Thermostable alpha-galactosidase and methods of use
AssigneesRutgers, The State University of New Jersey, The Board of Trustees of the University of Illinois
InventorsDinesh Yernool, Douglas E. Eveleigh, Michael R. King, Bruce Chassy
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Abstract
An isolated nucleic acid from the bacterium Thermotoga neapolitana, encoding a thermostable .alpha.-galactosidase is provided. The Thermotoga gene is cloned into a high expression vector to provide large quantities of a purified thermostable .alpha.-galactosidase. The thermostable enzyme is used in high-temperature processing of soy products to remove .alpha.-galactosides.
Claims12
An isolated nucleic acid molecule from Thermotoga neapolitana that comprises an open reading frame encoding a thermostable .alpha.-galactosidase and possess the same enzymatic activity as the protein having the amino acid sequence of SEQ ID NO:2.
An isolated nucleic acid molecule having a sequence selected from the group consisting of:
- a) SEQ ID NO:1;
- b) sequence hybridizing with SEQ ID NO:1 or its complement under hybridization conditions calculated to achieve hybridization between two single-stranded DNA molecules of greater than 80% homology, using a formula of:
- T.sub.m =81.5.degree. C.+16.6Log[Na+]+0.41 (% G+C)-0.63(% formamide)-600/#bp in duplex,
- wherein the T.sub.m of a DNA duplex decreases by 1-1.5.degree. C. with every 1% decrease in homology;
- and
- c) a sequence encoding a polypeptide having amino acid SEQ ID NO:2;
- wherein said isolated nucleic acid molecule has encodes a protein having the amino acid sequence of SEQ ID NO:2.
A method of depleting soybeans of .alpha.-galactosides, which comprises providing transgenic, fertile soybean plants that produce an enzymatically active Thermotoga neapolitana .alpha.-galactosidase possessing the same enzymatic activity as the protein having the amino acid sequence of SEQ ID NO:2, growing the plants, harvesting soybeans from the plants, and heating the soybeans for a time and at a temperature effective to deplete the .alpha.-galactosides from the soybeans.
Dependencies are inferred from the claim text.