Quantitative Analysis of Parasporal Crystal Protein from Bacillus thuringiensis by Capillary Electrophoresis

Cheng Ming Liu, Yew M. Tzeng

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The amount of parasporal crystal protein (δ-endotoxin) from the fermentation broth of the Bacillus thuringiensis is the best indication to assess the efficiency of the fermentation process or strain selection. Traditional methods for the assessment of insecticidal effect, such as bioassay or HPLC, were either time-consuming, inaccurate or inefficient. In this study, capillary electrophoresis (CE) was used for analyzing the amount of parasporal crystal protein after it was dissolved by adding a reducing agent, such as β-mercaptoethanol, to break the disulfide bonds. This soluble protein, δ-endotoxin, was then subjected to quantitative analysis by CE. The running buffer contained 300 mM boric acid, and pH was adjusted to 10.0 with 1 N NaOH. The dimensions of the capillary were 47 cm × 50 μm I.D. without coating. Lysozyme was used as internal standard for the quantitative assay of the δ-endotoxin. The migration time of the lysozyme peak was approximately 2 minutes earlier than that of the δ-endotoxin peak. The correlation between the concentration of δ-endotoxin and the ratio of the peak area of δ-endotoxin and the peak of lysozyme was calculated. The linear regression analysis showed that the correlation coefficient is equal to 0.9994, the slope is 0.4095 and the intercept is +0.0025. From this standard regression equation, the concentration of δ-endotoxin in fermentation broth or solution can be estimated easily by CE analysis.

Original languageEnglish
Pages (from-to)79-83
Number of pages5
JournalJournal of Food and Drug Analysis
Volume9
Issue number2
Publication statusPublished - Jun 2001

Fingerprint

crystal proteins
capillary electrophoresis
Capillary Electrophoresis
endotoxins
Bacillus thuringiensis
Endotoxins
quantitative analysis
Muramidase
lysozyme
Fermentation
fermentation
Proteins
boric acid
reducing agents
Mercaptoethanol
disulfide bonds
Reducing Agents
Bacillus Thuringiensis insecticidal crystal protein
Disulfides
Biological Assay

Keywords

  • Bacillus thuringiensis
  • Capillary electrophoresis
  • Parasporal crystal protein
  • Quantitative analysis

ASJC Scopus subject areas

  • Food Science
  • Pharmacology

Cite this

Quantitative Analysis of Parasporal Crystal Protein from Bacillus thuringiensis by Capillary Electrophoresis. / Liu, Cheng Ming; Tzeng, Yew M.

In: Journal of Food and Drug Analysis, Vol. 9, No. 2, 06.2001, p. 79-83.

Research output: Contribution to journalArticle

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