Simple Methods for the Preparation of Colloidal Chitin, Cell Free Supernatant and Estimation of Laminarinase

BIO-PROTOCOL ◽  
2021 ◽  
Vol 11 (19) ◽  
Author(s):  
Ananthamurthy Koteshwara
Keyword(s):  
Colloidal Chitin ◽  
Cell Free Supernatant

Characteristics and Function of the Chitin Binding Protein from Xenorhabdus nematophila

2019 ◽  
Vol 26 (6) ◽  
pp. 414-422
Author(s):  
Jia Liu ◽  
Ping Song ◽  
Jie Zhang ◽  
Ziyan Nangong ◽  
Xiaobei Liu ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Growth Inhibition ◽  
Insecticidal Activity ◽  
Spore Germination ◽  
Binding Protein ◽  
Colloidal Chitin ◽  
Inhibition Rate ◽  
Xenorhabdus Nematophila ◽  
Chitin Binding Protein ◽  
And Function

Background: Genome sequence analysis (GenBank access No.: FN667742.1) shows that Xenorhabdus nematophila ATCC19061 contains one gene (Xn-cbp) encoding chitin binding protein (Xn-CBP). Objective: The present work aims to clarify the characteristics and function of Xn-CBP from X. nematophila HB310. Methods: In this study, the Xn-cbp gene was cloned and expressed in Escherichia coli BL21 (DE3). Substrate binding assays were performed to explain the ability of Xn-CBP combined with the polysaccharide. The insecticidal toxicity of Xn-CBP against the second-instar larvae of Helicoverpa armigera was determined by feeding method. Besides, the antifungal activity of Xn-CBP against Coniothyrium diplodiella, Verticillium dahlia, and Fusarium oxysporum was tested by spore germination assay and hyphal extension assay. Results: Xn-CBP encoded 199 amino acids with a calculated mass of 28 kDa, which contained a signal peptide and a chitin binding domain. The Bmax and Kd values of Xn-CBP to colloidal chitin were 2.46 and 4.08, respectively. Xn-CBP had insecticidal activity against the H. armigera with a growth inhibition rate of 84.08%. Xn-CBP had the highest spore germination inhibitory effect on C. diplodiella with the inhibition rate of 83.11%. The hyphal growth inhibition rate of Xn-CBP to F. oxysporum, 41.52%, was higher than the other two fungi. Conclusion: The Xn-CBP had the highest binding ability to colloidal chitin and it showed insecticidal activity and antifungal activity. The present study laid a foundation for further exploitation and utilization of X. nematophila.

scholarly journals PRODUCTION AND CHARACTERIZATION OF CHITINASE ENZYMES FROM SULILI HOT SPRING IN SOUTH SULAWESI, Bacillus sp. HSA,3-1a

2010 ◽  
Vol 10 (2) ◽  
pp. 256-260 ◽  
Author(s):  
Hasnah Natsir ◽  
Abd. Rauf Patong ◽  
Maggy Thenawidjaja Suhartono ◽  
Ahyar Ahmad
Keyword(s):  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Extracellular Enzyme ◽  
Colloidal Chitin ◽  
Bacillus Sp ◽  
Optimum Temperature ◽  
South Sulawesi ◽  
Sds Page ◽  
Physiological Analysis

Chitinase is an extracellular enzyme which is capable in hydrolyzing insoluble chitin to its oligomeric and monomeric components. The enzyme produced by thermophilic bacteria was screened and isolated from Sulili hot spring in Pinrang, South Sulawesi, Indonesia. The gram positive spore forming rod shape bacteria was identified as Bacillus sp. HSA,3-1a through morphological and physiological analysis. The production of chitinase enzyme was conducted at various concentration of colloidal chitin at a pH of 7.0 and a temperature of 55 °C. The bacteria optimally was produced the enzyme at a colloidal chitin concentration of 0.5% after 72 h of incubation. The optimum temperature, pH and substrate concentration of chitinase were 60 °C, 7.0 and 0.3%, respectively. The enzyme was stable at a pH of 7.0 and a temperature of 60 °C after 2 h of incubation. The chitinase activities was increased by addition of 1 mM Mg2+, Ca2+ and Mn2+ ions, whereas the activities were  decreased by 1 mM Co2+, Fe2+ and Zn2 ions. The molecular weight of the crude enzyme was determined using SDS-PAGE analysis. Five protein fractions were obtained from SDS-PAGE, with MWs of 79, 71, 48, 43 and 22 kDa.   Keywords: colloidal chitin, thermophilic bacteria, chitinase

Effectiveness of Lactobacilli cell-free supernatant and propolis extract microcapsules on oxidation and microbiological growth in sardine burger

2021 ◽  
pp. 101417
Author(s):  
Esmeray KULEY ◽  
Hatice YAZGAN ◽  
Yesim OZOGUL ◽  
Yilmaz UCAR ◽  
Mustafa DURMUS ◽  
...  
Keyword(s):  
Propolis Extract ◽  
Cell Free Supernatant

scholarly journals Optimization of Agitation Rate in Bioreactor Increases Chitinase Activity of Serratia marcescens PT6

2020 ◽  
Vol 147 ◽  
pp. 03019
Author(s):  
Amara Faiz Wriahusna ◽  
Niswah Umhudloh Dzakiyya ◽  
Indun Dewi Puspita ◽  
Sri Pudjiraharti
Keyword(s):  
Serratia Marcescens ◽  
Bacterial Growth ◽  
Agar Medium ◽  
Chitinase Activity ◽  
Colloidal Chitin ◽  
Gram Negative Bacteria ◽  
Agitation Rate ◽  
Anova Analysis ◽  
Gram Negative ◽  
Shrimp Pond

Serratia marcescens PT6 is a Gram-negative bacteria isolated from shrimp pond sediment that capable of producing chitinase. This study aimed to observe the effect of agitation rate on growth and chitinase activity of S. marcescens PT-6 in a bioreactor. The production of chitinase was done in 1.5 l bioreactor using colloidal chitin broth at the condition of pH 7, the temperature of 30°C, aeration of 0.04 vvm, and variation of agitation rate (200, 350, 500 rpm). Bacterial growth was measured by colonies counting in agar medium, while chitinase activity was measured by means of colorimetric every day for four days incubation. The results of ANOVA analysis show that the agitation rate had no effect on bacterial growth, but a significant effect (P<0.05) was observed on chitinase activity. The highest growth and chitinase activity were obtained at 200 rpm, with the highest chitinase activity of 0.006 ± 0.001 U/ml was at day-2. This study implies that the optimized agitation rate in the bioreactor increased the chitinase activity produced by S. marcescens PT-6.

scholarly journals Potential of Cell-Free Supernatant from Lactobacillus plantarum NIBR97, Including Novel Bacteriocins, as a Natural Alternative to Chemical Disinfectants

2020 ◽  
Vol 13 (10) ◽  
pp. 266
Author(s):  
Sam Woong Kim ◽  
Song I. Kang ◽  
Da Hye Shin ◽  
Se Yun Oh ◽  
Chae Won Lee ◽  
...  
Keyword(s):  
Lactobacillus Plantarum ◽  
Influenza A ◽  
Antibacterial Activities ◽  
Proteinase K ◽  
Bacterial Membranes ◽  
Cell Free Supernatant ◽  
Antiviral Activities ◽  
Viral Envelopes ◽  
Proteinase K Treatment ◽  
Cellular Lysis

The recent pandemic of coronavirus disease 2019 (COVID-19) has increased demand for chemical disinfectants, which can be potentially hazardous to users. Here, we suggest that the cell-free supernatant from Lactobacillus plantarum NIBR97, including novel bacteriocins, has potential as a natural alternative to chemical disinfectants. It exhibits significant antibacterial activities against a broad range of pathogens, and was observed by scanning electron microscopy (SEM) to cause cellular lysis through pore formation in bacterial membranes, implying that its antibacterial activity may be mediated by peptides or proteins and supported by proteinase K treatment. It also showed significant antiviral activities against HIV-based lentivirus and influenza A/H3N2, causing lentiviral lysis through envelope collapse. Furthermore, whole-genome sequencing revealed that NIBR97 has diverse antimicrobial peptides, and among them are five novel bacteriocins, designated as plantaricin 1 to 5. Plantaricin 3 and 5 in particular showed both antibacterial and antiviral activities. SEM revealed that plantaricin 3 causes direct damage to both bacterial membranes and viral envelopes, while plantaricin 5 damaged only bacterial membranes, implying different antiviral mechanisms. Our data suggest that the cell-free supernatant from L. plantarum NIBR97, including novel bacteriocins, is potentially useful as a natural alternative to chemical disinfectants.

scholarly journals Induction of Apoptosis in Macrophage Cell Line, J774, by the Cell-Free Supernatant fromPseudomonas aeruginosa

2003 ◽  
Vol 47 (3) ◽  
pp. 199-206 ◽  
Author(s):  
Jianling Zhang ◽  
Hisao Takayama ◽  
Takashi Matsuba ◽  
Ru Jiang ◽  
Yoshinori Tanaka
Keyword(s):  
Cell Line ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Cell Free Supernatant ◽  
Induction Of Apoptosis
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