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Optimization of Asparaginase Enzyme in Bioreactor

Momeni, Vahid | 2014

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 46253 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Vosoughi, Manouchehr; Alemzadeh, Iran
  7. Abstract:
  8. L-asparaginase enzyme is a well known enzyme due to itschemotherapeutic properties. However, this is not the only feature of the enzyme. L- asparaginase can be also used in food processing to reduce of acrylamide which is a carcinogenic substance. furthermore, acrylamide formation, is prevented by addition of the enzyme before cooking process of some foods such as chips, which contains sugar and asparagine amino acid. Due to use of the enzyme in food industry, we need to produce enzyme with high activity and cheaper raw material. In the present study, Candida utilis ATCC 9950 was used to produce asparaginase enzyme, and beet molasses and yeast extract was used as a carbohydrate and nitrogen surces. Optimization of three parameters, yeastextract concentration (3.37 gr/ml),beet molasses concentration (86.93) and fermentation time (18.49) was done with response surface methodology (RSM) in Erlenmeyer flask. After that, these values were used in 13L labratoary fermentor with 3L production meduim to optimize aeration rate and agitation speed, which are effective parameters in fermentor to produce enzyme. The optimal codition of aeration rate and agitation speed in fermentor was 1.25 vvm and 300 rpm, which in this condition, the enzyme activity was 245.6 (IU/ml). In the present study, the purification of enzyme was also done using pricipptation with aceton and Q-seoharose coromatogeraphy column , which the best specific activity was observed in 60% saturated solution of acetone (6998.95 U/g) and this value was increaased to 7853 U/mg in chromatogeraphy culumn
  9. Keywords:
  10. Acryle Amide ; Candida Utilis Yeast ; Optimization ; L-Asparginase Enzyme ; Aeration and Agitation

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