Improved cell disruption of Pichia pastoris utilizing aminopropyl magnesium phyllosilicate (AMP) clay.
An efficient method for Pichia cell disruption that employs an aminopropyl magnesium phyllosilicate (AMP) clay-assisted glass beads mill is presented. AMP clay is functionalized nanocomposite resembling the talc parent structure Si8Mg6O20(OH)4 that has been proven to permeate the bacterial membrane and cause cell lysis. The recombinant capsid protein of cowpea chlorotic mottle virus (CCMV) expressed in Pichia pastoris GS115 was used as demonstration system for their ability of self-assembly into icosahedral virus-like particles (VLPs). The total protein concentration reached 4.24 mg/ml after 4 min treatment by glass beads mill combined with 0.2 % AMP clay, which was 11.2 % higher compared to glass beads mill only and the time was half shortened. The stability of purified CCMV VLPs illustrated AMP clay had no influence on virus assembly process. Considering the tiny amount added and simple approach of AMP clay, it could be a reliable method for yeast cell disruption.
Department of Chemical and Biochemical Engineering, Chosun University, #375 Seosuk-dong, Dong-gu, Gwangju 501-759, Republic of Korea.
Sun-Il
SI
Kim
Yuanzheng
Y
Wu
Ka-Lyun
KL
Kim
Geun-Joong
GJ
Kim
Hyun-Jae
HJ
Shin
D001709:Biotechnology / Q000379:methods*; D017795:Bromovirus / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification*; D008967:Molecular Biology / Q000379:methods*; D010843:Pichia / Q000737:chemistry* / Q000187:drug effects; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D017640:Silicates / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods*; D013312:Stress, Physiological
D036022:Capsid Proteins; D005656:Fungal Proteins; D011994:Recombinant Proteins; D017640:Silicates