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KMID : 1094720080130030366
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Biotechnology and Bioprocess Engineering
2008 Volume.13 No. 3 p.366 ~ p.371
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The effect of the cspA 5′-untranslated region on recombinant protein production at low temperature
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Kim Su-Hyun
Heo Mi-Ae
Kim Yu-Jin
Kim So-Yeon
Neelamegam Rameshkumar
Lee Sun-Gu
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Abstract
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Expression of recombinant proteins in Escherichia coli is often carried out at low temperatures so as to improve the solubility and stability of the expressed proteins. However, the use of low temperature significantly limits the growth rate of the expression host, leading to a reduction in protein productivity. Here, we show that the introduction of the cspA 5′-untranslated region (cspA 5′-UTR) onto the transcript of a target gene enhances the expression levels via the T7 promoter at low temperatures. First, the effects of the cspA 5′-UTR sequence and CspA coexpression on green fluorescence protein (GFP) production were investigated. While the cspA 5′-UTR sequence had no influence on the expression rate of GFP at 37°C, it improved the expression level of GFP by approximately 1.5 fold at 15°C. Coexpression of the CspA protein further increased the expression level of GFP only slightly in the presence of the cspA 5′-UTR. Subsequently, the effect of the cspA 5′-UTR on protein production was examined for six different proteins (CysS, HisS, ProS, ThrS, TrpS, and YadB) at 15°C. The introduction of cspA 5′-UTR sequence led to approximate two-fold improvement for HisS and ThrS production, a slight improvement for TrpS and YadB, and no influence on CysS and ProS protein production. Therefore, the effect of the cspA 5′-UTR on the expression level at low temperature varied depending on the target gene.
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KEYWORD
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cspA 5′-UTR, CspA, low temperature, T7 promoter, gene expression
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