Expression of human lysozyme in an insoluble form in yeast.
A high level of expression in yeast of a chemically synthesized human lysozyme (hL) gene was achieved by introducing an A-rich DNA fragment just upstream from the ATG start codon. The synthesized recombinant human lysozyme (r-hL) was insoluble and biologically inactive. It was solubilized with 7 M urea (pH 9) from yeast cells and its lytic activity was efficiently regenerated by oxidative renaturation. This renaturation experiment and Western blotting analysis under reducing and non-reducing conditions indicate that the insoluble form might be caused by the formation of incorrect intra- or intermolecular disulfide bonds. The N-terminal amino acid sequence of the purified r-hL was identical with that of authentic hL.
Hayakawa
T
T
Central Research Division, Takeda Chemical Industries, Ltd., Osaka, Japan.
Toibana
A
A
Marumoto
R
R
Nakahama
K
K
Kikuchi
M
M
Fujimoto
K
K
Ikehara
M
M
D000595:Amino Acid Sequence; D005786:Gene Expression Regulation; D006801:Humans; D009113:Muramidase / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids; D011489:Protein Denaturation; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012995:Solubility
D009113:Muramidase