Cloning and expression of a glucoamylase gene from Lactobacillus amylovorus ATCC 33621 in Escherichia coli |
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Authors: | Jennylynd A James Normand Robert Byong H Lee |
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Institution: | (1) Department of Food Science and Agricultural Chemistry, Macdonald Campus of McGill University, 21111 Lakeshore Rd., H9X 3V9 Ste-Anne de Bellevue, Québec, Canada;(2) Food Research and Development Centre, Agriculture Canada, J2S 8E3 St. Hyacinthe, Québec, Canada |
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Abstract: | Summary The glucoamylase gene from Lactobacillus amylovorus was cloned and expressed in Escherichia coli. A genomic DNA library from Lactobacillus amylovorus was prepared by partially digesting genomic DNA with EcoRI and ligating random fragments to the EcoRI digested cloning vector, pZErO-1.1. Three E. coli transformants expressing glucoamylase were identified using a probe prepared from the STA2 glucoamylase gene from Saccharomyces cerevisiae var. diastaticus. The physical maps of the recombinant plasmids were constructed. These plasmids contained inserts of about 5.2 Kb, 5.9 Kb and 6.4 Kb respectively. Temperature and pH optima of 45°C and 6.0, respectively, were obtained for both recombinant and purified wild type glucoamylases. Also, the enzymes were found to be thermolabile at temperatures above 50°C. |
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