Center of Innovative and Applied Bioprocessing (CIAB), Sector-81, Knowledge City, Mohali, 140306, India.
Center of Innovative and Applied Bioprocessing (CIAB), Sector-81, Knowledge City, Mohali, 140306, India; CSIR-Institute of Himalayan Bioresource Technology, Palampur, 176061, (HP), India.
Carbohydr Res. 2024 Jul;541:109150. doi: 10.1016/j.carres.2024.109150. Epub 2024 May 21.
Aim of the study was to optimize and produce beta-mannanase at fermenter scale by using cheaper minimal media. Increased production of beta-mannanase from Microbacterium camelliasinensis CIAB417 was achieved by heterologous expression in E. coli BL21 (DE3). The scale-up production of beta-mannanase was optimized from shake flask to 5-L fermenter. The cost-effective minimal media (M9+e) without any vitamins was found to be most effective and optimized for culturing the cells. The same media displayed no significant fluctuation in the pH while culturing the cells for the production of beta-mannanase both at shake flask and fermenter level. Additionally, E. coli cells were able to produce similar amount of dry cell weight and recombinant beta-mannanase both in the presence of micro and macro-oxygen environment. The optimized media was demonstrated to show no significant drop in pH throughout the recombinant protein production process. In one litre medium, 2.0314 g dry weight of E. coli cells yielded 1.8 g of purified recombinant beta-mannanase. The purified enzyme was lyophilized and demonstrated to hydrolyse locust bean gum to release mannooligosaccharides.
本研究旨在通过使用更廉价的最小培养基优化并生产β-甘露聚糖酶。通过在大肠杆菌 BL21（DE3）中异源表达，提高了对生花单胞菌 CIAB417 的β-甘露聚糖酶的产量。从摇瓶到 5-L 发酵罐优化了β-甘露聚糖酶的放大生产。发现不含任何维生素的经济型最小培养基（M9+e）对细胞培养最有效且经过优化。在摇瓶和发酵罐水平培养细胞生产β-甘露聚糖酶时，该培养基的 pH 值没有明显波动。此外，大肠杆菌细胞在微氧和大氧环境下均能产生相似量的干重和重组β-甘露聚糖酶。优化后的培养基在整个重组蛋白生产过程中 pH 值均无明显下降。在 1 升培养基中，2.0314 克干重的大肠杆菌细胞产生了 1.8 克纯化的重组β-甘露聚糖酶。纯化的酶经冻干后可水解刺槐豆胶释放甘露寡糖。
