Production of the kringle fragments of human apolipoprotein(a) by continuous lactose induction strategy.
A novel lactose induction strategy for the production of rhLK68, the kringle fragments of human apolipoprotein(a) (apo(a)) as a novel anti-angiogenic protein, was investigated. A scale-up of the production was accompanied by a decrease in expression level, and severe aggregation occurred during the solubilization of rhLK68 from the inclusion body during a conventional single introduction of lactose. To overcome this problem, a continuous induction strategy was applied where lactose was mixed with glycerol and fed continuously in a dissolved oxygen (DO)-stat manner. With the sub-optimal feed medium consisted of 1:50 of lactose/glycerol (w/w), the expression level reached 16% of the total cellular protein, which was 1.6-fold higher than that obtained from the conventional lactose induction. Moreover, the solubilization yield of rhLK68 from the inclusion body increased from 30 +/- 5 to 85 +/- 3% compared to the conventional single introduction of lactose. This result suggests that the continuous lactose induction strategy beneficially influenced the expression level of rhLK68 and the quality of its inclusion body.
Lim
Hyung-Kwon
HK
Mogam Biotechnology Research Institute, 341 Pojung-Ri, Koosung-Myun, Yongin, Kyonggi 449-913, Republic of Korea.
Kim
Sung-Geun
SG
Jung
Kyung-Hwan
KH
Seo
Jin-Ho
JH
D001053:Apolipoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D001054:Apolipoproteins A / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D053241:Apoprotein(a); D018929:Cell Culture Techniques / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism*; D007785:Lactose / Q000378:metabolism*; D017270:Lipoprotein(a) / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis
D001053:Apolipoproteins; D001054:Apolipoproteins A; D017270:Lipoprotein(a); D010446:Peptide Fragments; D011994:Recombinant Proteins; C487326:rhLK68 protein; D053241:Apoprotein(a); D007785:Lactose