Expression and Characterization of Two Chitinases with Synergistic Effect and Antifungal Activity from Xenorhabdus Nematophila

Overview

Journal World J Microbiol Biotechnol

Publisher Springer

Specialties Biotechnology
Microbiology

Date 2019 Jul 4

PMID 31267229

Citations 3

Authors

Jia Liu

Ziyan Nangong

Jie Zhang

Ping Song

Yin Tang

Yue Gao

Qinying Wang

Affiliations

Soon will be listed here.

Abstract

Xenorhabdus nematophila HB310 secreted the insecticidal protein toxin complex. Two chitinase genes, chi60 and chi70, were found in X. nematophila toxin complex locus. In order to clarify the function of two chitinases, chi60 and chi70 genes were cloned and expressed in Escherichia coli Transetta (DE3). As a result, we found that the Chi60 and Chi70 belonged to glycoside hydrolases (GH) family 18 with a molecular mass of 65 kDa and 78 kDa, respectively. When colloidal chitin was treated as the substrate, Chi60 and Chi70 were proved to have the highest enzymatic activity at pH 6.0 and 50 °C. Chi60 and Chi70 had obvious growth inhibition effect against the second larvae of Helicoverpa armigera with growth inhibiting rate of 81.99% and 90.51%. Chi70 had synergistic effect with the insecticidal toxicity of Bt Cry 1Ac, but the Chi60 had no synergistic effect with Bt Cry 1Ac. Chi60 and Chi70 showed antifungal activity against Alternaria brassicicola, Verticillium dahliae and Coniothyrium diplodiella. The results increased our understanding of the chitinases produced by X. nematophila and laid a foundation for further studies on the mechanism of the chitinases.

Citing Articles

Functional Comparison of Three Chitinases from Symbiotic Bacteria of Entomopathogenic Nematodes.

Son D, Kim G, Choo H, Chung N, Choo Y Toxins (Basel). 2024; 16(1).

PMID: 38251242 PMC: 10821219. DOI: 10.3390/toxins16010026.

Insecticidal Activity of Chitinases from HB310 and Its Relationship with the Toxin Complex.

Liu J, Bai H, Song P, Nangong Z, Dong Z, Li Z Toxins (Basel). 2022; 14(9).

PMID: 36136584 PMC: 9505380. DOI: 10.3390/toxins14090646.

Microbial chitinases: properties, current state and biotechnological applications.

Le B, Yang S World J Microbiol Biotechnol. 2019; 35(9):144.

PMID: 31493195 DOI: 10.1007/s11274-019-2721-y.

References

Patil , Ghormade V , Deshpande . Chitinolytic enzymes: an exploration. Enzyme Microb Technol. 2000; 26(7):473-483. DOI: 10.1016/s0141-0229(00)00134-4. View

Downing K, Thomson J . Introduction of the Serratia marcescens chiA gene into an endophytic Pseudomonas fluorescens for the biocontrol of phytopathogenic fungi. Can J Microbiol. 2000; 46(4):363-9. DOI: 10.1139/w99-147. View

Wiwat C, Thaithanun S, Pantuwatana S, Bhumiratana A . Toxicity of chitinase-producing Bacillus thuringiensis ssp. kurstaki HD-1 (G) toward Plutella xylostella. J Invertebr Pathol. 2000; 76(4):270-7. DOI: 10.1006/jipa.2000.4976. View

Morgan J, Sergeant M, Ellis D, OUSLEY M, Jarrett P . Sequence analysis of insecticidal genes from Xenorhabdus nematophilus PMFI296. Appl Environ Microbiol. 2001; 67(5):2062-9. PMC: 92837. DOI: 10.1128/AEM.67.5.2062-2069.2001. View

Liu M, Cai Q, Liu H, Zhang B, Yan J, Yuan Z . Chitinolytic activities in Bacillus thuringiensis and their synergistic effects on larvicidal activity. J Appl Microbiol. 2002; 93(3):374-9. DOI: 10.1046/j.1365-2672.2002.01693.x. View

Arora N, Ahmad T, Rajagopal R, Bhatnagar R . A constitutively expressed 36 kDa exochitinase from Bacillus thuringiensis HD-1. Biochem Biophys Res Commun. 2003; 307(3):620-5. DOI: 10.1016/s0006-291x(03)01228-2. View

Lertcanawanichakul M, Wiwat C, Bhumiratana A, Dean D . Expression of chitinase-encoding genes in Bacillus thuringiensis and toxicity of engineered B. thuringiensis subsp. aizawai toward Lymantria dispar larvae. Curr Microbiol. 2004; 48(3):175-81. DOI: 10.1007/s00284-003-4119-0. View

Singh Ajit N, Verma R, Shanmugam V . Extracellular chitinases of fluorescent pseudomonads antifungal to Fusarium oxysporum f. sp. dianthi causing carnation wilt. Curr Microbiol. 2006; 52(4):310-6. DOI: 10.1007/s00284-005-4589-3. View

Isaac Kirubakaran S, Sakthivel N . Cloning and overexpression of antifungal barley chitinase gene in Escherichia coli. Protein Expr Purif. 2006; 52(1):159-66. DOI: 10.1016/j.pep.2006.08.012. View

10.

Lee S, Stoilova-McPhie S, Baxter L, Fulop V, Henderson J, Rodger A . Structural characterisation of the insecticidal toxin XptA1, reveals a 1.15 MDa tetramer with a cage-like structure. J Mol Biol. 2007; 366(5):1558-68. DOI: 10.1016/j.jmb.2006.12.057. View

11.

Bhattacharya D, Nagpure A, Gupta R . Bacterial chitinases: properties and potential. Crit Rev Biotechnol. 2007; 27(1):21-8. DOI: 10.1080/07388550601168223. View

12.

Wang S, Peng J, Liang T, Liu K . Purification and characterization of a chitosanase from Serratia marcescens TKU011. Carbohydr Res. 2008; 343(8):1316-23. DOI: 10.1016/j.carres.2008.03.030. View

13.

Joshi M, Sharma A, Kant S, Birah A, Gupta G, Khan S . An insecticidal GroEL protein with chitin binding activity from Xenorhabdus nematophila. J Biol Chem. 2008; 283(42):28287-96. PMC: 2661396. DOI: 10.1074/jbc.M804416200. View

14.

Bravo A, Soberon M . How to cope with insect resistance to Bt toxins?. Trends Biotechnol. 2008; 26(10):573-9. DOI: 10.1016/j.tibtech.2008.06.005. View

15.

Cantarel B, Coutinho P, Rancurel C, Bernard T, Lombard V, Henrissat B . The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics. Nucleic Acids Res. 2008; 37(Database issue):D233-8. PMC: 2686590. DOI: 10.1093/nar/gkn663. View

16.

Mehmood M, Xiao X, Hafeez F, Gai Y, Wang F . Molecular characterization of the modular chitin binding protein Cbp50 from Bacillus thuringiensis serovar konkukian. Antonie Van Leeuwenhoek. 2011; 100(3):445-53. DOI: 10.1007/s10482-011-9601-2. View

17.

Busby J, Landsberg M, Simpson R, Jones S, Hankamer B, Hurst M . Structural analysis of Chi1 Chitinase from Yen-Tc: the multisubunit insecticidal ABC toxin complex of Yersinia entomophaga. J Mol Biol. 2011; 415(2):359-71. DOI: 10.1016/j.jmb.2011.11.018. View

18.

Landsberg M, Jones S, Rothnagel R, Busby J, Marshall S, Simpson R . 3D structure of the Yersinia entomophaga toxin complex and implications for insecticidal activity. Proc Natl Acad Sci U S A. 2011; 108(51):20544-9. PMC: 3251104. DOI: 10.1073/pnas.1111155108. View

19.

Zarei M, Aminzadeh S, Zolgharnein H, Safahieh A, Daliri M, Noghabi K . Characterization of a chitinase with antifungal activity from a native Serratia marcescens B4A. Braz J Microbiol. 2013; 42(3):1017-29. PMC: 3768790. DOI: 10.1590/S1517-838220110003000022. View

20.

Itoh T, Hibi T, Fujii Y, Sugimoto I, Fujiwara A, Suzuki F . Cooperative degradation of chitin by extracellular and cell surface-expressed chitinases from Paenibacillus sp. strain FPU-7. Appl Environ Microbiol. 2013; 79(23):7482-90. PMC: 3837743. DOI: 10.1128/AEM.02483-13. View