Efficient Extraction and Optimization of Amylase and Protease from Bacillus Species: A Comprehensive Study

Authors

  • Aastha Acharya Department of Biology and Microbiology, South Dakota State University, Brookings, SD, 57007
  • Samiran Subedi Central Department of Biotechnology, Tribhuwan University, Kirtipur, Nepal

DOI:

https://doi.org/10.38211/joarps.2025.06.304

Keywords:

Bacillus, amylase, protease, rhizosphere, fermentation substrates, microbial enzymes

Abstract

This review paper provides a comprehensive analysis of the crucial role of Bacillus species in the production of amylase and protease enzymes. Bacillus is one of the most important bacterial sources of enzymes, known for producing high yields of neutral and alkaline amylases and proteases. By breaking down complex starches and proteins into simpler, functional components, these enzymes play a critical role in enhancing efficiency and product quality across various industries, including food processing and textiles. These enzymes exhibit remarkable stability even under extreme conditions, such as high temperatures, fluctuating pH levels, and exposure to organic solvents, detergents, and oxidizing agents. Some Bacillus strains are highly efficient at secreting large quantities of extracellular enzymes, making them essential for industrial enzyme production. The primary objective of this review is to examine the processes involved in the production, purification, and characterization of amylases and proteases from Bacillus species, with a particular focus on those found in the rhizosphere. The first section of the paper discusses the properties and functions of these exoenzymes and highlights key factors that influence their optimization. The second section explores the use of cost-effective substrates for fermentation, aiming to enhance enzyme activity and stability. Given the extensive applications of amylases and proteases in various sectors, significant research has been dedicated to improving their production and efficiency. This review aims to advance our understanding of microbial enzyme systems and support the development of enhanced enzyme production strategies for emerging biotechnological applications.

Views: 1346Downloads: 822

Downloads

Download data is not yet available.

References

Abd-Elgawad, M. M. (2024). Revolutionizing Nematode Management–Nanomaterials as a Promising Approach for Managing Economically Important Plant-Parasitic Nematodes: Current Knowledge and Future Challenges. Nanotechnology and Plant Disease Management, 23-44. DOI: https://doi.org/10.1201/9781003256762-2

Abo-Kamer, A. M., Abdelaziz, A. A., Elkotb, E. S., & Al-Madboly, L. A. (2025). Production and characterization of a promising microbial-derived lipase enzyme targeting BCL-2 gene expression in hepatocellular carcinoma. Microbial Cell Factories, 24, 58. DOI: https://doi.org/10.1186/s12934-025-02671-7

Acharya, A., Khanal, A., Bajracharya, M. R., Timalsina, A., Bishwokarma, A., & Basnet, A. . (2019). Production, purification and optimisation of amylase by submerged fermentation using Bacillus subtilis. Int. J. Sci. Res. Sci. Eng. Technol, 6, 265-275. DOI: https://doi.org/10.32628/IJSRSET196122

Acharya, A., & Subedi, S. Efficient Extraction and Optimization of Amylase and Protease from Bacillus Species: A Comprehensive Study.

Al-Sharify, Z. T., Al-Najjar, S. Z., Naser, Z. A., Alsherfy, Z. A. I., & Onyeaka, H. (2025). The Impact of Fluid Flow on Microbial Growth and Distribution in Food Processing Systems. Foods, 14(3), 401. DOI: https://doi.org/10.3390/foods14030401

Andersen, P., & Doherty, T. M. (2005). The success and failure of BCG—implications for a novel tuberculosis vaccine. Nature Reviews Microbiology, 3(8), 656-662. DOI: https://doi.org/10.1038/nrmicro1211

Aygan, A., Arikan, B., Korkmaz, H., Dinçer, S., & Çolak, Ö. (2008). Highly thermostable and alkaline α-amylase from a halotolerant-alkaliphilic Bacillus sp. AB68. Brazilian Journal of Microbiology, 39, 547-553. DOI: https://doi.org/10.1590/S1517-83822008000300027

Balser, T. C., Wixon, D., Moritz, L. K., & Lipps, L. (2010). The microbiology of natural soils. Soil microbiology and sustainable crop production, 27-57. DOI: https://doi.org/10.1007/978-90-481-9479-7_2

Banerjee, R., & Bhattacharyya, B. (1992). Extracellular alkaline protease of newly isolated Rhizopus oryzae. Biotechnology letters, 14, 301-304. DOI: https://doi.org/10.1007/BF01022328

Bareither, R., & Pollard, D. (2011). A review of advanced small‐scale parallel bioreactor technology for accelerated process development: Current state and future need. Biotechnology progress, 27(1), 2-14. DOI: https://doi.org/10.1002/btpr.522

Bernfeld, P. (1986). Amylases, A and B methodology. Enzymology, 1, 149-158. DOI: https://doi.org/10.1016/0076-6879(55)01021-5

Blanco, A. S., Durive, O. P., Pérez, S. B., Montes, Z. D., & Guerra, N. P. (2016). Simultaneous production of amylases and proteases by Bacillus subtilis in brewery wastes. Brazilian Journal of Microbiology, 47, 665-674. DOI: https://doi.org/10.1016/j.bjm.2016.04.019

Boominadhan, U., Rajakumar, R., Sivakumaar, P. K. V., & Joe, M. M. (2009). Optimization of protease enzyme production using Bacillus sp. isolated from different wastes. Bot. Res. Int, 2(2), 83-87.

Boyno, G., Ansari, R. A., Durak, E. D., Güneş, H., Çevik, R., & Demir, S. (2024). Arbuscular mycorrhizal technology in sustainable agriculture: Current knowledge and challenges in agroforestry. Mycorrhizal Symbiosis and Agroecosystem Restoration, 173-195. DOI: https://doi.org/10.1007/978-981-99-5030-0_8

Cercignani, G., Serra, M. C., Fini, C., Natalini, P., Palmerini, C. A., Magni, G., & Ipata, P. L. (1974). Properties of 5'-nucleotidase from Bacillus cereus obtained by washing intact cells with water. Biochemistry, 13(17), 3628-3634. DOI: https://doi.org/10.1021/bi00714a035

Chahal, D. (1983). Growth characteristics of microorganisms in solid state fermentation for upgrading of protein values of lignocelluloses and cellulase production. In. ACS Publications. DOI: https://doi.org/10.1021/bk-1983-0207.ch020

Cherdyntseva, T., Rasinkov, V., & Egorov, N. (1982). An Exocellular Protease With the Activity of Coagulase Synthesized by Bacillus subtilis var. amyloliquefaciens in the Absence of Nitrogen Sources in the Medium. MIKROBIOLOGIYA., 51(3), 431-435.

Choudhary, D. K., & Johri, B. N. (2009). Interactions of Bacillus spp. and plants–with special reference to induced systemic resistance (ISR). Microbiological research, 164(5), 493-513. DOI: https://doi.org/10.1016/j.micres.2008.08.007

Claus, D., & Berkeley, R. (1986). Genus Bacillus in Bergeys manual of systematic bacteriology. The Willams and Wikins company Baltimore.

Coleman, G., & Elliott, W. (1962). Studies on α-amylase formation by Bacillus subtilis. Biochemical Journal, 83(2), 256. DOI: https://doi.org/10.1042/bj0830256

Danilova, I., & Sharipova, M. (2020). The practical potential of bacilli and their enzymes for industrial production. Frontiers in Microbiology, 11, 1782. DOI: https://doi.org/10.3389/fmicb.2020.01782

Das, G., & Prasad, M. (2010). Isolation, purification & mass production of protease enzyme from Bacillus subtilis. Int. Res. J. Microbiol, 1(2), 26-31.

Deb, P., Talukdar, S. A., Mohsina, K., Sarker, P. K., & Sayem, S. A. (2013). Production and partial characterization of extracellular amylase enzyme from Bacillus amyloliquefaciens P-001. SpringerPlus, 2, 1-12. DOI: https://doi.org/10.1186/2193-1801-2-154

Devi, S., Khaund, P., & Joshi, S. (2010). Thermostable α-amylase from natural variants of Bacillus spp. prevalent in eastern Himalayan range. African Journal of Microbiology Research, 4(23), 2534-2542.

Devi, T., Sistla, S., Khan, R. T., Kailoo, S., Bhardwaj, M., & Rasool, S. (2025). Purification and characterization of detergent stable alkaline lipase from Bacillus safensis TKW3 isolated from Tso Kar brackish water lake. PeerJ, 13, e18921. DOI: https://doi.org/10.7717/peerj.18921

Duckworth, A. W., Grant, W. D., Jones, B. E., & Van Steenbergen, R. (1996). Phylogenetic diversity of soda lake alkaliphiles. FEMS microbiology ecology, 19(3), 181-191. DOI: https://doi.org/10.1111/j.1574-6941.1996.tb00211.x

Erkılıç, T. O., & Bayraktar, B. (2025). Effect of Jackfruıt Pulp Extract on Serum Nesfatın-1 and Vısfatın Levels in Streptozotocın-induced Rats. Indian Journal of Animal Research, 59(2). DOI: https://doi.org/10.18805/IJAR.BF-1902

Esmeralda-Guzmán, M., Juan, J. B.-F., Aguilar, C. N., Juan, A. A.-V., & Sepúlveda, L. (2024). Parameters that influence the fermentation submerged process: bioprocess development in biotechnology. Bioresources and Bioprocess in Biotechnology for a Sustainable Future, 179-200. DOI: https://doi.org/10.1201/9781003410041-12

Farrar, J., Hawes, M., Jones, D., & Lindow, S. (2003). HOW ROOTS CONTROL THE FLUX OF CARBON TO THE RHIZOSPHERE. Ecology, 84(4), 827-837. https://doi.org/https://doi.org/10.1890/0012-9658(2003)084[0827:HRCTFO]2.0.CO;2 DOI: https://doi.org/10.1890/0012-9658(2003)084[0827:HRCTFO]2.0.CO;2

García-Moyano, A., Larsen, Ø., Gaykawad, S., Christakou, E., Boccadoro, C., Puntervoll, P., & Bjerga, G. E. K. (2020). Fragment exchange plasmid tools for CRISPR/Cas9-mediated gene integration and protease production in Bacillus subtilis. Applied and Environmental Microbiology, 87(1), e02090-02020. DOI: https://doi.org/10.1128/AEM.02090-20

Gorrab, A., Ouertani, R., Hammami, K., Souii, A., Kallel, F., Masmoudi, A. S.,…Neifar, M. (2024). In silico and experimental characterization of a new polyextremophilic subtilisin-like protease from Microbacterium metallidurans and its application as a laundry detergent additive. 3 Biotech, 14(9), 200. DOI: https://doi.org/10.1007/s13205-024-04043-1

Gupta, R., Beg, Q., Khan, S., & Chauhan, B. (2002). An overview on fermentation, downstream processing and properties of microbial alkaline proteases. Applied microbiology and biotechnology, 60, 381-395. DOI: https://doi.org/10.1007/s00253-002-1142-1

Horikoshi, K. (1971). Production of alkaline enzymes by alkalophilic microorganisms: part i. alkaline protease produced by Bacillus no. 221. Agricultural and Biological Chemistry, 35(9), 1407-1414. DOI: https://doi.org/10.1080/00021369.1971.10860094

Ibrahim, S. S., Al-dulaimi, F., & Abdulrahman, H. (2025). Evaluation of Saccharomyces Cerevisiae Efficiency in Producing Ethanol and CO2 from Maize and Sorghum. IOP Conference Series: Earth and Environmental Science, DOI: https://doi.org/10.1088/1755-1315/1449/1/012069

Jaizme-Vega, M., Rodríguez-Romero, A., Hermoso, C. M., & Declerck, S. (2003). Growth of micropropagated bananas colonized by root-organ culture produced arbuscular mycorrhizal fungi entrapped in Ca-alginate beads. Plant and Soil, 254, 329-335. DOI: https://doi.org/10.1023/A:1025523632413

Janiga-MacNelly, A., & Lavado, R. (2024). Unveiling the mysteries: a deep dive into extracellular polymeric substances in activated sludge systems of wastewater treatment plants. Environmental Technology Reviews, 13(1), 572-596. DOI: https://doi.org/10.1080/21622515.2024.2391079

Jisha, V. N., Smitha, R. B., Pradeep, S., Sreedevi, S., Unni, K. N., Sajith, S.,…Benjamin, S. (2013). Versatility of microbial proteases. Advances in enzyme research, 1(3), 39-51. DOI: https://doi.org/10.4236/aer.2013.13005

Jones, D. L., Hodge, A., & Kuzyakov, Y. (2004). Plant and mycorrhizal regulation of rhizodeposition. New phytologist, 163(3), 459-480. DOI: https://doi.org/10.1111/j.1469-8137.2004.01130.x

Joo, H.-S., & Chang, C.-S. (2005). Production of protease from a new alkalophilic Bacillus sp. I-312 grown on soybean meal: optimization and some properties. Process Biochemistry, 40(3-4), 1263-1270. DOI: https://doi.org/10.1016/j.procbio.2004.05.010

Joshi, A., & Ball, B. (1993). Extracellular alkaline enzymes of facultative bacteria of CaCO~ 3 kilns near Jabalpur. Indian Journal of Microbiology, 33, 179-179.

Kaissar, F. Z., Benine, M. L., Abbouni, B., Saadi, S. A., Emiliani, G., Barberini, S., & Lebouachera, S. E. I. (2025). Revealing of multifunctional newly isolated Bacillus strains from Algerian wastes with high pectinase activity for sustainable biotechnological application. Biologia, 1-16. DOI: https://doi.org/10.1007/s11756-025-01880-0

Kanekar, P., Nilegaonkar, S., Sarnaik, S., & Kelkar, A. (2002). Optimization of protease activity of alkaliphilic bacteria isolated from an alkaline lake in India. Bioresource technology, 85(1), 87-93. DOI: https://doi.org/10.1016/S0960-8524(02)00018-4

Kapoor, R., Sharma, D., & Bhatnagar, A. (2008). Arbuscular mycorrhizae in micropropagation systems and their potential applications. Scientia Horticulturae, 116(3), 227-239. DOI: https://doi.org/10.1016/j.scienta.2008.02.002

Kochi, A. (1994). Tuberculosis: distribution, risk factors, mortality. Immunobiology, 191(4-5), 325-336. DOI: https://doi.org/10.1016/S0171-2985(11)80437-7

Levine, N. D. (1975). Buchanan, RE & Gibbons, NE, eds. 1974. Bergey's Manual of Determinative Bacteriology. Williams & Wilkins Co., Baltimore, Md. 21202. xxvi+ 1246 pp. $45.00. In: Wiley Online Library. DOI: https://doi.org/10.1111/j.1550-7408.1975.tb00935.x

Liu, X., Wanika, L., Chappell, M., & Branke, J. Efficiently Establishing Practical Identifiability Via Active Learning. Available at SSRN 5125554.

Lyngwi, N. A., & Joshi, S. (2014). Economically important Bacillus and related genera: a mini review. Biology of useful plants and microbes, 3, 33-43.

Mathur, P., & Mukherjee, S. (2025). Rhizosphere Engineering and Stress Resilience in Plants: Concepts and Applications. DOI: https://doi.org/10.1201/9781003354109

Mishra, S., & Behera, N. (2008). Amylase activity of a starch degrading bacteria isolated from soil receiving kitchen wastes. African journal of Biotechnology, 7(18).

Mrudula, S. (2024). A Review on Microbial Alkaline Proteases: Optimization of Submerged Fermentative Production, Properties, and Industrial Applications. Applied Biochemistry and Microbiology, 60(3), 383-401. DOI: https://doi.org/10.1134/S0003683823602767

Mukendi, M. D., Salami, O. S., & Mketo, N. (2025). An In-Depth Review of Molecularly Imprinted Electrochemical Sensors as an Innovative Analytical Tool in Water Quality Monitoring: Architecture, Principles, Fabrication, and Applications. Micromachines, 16(3), 251. DOI: https://doi.org/10.3390/mi16030251

Mulimani, V., & GN Patil, R. (2000). α-Amylase production by solid state fermentation: a new practical approach to biotechnology courses. Biochemical Education, 28(3), 161-163. DOI: https://doi.org/10.1111/j.1539-3429.2000.tb00056.x

Nagata, Y., Yamaguchi, K., & Maruo, B. (1974). Genetic and biochemical studies on cell-bound α-amylase in Bacillus subtilis Marburg. Journal of Bacteriology, 119(2), 425-430. DOI: https://doi.org/10.1128/jb.119.2.425-430.1974

Nannipieri, P., GRECO, S., & Ceccanti, B. (2017). Ecological significance of the biological activity in soil. Soil biochemistry, 293-356. DOI: https://doi.org/10.1201/9780203739389-6

Negi, R., Yadav, N., & Yadav, A. N. (2025). Microbial biofertilizers: A paradigm shift towards agricultural sustainability. Biologia, 1-26. DOI: https://doi.org/10.1007/s11756-024-01848-6

Nie, M., Khalid, F., Hu, Q., Khalid, A., Wu, Q., Huang, S., & Wang, Z. (2024). Site-Directed Mutagenesis: Improving the Acid Resistance and Thermostability of Bacillus velezensis α-Amylase and Its Preliminary Feed Application. Journal of Agricultural and Food Chemistry, 72(18), 10487-10496. DOI: https://doi.org/10.1021/acs.jafc.4c01630

Nielsen, P., Fritze, D., & Priest, F. G. (1995). Phenetic diversity of alkaliphilic Bacillus strains: proposal for nine new species. Microbiology, 141(7), 1745-1761. DOI: https://doi.org/10.1099/13500872-141-7-1745

Niyomukiza, S., Owino, W., Maina, J. M., Maina, N., & Issifu, M. (2022). Concomitant production of α-amylase and protease by Bacillus Aerius strain FPWSHA isolated from food wastes. Biointerface Res Appl Chem, 13, 1-15. DOI: https://doi.org/10.33263/BRIAC134.310

Ogino, H., Otsubo, T., & Ishikawa, H. (2008). Screening, purification, and characterization of a leather-degrading protease. Biochemical Engineering Journal, 38(2), 234-240. DOI: https://doi.org/10.1016/j.bej.2007.07.008

Osunde, M., ABIOLA, O., PHILIP, S., ADEWALE, O., & BAKARE, M. Extracellular Thermophilic Α-Amylase from Bacillus Substilis Derived from Cassava Waste Water. Available at SSRN 5134155.

Outtrup, H., & Jørgensen, S. T. (2002). The importance of Bacillus species in the production of industrial enzymes. Applications and systematics of Bacillus and relatives, 206-218. DOI: https://doi.org/10.1002/9780470696743.ch14

Pant, B. D., Pant, P., Erban, A., Huhman, D., Kopka, J., & SCHEIBLE, W. R. (2015). Identification of primary and secondary metabolites with phosphorus status‐dependent abundance in A rabidopsis, and of the transcription factor PHR 1 as a major regulator of metabolic changes during phosphorus limitation. Plant, cell & environment, 38(1), 172-187. DOI: https://doi.org/10.1111/pce.12378

Petit, C., Caudal, F., Taupin, L., Dufour, A., Le Ker, C., Giudicelli, F.,…Bazire, A. (2024). Antibiofilm Activity of the Marine Probiotic Bacillus subtilis C3 Against the Aquaculture-Relevant Pathogen Vibrio harveyi. Probiotics and Antimicrobial Proteins, 1-12. DOI: https://doi.org/10.1007/s12602-024-10229-z

Priest, F. G. (1977). Extracellular enzyme synthesis in the genus Bacillus. Bacteriological reviews, 41(3), 711-753. DOI: https://doi.org/10.1128/br.41.3.711-753.1977

Radhakrishnan, R., Hashem, A., & Abd_Allah, E. F. (2017). Bacillus: A biological tool for crop improvement through bio-molecular changes in adverse environments. Frontiers in physiology, 8, 667. DOI: https://doi.org/10.3389/fphys.2017.00667

Rani, T. S., Takahashi, D., Mukherjee, S., Uemura, M., Madhuprakash, J., & Podile, A. R. (2025). Secretome analysis of the chitinolytic machinery of Chitiniphilus shinanonensis and its implication in chitooligosaccharide production. Carbohydrate Polymers, 353, 123272. DOI: https://doi.org/10.1016/j.carbpol.2025.123272

Rashid, N., Imanaka, H., Fukui, T., Atomi, H., & Imanaka, T. (2004). Presence of a novel phosphopentomutase and a 2-deoxyribose 5-phosphate aldolase reveals a metabolic link between pentoses and central carbon metabolism in the hyperthermophilic archaeon Thermococcus kodakaraensis. Journal of Bacteriology, 186(13), 4185-4191. DOI: https://doi.org/10.1128/JB.186.13.4185-4191.2004

Rawal, P., Pandey, B., Yadav, R. K., & Panta, S. (2025). Antioxidant, Alpha‐Amylase Inhibitory and Hypoglycemic Activity of Smallanthus sonchifolius Leaves from Nepal: An Integrated In Vitro, In Vivo, and In Silico Approach. Food Science & Nutrition, 13(1), e4672. DOI: https://doi.org/10.1002/fsn3.4672

Reed Rodrigues Coelho, R., & Drozdowicz, A. (1978). The occurrence of Actinomycetes in a cerrado soil in Brazil.

Saeed, H., Ragaey, A., Samy, Z., Ashraf, V., ElMostafa, A., Ahmad, N.,…Bakry, A. (2025). Optimization and characterization studies of poultry waste valorization for peptone production using a newly Egyptian Bacillus subtilis strain. AMB Express, 15(1), 9. DOI: https://doi.org/10.1186/s13568-024-01794-1

Sandler, N., & Keynan, A. (1988). Membrane binding and release of Bacillus subtilis DNA as a function of the cell cycle. Microbiology, 134(5), 1155-1163. DOI: https://doi.org/10.1099/00221287-134-5-1155

Sargent, M., & Lampen, J. (1970). A mechanism for penicillinase secretion in Bacillus licheniformis. Proceedings of the National Academy of Sciences, 65(4), 962-969. DOI: https://doi.org/10.1073/pnas.65.4.962

Schäfer, M., Schmitz, C., Facius, R., Horneck, G., Milow, B., Funken, K. H., & Ortner, J. (2000). Systematic study of parameters influencing the action of rose bengal with visible light on bacterial cells: comparison between the biological effect and singlet‐oxygen production. Photochemistry and photobiology, 71(5), 514-523. DOI: https://doi.org/10.1562/0031-8655(2000)071<0514:SSOPIT>2.0.CO;2

Schallmey, M., Singh, A., & Ward, O. P. (2004). Developments in the use of Bacillus species for industrial production. Canadian journal of microbiology, 50(1), 1-17. DOI: https://doi.org/10.1139/w03-076

Seidler, Y., Rimbach, G., Lüersen, K., Vinderola, G., & Ipharraguerre, I. R. (2024). The postbiotic potential of Aspergillus oryzae–a narrative review. Frontiers in Microbiology, 15, 1452725. DOI: https://doi.org/10.3389/fmicb.2024.1452725

Sen, S., Mandal, B., & Dasu, V. (2009). Alkaline protease: a tool to clean environment. AIChE Spring National Meeting,

Septian, P. F., Budiyanto, D., & Sumaryam, S. (2025). Pengaruh Pemberian Probiotik Biolacto dengan Dosis yang Berbeda pada Pakan terhadap Pertumbuhan Berat Mutlak Udang Vaname (Lithopenaeus Vannamei) DOC 80 Hari. Mikroba: Jurnal Ilmu Tanaman, Sains Dan Teknologi Pertanian, 2(1), 210-224. DOI: https://doi.org/10.62951/mikroba.v2i1.290

Silva, A. S., Neves, K. M., Freitas, R. F., das Chagas, T. P., Salay, L. C., da Silva, E. G. P.,…da Costa, A. M. (2025). Valorization of Cocoa and Peach-Palm Wastes for the Production of Amylases by Pleurotus pulmonarius CCB19 and Its Application as an Additive in Commercial Detergents. Applied Biochemistry and Biotechnology, 1-24. DOI: https://doi.org/10.1007/s12010-024-05147-3

Su, Y., Liu, C., Fang, H., & Zhang, D. (2020). Bacillus subtilis: a universal cell factory for industry, agriculture, biomaterials and medicine. Microbial Cell Factories, 19, 1-12. DOI: https://doi.org/10.1186/s12934-020-01436-8

Sun, B., Zou, K., Zhao, Y., Tang, Y., Zhang, F., Chen, W.,…Zheng, Y. (2023). The fermentation optimization for alkaline protease production by Bacillus subtilis BS-QR-052. Frontiers in Microbiology, 14, 1301065. DOI: https://doi.org/10.3389/fmicb.2023.1301065

Switzer, R. L., & Garrity, L. F. (1999). Experimental biochemistry. Macmillan.

Syarifah, S. M., Aripin, A. M., Ishak, N., Anindita, N. S., Abdul-Wahab, M. F., & Kassim, A. S. M. (2025). Partial Purification of Bacillus cereus Enzyme Expression for Bio-Pulping of Lignin Degraders Isolated from Coptotermus curvignathus. Engineering Proceedings, 84(1), 41. DOI: https://doi.org/10.3390/engproc2025084041

Taipa, M. Â., Fernandes, P., & de Carvalho, C. C. (2019). Production and purification of therapeutic enzymes. Therapeutic Enzymes: Function and Clinical Implications, 1-24. DOI: https://doi.org/10.1007/978-981-13-7709-9_1

Tejnecký, V., Luláková, P., Šantrůčková, H., Křížová, P., Lehejček, J., Hájek, T.,…Drábek, O. (2025). Arctic willow (Salix polaris) exudation as a driver of microbial activity and soil formation in the high arctic tundra. Biogeochemistry, 168(2), 1-22. DOI: https://doi.org/10.1007/s10533-025-01222-x

Terui, G. (1973). Kinetics of hydrolase production by microorganisms. In Microbial Engineering (pp. 377-395). Elsevier. DOI: https://doi.org/10.1016/B978-0-408-70549-3.50012-3

Tsotetsi, T., Nephali, L., Malebe, M., & Tugizimana, F. (2022). Bacillus for plant growth promotion and stress resilience: what have we learned? Plants, 11(19), 2482. DOI: https://doi.org/10.3390/plants11192482

Vasantha-Srinivasan, P., Park, K. B., Kim, K. Y., Jung, W.-J., & Han, Y. S. (2025). The role of Bacillus species in the management of plant-parasitic nematodes. Frontiers in Microbiology, 15, 1510036. DOI: https://doi.org/10.3389/fmicb.2024.1510036

Vijayalakshmi, K., Abha, S., & Chander, P. (2012). Isolation and characterization of Bacillus subtilis KC3 for amylolytic activity. International journal of bioscience, biochemistry and bioinformatics, 2(5), 336-341. DOI: https://doi.org/10.7763/IJBBB.2012.V2.128

Vijayalakshmi, S., Ranjitha, J., & Rajeswari, V. D. (2013). Enzyme production ability by Bacillus subtilis and Bacillus licheniformis-A comparative study. Asian J. Pharm. Clin. Res, 6(4), 29-32.

Wang, B., Kes, M. B., van Saparoea, A. C. v. d. B., Dugar, G., Luirink, J., & Hamoen, L. W. (2024). Inactivation of the conserved protease LonA increases production of xylanase and amylase in Bacillus subtilis. Microbial Cell Factories, 23(1), 335. DOI: https://doi.org/10.1186/s12934-024-02616-6

Wang, M., Zhao, W., Zhou, D., & Huang, J. (2025). Bacillus subtilis Contributes to Amylase Production in the Honey Sac of Apis mellifera. Insects, 16(2), 221. DOI: https://doi.org/10.3390/insects16020221

Webb, C. D., Teleman, A., Gordon, S., Straight, A., Belmont, A., Lin, D. C.-H.,…Losick, R. (1997). Bipolar localization of the replication origin regions of chromosomes in vegetative and sporulating cells of B. subtilis. Cell, 88(5), 667-674. DOI: https://doi.org/10.1016/S0092-8674(00)81909-1

Xiang, M., Kang, Q., & Zhang, D. (2020). Advances on systems metabolic engineering of Bacillus subtilis as a chassis cell. Synthetic and systems biotechnology, 5(4), 245-251. DOI: https://doi.org/10.1016/j.synbio.2020.07.005

Xin, X., Li, L., Cheng, J., Wang, Y., Lu, B., Yang, Y.,…Wong, J. W. (2025). Synchronous production of bioethanol and short-chain fatty acids associated with microbial mechanisms via the short-term cultivation of waste molasses inoculated with Aspergillus oryzae. Journal of Environmental Management, 380, 124888. DOI: https://doi.org/10.1016/j.jenvman.2025.124888

Xu, X., & Kovács, Á. T. (2024). How to identify and quantify the members of the Bacillus genus? Environmental Microbiology, 26(2), e16593. DOI: https://doi.org/10.1111/1462-2920.16593

Yabuki, M., & Fukui, S. (1970). Presence of binding site for α-amylase and of masking protein for this site on mycelial cell wall of Aspergillus oryzae. Journal of Bacteriology, 104(1), 138-144. DOI: https://doi.org/10.1128/jb.104.1.138-144.1970

Yadav, P., Anu, Kumar Tiwari, S., Kumar, V., Singh, D., Kumar, S.,…Singh, B. (2024). Sugarcane bagasse: An important lignocellulosic substrate for production of enzymes and biofuels. Biomass Conversion and Biorefinery, 14(5), 6111-6142. DOI: https://doi.org/10.1007/s13399-022-02791-9

Yang, H., Qu, J., Zou, W., Shen, W., & Chen, X. (2021). An overview and future prospects of recombinant protein production in Bacillus subtilis. Applied microbiology and biotechnology, 105(18), 6607-6626. DOI: https://doi.org/10.1007/s00253-021-11533-2

Yousten, A. A., & Nelson, K. D. (1976). A single form of superoxide dismutase found in Bacillus popilliae and in some other Gram-positive bacteria. The Journal of General and Applied Microbiology, 22(3), 161-164. DOI: https://doi.org/10.2323/jgam.22.161

Zhang, J., Zhu, B., Li, X., Xu, X., Li, D., Zeng, F.,…Lu, F. (2022). Multiple modular engineering of Bacillus amyloliquefaciens cell factories for enhanced production of alkaline proteases from B. clausii. Frontiers in Bioengineering and Biotechnology, 10, 866066. DOI: https://doi.org/10.3389/fbioe.2022.866066

Downloads

Published

30-05-2025

How to Cite

Efficient Extraction and Optimization of Amylase and Protease from Bacillus Species: A Comprehensive Study (A. Acharya & S. Subedi, Trans.). (2025). Journal of Applied Research in Plant Sciences, 6(1), 34-47. https://doi.org/10.38211/joarps.2025.06.304

Similar Articles

1-10 of 19

You may also start an advanced similarity search for this article.