Short CommunicationHigh butanol production from glycerol by using Clostridium sp. strain CT7 integrated with membrane assisted pervaporation
Introduction
Biobutanol can be specifically synthesized through traditional acetone-butanol-ethanol (ABE) fermentation with a mass ratio of 3:6:1 by using solventogenic Clostridium species, such as C. acetobutylicum, C. beijerinckii, and C. pasteurianum (Jones and Woods, 1986). However, the sustainable large scaling biobutanol production is still impeded by the high cost of traditional feedstocks, mainly the starchy based materials (Gu et al., 2011, Zhang, 2015). Accordingly, the adoption of low cost substrates, such as organic wastes would add on the economical values for biobutanol production (Dürre, 2007, Shanmugam et al., 2019). Glycerol, the by-product in biodiesel industry with production of approximate ten percent of biodiesel in weight has gained great attention owning to its low price and availability (Dobson et al., 2012). The abundance and cost competitiveness of glycerol have made it an excellent alternative for biochemicals production including biobutanol, 1,3-propanol et al. compared with other carbon substrates (Taconi et al., 2009). In addition, glycerol could provide more reducing powers mainly NADH than monosaccharides such as glucose for reduced products synthesis including butanol, which will attribute to a high product yield (González-Pajuelo et al., 2005).
As a lipophilic fuel, butanol would cause the lysis of cell membrane and finally lead to the low butanol titer (7–13 g/L) (Kumar and Gayen, 2011). Alternatively, separation techniques with in situ butanol recovery including adsorption, liquid-liquid extraction, gas stripping, and pervaporation could be coupled with ABE fermentation to alleviate the butanol toxicity to cells, improving substrate utilization and enhancing butanol productivity (Liu et al., 2011, Nielsen and Prather, 2009, Xue et al., 2016). Recently, a novel close-circulating vapor stripping-vapor permeation technique has been developed as a one-step process for online butanol recovery (Yang et al., 2018, Zhu et al., 2018). Membrane coupled pervaporation (PV) process has been considered to be the greatest potential separation technology owning to its energy-saving and high separation efficiency, as well as no harm to microorganisms (Xue et al., 2017, Yang et al., 2019). Thin polydimethylsiloxane (PDMS) layer has shown to improve the permeate flux and separation factor owning to its negligible transportation resistance of the support (Xiangli et al., 2007). Especially, PDMS coupled ABE fermentation could efficiently relieve the toxicity of butanol to microorganisms by continuous butanol removal from the fermenter, so that cells could grow vegetatively instead of sporulation.
Currently, C. pasteurianum is the best candidate for butanol production from glycerol (Xin et al., 2016). In our previous study, a novel Clostridium sp. strain CT7 with the capabilities of glycerol-utilizing and butanol-generating was isolated and characterized (Xin et al., 2017). Moreover, this strain possessed a unique metabolic pathway with butanol-ethanol (BE) production uncoupled with acetone formation. Therefore, PDMS/ceramic composite membrane was further employed in this study to improve the final butanol titer and yield.
Section snippets
Strain and media
Clostridium sp. strain CT7 was isolated by our lab (Xin et al., 2017). The defined mineral salts medium contained (per liter of distilled water): KH2PO4, 0.75 g; K2HPO4, 0.75 g; CH3COONH4, 2 g; yeast extract, 5 g. Moreover, 1 mL of trace element solution, 1 mL of Na2SeO3-Na2WO4 solution and 10 mg of resazurin were added to 1 L of the medium. Subsequently, reductants Na2S, L-cysteine, and DL-Dithiothreitol under N2 were added to a final concentration of 0.2, 0.2, and 0.5 mM, respectively. 20 mM
Results and discussion
When batch fermentation using 60 g/L of glycerol as the sole carbon source was carried out by using Clostridium sp. strain CT7, 10.4 g/L of butanol, 0.4 g/L of ethanol and total 5.8 g/L of volatile fatty acids (VFAs) were produced after 120 h of fermentation, along with 32.1 g/L of glycerol leftover in the fermentation broth, which corresponded to the butanol yield of 0.32 g/g (Data not shown). It should be noticed that an obvious inhibition was observed after 56 h due to the toxicity caused by
Conclusion
PDMS/ceramic composite membrane was applied in the PV coupled butanol-ethanol fermentation process from glycerol to in situ remove butanol by the novel Clostridium sp. strain CT7. The PV coupled fed-batch fermentation process was considered the optimal operation to achieve butanol of 41.9 g/L and butanol productivity of 0.21 g/L/h with a total flux and butanol separation factor of 546.9 g/m2/h and 24.4, respectively. In conclusion, our work indicated the PV coupled fed-batch fermentation
Acknowledgements
This work was supported by the Jiangsu Province Natural Science Foundation for Youths (No. BK20170993), Jiangsu Key Laboratory of Biomass Energy and Materials (JSBEM201908), the Key Science and Technology Project of Jiangsu Province (BE2016389), the National Natural Science Foundation of China (No. 21706125, No. 21727818, No. 21706124, No. 31700092), The Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture of China. We also thank Prof. Wanqin Jin, Dr. Gongping Liu and Ph.D. Haipeng
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