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KMID : 1160620010060010079
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Preventive Nutrition and Food Science
2001 Volume.6 No. 1 p.79 ~ p.81
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Protein Engineering of Deoxynucleoside Kinase from Lactobacillus acidophilus: Effect of Site-Directed Mutagenesis on Microbial Growth
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Park In-Shik
Kim Eun-Ae
Bang Keuk-Seung
Kim Seok-Hwan
Kim Gi-Nahm
Lee Min-Kyung
Kil Ji-Oeun
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Abstract
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Deoxynucleoside kinases exist as heterodimeric pairs specific for deoxyadenosine/deoxyguanosine kinase (dAK/dGK) and deoxyadenosine/deoxycytidine kinase (dAK/dCK). The aspartic acid-84 in dGK was mutated to alanine, asparagine and glutamic acid by site-directed mutagenesis. The mutation resulted in a drastic decease in dGK activity compared to the unmodified cloned enzyme while it increased production of dAK activity. The mutated dak/dgk genes, which synthesize tandem deoxyadenosine/deoxyguanosine kinase, were inserted back to the Lactobacillus acidophilus and Lactococcus lactis by electroporation to determine the effect of site-directed mutation of he enzymes on the microbial growth. However, no significant change was observed in cell growth and lactic acid production between wild type and mutant lactic acid bacteria.
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KEYWORD
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deoxyguanosine kinase, site-directed mutagenesis, electroporation, Lactobacillus acidophilus
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