Purification sucrolytic produced from Bacillus licheniformis

Vivian Daifallah

Abstract
Sucrolytic produced by Bacillus licheniformis using submerged fermentation under optimized conditions was purified. The purification process included the following steps: precipitation of protein by 60% ammonium sulphate salts followed by sephadex G-100 gel filtration column. Folds of purification of sucrolytic reached 2.66 fold with specific activity of 0.612IU/mL and recovery 4.27%.

Keywords
Sucrolytic; Purification; Bacillus licheniformis; Submerged fermentation.

Resumo
Purificação da enzima sacrolítica produzida por Bacillus licheniformis. A enzima sacrolítica produzida por Bacillus licheniformis utilizando fermentação submersa em condições otimizadas foi purificada. O processo de purificação incluiu as seguintes etapas: precipitação da proteína com sulfato de amônio a 60%, seguida por filtração em coluna de gel Sephadex G-100. O fator de purificação da enzima sacrolítica atingiu 2,66 vezes, com atividade específica de 0,612 UI/mL e recuperação de 4,27%.

Palavras-chave
Sucrolítica; Purificação; Bacillus licheniformis; Fermentação submersa.

DOI
10.21438/rbgas(2026)133308


Full text

 Baixar este arquivo PDF

 


References
Ackerman, D. L.; Craft, K. M.; Townsend, S. D. Infant food applications of complex carbohydrates: Structure, synthesis, and function. Carbohydrate Research, v. 437, p. 16-27. 2017. https://doi.org/10.1016/j.carres.2016.11.007

Ammar, Y. B.; Matsubara, T.; Ito, K.; Iizuka, M.; Limpaseni, T.; Pongsawasdi, P.; Minamiura, N. Characterization of a thermostable levansucrase from Bacillus sp. TH4-2 capable of producing high molecular weight levan at high temperature. Journal of Biotechnology, v. 99, n. 2, p. 111-119, 2002. https://doi.org/10.1016/S0168-1656(02)00160-8

Clements, L. D.; Miller, B. S.; Streips, U. N. Comparative growth analysis of the facultative anaerobes Bacillus subtilis, Bacillus licheniformis, and Escherichia coli. Systematic and Applied Microbiology, v. 25, n. 2, p. 284-286, 2002. https://doi.org/10.1078/0723-2020-00108

De Boer, A. S.; Priest, F.; Diderichsen, B. On the industrial use of Bacillus licheniformis: A review. Applied Microbiology and Biotechnology, v. 40, p. 595-598, 1994. https://doi.org/10.1007/BF00173313

Deng, W.; Dong, X. F.; Tong, J. M.; Zhang, Q. The probiotic Bacillus licheniformis ameliorates heat stress-induced impairment of egg production, gut morphology, and intestinal mucosal immunity in laying hens. Poultry Science, v. 91, n. 3, p. 575-582, 2012. https://doi.org/10.3382/ps.2010-01293

Doan, C. T.; Tran, T. N.; Nguyen, M. T.; Nguyen, V. B.; Nguyen, A. D.; Wang, S. L. Anti-α-glucosidase activity by a protease from Bacillus licheniformis. Molecules, v. 24, n. 4, 691, 2019. https://doi.org/10.3390/molecules24040691

Doan, C. T.; Tran, T. N.; Nguyen, M. T.; Nguyen, V. B.; Nguyen, A. D.; Wang, S. L. Production of sucrolytic enzyme by Bacillus licheniformis by the bioconversion of Pomelo Albedo as a carbon source. Polymers, v. 13, n. 12, 1959, 2021. https://doi.org/10.3390/polym13121959

González-Garcinuño, A.; Tabernero, A.; Domínguez, A.; Galán, M. A.; Del Valle, E. M. M. Levan and levansucrases: Polymer, enzyme, micro-organisms and biomedical applications. Biocatalysis and Biotransformation, v. 36, n. 3, p. 233-244, 2018. https://doi.org/10.1080/10242422.2017.1314467

Li, X.; Wang, X.; Meng, X.; Dijkhuizen, L.; Liu, W. Structures, physicochemical properties, production and (potential) applications of sucrose-derived α-D-glucans synthesized by glucansucrases. Carbohydrate Polymers, v. 249, 116818, 2020. https://doi.org/10.1016/j.carbpol.2020.116818

Lide, D. R. CRC Handbook of Chemistry and Physics. 87. ed. Boca Raton: CRC Press, 2006.

Lincoln, L.; More, S. S. Bacterial invertases: Occurrence, production, biochemical characterization, and significance of transfructosylation. Journal of Basic Microbiology, v. 57, n. 10, p. 803-813, 2017. https://doi.org/10.1002/jobm.201700269

Lincoln, L.; More, S. S.; Reddy, S. V. Purification and biochemical characterization of β-D-fructofuranosidase from Bacillus subtilis LYN12. Journal of Food Biochemistry, v. 42, e12592, 2018. https://doi.org/10.1111/jfbc.12592

Lowry, O. H.; Rosebrough, N. J.; Farr, A. L.; Randall, R. J. Protein measurement with the Folin phenol reagent. World Journal of Microbiology and Biotechnology, v. 193, n. 1, p. 265-275, 1951. https://doi.org/10.1016/S0021-9258(19)52451-6

Manoochehri, H.; Hosseini, N. F.; Saidijam, M.; Taheri, M.; Rezaee, H.; Nouri, F. A review on invertase: Its potentials and applications. Biocatalysis and Agricultural Biotechnology, v. 25, 101599, 2020. https://doi.org/10.1016/j.bcab.2020.101599

Mariz, B. P.; Carvalho, S.; Batalha, I. L.; Pina, A. S. Arti?cial enzymes bringing together computational design and directed evolution. Organic & Biomolecular Chemistry, v. 19, n. 9, p. 1915-1925, 2021. https://doi.org/10.1039/D0OB02143A

Mehta, P. K.; Sehgal, S. Microbial enzymes in food processing. In: Husain, Q.; Ullah, M. (Eds.). Biocatalysis. Cham: Springer, 2019. p. 255-272.

Muras, A.; Romero, M.; Mayer, C.; Otero, A. Biotechnological applications of Bacillus licheniformis. Critical Reviews in Biotechnology, v. 41, n. 4, p. 609-627, 2021. https://doi.org/10.1080/07388551.2021.1873239

Nakapong, S.; Pichyangkura, R.; Ito, K.; Iizuka, M.; Pongsawasdi, P. High expression level of levansucrase from Bacillus licheniformis RN-01 and synthesis of levan nanoparticles. International Journal of Biological Macromolecules, v. 54, p. 30-36, 2013. https://doi.org/10.1016/j.ijbiomac.2012.11.017

Parrado, J.; Rodriguez-Morgado, B.; Tejada, M.; Hernandez, T.; Garcia, C. Proteomic analysis of enzyme production by Bacillus licheniformis using different feather wastes as the sole fermentation media. Enzyme and Microbial Technology, v. 57, p. 1-7, 2014. https://doi.org/10.1016/j.enzmictec.2014.01.001

Pham, J. V.; Yilma, M. A.; Feliz, A.; Majid, M. T.; Maffetone, N.; Walker, J. R.; Kim, E.; Cho, H. J.; Reynolds, J. M.; Song, M. C.; Park, S. R.; Yoon, Y. J. A review of the microbial production of bioactive natural products and biologics. Frontiers in Microbiology, v. 10, 1404, 2019. https://doi.org/10.3389/fmicb.2019.01404

Rasbold, L. M.; Heinen, P. R.; Silva, J. L. C.; Simão, R. C. G.; Kadowaki, M. K.; Maller, A. Cunninghamella echinulata PA3S12MM invertase: Biochemical characterization of a promiscuous enzyme. Journal of Food Biochemistry, v. 45, e13654, 2021. https://doi.org/10.1111/jfbc.13654

Sellami-Kamoun, A.; Haddar, A.; Ne, H. A.; Ghorbel-Frikha, B.; Kanoun, S.; Nasri, M. Stability of thermostable alkaline protease from Bacillus licheniformis RP1 in commercial solid laundry detergent formulations. Microbiological Research, v. 163, n. 1, p. 299-306, 2008. https://doi.org/10.1016/j.micres.2006.06.001

Taskin, M.; Ortucu, S.; Unver, Y.; Canli, O. Invertase production and molasses decolourization by cold-adapted filamentous fungus Cladosporium herbarum ER-25 in non-sterile molasses medium. Process Safety and Environmental Protection, v. 103, Part A, p. 136-143, 2016. https://doi.org/10.1016/j.psep.2016.07.006

Wang, J.; Xiao, H.; Zhao, F.; Zhao, B.; Xu, M.; Zhou, Z.; Han, Y. A fructansucrase secreted extracellular and purified in one-step by gram-positive enhancer matrix particles. Processes, v. 9, n. 1, 95, 2021. https://doi.org/10.3390/pr9010095

Xavier, J. R.; Ramana, K. V. Optimization of levan production by cold active Bacillus licheniformis ANT179 and fructooligosaccharide synthesis by its levansucrase. Applied Biochemistry and Biotechnology, v. 181, p. 986-1006, 2017. https://doi.org/10.1007/s12010-016-2264-8<

Xu, Z.-W.; Li, Y.-Q.; Wang, Y.-H.; Yang, B.; Ning, Z.-X. Production of β-fructofuranosidase by Arthrobacter sp. and its application in the modification of stevioside and rebaudioside A. Food Technology & Biotechnology, v. 47, n. 2, p. 137-143, 2009.

Yoon, M.; Choi, W.; Kwon, S.; Yi, S.; Lee, D.; Lee, J. Purification and properties of intracellulaseinvertase from alkalophilic and thermophilic Bacillus cereus TA-11. Journal of Applied Biological Chemistry, v. 50, n. 4, p. 196-201, 2007.


 

ISSN 2359-1412