title: Secretory Expression and Characterization of Chinese Narcissus GNA-Like Lectin in Pichia pastoris.
abstract: Narcissus tazetta lectin (NTL) is a GNA-like lectin, which has a wide potential application in medicine, agriculture, and glycobiology. In the present paper, the codon-optimized ntl gene was transformed into the yeast Pichia pastoris; SDS-PAGE gel and western blotting analysis revealed that the recombinant lectin was expressed successfully in Pichia yeast. The similarity between the recombinant NTL and the native NTL was confirmed by circular dichroism (CD) and hemagglutination assay further. In the 5-L scale fermentator, the protein yield was as high as 1.2 g/L after fermentation for 96 h. In addition, the effect of metal ions (K+, Mg2+, Ca2+, and Cu2+), acid, and alkaline on hemagglutinating activity of NTL was tested, which provided biochemical characterizations of the mannose-binding lectin from Chinese Narcissus.
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.
authors.forename: Xinyu
authors.initials: X
authors.lastname: Li
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.
authors.forename: Pengpeng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liang
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China. wangwy@mail.buct.edu.cn.
authors.forename: Wenya
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.
authors.forename: Qipeng
authors.initials: Q
authors.lastname: Yuan
authors.affiliation: Department of Chemical Engineering, Tsinghua University, Beijing, 100084, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D015870:Gene Expression; D006384:Hemagglutination / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D031427:Narcissus / Q000235:genetics*; D010843:Pichia / Q000235:genetics*; D037121:Plant Lectins / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D008670:Metals; D037121:Plant Lectins; D011994:Recombinant Proteins
keywords: Chinese Narcissus; Mannose-binding lectin; Metal ions; Pichia pastoris; Secretory expression