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Jie Sun,Jie Jiang,Xinyang Zhai,Shaoming Zhu,Zhenzhen Qu,Wei Yuan,Zhao Wang,Chun Wei 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.6
The large-scale production of functional recombinant lactoferrin has become a major goal because of its medicinal value and global demand. Secreting recombinant proteins into a culture medium offers a way to simplify protein purification and avoid toxicity from intracellularly accumulated materials. In this study, after 84 h of induction with methanol in a shaking flask, the recombinant bovine lactoferrin (rbLf) titer in the culture supernatant of the strain that integrated two copies of the rbLf gene was only 121.6 μg/L. A bottleneck might have existed in the folding and secretion pathways of rbLf. We then attempted to further improve the rbLf titer by overexpressing the transcription factor Hac1p and α-signal peptide-cutting protease Kex2p with different promoters. Results showed that the inducible coexpression of Hac1p and Kex2p linked with the 2A sequence improved the rbLf titer 5.0-fold (735.8 μg/L) after 84 h of induction with methanol. The maximal titer in a shaking flask was 1,150.5 μg/L after 120 h of induction. The rbLf titer achieved 35.6 mg/L in a 5 L fed-batch fermenter. Thus, Kex2 and Hac1 overexpression driven by methanol-induced promoter alleviated the bottleneck in the folding and secretion pathways and greatly improved the secretory expression of rbLf in Pichia pastoris.
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Sisi Ma,Ligang Lin,Xinyang Li,Wenying Shi,Xiaofei Zhai,Jing Yang 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.112 No.-
Developing multifunctional, efficient and durable membrane for treating complex oily wastewater ishighly desirable but still a challenge due to the severe membrane fouling. Herein, nanosponge membranewith 3D-macrocycle b-cyclodextrin (b-CDs) as molecular cage was manufactured by azide-alkyne clickreaction for oil/water treatment and antifouling properties simultaneously. The macrocyclic ‘molecularcage’ geometry of b-CDs can induce various guest molecules into their cavities. When clickable b-CDN3was fixed onto a clickable EVAL- membrane surface, the hydrophilicity of the membrane was greatlyimproved. Furthermore, the molecular cage–grafted membrane (EVAL-g-CD) showed better antifoulingperformance than a pure EVAL membrane, with lower water flux decline (15%) and higher water fluxrecovery (91%). The flux and separation efficiency values of the EVAL-g-CD membrane were higher than120 Lm2h1 and 99%, respectively. The EVAL-g-CD membrane also exhibited good adsorption performancefor organic pollutants owing to its cavity structure. Furthermore, the membrane showed desirablestability and its rejection remained at 99% after filtration. This proposed 3D membrane strategy based onmolecular cages sheds light on the formation of hydrophilic membrane surfaces and shows great promisefor potential applications such as the separation of oil-in-water emulsions.
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