The Use of Supercritical CO? with Additives to Enhance Starch-Digesting Enzyme Activities in Bacillus subtilis

Hongde Yan; Zhijiang  Wang; Lihui Dong

doi:10.4025/actascitechnol.v48i1.74265

Authors

Hongde Yan Zhejiang Pharmaceutical University https://orcid.org/0000-0002-8099-5225
Zhijiang Wang Zhejiang Pharmaceutical University
Lihui Dong Zhejiang Pharmaceutical University

DOI:

https://doi.org/10.4025/actascitechnol.v48i1.74265

Keywords:

mutation breeding, dimethyl sulfoxide, hydrogen peroxide, spore.

Abstract

Supercritical carbon dioxide (SC-CO₂) has been proposed as a sterilization technique and utilized as a breeding method to enhance the activity of enzymes produced by microorganisms by treating their vegetative cells. This study explores whether this breeding method could be applicable to bacterial spores. Spores of Bacillus subtilis were treated with SC-CO₂ in the presence of additives such as dimethyl sulfoxide (DMSO) or hydrogen peroxide (H₂O₂). Various concentrations of these additives were employed to screen for strains exhibiting increased starch-digesting enzyme activity. The resulting strains, D2-5 and H2-1, demonstrated improved starch-digesting enzyme activities after SC-CO₂ treatment with 2% DMSO or 0.6% H₂O₂, respectively, which were 74.7% and 67.3% higher than the wild-type strain. Eight successive subcultures of these two strains indicated their hereditary stability. The findings demonstrated that SC-CO₂, in combination with specific additives, could effectively increase the activity of enzymes produced by microorganisms when their spores were treated.

Downloads

Download data is not yet available.

References

Brunner, G. (2005). Supercritical fluids: Technology and application to food processing. Journal of Food Engineering, 67(1), 21–33. https://doi.org/10.1016/j.jfoodeng.2004.05.060

Calvo, L., & Casas, J. (2018). Sterilization of biological weapons in technical clothing and sensitive material by high-pressure CO2 and water. Industrial & Engineering Chemistry Research, 57(13), 4680–4687. https://doi.org/10.1021/acs.iecr.7b04794

Cheftel, J. C. (1995). Review: High-pressure, microbial inactivation and food preservation. Food Science and Technology International, 1(2-3), 75–90. https://doi.org/10.1177/108201329500100203

Damar, S., & Balaban, M. O. (2006). Review of dense phase CO2 technology: Microbial and enzyme inactivation, and effects on food quality. Journal of Food Science, 71(1), R1–R11. https://doi.org/10.1111/j.1365-2621.2006.tb12397.x

da Silva, M. A., de Araujo, A. P., de Souza Ferreira, J., & Kieckbusch, T. G. (2016). Inactivation of Bacillus subtilis and Geobacillus stearothermophilus inoculated over metal surfaces using supercritical CO2 process and nisin. The Journal of Supercritical Fluids, 109, 87–94. https://doi.org/10.1016/j.supflu.2015.11.013

Dillow, A. K., Dehghani, F., Hrkach, J. S., Foster, N. R., & Langer, R. (1999). Bacterial inactivation by using near- and supercritical carbon dioxide. Proceedings of the National Academy of Sciences, 96(18), 10344–10348. https://doi.org/10.1073/pnas.96.18.10344

Efaq, A. N., Rahman, N. N. N. A., Nagao, H., Al-Gheethi, A. A., & Kadir, M. O. A. (2017). Inactivation of Aspergillus spores in clinical wastes by supercritical carbon dioxide. Arabian Journal for Science and Engineering, 42(1), 39–51. https://doi.org/10.1007/s13369-016-2087-5

Foster, N., Mammucari, R., Dehghani, F., Barrett, A., Bezanehtak, K., Coen, E., Combes, G., Meure, L., Ng, A., Regtop, H. L., & Tandya, A. (2003). Processing pharmaceutical compounds using dense gas technology. Industrial & Engineering Chemistry Research, 42(25), 6476–6493. https://doi.org/10.1021/ie030219x

Furukawa, S., Watanabe, T., Koyama, T., Hirata, J., Narisawa, N., Ogihara, H., & Yamasaki, M. (2006). Effect of high pressure carbon dioxide on the clumping of the bacterial spores. International Journal of Food Microbiology, 106(1), 95–98. https://doi.org/10.1016/j.ijfoodmicro.2005.05.016

Garcia-Gonzalez, L., Geeraerd, A. H., Spilimbergo, S., Elst, K., van Ginneken, L., Debevere, J., & Devlieghere, F. (2007). High pressure carbon dioxide inactivation of microorganisms in foods: The past, the present and the future. International Journal of Food Microbiology, 117(1), 1–28. https://doi.org/10.1016/j.ijfoodmicro.2007.02.018

Gong, Z., Yue, P., Chen, Y., Li, W., Zhao, L., & Hu, D. (2023). Structure-tunable polyether block amide/polylactic acid foams with shape memory performance using supercritical CO2 foaming. The Journal of Supercritical Fluids, 203, Artigo 106090. https://doi.org/10.1016/j.supflu.2023.106090

Haas, G. J., Prescott, J. R., Dudley, E., Dik, R., Hintlian, C., & Keane, L. (1989). Inactivation of microorganisms by carbon dioxide under pressure. Journal of Food Safety, 9(4), 253–265. https://doi.org/10.1111/j.1745-4565.1989.tb00525.x

Hemmer, J. D., Drews, M. J., LaBerge, M., & Matthews, M. A. (2007). Sterilization of bacterial spores by using supercritical carbon dioxide and hydrogen peroxide. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 80B(2), 511–518. https://doi.org/10.1002/jbm.b.30625

Ishikawa, H., Shimoda, M., Tamaya, K., Yonekura, A., Kawano, T., & Osajima, Y. (1997). Inactivation of Bacillus spores by the supercritical carbon dioxide micro-bubble method. Bioscience, Biotechnology, and Biochemistry, 61(6), 1022–1023. https://doi.org/10.1271/bbb.61.1022

Kamihira, M., Taniguchi, M., & Kobayashi, T. (1987). Sterilization of microorganisms with supercritical and liquid carbon dioxide. Agricultural and Biological Chemistry, 51(2), 407–412. https://doi.org/10.1080/00021369.1987.10858139

Obaidullah, A. J., & Almehizia, A. A. (2023). Machine learning-based prediction and mathematical optimization of capecitabine solubility through the supercritical CO2 system. Journal of Molecular Liquids, 391, Artigo 123229. https://doi.org/10.1016/j.molliq.2023.123229

Seok, P. H., & Kim, K. H. (2013). Enhancement of supercritical CO2 inactivation of spores of Penicillium oxalicum by ethanol cosolvent. Journal of Microbiology and Biotechnology, 23(6), 833–836. https://doi.org/10.4014/jmb.1211.11072

Shieh, E., Paszczynski, A., Wai, C. M., Lang, Q., & Crawford, R. L. (2009). Sterilization of Bacillus pumilus spores using supercritical fluid carbon dioxide containing various modifier solutions. Journal of Microbiological Methods, 76(3), 247–252. https://doi.org/10.1016/j.mimet.2008.11.005

Soares, G. C., Learmonth, D. A., Vallejo, M. C., Davila, S. P., González, P., Sousa, R. A., & Oliveira, A. L. (2019). Supercritical CO2 technology: The next standard sterilization technique? Materials Science and Engineering: C, 99, 520–540. https://doi.org/10.1016/j.msec.2019.01.121

Spilimbergo, S., & Bertucco, A. (2003). Non-thermal bacteria inactivation with dense CO2. Biotechnology and Bioengineering, 84(6), 627–638. https://doi.org/10.1002/bit.10783

White, A., Burns, D., & Christensen, T. W. (2006). Effective terminal sterilization using supercritical carbon dioxide. Journal of Biotechnology, 123(4), 504–515. https://doi.org/10.1016/j.jbiotec.2005.12.033

Yan, H. D., ShenTu, Z. P., & Qian, J. Q. (2013). Mutation breeding of amylase-producing strain Bacillus subtilis by supercritical CO2. Bulletin of Fermentation Science and Technology, 42(3), 5–8.

Yoo, Y. J., Hong, J., & Hatch, R. T. (1987). Comparison of ?-amylase activities from different assay methods. Biotechnology and Bioengineering, 30(1), 147–151. https://doi.org/10.1002/bit.260300120

Zhang, J., Burrows, S., Gleason, C., Matthews, M. A., Drews, M. J., LaBerge, M., & An, Y. H. (2006b). Sterilizing Bacillus pumilus spores using supercritical carbon dioxide. Journal of Microbiological Methods, 66(3), 479–485. https://doi.org/10.1016/j.mimet.2006.01.012

Zhang, J., Dalal, N., Gleason, C., Matthews, M. A., Waller, L. N., Fox, K. F., Fox, A., & An, Y. H. (2006c). On the mechanisms of deactivation of Bacillus atrophaeus spores using supercritical carbon dioxide. The Journal of Supercritical Fluids, 38(2), 268–273. https://doi.org/10.1016/j.supflu.2006.02.015

Zhang, J., Dalal, N., Matthews, M. A., Waller, L. N., Saunders, C., Fox, K. F., & Fox, A. (2007). Supercritical carbon dioxide and hydrogen peroxide cause mild changes in spore structures associated with high killing rate of Bacillus anthracis. Journal of Microbiological Methods, 70(3), 442–451. https://doi.org/10.1016/j.mimet.2007.05.019

Zhang, J., Davis, T. A., Matthews, M. A., Drews, M. J., LaBerge, M., & An, Y. H. (2006a). Sterilization using high-pressure carbon dioxide. The Journal of Supercritical Fluids, 38(3), 354–372. https://doi.org/10.1016/j.supflu.2005.05.005

Zhang, Q. Y., Qian, J. Q., Guo, H., & Yang, S. L. (2008). Supercritical CO2: A novel environmentally friendly mutagen. Journal of Microbiological Methods, 75(1), 25–28. https://doi.org/10.1016/j.mimet.2008.04.007

Zhang, Q. Y., Qian, J. Q., & Ma, L. Z. (2013). Mutation breeding of lipase-producing strain Flavobacterium sp. by supercritical CO2 with hydrazine hydrate. Brazilian Archives of Biology and Technology, 56(4), 541–546. https://doi.org/10.1590/S1516-89132013000400003