This work reports a simple and facile method to prepare novel magnetic mesoporous silica (MMS) materials with high magnetic strength for the convenient and high throughput detection of histidine-tagged enzymes with Ni-doped surfaces. These materials are designed by the incorporation of high-abundance and homogeneously dispersed iron nanoparticles within the mesopores by thermal hydrogen reduction after the incorporation of ferrous ions and demonstrated the selective enrichment and high-throughput recognition of His-tagged enzymes with multi-point anchoring by selective conjugation between the His-tag and Ni ions. Selective His-tagged enzyme enrichment efficiency was compared with nickel-based MMS materials, such as Ni 2+ -MMS and Ni-MMS, and nickel ion doped silica-coated magnetic nanoparticles (Ni 2+ -MNPs). The efficiency was calculated to be 100 AE 1.93%, 70.94 AE 1.95%, and 37.03 AE 5.93% for Ni 2+ -MMS, Ni-MMS, and Ni 2+ -MNPs, respectively. This method enables a highthroughput and advanced systematic approach for the separation and immobilization of proteins which cover a broad spectrum of polyhistidine-tagged proteins.
Here we proposed clean and acceptable pretreatment methods for the pretreatment of lignlicellulosic biomass. To prove the feasibility of it, we compared three modes of mills such as ball, attrition and planetary mills. Among the mill modes, attrition and planetary mills are more effective to reduce biomass size when compared to ball mill. In enzymatic saccharification process, planetary mill treated crystalline cellulose produced higher amount of glucose and galactose than attrition mill treated. The mill pretreatments did not cause toxic compounds, such as hydroxymethylfurfuraldehyde (HMF) and levulinic acid. It was performed in enzymatic process buffer. Hence, it is not needed to exchange buffer for further process. It is convenient and economic process for cellulosic biomass pretreatment and can be applied to mass biofuel production
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