本研究探討利用地衣芽孢桿菌（Bacillus licheniformis）建立一套可有效分解豆渣的綠色循環模組，以促進食品副產物的資源化與永續利用。豆渣為黃豆製品加工過程中的副產物，含有豐富的蛋白質與纖維素，具備再利用潛力，惟目前多以堆肥或棄置方式處理，不僅效率不彰，亦可能造成環境污染。本研究核心在於利用B. licheniformis所分泌的酵素（蛋白酶與纖維素酶）加速豆渣的生物降解，並構建一套具應用潛力的循環系統。

研究首先針對菌株的最適生長條件進行測試，發現B. licheniformis在pH 9與45°C條件下具有最佳生長速率。接著設計含豆渣之液體培養基，搭配不同濃度的PBS與HEPES緩衝溶液，模擬適宜微生物生長的環境，以評估其對分解效率的影響。結果顯示，在添加40 mM HEPES與1X PBS之組別中，菌體生長穩定且豆渣降解速度顯著，經10天培養後，可將近95%的豆渣降解為粒徑低於1μm的小分子，顯示出良好的水解能力。

為驗證模組可行性，研究亦進行循環培養測試，將發酵液再次引入新豆渣培養基中進行分解反應，結果雖顯示具一定降解效果，但亦受到菌液濃度與豆渣前處理條件影響，顯示後續操作仍需標準化與優化。

整體而言，本研究成功建立以地衣芽孢桿菌為核心的豆渣分解循環模組，展示其於食品副產物資源化處理的應用潛力，並具發展為益生菌製品、生物飼料或再生能源等方向的可能性。未來建議進一步進行酵素活性分析、代謝產物鑑定及中試規模放大測試，以驗證其於實際廢棄物處理系統中的可行性與經濟效益，邁向低成本、高效能的永續資源再利用解方。

This study investigates the use of Bacillus licheniformis to establish a green recycling module capable of effectively decomposing okara, a by-product of soybean processing, in order to promote resource recovery and sustainable utilization of food by-products. Okara is rich in protein and fiber, making it a potentially valuable resource. However, it is commonly disposed of through composting or landfill, which are inefficient and environmentally problematic. This research focuses on utilizing enzymes secreted by B. licheniformis—specifically proteases and cellulases—to accelerate the biodegradation of okara, aiming to construct a recycling system with practical application potential.

The study first explored the optimal growth conditions for the bacterium, identifying pH 9 and a temperature of 45°C as ideal for rapid proliferation. Liquid culture media containing okara were then prepared, supplemented with different concentrations of PBS and HEPES buffer solutions to simulate conditions favorable for microbial activity and to assess degradation efficiency. The results showed that the combination of 40 mM HEPES and 1X PBS supported stable bacterial growth and significantly enhanced okara breakdown. After 10 days of cultivation, nearly 95% of the okara was degraded into molecules smaller than 1 μm, demonstrating strong hydrolytic capacity.

To verify the feasibility of the recycling module, the study also conducted a reuse test by transferring the fermented broth into freshly prepared okara media. While the system still exhibited some degradation capability, results were influenced by variables such as initial bacterial concentration and the pre-treatment of okara, suggesting that further process standardization and optimization are necessary.

Overall, this study successfully developed a microbial-based recycling module centered around B. licheniformis, showcasing its potential in the valorization of food by-products. The system could be further developed into applications such as probiotic fermentation products, animal feed, or bioenergy. Future research is recommended to include enzyme activity assays, metabolic product identification, and pilot-scale testing to verify the system’s applicability and cost-efficiency in real-world waste management scenarios, contributing to a low-cost, high-efficiency, and sustainable solution for food waste recycling.

Ansari, S. A., Kumar, T., Sawarkar, R., Gobade, M., Khan, D., & Singh, L. (2024). Valorization of food waste: A comprehensive review of individual technologies for producing bio-based products. Journal of Environmental Management, 364, 121439.
Asghar, A., Afzaal, M., Saeed, F., Ahmed, A., Ateeq, H., Shah, Y. A., Islam, F., Hussain, M., Akram, N., & Shah, M. A. (2023). Valorization and food applications of okara (soybean residue): A concurrent review. Food science & nutrition, 11(7), 3631-3640.
Bajaj, B. K., & Manhas, K. (2012). Production and characterization of xylanase from Bacillus licheniformis P11 (C) with potential for fruit juice and bakery industry. Biocatalysis and Agricultural Biotechnology, 1(4), 330-337.
Choi, I. S., Kim, Y. G., Jung, J. K., & Bae, H.-J. (2015). Soybean waste (okara) as a valorization biomass for the bioethanol production. Energy, 93, 1742-1747.
Ekiz, D. O., Comlekcioglu, U., Comlekcioglu, N., & Aygan, A. (2024). Cloning, characterization and functional analysis of lichenase produced by Bacillus licheniformis RB16 isolated from cattle faeces. Anais da Academia Brasileira de Ciências, 96(suppl 1), e20231156.
FAO. (2011). Global food losses and food waste – Extent, causes and prevention. https://www.fao.org/4/mb060e/mb060e00.htm
Ghaly, A., Arab, F., Mahmoud, N., & Higgins, J. (2007). Production of levan by Bacillus licheniformis for use as a soil sealant in earthen manure storage structures. Am. J. Biochem. Biotechnol, 3, 47-54.
Girotto, F., Alibardi, L., & Cossu, R. (2015). Food waste generation and industrial uses: A review. Waste management, 45, 32-41.
Guimarães, R. M., Silva, T. E., Lemes, A. C., Boldrin, M. C. F., da Silva, M. A. P., Silva, F. G., & Egea, M. B. (2018). Okara: A soybean by-product as an alternative to enrich vegetable paste. Lwt, 92, 593-599.
Guo, J., Zhang, J., Zhang, K., Li, S., & Zhang, Y. (2023). Effect of γ-PGA and γ-PGA SAP on soil microenvironment and the yield of winter wheat. Plos one, 18(7), e0288299.
Hall, F. F., Kunkel, H., & Prescott, J. (1966). Multiple proteolytic enzymes of Bacillus licheniformis. Archives of Biochemistry and Biophysics, 114(1), 145-153.
He, H., Yu, Q., Ding, Z., Zhang, L., Shi, G., & Li, Y. (2023). Biotechnological and food synthetic biology potential of platform strain: Bacillus licheniformis. Synthetic and Systems Biotechnology, 8(2), 281-291.
Ivanova, V. N., Dobreva, E. P., & Emanuilova, E. I. (1993). Purification and characterization of a thermostable alpha-amylase from Bacillus licheniformis. Journal of biotechnology, 28(2-3), 277-289.
Kaszycki, P., Głodniok, M., & Petryszak, P. (2021). Towards a bio-based circular economy in organic waste management and wastewater treatment–The Polish perspective. New Biotechnology, 61, 80-89.
Kaur, P. S., Kaur, S., Kaur, H., Sharma, A., Raj, P., & Panwar, S. (2015). Solid substrate fermentation using agro industrial waste: new approach for amylase production by Bacillus licheniformis. Int J Curr Microbiol App Sci, 4(12), 712-717.
Kiran, E. U., Trzcinski, A. P., Ng, W. J., & Liu, Y. (2014). Bioconversion of food waste to energy: A review. Fuel, 134, 389-399.
Kuribayashi, L. M., do Rio Ribeiro, V. P., De Santana, R. C., Ribeiro, E. J., Dos Santos, M. G., Falleiros, L. N. S. S., & Guidini, C. Z. (2021). Immobilization of β-galactosidase from Bacillus licheniformis for application in the dairy industry. Applied Microbiology and Biotechnology, 105, 3601-3610.
Lardinois, I., & Van de Klundert, A. (1993). Organic waste. Options for small-scale resource recovery.
Li, B., Qiao, M., & Lu, F. (2012). Composition, nutrition, and utilization of okara (soybean residue). Food Reviews International, 28(3), 231-252.
Mak, T. M., Xiong, X., Tsang, D. C., Yu, I. K., & Poon, C. S. (2020). Sustainable food waste management towards circular bioeconomy: Policy review, limitations and opportunities. Bioresource Technology, 297, 122497.
Marco, É. G. D., Heck, K., Martos, E. T., & Van Der Sand, S. T. (2017). Purification and characterization of a thermostable alkaline cellulase produced by Bacillus licheniformis 380 isolated from compost. Anais da Academia Brasileira de Ciências, 89(3 Suppl), 2359-2370.
Mo, W. Y., Man, Y. B., & Wong, M. H. (2018). Use of food waste, fish waste and food processing waste for China's aquaculture industry: Needs and challenge. Science of the Total Environment, 613, 635-643.
Muras, A., Romero, M., Mayer, C., & Otero, A. (2021). Biotechnological applications of Bacillus licheniformis. Critical Reviews in Biotechnology, 41(4), 609-627.
Paritosh, K., Kushwaha, S. K., Yadav, M., Pareek, N., Chawade, A., & Vivekanand, V. (2017). Food waste to energy: an overview of sustainable approaches for food waste management and nutrient recycling. BioMed research international, 2017(1), 2370927.
Rahman, M. M., Mat, K., Ishigaki, G., & Akashi, R. (2021). A review of okara (soybean curd residue) utilization as animal feed: Nutritive value and animal performance aspects. Animal Science Journal, 92(1), e13594.
Redondo-Cuenca, A., Villanueva-Suárez, M. J., & Mateos-Aparicio, I. (2008). Soybean seeds and its by-product okara as sources of dietary fibre. Measurement by AOAC and Englyst methods. Food Chemistry, 108(3), 1099-1105.
Sher, S., Sultan, S., & Rehman, A. (2021). Characterization of multiple metal resistant Bacillus licheniformis and its potential use in arsenic contaminated industrial wastewater. Applied Water Science, 11, 1-7.
Shleeva, M. O., Kondratieva, D. A., & Kaprelyants, A. S. (2023). Bacillus licheniformis: a producer of antimicrobial substances, including antimycobacterials, which are feasible for medical applications. Pharmaceutics, 15(7), 1893.
Vong, W. C., & Liu, S.-Q. (2016). Biovalorisation of okara (soybean residue) for food and nutrition. Trends in Food Science & Technology, 52, 139-147.
Xavier, J. R., Madhan Kumarr, M. M., Natarajan, G., Ramana, K. V., & Semwal, A. D. (2020). Optimized production of poly (γ‐glutamic acid)(γ‐PGA) using Bacillus licheniformis and its application as cryoprotectant for probiotics. Biotechnology and Applied Biochemistry, 67(6), 892-902.
Xu, F., Li, Y., Ge, X., Yang, L., & Li, Y. (2018). Anaerobic digestion of food waste–Challenges and opportunities. Bioresource Technology, 247, 1047-1058.
水中總菌落數檢測方法－混合稀釋法. https://www.nera.gov.tw/upload/old/27b878e2-a788-4899-aeec-42de25240d01/E20455b.pdf
水中總溶解固體及懸浮固體檢測方法─103～105℃乾燥. https://www.nera.gov.tw/upload/old/799e7511-8ad4-4f54-90e6-e46cd049fc26/W21058A.pdf