Modification of adsorption of CBM onto lignin or microcelullose using rational mutation and additive addition


Baojie Jiang;Xu fang;


Using the lignocelluloses as raw materials to produce biofuel is considered to be an attractive alternative to fossil fuel.However,the presence of lignin hinders the hydrolysis of lignocelluloses,reducing high cost of cellulase.Two methods are generally applied to improve the activity of cellulase:enhance binding affinity of cellulase to cellulose or decrease adsorption of cellulase to lignin.In this paper,CBM(carbohydrate-binding module)from CBHI of Trichoderma reesei was confused with GFP.The fused protein was named GLC and mutated to increase the negative charge using the site-directed mutagenesis method.Then the adsorption rates of GLC and its variants were determined.Our result showed that the negative charge increase of CBM has a significant influence on the adsorption when GLC binding to lignin or microcellulose.Meanwhile,the additive-Dodecyltrimethylammonium Bromide(DTAB)or Poly(Dimethyl Diallyl Ammonium Chloride)(PolyDADMAC),which has positive charge,was added into the system of GLC and microcellulose or lignin.Our results showed that both DTAB and PolyDADMAC enhance binding affinity of GLC to microcellulose.However,the adsorption of GLC binding to lignin was decreased when DTAB or PolyDADMAC was added.The addition of DTAB or PolyDADMAC improved the degradation efficiency of CBHI purified from T.reesei when presence of lignin in the hydrolysis system.




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