Cloning and expression of chitinase A, B, and C (chiA, chiB, chiC) genes from Serratia marcescens originating from Helicoverpa armigera and determining their activities

Danismazoglu, Mehtap; Demir, İsmail; Sezen, Kazım; Muratoglu, Hacer; Nalçacıoğlu, Remziye

doi:10.3906/biy-1404-31

Cited by 27 publications

(12 citation statements)

References 78 publications

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“…Also, domain positions were opposite in S. marcescens-ChiC and viral chitinases, while the glyco_18 and GH_18 domains located at the C terminal of viral chitinases showed similar domains located at the N terminal of S. marcescens-ChiC. S.marcescens-ChiA, structurally analyzed previously by Danişmazoğlu et al (2015), had a similar domain organization to those of AcNPV-Chi and CpGV-Chi.…”

Section: Potential Activity Of Chitinase Proteinsmentioning

confidence: 65%

“…Bacterial chitinase protein (chitinase C) belongs to Serratia marcescens that was previously isolated from dead Helicoverpa armigera (Hubner, 1808) larvae. The S. marcescens chitinase gene (accession number: KF823632) was provided cloned into the pET-28a (+) expression vector (Danişmazoğlu et al 2015). Labreared culture of Galleria mellonella (Linnaeus, 1758).…”

Section: Viral and Bacterial Strains And Insect Culturementioning

confidence: 99%

“…The potential activity of S. marcescens ChiC protein, used in this study, was previously studied by Danişmazoğlu et al (2015), who tested the mortality not only on ChiC protein but also chiA and chiB proteins of S. marcescens on Malacosoma neustria (Linnaeus, 1758) and Helicoverpa armigera larvae. Among these 3 chitinase proteins, ChiC produced the highest potential activity against used insect larvae.…”

Section: Potential Activity Of Chitinase Proteinsmentioning

confidence: 99%

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Comparison of the potential activities of viral and bacterial chitinases

Abulikemu

Yesilyurt

Gencer

et al. 2021

Egypt J Biol Pest Control

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Background Chitin, a long-chain polymer of N-acetylglucosamine, is a major structural component of the insect exoskeleton and the peritrophic membrane (PM). Chitinases are able to effectively break down glycosidic bonds of chitin polymer thus can be used in agriculture to control plant pathogen insects. These enzymes can be synthesized by higher plants, animals, protista, bacteria, and viruses. Results In this study, viral and bacterial chitinases were compared for their potential activity on a laboratory test insect. The genes encoding chitinases of Autographa californica nucleopolyhedrovirus (AcNPV) and Cydia pomonella granulovirus (CpGV) were amplified from genomic DNAs by PCR and cloned into the pET-28a (+) expression vector. The chitinase proteins of these 2 viruses (AcNPV-Chi, CpGV-Chi) and Serratia marcescens chitinase C (ChiC) protein which was previously cloned were overexpressed in Escherichia coli. Expressed proteins were purified and confirmed by western blot analysis as 50, 63, and 68 kDa for AcNPV, CpGV, and S. marcescens chitinases, respectively. Enzyme activities of the chitinases were confirmed. Chitinases were also compared to each other in silico. The insecticidal effects of these proteins were evaluated on Galleria mellonella L. larvae. Bioassays were performed on the 3rd instar larvae for each chitinase protein in triplicate. The results showed that although there were differences in enzymatic activities and domain organizations, all 3 microbial chitinases produced almost the same level of insecticidal activity on the test insect. LC50 and LT50 values were compatible with the mortality results. These results were a preanalysis for comparing the effects of microbial chitinases. Conclusion Potential activity experiments should be carried out on more insects to provide detailed information on the insecticidal effects of bacterial and viral chitinases.

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Section: Potential Activity Of Chitinase Proteinsmentioning

confidence: 65%

Section: Viral and Bacterial Strains And Insect Culturementioning

confidence: 99%

Section: Potential Activity Of Chitinase Proteinsmentioning

confidence: 99%

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Comparison of the potential activities of viral and bacterial chitinases

Abulikemu

Yesilyurt

Gencer

et al. 2021

Egypt J Biol Pest Control

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“…Among hydrolytic enzymes, chitinolytic enzymes are the most important for the biocontrol of fungal phytopathogens. Chitinases are encoded by a number of genes that include, e.g., chiA , chiB , chiC and chiD , chiF , chiG , chiR , chiW , and chiX ( Alam et al, 1996 ; Hamilton et al, 2014 ; Danişmazoğlu et al, 2015 ; Martínez-Zavala et al, 2020 ; Azizoglu et al, 2021 ; Kumar et al, 2022 ). The efficacy of chitinases produced by B. pumilus against fungi has been documented in a few studies.…”

Section: Hydrolytic Enzymes Produced By Bacillus Pumilusmentioning

confidence: 99%

Biocontrol of fungal phytopathogens by Bacillus pumilus

Dobrzyński¹,

Jakubowska²,

Kulkova³

et al. 2023

Front. Microbiol.

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Plant growth-promoting bacteria are one of the most interesting methods of controlling fungal phytopathogens. These bacteria can participate in biocontrol via a variety of mechanisms including lipopeptide production, hydrolytic enzymes (e.g., chitinase, cellulases, glucanase) production, microbial volatile organic compounds (mVOCs) production, and induced systemic resistance (ISR) triggering. Among the bacterial genera most frequently studied in this aspect are Bacillus spp. including Bacillus pumilus. Due to the range of biocontrol traits, B. pumilus is one of the most interesting members of Bacillus spp. that can be used in the biocontrol of fungal phytopathogens. So far, a number of B. pumilus strains that exhibit biocontrol properties against fungal phytopathogens have been described, e.g., B. pumilus HR10, PTB180, B. pumilus SS-10.7, B. pumilus MCB-7, B. pumilus INR7, B. pumilus SE52, SE34, SE49, B. pumilus RST25, B. pumilus JK-SX001, and B. pumilus KUDC1732. B. pumilus strains are capable of suppressing phytopathogens such as Arthrobotrys conoides, Fusarium solani, Fusarium oxysporum, Sclerotinia sclerotiorum, Rhizoctonia solani, and Fagopyrum esculentum. Importantly, B. pumilus can promote plant growth regardless of whether it alters the native microbiota or not. However, in order to increase its efficacy, research is still needed to clarify the relationship between the native microbiota and B. pumilus. Despite that, it can already be concluded that B. pumilus strains are good candidates to be environmentally friendly and commercially effective biocontrol agents.

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“…Vì vậy, chitinase tái tổ hợp được tạo ra bằng kỹ thuật DNA nhằm sản xuất lượng lớn enzyme này đã được quan tâm nghiên cứu. Các vật chủ thường được các nhà nghiên cứu chọn để biểu hiện gen chitinase là các vi sinh vật có khả năng sinh trưởng và phát triển nhanh như E. coli (Lobo et al, 2013;Mehtap et al, 2015). Quá trình lên men chủng tái tổ hợp chịu ảnh hưởng của nhiều yếu tố khác nhau như: nồng độ chất cảm ứng, thời gian cảm ứng, mật độ tế bào tại thời điểm cảm ứng, nhiệt độ cảm ứng.…”

Section: Mở đầUunclassified

Optimization and improvement of recombinant chitinase biosynthesis in E. coli using response surface method

Hà¹,

Quyền²,

Binh³

2017

V J Bio

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Chitinases (EC 3.2.1.14) are enzymes that degrade chitin. These enzymes are gaining importance for their wide-ranging applications including the preparation of pharmaceutically important chitooligosaccharides and N-acetyl D glucosamine, preparation of single-cell protein, isolation of protoplasts from fungi and yeast, control of pathogenic fungi, treatment of chitinous waste, mosquito control and morphogenesis and especially in agriculture fields to control pathogens. Previously, we have cloned chiA gene encoding chitinase for overexpresion in E. coli. Herein, we optimized the expression conditions for improvement of chitinase biosynthesis in recombinant E. coli strain using response surface method (RSM) based on Design-Expert 10.1 software (Stat-Ease, Inc., Minneapolis, USA. Fermentation conditions such as inducer concentrate, induction time and cell density at the time of induction and their effects on the chitinase biosynthesis were investigated. The RMS analysis suggested that the optimal conditions for production of recombinant ChiA are at lactose concentration of 6,15 mM, 8,12 hours after induction and cell density OD600nm = 0,87 with predicted enzyme activity of 7,49 U/ml. The model obtained was used for fermentation of recombinant strain and the real chitinase activity achieved 7,54 U/ml which is 1.32 times higher compared to that of the pre-optimization (5.71 U/ml). The RSM was found to be useful in optimizing and determining the interactions among process variables in ChiA enzyme production. Our data also indicated that using lactose as the inducer instead of ITPG for ChiA expression can also reduce product costs. Citation: Trinh Thi Thu Ha, Dong Van Quyen, Ngo Dinh Binh, 2018. Optimization and improvement of recombinant chitinase biosynthesis in E. coli using response surface method. Tap chi Sinh hoc, 40(1): x-xx. DOI: 10.15625/0866-7160/v40n1.10495. *Corresponding author: binh.gen@gmail.com Received 5 July 2017, accepted 20 December 2017

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Cloning and expression of chitinase A, B, and C (chiA, chiB, chiC) genes from Serratia marcescens originating from Helicoverpa armigera and determining their activities

Cited by 27 publications

References 78 publications

Comparison of the potential activities of viral and bacterial chitinases

Comparison of the potential activities of viral and bacterial chitinases

Biocontrol of fungal phytopathogens by Bacillus pumilus

Optimization and improvement of recombinant chitinase biosynthesis in E. coli using response surface method

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