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KMID : 1094720000050050320
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Biotechnology and Bioprocess Engineering
2000 Volume.5 No. 5 p.320 ~ p.326
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Comparison of promoters suitable for regulated overexpression of β-galactosidase in the alkane-utilizing yeast Yarrowia lipolytica
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Juretzek Thomas
Wang Hui-Jie
Nicaud Jean-Marc
Mauersberger Stephan
Barth Gerold
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Abstract
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Promoters of the genesG3P, ICL1, POT1, POX1, POX2 andPOX5 of the yeastY. lipolytica were studied in respect to their regulations and activities during growth on different carbon sources. The aim of this study was to select suitable promoters for high expression of heterologous genes in this yeast. For this purpose the promoters were fused with the reporter genelacZ ofE. coli and integrated as single copies into the genome ofY. lipolytica strain PO1d. The measurement of expressed activities of β-galactosidase revealed thatpICL1, pPOX2 andpPOT1 are the strongest regulable promoters available forY. lipolytica, at present.pPOX2 andpPOT4 were highly induced during growth on oleic acid and were completely repressed by glucose and glycerol.pICL1 was strongly inducible by ethanol besides alkanes and fatty acids, however, not completely repressible by glucose or glycerol. Ricinoleic acid methyl ester appeared as a very strong inducer forpPOT1 andpPOX2, in spite of that it inhibited growth ofY. lipolytica transformants.
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KEYWORD
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Yarrowia lypolytica, expression vectors, regulated promoters, lacZ expression
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