Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound
Patent
1994-03-25
1997-06-24
Vogel, Nancy T.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing oxygen-containing organic compound
4351723, 4352542, 43525421, 43525423, 536 241, C12P 706, C12N 1500, C12N 119, C12N 116
Patent
active
056416610
ABSTRACT:
Two novel genomic clones, ZZA1 and ZZA2, were isolated which encode for Pichia pastoris alcohol oxidase isozymes. The 5' non-coding region of ZZA1 contains common structural features involved in the transcription and translation of eukaryotic genes. Comparison of the nucleotide sequences of the ZZA1 and AOX1 51 noncoding regions showed that they are 66% similar to each other.
The rice .alpha.-amylase gene OS103 was placed under the transcriptional control of the ZZA1 promoter. The nucleotide sequences of ZZA1 and other methanol-regulated promoters were analyzed. A highly conserved sequence (TTGNNNGCTTCCAANNNNNTGGT) (SEQ ID NO:2) was found in the 51 flanking region. A yeast strain containing the ZZA1 -OS103 fusion and secreting biologically active a-amylase into the culture media while converting starch to ethanol was produced. The ZZA1 and ZZA2 regulatory sequences may be used to contol the expression of other heterologous proteins in multiple yeast species.
Methods of purifying proteins that are regulators of alcohol oxidase expression (referred to as AOER proteins) and methods of isolating these proteins are also provided.
The invention also provides for the efficient conversion of starch into ethanol.
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Cregg et al., Mol. Cell. Biol., 9:1316-1323 (1989).
Koutz et al., Yeast 5:167-177 (1989).
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Cohen et al., Mol. Cell. Biol., 7:2753-2761 (1987).
Kumagai Monto H.
Sverlow Genadie G.
Biosource Technologies, Inc.
Halluin Albert P.
Vogel Nancy T.
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