PENGARUH KONSENTRASI PENGINDUKSI METANOL SERTA"SUMBER KARBON SORBITOL DAN MONITOL TERHADAP PRODUKSI a-AMILASE Saccharomycopsis fibuligera R64 DALAM Pichia pastoris
Abstract: Pichia pastoris has
been widely used as host for heterologous protein expression for commercial
purposes. The advantages of P. pastoris as protein expression host is its
ability to grow with high cell density and the presence of a gene promoter that
can be induced tightly, named AOXI encoding alcohol oxidase. Improvement of
recombinant protein production under control of AOXI promoter in P. pastoris
expression system is still a major concern. Optimization of methanol
concentration as inducer and addition of carbon sources, is one of the
strategies to improve the expression level. This research aims to study the
effect of methanol as inducer as well as sorbitol and mannitol as additional
carbon source to the expression level of recombinant Saccharomycopsis
fibuligera a-amylase (Sfamy) by P. pastoris (Mut). Sorbitol and manuot known as
non-repressive carbon source, to increase the growth ofP. past oris, but not
inhibit the AOX1 promoter and foreign proteins expression. Sfamy was expressed
in P. pastoris GS115 (His-, Mut) with addition of carbon source, sorbitol and
mannitol saparately to the expression medium. The result showed that, the
optimum concentration" of methanol inducer for Sfamy production is 0.75%.
The addition of sorbitol or mannitol increased Sfamy production. Concentrations
of sorbitol and mannitol 2% with 0.75% methanol inducer increase the secretion
level of recombinant Sfamy 2.13 times and 1.94 times respectively, compared
with no additional carbon source. This result indicated that addition of carbon
source can improved recombinant protein production by P. pastoris, and the used
of sorbitol as additional carbon source is more effective compared to mannitol.
Penulis: Shabarni Gaffar, Dani
Permana, Diana P Rahmawati, Triana N Meirina, Abu Bakar M.I. Syihab, Safrl
Ismayana, Toto Subroto, O Suprijana
Kode Jurnal: jpkimiadd110186
