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논문 기본 정보
- 자료유형
- 학술저널
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제20권 제6호
- 발행연도
- 2010.1
- 수록면
- 1,018 - 1,021 (4page)
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초록· 키워드
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Metallic and non-metallic isomerases can be used to produce commercially important monosaccharides. To determine which category of isomerase is more suitable as a template for directed evolution to improve enzymes for galactose isomerization, L-arabinose isomerase from Escherichia coli (ECAI; E.C. 5.3.1.4) and tagatose-6-phosphate isomerase from Staphylococcus aureus (SATI; E.C. 5.3.1.26) were chosen as models of a metallic and non-metallic isomerase,respectively. Random mutations were introduced into the genes encoding ECAI and SATI at the same rate, resulting in the generation of 515 mutants of each isomerase. The isomerization activity of each of the mutants toward a non-natural substrate (galactose) was then measured. With an average mutation rate of 0.2 mutations/kb, 47.5% of the mutated ECAIs showed an increase in activity compared with wild-type ECAI, and the remaining 52.5% showed a decrease in activity. Among the mutated SATIs, 58.6%showed an increase in activity, whereas 41.4% showed a decrease in activity. Mutant clones showing a significant change in relative activity were sequenced and specific increases in activity were measured. The maximum increase in activity achieved by mutation of ECAI was 130%, and that for SATI was 190%. Based on these results, the characteristics of the different isomerases are discussed in terms of their usefulness for directed evolution of nonnatural substrate isomerization.
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참고문헌 (16)
참고문헌 신청
Asboth, B / 2000 / Mechanism of action of D-xylose isomerase / Curr. Protein Pept. Sci 1 : 237 ~ 254
Bhosale, S. H / 1996 / Molecular and industrial aspects of glucose isomerase / Microbiol. Rev 60 : 280 ~ 300
Choi, Ki-Won / 2008 / Modulation of the Regioselectivity of a Thermotoga neapolitana β-Glucosidaseby Site-Directed Mutagenesis / Journal of Microbiology and Biotechnology 18
Dische, Z / 1951 / A new spectrophotometric method for the detection and determination of keto sugars and trioses / J. Biol. Chem 192 : 583 ~ 587
Kim, P / 2001 / Improvement of tagatose conversion rate by genetic evolution of thermostable galactose isomerase / Biotechnol. Appl. Biochem 34 : 99 ~ 102
Bhosale, S. H / 1996 / Molecular and industrial aspects of glucose isomerase / Microbiol. Rev 60 : 280 ~ 300
Choi, Ki-Won / 2008 / Modulation of the Regioselectivity of a Thermotoga neapolitana β-Glucosidaseby Site-Directed Mutagenesis / Journal of Microbiology and Biotechnology 18
Dische, Z / 1951 / A new spectrophotometric method for the detection and determination of keto sugars and trioses / J. Biol. Chem 192 : 583 ~ 587
Kim, P / 2001 / Improvement of tagatose conversion rate by genetic evolution of thermostable galactose isomerase / Biotechnol. Appl. Biochem 34 : 99 ~ 102
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