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Molecular Orbital Theory on Cellulolytic Reactivity Between pNP-Cellooligosccharides and ${\beta}$-Glucosidase from Cellulomonas uda CS1-1

Yoon, Min-Ho   (Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Chungnam National UniversityUU0001302  ); Nam, Yun-Kyu   (Department of Bioenvironment, Agricultural Research & Extension Services of Chungcheongnam-Do Province  ); Choi, Woo-Young   (Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Chungnam National UniversityUU0001302  ); Sung, Nack-Do   (Division of Applied Biology and Chemistry, College of Agriculture and Life Sciences, Chungnam National UniversityUU0001302  );
  • 초록

    A ${\beta}$ -glucosidase with the molecular mass of 160,000 Da was purified to homogeneity from cell extract of a cellulolytic bacterium, Cellulomonas uda CS1-1. The kinetic parameters ( $K_m$ and $V_{max}$ ) of the enzyme were determined with pNP-cellooligosccharides (DP 1-5) and cellobiose. The molecular orbital theoretical studies on the cellulolytic reactivity between the pNP-cellooligosaccharides as substrate (S) molecules and the purified ${\beta}$ -glucosidase (E) were conducted by applying the frontier molecular orbital (FMO) interaction theory. The results of the FMO interaction between E and S molecules verified that the first stage of the reaction was induced by exocyclic cleavage, which occurred in an electrophilic reaction based on a strong charge-controlled reaction between the highest occupied molecular orbital (HOMO) energy of the S molecule and the lowest occupied molecular orbital (LUMO) energy of the hydronium ion ( $H_3O^+$ ), more than endocyclic cleavage, whereas a nucleophilic substitution reaction was induced by an orbital-controlled reaction between the LUMO energy of the oxonium ion ( $SH^+$ ) protonated to the S molecule and the HOMO energy of the $H_2O_2$ molecule. A hypothetic reaction route was proposed with the experimental results in which the enzymatic acid-catalyst hydrolysis reaction of E and S molecules would be progressed via $SN_1$ and $SN_2$ reactions. In addition, the quantitative structure-activity relationships (QSARs) between these kinetic parameters showed that $K_m$ has a significant correlation with hydrophobicity (logP), and specific activity has with dipole moment, respectively.


  • 주제어

    Cellulolytic reactivity .   Cellulomonas uda CS1-1 .   ${\beta}$-glucosidase .   pNP- ${\beta}$-cellooligosaccharides .   MO theory.  

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