Proteinase B disruption is required for high level production of human mechano-growth factor in Saccharomyces cerevisiae.
Mechano-growth factor (MGF) is a product of a unique muscle-specific splice variant of the insulin-like growth factor I gene. Potential use of MGF to improve regenerative capability of skeletal muscle as well as to prevent neuronal damage has been widely discussed. Human MGF was expressed in Saccharomyces cerevisiae, but the yield of the recombinant protein was low due to its rapid degradation. The proteinase B was identified as the enzyme responsible for MGF processing. A yeast strain with Deltaprb1 deletion was created resulting in a fivefold increase of MGF yield that reached 50 mg/l. The biological activity of recombinant MGF was verified in a proliferation assay employing human postnatal myoblasts.
Morozkina
Elena V
EV
A.N. Bach Institute of Biochemistry, Moscow, Russian Federation. morozkina@inbi.ras.ru
Marchenko
Alexey N
AN
Keruchenko
Jan S
JS
Keruchenko
Irina D
ID
Khotchenkov
Vyacheslav P
VP
Popov
Vladimir O
VO
Benevolensky
Sergei V
SV
D017353:Gene Deletion; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000096:biosynthesis* / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C523997:mechano-growth factor E, human; D007334:Insulin-Like Growth Factor I; D012697:Serine Endopeptidases; C023302:yeast proteinase B