, Volume 30, Issue 4, pp 351-357

The fermentation of xylose —an analysis of the expression of Bacillus and Actinoplanes xylose isomerase genes in yeast

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Summary

The genes encoding xylose isomerase from Bacillus subtilis and Actinoplanes missouriensis have been isolated by complementation of a xylose isomerase defective Escherichia coli mutant. The xylose isomerase gene from A. missouriensis could be expressed in E. coli under the control of its own promoter, whereas the cloned Bacillus gene was expressed in E. coli only after the spontaneous integration of the E. coli IS5 element. After fusion of the Bacillus gene to the yeast PDC1 promoter, transformants of Saccharomyces cerevisiae contained the xylose isomerase protein. Approx. 5% of the total cellular protein of transformants consisted of xylose isomerase that was found to be at least partly insoluble. Neither the insoluble protein nor Triton X-114 solubilized isomerase was catalytically active. To investigate whether the xylose isomerase of A. missouriensis can be expressed in S. cerevisiae the coding region was fused to the yeast GAL1 promoter. Analysis of total RNA from yeast transformants containing this construction showed a xylose isomerase specific mRNA.

Dedicated to Professor Karl Esser on the occasion of his 60th birthday