Cryoprotective effect of the serine-rich repetitive sequence in silk protein sericin.
The silk proteins, fibroin and sericin, are produced in the silk gland of Bombyx mori, and hydrophilic sericin envelops fibroin with successive sticky layers in the formation of a cocoon. To study the biological functions of sericin, we focused on the serine-rich sericin peptide consisting of 38 amino acids, which is a highly conserved and internally repetitive sequence of a sericin protein. The corresponding gene was chemically synthesized, and the PCR-amplified gene was ligated to oligomerize sericin peptide and fused at the amino terminus to a His-tagged and proteolytic cleavage sequence in an inducible expression vector. When the dimers of sericin peptides were overexpressed in Escherichia coli, the transformants showed a prominent increase in cell viability after freezing in medium. Further, the purified dimeric sericin peptide from E. coli was found to be effective in protecting lactate dehydrogenase from denaturation caused by freeze-thaw. Both of these protective effects against freezing stress in cells and proteins were also observed with sericin hydrolysate. These results indicate that this unique sericin peptide, like sericin, has a high cryoprotective activity and will be valuable as a new biomaterial for industrial use.
Tsujimoto
K
K
Technology Department, Seiren Co. Ltd., Keya, Fukui 918-8560, Japan.
Takagi
H
H
Takahashi
M
M
Yamada
H
H
Nakamori
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods; D012831:Bombyx; D003451:Cryoprotective Agents / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005615:Freezing; D006868:Hydrolysis; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011489:Protein Denaturation / Q000187:drug effects; D047030:Sericins; D012694:Serine / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014169:Transformation, Bacterial
D003451:Cryoprotective Agents; D010446:Peptide Fragments; D010456:Peptides, Cyclic; D047030:Sericins; D012694:Serine; D007770:L-Lactate Dehydrogenase