Human α-amylase (EC 3.2.1.1) genes are located in a cluster on the chromosome that includes salivary amylase genes (AMY1), two pancreatic α-amylase genes (AMY2A and AMY2B). Various studies have been conducted on the structure, mutagenesis and kinetic analysis for chemically synthesized substrate of AMY2A. However, characterization of AMY2B has received less attention. In this study, human pancreatic α-amylase (AMY2B) gene was cloned and expressed in Pichia pastoris using the methanol-induced alcohol oxidase (AOX1) promoter. After 48 hours for incubation, the activity of recombinant AMY2B in the culture supernatant reached the peak and purified by Ni-NTA affinity chromatography. The optimum temperature and pH of purified enzymes were similar to those of AMY2A. Furthermore that is consistent with the characteristics of AMY2A. Kinetic analysis for various native starches revealed that chloride ion was required for maximum activity of recombinant AMY2B and increase in kcat values in the presence of NaCl was mostly contributed to the maximum activity.
저자
Min-Kyu Kim [ Department of Food and Biotechnology, Korea University ]
Young-Jong Kim [ Department of Food and Biotechnology, Korea University ]
Young-Wan Kim [ Department of Food and Biotechnology, Korea University ]
본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.