title: Immobilization of trypsin on miniature incandescent bulbs for infrared-assisted proteolysis.
abstract: A novel efficient proteolysis approach was developed based on trypsin-immobilized miniature incandescent bulbs and infrared (IR) radiation. Trypsin was covalently immobilized in the chitosan coating on the outer surface of miniature incandescent bulbs with the aid of glutaraldehyde. When an illuminated enzyme-immobilized bulb was immersed in protein solution, the emitted IR radiation could trigger and accelerate heterogeneous protein digestion. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of hemoglobin (HEM), cytochrome c (Cyt-c), lysozyme (LYS), and ovalbumin (OVA) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF-MS with the sequence coverages of 91%, 77%, 80%, and 52% for HEM, Cyt-c, LYS, and OVA (200 ng μL(-1) each), respectively. The suitability of the prepared bulb bioreactors to complex proteins was demonstrated by digesting human serum.
authors.affiliation: School of Pharmacy, Department of Chemistry, Fudan University, 220 Handan Road, Shanghai 200433, China.
authors.forename: Huimin
authors.initials: H
authors.lastname: Ge
authors.affiliation: School of Pharmacy, Department of Chemistry, Fudan University, 220 Handan Road, Shanghai 200433, China.
authors.forename: Huimin
authors.initials: H
authors.lastname: Bao
authors.affiliation: School of Pharmacy, Department of Chemistry, Fudan University, 220 Handan Road, Shanghai 200433, China.
authors.forename: Luyan
authors.initials: L
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Department of Chemistry, Fudan University, 220 Handan Road, Shanghai 200433, China. Electronic address: gangchen@fudan.edu.cn.
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
mesh_terms: D019149:Bioreactors; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007259:Infrared Rays; D008029:Lighting; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism; D059748:Proteolysis; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D006454:Hemoglobins; D010047:Ovalbumin; D045304:Cytochromes c; D009113:Muramidase; D014357:Trypsin
keywords: Infrared; Mass spectra; Miniature incandescent bulb; Peptide mass mapping; Proteolysis