Enzymatic Transgalactosylation of Ascorbic Acid by ${\beta}-Galactosidase$

${\beta}-Galactosidase$에 의한 Ascorbic Acid의 효소적 배당화

  • Jang, So-Young (Dept. of Food Science and Technology, Chonbuk National University) ;
  • Kim, Young-Hoi (Korea Ginseng and Tobacco Research Institute) ;
  • Kim, Myung-Kon (Iri Nat'l College of Agriculture and Technology) ;
  • Kim, Young-Soo (Dept. of Food Science and Technology, Chonbuk National University) ;
  • Hong, Jai-Sik (Dept. of Food Science and Technology, Chonbuk National University)
  • Published : 1998.12.31

Abstract

The enzymatic transgalactosylation of L-ascorbic acid was investigated to synthesize a chemically stable form of L-ascorbic acid by using commercially available ${\beta}-galactosidases$. Among various enzymes examined, Aspergillus oryzae ${\beta}-galactosidase$ was found to be formed the derivative of ascorbic acid in a high yield from ascorbic acid and lactose. The reaction product was isolated by ion exchange chromatography on a $Dowex\;1\;{\times}\;8$ (Cl - form) resin and Toyopearl HW-40S gel chromatography. The product was identified as $6-O-{\beta}-_D-galactopyranosyl-_L-ascorbic\;acid$ on the basis of various experimental results, viz., UV, IR, $^1H-NMR,\;^{13}C-NMR$ and mass spectral data.

Ascorbic acid(AsA)의 산화 안정성 증진을 목적으로 효소적 배당화를 시도하였다. 본 실험에서 사용한 6종의 ${\beta}-galactosidase$에 대한 당 전이 생성물의 수율을 분석 비교한 결과 Asp. oryzae에서 분리한 ${\beta}-galactosidase$의 당 전이활성이 가장 높았으며, 당 공여체인 lactose와 당 수용체인 AsA 함유용액에서 당 전이 수율은 약 30% 수준이었다. 당 전이 생성물은 $Dowex\;1{\times}8$(Cl-형) 수지에 의한 ion exchange chromatography와 Toyopearl 40S gel chromatography에 의해 분리한 다음 동결건조하였다. Asp. oryzae 유래의 ${\beta}-galactosidase$ 존재 하에서 AsA와 lactose의 반응액에서 분리한 생성물은 UV, IR, CI-MS, $^1H-NMR,\;^{13}C-NMR$ 등을 이용한 기기분석, 산 및 ${\beta}-galactosidase$에 의한 가수분해 특성을 조사한 결과 AsA에 galactose 1분자가 결합된 $6-O-{\beta}-_D-galactopyranosyl-_L-ascorbic\;acid$로 판명되었다.

Keywords