Zhou Jiushun, Cai Menghao, Jiang Tao, Zhou Weiqiang, Shen Wei, Zhou Xiangshan, Zhang Yuanxing
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
Bioprocess Biosyst Eng. 2014 Mar;37(3):575-84. doi: 10.1007/s00449-013-1024-1. Epub 2013 Aug 2.
Medium and culture conditions for alginate lyase production by marine Vibrio sp. QY102 were first optimized using statistical methods including Plackett-Burman design and central composite design. Then, fermentation in 5-L bioreactor showed that alginate acted as easily used carbohydrate for Vibrio sp. QY102, while starch extended its growth phase and stabilized pH variations. Thus, a novel strategy using mixed carbon sources was proposed that starch supported growth while enzyme synthesis was induced by pulse feedings of solid alginate. The optimized process followed that Vibrio sp. QY102 grew on starch until the end of the logarithmic growth phase, and then solid alginate was added as 1 g/L every 3 h. Meanwhile, initial pH 5.0 and natural pH during fermentation was favorable for alginate lyase production. After optimization, the highest alginate lyase production reached 52.8 U/mL, which was 329 % higher than the control. Finally, fermentation scale-up was performed in 30-L bioreactor and the maximum alginate lyase production was obtained as 46.8 U/mL.
首先使用包括Plackett-Burman设计和中心复合设计在内的统计方法优化了海洋弧菌属菌株QY102产生海藻酸盐裂解酶的培养基和培养条件。然后,在5升生物反应器中进行的发酵表明,海藻酸盐是弧菌属菌株QY102易于利用的碳水化合物,而淀粉延长了其生长阶段并稳定了pH值变化。因此,提出了一种使用混合碳源的新策略,即淀粉支持生长,同时通过脉冲添加固体海藻酸盐诱导酶的合成。优化后的工艺如下:弧菌属菌株QY102在淀粉上生长至对数生长期结束,然后每3小时添加1克/升固体海藻酸盐。同时,初始pH值为5.0且发酵过程中的自然pH值有利于海藻酸盐裂解酶的产生。优化后,海藻酸盐裂解酶的最高产量达到52.8 U/mL,比对照高329%。最后,在30升生物反应器中进行发酵放大,获得的海藻酸盐裂解酶最大产量为46.8 U/mL。