Ethanolic fermentation of xylose with Saccharomyces cerevisiae harboring the Thermus thermophilus xylA gene, which expresses an active xylose (glucose) isomerase

The Thermus thermophilus xylA gene encoding xylose (glucose) isomerase was cloned and expressed in Saccharomyces cerevisiae under the control of the yeast PGK1 promoter. The recombinant xylose isomerase showed the highest activity at 85 degrees C with a specific activity of 1.0 U mg-1. A new functional metabolic pathway in S. cerevisiae with ethanol formation during oxygen-limited xylose fermentation was demonstrated. Xylitol and acetic acid were also formed during the fermentation.

[1]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[2]  B. Hahn-Hägerdal,et al.  HPLC determination of xylulose formed by enzymatic xylose isomerization in lignocellulose hydrolysates , 1989 .

[3]  B. Hahn-Hägerdal,et al.  Expression of different levels of enzymes from the Pichia stipitis XYL1 and XYL2 genes in Saccharomyces cerevisiae and its effects on product formation during xylose utilisation , 1997, Applied Microbiology and Biotechnology.

[4]  T. Jeffries Utilization of xylose by bacteria, yeasts, and fungi. , 1983, Advances in biochemical engineering/biotechnology.

[5]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[6]  B. Hall,et al.  Expression of the Escherichia coli xylose isomerase gene in Saccharomyces cerevisiae , 1987, Applied and environmental microbiology.

[7]  John N. Saddler,et al.  Bioconversion of Forest and Agricultural Plant Residues , 1993 .

[8]  B. Hahn-Hägerdal,et al.  Xylulose and glucose fermentation by Saccharomyces cerevisiae in chemostat culture , 1996, Applied and environmental microbiology.

[9]  Gerald R. Fink,et al.  Methods in Yeast Genetics: Laboratory Manual , 1981 .

[10]  V. Rich Personal communication , 1989, Nature.

[11]  Li Fu Chen,et al.  Purification and properties of glucose isomerase of Actinoplanes missouriensis , 1980 .

[12]  A. Kingsman,et al.  Efficient synthesis of enzymatically active calf chymosin in Saccharomyces cerevisiae. , 1983, Gene.

[13]  M. Penttilä,et al.  Xylose-metabolizing Saccharomyces cerevisiae strains overexpressing the TKL1 and TAL1 genes encoding the pentose phosphate pathway enzymes transketolase and transaldolase , 1995, Applied and environmental microbiology.

[14]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[15]  S. Udaka,et al.  Xylose (glucose) isomerase gene from the thermophile Thermus thermophilus: cloning, sequencing, and comparison with other thermostable xylose isomerases , 1991, Journal of bacteriology.

[16]  John D. Wright,et al.  Xylose fermentation , 1989 .

[17]  Toshiomi Yoshida,et al.  Construction of xylose-assimilating Saccharomyces cerevisiae , 1993 .

[18]  P. Trinder Determination of Glucose in Blood Using Glucose Oxidase with an Alternative Oxygen Acceptor , 1969 .