Av. Sahasrabudhe et al., Production of recombinant human bile salt stimulated lipase and its variant in Pichia pastoris, PROT EX PUR, 14(3), 1998, pp. 425-433
hBSSL and its truncated variant hBSSL-C cDNA clones were expressed in Pichi
a pastoris using two different signal peptides, native signal peptide and i
nvertase signal peptide, respectively, to facilitate secretion of the recom
binant proteins into the culture medium. Both recombinant proteins were sec
reted into the culture medium to a level of 45-50 mg/liter in shake flask c
ultures. Native signal peptide of hBSSL was recognized in P, pastoris and w
as cleaved at the same site as in humans. The level of expression of the hB
SSL gene was found to be dependent on the number of its copies integrated i
nto the host chromosome. The multicopy transformant clone was found to be v
ery stable. When grown and induced in a fermentor, the level of accumulatio
n of the recombinant hBSSL in the culture medium improved from 50 mg/liter
in shake flask cultures to 300 mg/liter. The recombinant hBSSL purified fro
m the culture supernatant was found to be similar to the native hBSSL in it
s biochemical properties except for the lectin-binding profile. (C) 1998 Ac
ademic Press.