KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Flow Injection Analysis for On-line Monitoring of Trehalose in Fermentation Processes

발효공정에서 트레할로스의 온라인 모니터링을 위한 흐름주입분석

  • Han, Kyung-Ah (Department of Material and Biochemical Engineering, Chonnam National University) ;
  • Rhee, Jong-Il (The Research Institute for Catalysis, Chonnam National University)
  • 한경아 (전남대학교 물질생물화학공학과) ;
  • 이종일 (전남대학교 촉매연구소)
  • Published : 2007.04.30
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Abstract

Trehalose is non-reducing disaccharide which is found in bacteria, fungi, plants and insects. Trehalose has been determined by several analysis methods. To monitor the concentrations of trehalose in a process, enzymatic methods have more advantage over others, e.g. more specific. In this work, trehalase was immobilized on VA-epoxy polymer and applied to FIA systems. The behaviours of these FIA systems were characterized and used to monitor the trehalose concentrations. Use of optical detection technique was chosen for trehalose-FIA system. On-line monitoring data and off-line data were measured by HPLC.

본 연구에서는 고가의 TreH를 형질전환한 대장균으로부터 생산하고 부분적으로 정제한 후, 에폭시에 고정화하고 트레할로스-FIA 시스템에 이용하였다. 산소 전극을 이용한 트레할로스 검출 방법과 분광학적 방법을 이용한 트레할로스 검출 방법을 비교하고, 공정에서의 온라인 모니터링의 가능성을 검토하였다. 산소 전극을 이용한 분석의 경우 다른 대사 물질로 인한 간섭현상이 상대적으로 적고 검출 가능한 농도가 0.1$\sim$1.0 g/L로 많은 량의 트레할로스를 생산하는 공정을 모니터링할 경우 적합한 시스템임을 알 수 있었다. 반면 분광학적 방법으로 트레할로스를 검출한 실험의 경우 TreH와 GOD/HRP를 고정하여 실험한 경우와 TreH만 고정하고 GOD와 HRP를 직접 주입하는 방식을 비교했을 때 검출 가능한 농도는 0.01$\sim$0.2 g/L로 낮은 농도의 트레할로스를 모니터링할 경우에 적합하다. 그러나 ABTS나 GOD, HRP의 주입량 등 경제성과 운전이 편이성 등을 고려할 경우 GOD와 HRP를 직접 주입하는 시스템이 가장 효과적임을 알 수 있었다. 이 시스템을 이용하여 소형 생물반응기 내 트레할로스의 농도를 온라인 모니터링한 결과는 오프라인 값과 매우 잘 일치함을 보였다. 생물공정에서 트레할로스의 온라인 모니터링이 FIA 기술에 의해 가능함을 확인하였다.

Keywords

References

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