Liu Sheng-Rong, Yang Xiao-Juan, Sun Duan-Fang
College of Life Science, and Fujian Higher Education Research Center for Local Biological Resources, Ningde Normal University, Ningde, 352100, Fujian, China.
Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, 510070, China.
Bioprocess Biosyst Eng. 2021 Oct;44(10):2109-2120. doi: 10.1007/s00449-021-02587-7. Epub 2021 May 28.
ε-Poly-L-lysine (ε-PL) is a naturally-occurring L-lysine homopolymer having a broad-spectrum antimicrobial activity and used widely as a food preservative. In the present study, the combined use of immobilization and in situ product removal (ISPR) was evaluated for the production of ε-PL by Streptomyces ahygroscopicus GIM8. Results showed that ε-PL production in the flask cultures decreased from 0.84 to 0.38-0.56 g/L upon immobilization on loofah sponge with different amounts (0.5-3 g in 50 mL medium in a flask). By applying continuous ISPR to the immobilized flask cultures, ε-PL production as high as 3.51 g/L was obtained compared to 0.51 g/L of the control. A satisfactory titer of 1.84 g/L ε-PL could also be achieved with intermittent ISRP (three cycles of ISPR operation during cultivation). Further investigation showed that low levels of ε-PL retained in the broth appeared to favor its biosynthesis. In the repeated-batch fermentation in a 5 L immobilized bioreactor, with continuous ISPR, the final average ε-PL concentration and productivity were 3.35 g/L and 0.797 g/L/day, respectively, and 3.18 g/L and 0.756 g/L/day for the alternative (intermittent ISPR), in comparison to 1.16 g/L and 0.277 g/L/day with no ISPR usage. In the fed-batch fermentation with immobilized cells, the combined use of intermittent ISPR and extra nutrient feeding increased ε-PL concentration and productivity up to 24.57 g/L and 9.34 g/L/day. The fermentation processes developed could serve as an effective approach for ε-PL production and, moreover, the combination could greatly simplify downstream processing for ε-PL separation and purification.
ε-聚-L-赖氨酸(ε-PL)是一种天然存在的L-赖氨酸同聚物,具有广谱抗菌活性,被广泛用作食品防腐剂。在本研究中,评估了固定化与原位产物去除(ISPR)相结合用于吸水链霉菌GIM8生产ε-PL的效果。结果表明,在烧瓶培养中,当以不同量(50 mL烧瓶培养基中0.5 - 3 g)固定在丝瓜海绵上时,ε-PL产量从0.84 g/L降至0.38 - 0.56 g/L。通过对固定化烧瓶培养物应用连续ISPR,与对照的0.51 g/L相比,获得了高达3.51 g/L的ε-PL产量。通过间歇ISRP(培养期间三个ISPR操作循环)也可以实现1.84 g/L ε-PL的满意滴度。进一步研究表明,肉汤中保留的低水平ε-PL似乎有利于其生物合成。在5 L固定化生物反应器的重复分批发酵中,采用连续ISPR时,最终平均ε-PL浓度和生产率分别为3.35 g/L和0.797 g/(L·天),交替(间歇ISPR)时分别为3.18 g/L和0.756 g/(L·天),而不使用ISPR时分别为1.16 g/L和0.277 g/(L·天)。在固定化细胞的补料分批发酵中,间歇ISPR和额外营养物补料相结合使ε-PL浓度和生产率分别提高到24.57 g/L和9.34 g/(L·天)。所开发的发酵工艺可作为生产ε-PL的有效方法,此外,这种组合可大大简化ε-PL分离和纯化的下游加工过程。