BioChem - DOS

Transcriptional analysis for carbon metabolism and kinetic modeling for heterologous proteins productions by Pichia pastoris in induction process with methanol/sorbitol co-feeding

Process biochemistry

Jia, Luqiang; Mpofu, Enock; Tu, Tingyong; Huai, Qiangqiang; Sun, Jiaowen; Chen, Shanshan; Ding, Jian; Shi, Zhongping

<P><B>Abstract</B></P> <P>It is difficult to control concentrations of methanol/dissolved oxygen at high levels simultaneously in heterologous proteins productions by <I>Pichia pastoris</I> during induction phase. Two strains, a methanol utilization slow (Mut<SUP>S</SUP>) type and a plus (Mut<SUP>+</SUP>) type were used with methanol/sorbitol co-feeding strategy to induce porcine interferon-&alpha; and human serum albumin-human granulocyte colony stimulating factor respectively, under the conditions of &ldquo;methanol sufficient-oxygen limited (MS-OL)&rdquo; and &ldquo;methanol limited-oxygen sufficient (ML-OS)&rdquo;. For the Mut<SUP>S</SUP>/Mut<SUP>+</SUP> strains, the target proteins titers under &ldquo;MS-OL&rdquo; were 6-fold/19.2% of those under &ldquo;ML-OS&rdquo;. The key genes in methanol metabolism of the Mut<SUP>S</SUP> strain were up-regulated under &ldquo;MS-OL&rdquo;, but they were not differently expressed in the Mut<SUP>+</SUP> strain. Methanol utilization rate (<I>r</I> <SUB>MeOH</SUB>) of the Mut<SUP>S</SUP> strain reduced when decreasing methanol concentration, but <I>r</I> <SUB>MeOH</SUB> of the Mut<SUP>+</SUP> strain unchanged unless methanol concentration decreased to a low-limit of 0.6g/L. Finally, kinetic models were designed to describe the methanol/sorbitol co-feeding process.</P> <P><B>Highlights</B></P> <P> <UL> <LI> &ldquo;Methanol sufficient-oxygen limited&rdquo; enhances the protein synthesis of Mut<SUP>S</SUP> strain. </LI> <LI> &ldquo;Methanol sufficient-oxygen limited&rdquo; enhances methanol metabolism in Mut<SUP>S</SUP> strain. </LI> <LI> &ldquo;Methanol sufficient-oxygen limited&rdquo; represses the protein synthesis of Mut<SUP>+</SUP> strain. </LI> <LI> &ldquo;Methanol sufficient-oxygen limited&rdquo; doesn&rsquo;t affect methanol metabolism in Mut<SUP>+</SUP> strain. </LI> <LI> Models for the relationship between methanol and sorbitol utilization were designed. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

2017

Elsevier

1359-5113

59

2

pp.159-166

10.1016/j.procbio.2017.05.011

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART78540727

P. pastoris; Methanol/sorbitol co-feeding; Methanol concentration; Dissolved oxygen; Transcriptome analysis; Kinetic model