Induction and purification of a thermophilic chitinase produced byAeromonas sp. DYU-too7 using glucosamine

Aims: To investigate the distribution of chitinase IO8 in Bacillus cereus strains, the enhancing effects of the chitinase-producing B. cereus strains on biocontrol potential by dual culture assay and in vivo assay against Botrytis cinerea and also the enhancing effects of the chiIO8 on disinfectant properties against seedborne diseases. Moreover, the application of chiIO8 treatment was also observed to improve the germinative energy. Methods and Results: The purification steps included ammonium sulfate precipitation, with columns of DEAE-Sepharose anion-exchange chromatography and Sephacryl S-400 high-resolution gel chromatography. The method gave a 5Á8-fold increase in the specific activity and had a yield of 17%. The molecular weight of the partially purified chitinase chiIO8 was found to be around 30 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal pH and optimal temperature of the partially purified chitinase were pH 6Á5 and 65°C, respectively. The thermostable chitinase still retained the activity after incubation for 100 min at 65°C, and it was increased about 1Á25 times than that of the control (before heating) when the enzyme solution heated at 65°C for 60 min. The partially purified chitinase chiIO8 displays a wide inhibitory spectrum towards all phytopathogenic fungi tested. chiIO8 also exhibited effective disinfectant properties against seed-borne diseases. Conclusion: The present investigation emphasizes the potential of chitinaseproducing micro-organism as promising biocontrol agents of fungal plant pathogens with chitinous cell wall. The novel chitinase chiIO8 proved an efficient, environmentally safe and user-friendly solution. Significance and Impact of the Study: This is the first investigation devoted exclusively to analyse the distribution of chitinase in B. cereus. It infers that the chitinase produced by B. cereus might play a role in the activity of the biopesticide.

show abstract

“…DYU‐Too7 (Lien et al . ), 53 KDa for ASCHI53 and 61 KDa for ASCHI61 form Aeromonas schubertii (Liu et al . ).…”

Section: Discussionmentioning

confidence: 99%

Partial purification and characterization of chiIO8, a novel antifungal chitinase produced by Bacillus cereus IO8

Hammami¹,

Siala

Jridi

et al. 2013

J Appl Microbiol

View full text Add to dashboard Cite

show abstract

“…The stability test [28] of the purified enzyme at different pH levels indicated that it was stable in the pH range of 3.0 to 5.0 for No. 6 chimeric enzyme and 3.0 to 7.0 for No.…”

Section: éE=pí~äáäáíó=~åç=léíáãìã=ée=mentioning

confidence: 97%

Construction and characterization of novel chimeric β-glucosidases with Cellvibrio gilvus (CG) and Thermotoga maritima (TM) by overlapping PCR

Deog

Jin

Hayashi

2009

Biotechnol Bioproc E

View full text Add to dashboard Cite

“…However, a different optimum pH was obtained by the chitinase enzyme isolated from Aeromonas sp. DYU-Too7 [32], Vibrio sp. 98CJ11027 [33] and Paenibacillus sp.…”

Section: Enzyme Purification and Characterizationmentioning

confidence: 99%

Production and Characterization of Chitinases from Thermophilic Bacteria Isolated from Prataan Hot Spring, East Java

Chrisnasari¹,

Yasaputera²,

Christianto³

et al. 2016

j.math.fund.sci.

View full text Add to dashboard Cite

Abstract. Thermophilic bacteria producing chitinase were collected from Prataan hot spring, East Java, Indonesia and screened. The isolated bacterium was analyzed using 16S rRNA gene sequencing analysis and identified as Paenibacillus sp. The molecular identification was confirmed through morphological and physiological analyses. The production of chitinase was conducted at various incubation times, temperatures, pH and concentrations of colloidal chitin. The optimum condition of the isolate to produce the highest chitinase was 0.9% (w/v) of colloidal chitin (pH 7.0) at 48 °C for 24 hours. The obtained chitinases were optimally active at 55 °C and pH 6.0-7.0. The chitinases were gradually purified by ammonium sulfate precipitation, Sephadex G-100 gel filtration, followed by DEAE-cellulose ion exchange chromatography (IEC). The purification method gave a purification factor of 9.43 and a yield of 2.68%. Two protein fractions were obtained from sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) with molecular weights of 68 and 82 kDa.

show abstract

Induction and purification of a thermophilic chitinase produced byAeromonas sp. DYU-too7 using glucosamine

Cited by 29 publications

References 29 publications

Partial purification and characterization of chiIO8, a novel antifungal chitinase produced by Bacillus cereus IO8

Partial purification and characterization of chiIO8, a novel antifungal chitinase produced by Bacillus cereus IO8

Construction and characterization of novel chimeric β-glucosidases with Cellvibrio gilvus (CG) and Thermotoga maritima (TM) by overlapping PCR

Production and Characterization of Chitinases from Thermophilic Bacteria Isolated from Prataan Hot Spring, East Java

Contact Info

Product

Resources

About