Studies on the Use of Carboxymethylcellulose Sodium Salt Matrix for the Immobilization of Photobacterium phosphoreum

Photobacterium phosphoreum을 고정화하기 위한 Carboxymethylcellulose Sodium Salt [CMC] 담체의 이용성 연구

  • 이용제 (경희대학교 생명과학부 식품가공학과) ;
  • 정성제 (경희대학교 생명과학부 식품가공학과) ;
  • 허문석 (경희대학교 생명과학부 식품가공학과) ;
  • 전억한 (경희대학교 생명과학부 식품가공학과)
  • Published : 2000.02.01

Abstract

Bioluminescence of Photobacterium phosphoreum has been used for the detection of pollutants in the environment. Immobilization method was used to maintain the stability of bioluminescence of P. phosphoreum. The carboxymethylcellulose was investigated to find out whether it was suitable for the immobilization of P. phosphoreum as a matrix without disturbing the bioluminescence emission. A maintenance of bioluminescence was determined from the P. phosphoreum immobilized on the various concentrations of carboxymethylcellulose. A relatively high bioluminescence intensity was shown with immobilized cells on 1%(w/v) carboxymethylcellulose. The effect of carboxymethylcellulose concentrations on the sensitivity of Crcompounds including $Na_{2}CrO_{4}$, $K_{2}CrO_{4}$, $CrO_{3}$, CrK$(SO_4)_{2}$ and $CrCl_{3}$ to the bioluminescence intensity. The calculated $EC_{50}$ showed that the linear relations between such substances and bioluminesence intensity were established.

1.0%(w/v)의 CMC 담체는 고정화를 유지할 수 있는 점도를 가지며 0.1 M 이하의 양이온과 이온가교결합을 할 수 있는 농도이다. Luminometer tube내의 시료에 유동을 최소화함으로써 산소의 공급을 일정하게 하여 P. phosphoreum을 고정화 30분 후 bioluminescence intensity가 안정되어 바로 측정할 수 있는 장점을 가지고 있었다. 1.0%(w/v) CMC담체는 pH 6.92로 최적조건인 pH 7.0에 근접했으며, 발광기작에 필요한 산소전달(oxygen transfer)이 1.5%(w/v)~3.0%(w/v) CMC 담체보다 뛰어나 Bioluminescence intensity의 안정성을 부여하였다. Cr-화합물인 $Na_{2}CrO_{4}$, $K_{2}CrO_{4}$, $CrO_{3}$, CrK$(SO_4)_{2}$$CrCl_{3}$의 CMC담체에 대한 민감도는 $\gamma$값을 이용해서 $EC_{50}$값으로 나타내었을 때 $Na_{2}CrO_{4}$, $K_{2}CrO_{4}$, CrK$(SO_4)_{2}$$CrCl_{3}$$EC_{50}$값이 1.0%(w/v)CMC에서는 5.4~16.3 g/L으로 1.5%(w/v)~3.0%(w/v) CMC에서는 6.2~555.9 g/L의 범위로 나타났다. 이것은 1.0%(w/v) CMC가 낮은 독성 농도에서 bioluminescence intensity가 50% 감소함을 알 수 있고, 상관계수($R^2$)가 0.911~0.990 으로 높게 산출되었다. 따라서 1.0%(w/v) CMC 담체가 P. phosphoreum의 biolumincsene에 안정성을 주었으며, 독성물질에 가장 민감하다는 것을 알 수 있었다.

Keywords

References

  1. Principles of Biochemistry(2nd ed.) Albert L.L.;D.L. Nelson;M.M. Cox
  2. Anal. Biochem. v.219 Chemiluminescent and Bioluminescent: Reporter Gene Assays Bronstein I.;J. Fortin;P.E. Stanley;G.S.A.B. Stewart;L.J. Kricka
  3. J. Food Prot. v.55 no.1 Bacterial Bioluminescence Applications in Food Microbiology Baker J.M.;M.W. Griffiths;K.L. Collins-Thompson
  4. Symbiosis v.4 Review, How are Growth and Luminescence Regulated Independently in Light Organ Symbionts? Hasting J.W.;J. Mademsom;D. Paul
  5. Korean J. Biotechnol. Bioeng. v.14 no.3 Response of Photobacterium phosphoreium to Heavy Metal Chung K.H.;H.S. Kim;E.S. Lee;S.J. Jung;J.G. Lee;E.K. Kim;U.H. Chun
  6. Bioprocess Engineering Michael L.S.;K. Fikiet
  7. pestic. sci. v.45 Stability and Controlled Release Properties of Carboxymethylcellulose-Encapsulated Bucillus thuringiensts var. isiaelensts Cumhur C.;E. Murat
  8. Carbohydrate Polumers v.26 Scanning Electron Microscopic Analysis and Swelling Behaviout of Ionotropically Crosslinked Carboxymethylcellulose and Carboxymethylcellulose-gelatin Matrices Piasad M.P.;M. Kalynansundaram
  9. Carbolryrate Polymers v.40 Enzyme-Acided Characterization of Carboxymethylcellulose Horner S.;J. Puls;B. Saake;E.A. Klohr;H. Thielking
  10. J. Economic Entomology v.88 no.4 Aluminum Carboxymethlycellulose Encapsulation of Bacillus sphaericus 2362 for control of Culex app. Larvae Elcin Y.M.;C. Cumhur;S.C. Sacilik
  11. Toxicity screening procedures using bacterial systems Determination of the Toxicity of Selected Metalworking Fluid Preservatives by Use of the Microtox System and an In Vitro Enzyme Assay Mallak F.P.;R.L. Brunker;Dickson Liu(ed.);B.J. Dutka(eds.)
  12. Korean J. Biotechnol. Bioeng. v.14 no.1 The Effect of Storage Temperatire on the Viability and Bioluminescence of Photobacterium phosphoreum Kim H.S.;S.J. Jung;K.H. Chung;U.H. Chun
  13. Dept. of Food science and Technology Studies on the Stability of Bioluminescence Produced from Photobacterium phosphoreum and Its Linear Relationship with Toxic Substances Lee H.J.;M.S. Thesis(ed.)