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Establishment of a Cryopreservation Method for the Industrial Use of D-Amino Acid Oxidase-Overexpressing Escherichia coli

Authors
Kim, Sae-JinPark, Hye WonShin, Chang-HunKim, Chan-Wha
Issue Date
2월-2009
Publisher
TAYLOR & FRANCIS LTD
Keywords
cryopreservation; glycerol; master cell bank; working cell bank
Citation
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, v.73, no.2, pp.299 - 303
Indexed
SCIE
SCOPUS
Journal Title
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
Volume
73
Number
2
Start Page
299
End Page
303
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/120689
DOI
10.1271/bbb.80507
ISSN
0916-8451
Abstract
A cryopreservation condition for D-amino acid oxidase (DAAO)-overexpressing Escherichia coli (E. coli BL21(DE3)/pET-DAAO) was established. Ten percent was the optimum concentration of glycerol as a cryoprotectant, and its diffusion into stationary phase cells was superior to that into log cells. The results also showed that rather than fast cooling, a slow cooling method was appropriate to our recombinant E. coli. In addition, 15min was the best equilibration period, at which higher than 90% of recovery rates were maintained at all test points. Most importantly, the relative recovery rates, product yield, and fermentation pattern of the cell banks (CBs) constructed according to our cryopreservation method did not change over 12 months, confirming that our method not only permits exceptional cryopreservation, but offers prolonged productivity. Taken together, our results demonstrating a cryopreservation method for E. coli BL21(DE3)/pET-DAAO provide insight into an improvement in the industrial production of DAAO.
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