|
|
|
| Optimization of Fermentation in Shake Flasks for Recombinant γ-CGTase Expression in E. coli |
| JI Li-ping1,2, WU Dan1,2, WU Jing1,2, CHEN Jian1,2 |
1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;
2. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China |
|
|
|
Abstract In order to achieve efficient extracellular production of γ-cyclodextrin glycosyltransferase(γ-CGTase), E. coli BL21(DE3) carrying the γ-cgt gene of Alkalophilic Bacillus clarkii 7364 was investigated. By comparison of different culture media and conditions, got the optimized medium: 5g/L glycerol, 6g/L peptone, 24g/L yeast extract, 6mmol/L Ca2+, 2mmol/L Mg2+, 0.75% glycine, the optimized conditions: cultivated in 25℃, 220r/min, 30ml/250ml, pH 6.5, induced by 5g/L lactose at 10h of culture, which contained 0.02% SDS, the enzyme activity rised from 5189.2U/ml to 14683.4U/ml.
|
|
Received: 09 May 2011
Published: 25 October 2011
|
|
|
|
|
|
[1] Szerman N,Schroh I, Rossi A L,et al.Cyclodextrin production by cyclodextrin glycosyltransferase from Bacillus circulans DF 9R. Bioresour Technol, 2007,98(15):2886-2891.
[2] Li Z F, Wang M, Wang F, et al.Gamma-cyclodextrin:a review on enzymatic production and applications.App Microbiol Biotechnol, 2007,77(2):245-255.
[3] Cucinotta V, Contino A, Giuffrida A, et al. Application of charged single isomer derivatives of cyclodextrins in capillary electrophoresis for chiral analysis. Journal of Chromatography A, 2010,1217(7):953-967.
[4] Loftsson T, Brewster M E. Pharmaceutical applications of cyclodextrins: basic science and product development. Journal of Pharmacy and Pharmacology, 2010,62(11):1607-1621.
[5] Szente L, Szejtli J. Cyclodextrins as food ingredients. Trends Food Sci Tech, 2004, 15(3-4): 137-142.
[6] Moldenhauer J, Cully J. Method for producing a coenzyme Q10/g-cyclodextrin complex. US Pat, 0012774A1.2003.
[7] 胡静,陈育如,魏霞,等. 高产环糊精葡萄糖基转移酶的枯草芽孢杆菌选育、产酶与酶学特性. 食品与生物技术学报, 2008,27(4):97-102. Hu J,Chen Y R,Wei X, et al. Journal of Food Science and Biotechnology, 2008, 27(4):97-102.
[8] 朱德艳. 一株产环糊精葡萄糖基转移酶的枯草芽孢杆菌的选育及产酶条件的研究. 荆楚理工学院学报, 2009,24(5):29-32. Zhu D Y. Breeding and Enzymatic Properties of a Bacillus subtilis producing cyclomaltodextrin glucanotransferase. Journal of JingChu University of Technology, 2009,24(5):29-32.
[9] 曹新志,金征宇.嗜碱芽孢杆菌产环糊精葡萄糖基转移酶发酵条件的优化.食品科学, 2005,26(2):122-126. Cao X Z,Jin Z Y. Food Science, 2005,26(2):122-126.
[10] Wu Q, Xu H, Ying H J, et al. Kinetic analysis and pH-shift control strategy for poly(gamma-glutamic acid) production with Bacillus subtilis CGMCC 0833. Biochemical Engineering Journal, 2010,50(2):24-28.
[11] Wei G, Li Y, Du G, et al. Application of a two-stage temperature control strategy for enhanced glutathione production in the batch fermentation by Candida utilis. Biotechnol Lett, 2003, 25: 887-890.
[12] Goyal D, Sahni G, Sahoo D K. Enhanced production of recombinant streptokinase in Escherichia coli using fed-batch culture. Bioresource Technology, 2009,100(19): 4468-4474.
[13] Sorensen H P, Mortensen K K. Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol, 2005, 115(2): 113-128.
[14] Reddy R M, Reddy P G, Seenayya G. Enhanced production of thermostable β-amylase and pullulanase in the presence of surfactants by Clostridium thermosulfurogenes SV2. Process Biochem, 1999, 34: 87-92.
[15] Vibhor S, Jeewon L, Dae Y K. Synthesis of recombinant human interleukin-2 via controlled feed of lactose-complex media in fed-batch cultures of Escherichia coli BL21(DE3). Biotechnology Letters, 2000,22(4): 261-265.
[16] Laxman S S, Savergave S D, Vitthal V J, et al. Production and single step purification of cyclodextrin glycosyltransferase from alkalophilic Bacillus firmus by ion exchange chromatography. Biochem Eng J, 2008, 39:510-515.
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
