Novel Fusion Protein Consisting of Metallothionein, Cellulose Binding Module, and Superfolder GFP for Lead Removal from the Water Decoction of Traditional Chinese Medicine

[Image: see text] Many methods have been used to detect heavy metals in herbal medicines, while few are developed to remove them. In this study, a novel genetically engineered fusion protein composed of metallothionein (MT), cellulose binding module (CBM), and superfolder GFP (sfGFP) was designed to remove heavy metals. MT, a kind of cysteine-rich protein, was used to chelate heavy metals with high specific affinity. The CBM facilitated the fusion protein MT-CBM-sfGFP binding to cellulose specifically, which made the purification and immobilization in one step. The sfGFP was used to detect the fusion protein MT-CBM-sfGFP easily during the process of expression and immobilization. The MT from Cancer pagurus (MT(Cap)) and the CBM from Cellulomonas fimi (CBM(Cef)) were used as an example and the fusion protein (MT(Cap)-CBM(Cef)-sfGFP) was expressed in Escherichia coli. Then, the cell lysates were mechanically mixed with cellulose to create biosorbent MT(Cap)-CBM(Cef)-sfGFP@cellulose. The efficiency of the biosorbent MT(Cap)-CBM(Cef)-sfGFP@cellulose for Pb(2+) removal was evaluated using the water decoction of Honeysuckle as a model. Results suggested that MT(Cap)-CBM(Cef)-sfGFP@cellulose had high efficiency for Pb(2+) removal from the water decoction of Honeysuckle without affecting its active ingredients. The low-cost, easy production, and high efficiency of the biosorbent enable it to have many applications in heavy metal removal from aqueous solutions of herbal medicines and food.