Overexpression of Chitinase A Gene from Serratia marcescens in Bacillus subtilis and Characterization of Enhanced Chitinolytic Activity

Abstract

Chitinase enzymes possess various usages in agriculture, biotechnology and medicine due to their chitin degrading property. Thus, efficient production of chitinase enzymes with desired properties has importance for its use. In this study, chitinase A (chiA) gene from Serratia marcescens Bn10 was cloned and heterologously overexpressed using pHT43 vector in Bacillus subtilis 168. The recombinant chitinase was characterized in terms of temperature, pH, and various effectors. The extracellular chitinase activity in recombinant B. subtilis was found 2.15-fold higher than the parental strain after 2 h of IPTG induction. Optimum temperature and pH for the extracellular chitinase activity in the recombinant B. subtilis were determined as 60 o C and pH 9.0, respectively. NaCl, Ca 2+ , Mn 2+ , Cu 2+ , Zn 2+ , sodium dodecyl sulfate (SDS), Tween-20, and ethanol increased the chitinase activity whereas Mg 2+ caused an inhibition. The most notable increment on the chitinase activity was provided by Zn 2+ (3.2 folds) and then by SDS (2.9 folds). The chitinase, overproduced by the recombinant B. subtilis 168 heterologously expressing chiA, was determined to have optimum activity at high temperature and alkaline conditions as well as various effectors increase its activity. The extracellular chitinase of recombinant B. subtilis might be a promising source for agricultural, biotechnological and medical applications.

Unveiling Novel Biocontrol Strategies: Serratia marcescens chiA Gene Against Myzus persicae

“…In a similar study, Okay and Alshehri (2020) found that the optimum pH for the chiA protein they isolated from S. marcescens Bn10 was 9 (alkaline). 44 marcescens and found the optimum pH to be 8. 46 Consequently, Babashpour et al…”

“…In a similar study, Okay and Alshehri (2020) found that the optimum pH for the chiA protein they isolated from S. marcescens Bn10 was 9 (alkaline). 44 Likewise, Aggarwal et al (2016) reported the optimum pH of the chitinase protein they isolated from S. marcescens as 9. 45 Similarly, Danışmanzoğlu et al (2015) isolated chiA from S. marcescens and found the optimum pH to be 8.…”

Bioformulados para mantener la viabilidad de rizobacterias y su aplicación en Theobroma cacao L. CCN-51

“…Se determinó la presencia del gen chiA en las cepas (PM3-8, PM3-14, BM2-12, CHA0) con el producto de amplificación 225 pb, que codifica el dominio catalítico a la producción de quitinasas bacterianas. Se han reportado cepas de los géneros Serratia, Pseudomonas, Acinetobacter, Enterobacter con presencia del gen chiA, como un indicativo para el control de enfermedades en cultivos agrícolas (Cucu et al, 2019;Medina-de la Rosa et al, 2016;Okay & Alshehri, 2020;Verma et al, 2023).…”

Heterologous Expression of chi42 Gene from Trichoderma asperellum in Bacillus subtilis

“…To date, only a few microbial chitinase genes have been successfully expressed on Bacillus hosts such as the chiA gene from S. marcescens (Okay et al 2008;Okay and Alshehri 2020) or the chitinase gene from B. pumilus (Ahmadian et al 2012;Rostami et al 2017). And most of them are promising sources for agricultural and biotechnological applications.…”

“…Although B. subtilis has become an increasingly popular host for recombinant protein expression due to its ability to directly secrete protein into culture media, it is amenable to medium-and large-scale fermentation, lack of codon bias and designation as a safe organism (FDA 2018). To date, only a few studies produced foreign chitinase in B. subtilis such as the chitinase derived from B. pumilus (Ahmadian et al 2012) and chitinase A from S. marcescens (Okay and Alshehri 2020). This study reports on the expression of the 42 kDa chitinase (GH family 18) from T. asperellum SH16 in B. subtilis BD170 and its characterizations and antifungal activity.…”