Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada, Kobe 657-8501, Japan.
Appl Microbiol Biotechnol. 2013 Aug;97(16):7165-72. doi: 10.1007/s00253-013-5009-4. Epub 2013 Jun 9.
We constructed beta-glucosidase (BGL)-displaying Corynebacterium glutamicum, and direct L-lysine fermentation from cellobiose was demonstrated. After screening active BGLs, Sde1394, which is a BGL from Saccharophagus degradans, was successfully displayed on the C. glutamicum cell surface using porin as an anchor protein, and cellobiose was directly assimilated as a carbon source. The optical density at 600 nm of BGL-displaying C. glutamicum grown on cellobiose as a carbon source reached 23.5 after 48 h of cultivation, which was almost the same as that of glucose after 24 h of cultivation. Finally, Sde1394-displaying C. glutamicum produced 1.08 g/l of L-lysine from 20 g/l of cellobiose after 4 days of cultivation, which was about threefold higher than the amount of produced L-lysine using BGL-secretory C. glutamicum strains (0.38 g/l after 5 days of cultivation). This is the first report on amino acid production using cellobiose as a carbon source by BGL-expressing C. glutamicum.
我们构建了展示β-葡萄糖苷酶(BGL)的谷氨酸棒杆菌,并证明了可以直接从纤维二糖发酵生产 L-赖氨酸。在筛选出活性 BGL 后,成功地将来自 Saccharophagus degradans 的 BGL Sde1394 作为锚定蛋白展示在谷氨酸棒杆菌的细胞表面上,并将纤维二糖直接作为碳源同化。以纤维二糖为碳源培养的展示 BGL 的谷氨酸棒杆菌在 48 小时后达到 23.5 的 600nm 吸光度,几乎与葡萄糖在 24 小时培养后的吸光度相同。最后,展示 Sde1394 的谷氨酸棒杆菌在 4 天的培养时间内从 20g/L 的纤维二糖中生产出了 1.08g/L 的 L-赖氨酸,比使用 BGL 分泌型谷氨酸棒杆菌菌株(培养 5 天后产生 0.38g/L 的 L-赖氨酸)生产的 L-赖氨酸产量高约三倍。这是首例报道通过表达 BGL 的谷氨酸棒杆菌利用纤维二糖作为碳源生产氨基酸。