Transforming global agricultural waste into eco-friendly products like industrial enzymes through bioconversion can help address sustainability challenges aligning with the United Nations’ Sustainable Development Goals. Present study explored the production of high-yield food-grade cellulolytic enzymes from MTCC 4876, using a novel media formulation with a combination of waste sorghum grass and cottonseed oil cake (3:1). Optimization of physical and environmental parameters, along with the screening and optimization of media components, led to an upscaled process in a novel 6-L solid-state fermentation (SSF)-packed bed reactor (PBR) with a substrate loading of 200 g. Saturated forced aeration proved crucial, resulting in high fungal biomass (31.15 ± 0.63 mg glucosamine/gm dry fermented substrate) and high yield cellulase (20.64 ± 0.36 FPU/g-ds) and xylanase (16,186 ± 912 IU/g-ds) production at an optimal airflow rate of 0.75 LPM. The PBR exhibited higher productivity than shake flasks for all the enzyme systems. Microfiltration and ultrafiltration of the crude cellulolytic extract achieved 94% and 71% recovery, respectively, with 13.54 FPU/mL activity in the cellulolytic enzyme concentrate. The concentrate displayed stability across wide pH and temperature ranges, with a half-life of 24.5-h at 50 °C. The cellulase concentrate, validated for food-grade safety, complies with permissible limits for potential pathogens, heavy metals, mycotoxins, and pesticide residue. It significantly improved apple juice clarity (94.37 T%) by reducing turbidity (21%) and viscosity (99%) while increasing total reducing sugar release by 63% compared with untreated juice. The study also highlighted the potential use of lignin-rich fermented end residue for fuel pellets within permissible SOx emission limits, offering sustainable biorefinery prospects. Utilizing agro wastes in a controlled bioreactor environment underscores the potential for efficient large-scale cellulase production, enabling integration into food-grade applications and presenting economic benefits to fruit juice industries.

中文翻译：

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在新型固态发酵生物反应器中利用高粱废物生产食品级纤维素分解酶的生物工艺优化，以增强苹果汁的澄清度

通过生物转化将全球农业废物转化为工业酶等环保产品，有助于应对可持续发展挑战，符合联合国可持续发展目标。本研究探索了使用废高粱草和棉籽油饼 (3:1) 组合的新型培养基配方，从 MTCC 4876 生产高产食品级纤维素分解酶。物理和环境参数的优化以及培养基成分的筛选和优化，在底物负载为 200 g 的新型 6 L 固态发酵 (SSF) 填充床反应器 (PBR) 中实现了升级工艺。事实证明，饱和强制通气至关重要，可产生高真菌生物量（31.15 ± 0.63 mg 葡萄糖胺/gm 干发酵底物）和高产纤维素酶（20.64 ± 0.36 FPU/g-ds）和木聚糖酶（16,186 ± 912 IU/g-ds）。最佳气流率为 0.75 LPM。对于所有酶系统，PBR 均表现出比摇瓶更高的生产率。粗制纤维素分解提取物的微滤和超滤回收率分别为 94% 和 71%，纤维素分解酶浓缩物的活性为 13.54 FPU/mL。该浓缩物在较宽的 pH 值和温度范围内表现出稳定性，50 °C 下的半衰期为 24.5 小时。该纤维素酶浓缩物经过食品级安全性验证，符合潜在病原体、重金属、霉菌毒素和农药残留的允许限值。与未经处理的果汁相比，它通过降低浊度（21%）和粘度（99%）显着提高了苹果汁的澄清度（94.37 T%），同时总还原糖释放量增加了 63%。该研究还强调了在允许的 SOx 排放限制内将富含木质素的发酵末残渣用于燃料颗粒的潜在用途，从而提供了可持续的生物炼制前景。在受控生物反应器环境中利用农业废物凸显了高效大规模纤维素酶生产的潜力，能够整合到食品级应用中，并为果汁行业带来经济效益。