Assay of $\beta$-Galactosidase Using High Performance Liquid Chromatography

고속액체크로마토그래피를 이용한 유당분해효소의 활성도 측정

  • Shin, Myung Gon (Rice Utilization Research Center, Korea Food Research Institute) ;
  • Chang, Pahn Shick (Department of Food Science and Technology, Seoul National Polytechnic University) ;
  • Min, Bong Kee (Rice Utilization Research Center, Korea Food Research Institute) ;
  • Kim, Sun Chang (Department of Food Science and Technology, Kyungnam University)
  • 신명곤 (한국식품개발연구원 쌀이용연구센터) ;
  • 장판식 (서울산업대학교 식품공학과) ;
  • 민봉기 (한국식품개발연구원 쌀이용연구센터) ;
  • 김선창 (경남대학교 식품공학과)
  • Received : 1992.08.14
  • Published : 1992.12.25

Abstract

An analytical procedure is presented for the quantitative determination of lactose, glucose, and galactose in the hydrolyzate of lactose by ${\beta}$-galactosidase with high-performance liquid chromatography. An Aminex HPX-87C column at $85^{\circ}C$ and refractive index detector were used to resolve lactose, glucose, and galactose in only 12 minutes with distilled and deionized water as a mobile phase. The validity of high-performance liquid chromatography as a method for the assay of ${\beta}$-galactosidase was supported by recovery experiments and comparision of results with those by ONPG method, a spectrophotometric assay. The procedure was appropriate for determination of sugars in the enzyme reaction mixture and could by applied to analysis of ${\beta}$-galactosidase activity.

Keywords

${\beta}$-Galactosidase;HPLC;ONPG;quantitative determination