes appeared over several minutes, compared to the inactivation time of the spores, which is less than a minute. We conclude that vibrational spectroscopy provides powerful means to detect changes in spores but it might be problematic to identify if spores are live or dead after a decontamination procedure.

Real-Time Detection on the Expression of Soluble Protein and Inclusion Body in the Recombinant Escherichia coli with Laser Tweezers Raman Spectroscopy

2014 ◽  
Vol 41 (12) ◽  
pp. 1215003
Author(s):  
黄庶识 Huang Shushi ◽  
卢明倩 Lu Mingqian ◽  
李冰 Li Bing ◽  
周冰 Zhou Bing ◽  
陈丽梅 Chen Limei
Keyword(s):  
Escherichia Coli ◽  
Raman Spectroscopy ◽  
Real Time ◽  
Inclusion Body ◽  
Soluble Protein ◽  
Recombinant Escherichia Coli ◽  
Laser Tweezers ◽  
Real Time Detection

scholarly journals Genome-Wide Identification of Cellulose-Like Synthase D Gene Family in Dendrobium Catenatum

2021 ◽  
Author(s):  
Hangxian Xi ◽  
Qing Li ◽  
Xueliang Chen ◽  
Chen Liu ◽  
Yuxue Zhao ◽  
...  
Keyword(s):  
Protein Structures ◽  
Protein Secondary Structure ◽  
Regulatory Elements ◽  
Cellulose Synthase ◽  
Random Coil ◽  
Promoter Regions ◽  
Genome Wide ◽  
Recovery Treatment ◽  
Stems And Leaves ◽  
Α Helix

Abstract Background: Dendrobium catenatum, which grows on the semi humid rocks in the mountains, has been at the top of the "Nine Immortals of China" since ancient times. It is a kind of yin tonic medicine and its main active component is polysaccharide. Cellulose synthase-like D(CslD) genes were predicted to catalyse the biosynthesis of 1,4-β-d-glycan backbone of hemicelluloses, which plays fundamental roles in plant development. Results: To investigate the role of CslD in the development of Dendrobium catenatum, eight CslD genes (DcCslD1,2a,2b,3a,3b,4a,4b,5) were identified. The results of protein prediction and analysis showed that CslD2a/2b/4a/4b proteins were acidic proteins, the rest were basic proteins; Leu, Ser, Ala, Gly, Arg, Pro and Asp were the main amino acids. All the proteins had obvious hydrophobic or hydrophilic regions, and had transmembrane structure. The main elements of protein secondary structure were α-helix, random coil and extended chain. Phylogenetic analysis revealed that the DcCslD family could be divided into Ⅰ, Ⅱ, Ⅲ and Ⅳ groups. DcCslD proteins had typical Cellulose synthase domain and similar protein structures to the CslDs of other plants. Their promoter regions contain cis regulatory elements related to stress and hormone. The results of qRT-PCR showed that the identified DcCslDs were differentially expressed in roots, stems and leaves. Most of them hightly expressed in stems and leaves. What’s more, the environmental stress1es examination showed that the expressions of DcCslD5 were closely associated with drought-recovery treatment, the expression of DcCslD1, DcCslD2a, DcCslD2b, DcCslD3a, and DcCslD5 were significantly influenced by low temperature. Conclusions: This study systematically analyzed the sequence characteristics of CslD protein of D. catenatum, which can provide reference for further study on the function of CslD protein in polysaccharide metabolism of D. catenatum.

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