Abstract
In this study, we isolated and optimized the culture conditions for β-1,3-glucanase biosynthesis by a highly enzyme-producing bacterial strains isolated in Thua Thien Hue province. Among the three isolates M6, DH2, and T5, strain M6 exhibited the strongest β-1,3-glucanase production with the strongest hydrolysis zone of 19.03±0.05 mm diameter. Strain M6 was identified through nucleotide sequencing by 16S rRNA region, showing a similarity of 99.72% with Paenibacillus polymyxa HGPJ-62 in the GenBank database (NCBI). The optimized medium composition and culture conditions for strain Paenibacillus polymyxa M6 were determined as follows: the β-1,3-glucanase induction medium comprised the following components: K₂HPO₄·3H₂O 1.3 g; MgSO₄·7H₂O 1 g; KCl 0.5 g; yeast extract 0.5 g; NaNO₃ 1 g; oat flour 5 g; pH = 6, incubation temperature 40–50°C for 24 hours, shaking speed 180 rpm, and inoculum ratio 1%. The β-1,3-glucanase activity reached 1.5 U/mL. The present study highlights the β-1,3-glucanase biosynthesis by strain M6, demonstrating its suitability for development and application for prevent gummosis disease in Thanh Tra pomelo trees as well as in controlling fungal pathogens with glucan-dominant cell walls.
References
- Gao, F., Liu, X., Dong, R., Liao, M., Qin, X., Wang, X. et al. (2023), Exploring the antifungal mechanism of β-1,3-glucanase for effectively inhibiting the food contamination by Aspergillus flavus and Aspergillus fumigatus, LWT, 187, 115342,https://doi.org/10.1016/j.lwt.2023.115342.
- Hoang Anh Nguyen, Thu-Ha Nguyen, Tien-Thanh Nguyen, Clemens K Peterbauer, Geir Mathiesen and Dietmar Haltrich (2012), Chitinase from Bacillus licheniformis DSM13: Expression in Lactobacillus plantarum WCFS1 and biochemical characterization, Protein Expr Purif, 81(2), 166–174. https://doi.org/10.1016/j.pep.2011.10.005.
- Ueki, A., Takehara, T., Ishioka, G., Kaku, N. and Ueki, K. (2020), β-1,3-Glucanase production as an anti-fungal enzyme by phylogenetically different strains of the genus Clostridium isolated from anoxic soil that underwent biological disinfestation, Appl Microbiol Biotechnol, 104(12), 5563–5578, https://doi.org/10.1007/s00253-020-10626-8.
- Sahgal, M. (2022), Fungal Enzymes in Biocontrol of Phytopathogens, 327–356, https://doi.org/10.1007/978-981-16-3307-2_12.
- Perrot, T., Pauly, M. and Ramírez, V. (2022), Emerging Roles of β-Glucanases in Plant Development and Adaptative Responses, 11(9), 1119, Plants, 11. https://doi.org/10.3390/plants11091119.
- Dobrzyński, J. and Naziębło, A. (2024), Paenibacillus as a Biocontrol Agent for Fungal Phytopathogens: Is P. polymyxa the Only One Worth Attention? Microb Ecol, 87(1), 134. https://doi.org/10.1007/s00248-024-02450-8.
- Borshchevskaya, L.N., Gordeeva, T.L., Kalinina, A.N., Fedorov, A.S. and Sineoky, S.P. (2019), Comparison of β-Glucanases from Bacillus pumilus, Paenibacillus polymyxa, Bacillus subtilis, and Bacillus amyloliquefaciens in the Expression System of Pichia pastoris: Biochemical Characteristics and Potential in Fodder Production, Appl Biochem Microbiol, 55(8), 780–787, https://doi.org/10.1134/S0003683819080040.
- Pham Thi Ly Thu, Nguyen Thi Hong Minh, Nguyen Duc Anh, Dao Thi Thu Hang, Nguyen Duc Thanh, Nguyen Thi Bich Ngoc et al. (2022), Isolation and molecular authentication of antagonistic Trichoderma fungi for controlling citrus leaf yellowing, root rot in provinces from the Mekong River Delta , Journal of Vietnam Agricultural Science and Technology, 5(138), 50–57.
- He, G.Q., Tang, X.J., Mukhtar, A.M.A. and Chen, Q.H. (2003), Optimization of cultural conditions for thermostable β-1,3-1,4-glucanase production by Bacillus subtilis ZJF-1A5, 4(6), 719–726, J Zhejiang Univ Sci, 4. https://doi.org/10.1631/jzus.2003.0719.
- C. Giese, E., F. H. Dekker, R. and M. Barbosa, A. (2012), Production of β-(1,3)-glucanases by Trichoderma harzianum Rifai: Optimization and Application to Produce Gluco-oligosaccharides from Paramylon and Pustulan, Ferment Technol, 1(1). https://doi.org/10.4172/2167-7972.1000102.
- Pham Thi Lich and Tran Thanh Thuy (2013), A research on the transplanting conditions for the production of antifungal chitinase enzyme from Trichoderma sp., Ho Chi Minh City university of education journal of science, 51, 117–29.
- Mahasneh, A.M. and Stewart, D.J. (1980), A Medium for Detecting β‐(1 →3) Glucanase Activity in Bacteria, J. Appl. Bacteriol, 48(3), 457–458. https://doi.org/10.1111/j.1365-2672.1980.tb01035.x.
- Tran Thi Hong, Nguyen Thi Kim Cuc, Pham Viet Cuong and Pham Thi Thuy Hoai. (2014), Isolation of antagonistic microorganisms against some plant fungal pathogens and evaluation of their activity in vitro and in vivo, Science and Technology, 52(4), 419–430.
- Vinche, M.H., Khanahmadi, M., Ataei, S. ahmad and Danafar, F. (2024), Purification and characterization of beta 1,3-1,4-glucanase from Aspergillus niger CCUG33991, Authorea, 1–15. https://doi.org/10.22541/au.170668361.11920777/v1.
- Zacky, F.A. and Ting, A.S.Y. (2013), Investigating the bioactivity of cells and cell-free extracts of Streptomyces griseus towards Fusarium oxysporum f. sp. cubense race 4, Biological Control, 66(3), 204–208, https://doi.org/10.1016/j.biocontrol.2013.06.001.
- Aktuganov, G.E., Galimzyanova, N.F., Melent’Ev, A.I. and Kuz’Mina, L.Y. (2007), Extracellular hydrolases of strain Bacillus sp. 739 and their involvement in the lysis of micromycete cell walls, Microbiology (N Y), 76(4), 413–420, https://doi.org/10.1134/S0026261707040054.
- Ueki, A., Takehara, T., Ishioka, G., Kaku, N. and Ueki, K. (2019), Production of β-1,3-glucanase and chitosanase from clostridial strains isolated from the soil subjected to biological disinfestation, AMB Express, 9(1), 114, https://doi.org/10.1186/s13568-019-0842-1.
- Thomas, C.R. and Geer, D. (2011), Effects of shear on proteins in solution. Biotechnol Lett, 33(3), 443–456, https://doi.org/10.1007/s10529-010-0469-4.
- Maa, Y.F. and Hsu, C.C. (1997), Protein denaturation by combined effect of shear and air-liquid interface. Biotechnol Bioeng, 54(6), 503–512, https://doi.org/10.1002/(SICI)1097-0290(19970620)54:6<503::AID-BIT1>3.0.CO;2-N.
- Tang, X.J., He, G.Q., Chen, Q.H., Zhang, X.Y. and Ali, M.A.M. (2004), Medium optimization for the production of thermal stable β-glucanase by Bacillus subtilis ZJF-1A5 using response surface methodology. Bioresour Technol, 93(2), 175–181, https://doi.org/10.1016/j.biortech.2003.10.013.
- Lee, K.C., Arai, T., Ibrahim, D., Kosugi, A., Prawitwong, P., Lan, D. et al. (2014), Purification and characterization of a thermostable laminarinase from Penicillium rolfsii c3-2(1) IBRL, Bioresources, 9(1), 1072–1084, https://doi.org/10.15376/biores.9.1.1072-1084.
- Sena, A.R., Júnior, G.L.V., Neto, A.G., Taranto, A.G., Pirovani, C.P., Cascardo, J.C.M. et al. (2011), Production, purification and characterization of a thermostable β-1,3-glucanase (laminarinase) produced by Moniliophthora perniciosa. An Acad Bras Cienc, 83(2), 599–609, https://doi.org/10.1590/S0001-37652011005000007.
- Santana, M.L., Maciel Paulo, E., Bispo, J.A., de Sena, A.R. and de Assis, S.A. (2018), Production and partial characterization of β-1,3-glucanase obtained from Rhodotorula oryzicola, Prep Biochem Biotechnol, 48(2), 165–171, https://doi.org/10.1080/10826068.2017.1421962.
- Zhang, S.B., Zhang, W.J., Zhai, H.C., Lv, Y.Y., Cai, J.P., Jia, F. et al. (2019), Expression of a wheat β-1,3-glucanase in Pichia pastoris and its inhibitory effect on fungi commonly associated with wheat kernel, Protein Expr Purif, 154, 134–139, https://doi.org/10.1016/j.pep.2018.10.011.
- Calloni, R.D., Muchut, R.J., Garay, A.S., Arias, D.G., Iglesias, A.A. and Guerrero, S.A. (2023), Functional and structural characterization of an endo-β-1,3-glucanase from Euglena gracilis, Biochimie, 208, 117–128, https://doi.org/10.1016/j.biochi.2022.12.016.
- Blättel, V., Larisika, M., Pfeiffer, P., Nowak, C., Eich, A., Eckelt, J. et al. (2011), β-1,3-Glucanase from delftia tsuruhatensis strain MV01 and its potential application in vinification, Appl Environ Microbiol, 77(3), 983–990, https://doi.org/10.1128/AEM.01943-10.