Expression and Fermentation Optimization of Candida antarctica Lipase B in Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2020-0529

Abstract

Abstract:

In order to achieve efficient soluble expression of Candida antarctica lipase B（CALB）in Escherichia coli at lower cost,CALB gene expression plasmids with different signal peptides were constructed and transformed into different E. coli hosts,and then a series of fermentation experiments were employed in shaking flasks to optimize the culture medium types,inducing conditions,components of medium and time course of cultivation. The results showed that the recombinant strain pET25b-CALB-1/Rosetta（DE3）with PelB signal peptide was induced to have the best expression in TY medium at 20℃ employing 0.5%（W/V）lactose. The optimal composition of fermentation medium was 1.75%（W/V）sorbitol,2.25%（W/V）fish peptone,1%（W/V）Angel yeast extract and 0.75%（W/V）Na₂HPO₄. The recombination system achieved a maximum CALB activity of 35.67 U/mL after induction for 60 h,which was 17.77 times higher than that before optimization,i.e.,the highest one of CALB produced by E. coli at present. In conclusion,the expression system in E. coli was successfully established,and the high-level soluble expression of CALB was obtained after system optimization.

Key words: Candida antarctica lipase B, Escherichia coli, soluble expression, fermentation optimization

WU Rong, CAO Jia-rui, CAO Jun, LIU Fei-xiang, YANG Meng, SU Er-zheng. Expression and Fermentation Optimization of Candida antarctica Lipase B in Escherichia coli[J]. Biotechnology Bulletin, 2021, 37(2): 138-148.

Figures/Tables 12

References 42

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Carbon sources （1.0%,W/V）	OD₆₀₀	Enzyme activity（U/mL）	Specific activity（U/mg）
Sucrose	4.72±0.12	1.59±0.02	2.83±0.05
Glucose	2.84±0.09	0.78±0.01	5.77±0.07
Glycerol	10.10±0.24	5.93±0.06	5.84±0.08
Sorbitol	13.20±0.21	12.12±0.11	7.61±0.08
Starch	8.82±0.17	2.45±0.02	2.58±0.04
Dextrin	12.70±0.43	7.43±0.07	6.07±0.05

Carbon sources （1.0%,W/V）	OD₆₀₀	Enzyme activity（U/mL）	Specific activity（U/mg）
Sucrose	4.72±0.12	1.59±0.02	2.83±0.05
Glucose	2.84±0.09	0.78±0.01	5.77±0.07
Glycerol	10.10±0.24	5.93±0.06	5.84±0.08
Sorbitol	13.20±0.21	12.12±0.11	7.61±0.08
Starch	8.82±0.17	2.45±0.02	2.58±0.04
Dextrin	12.70±0.43	7.43±0.07	6.07±0.05

Nitrogen sources （2.0%,W/V）	OD₆₀₀	Enzyme activity （U/mL）	Specific activity （U/mg）
Tryptone	12.69±0.32	19.41±0.20	12.15±0.09
SM	15.16±0.37	9.50±0.07	4.22±0.06
BBP	12.88±0.25	20.99±0.19	6.66±0.07
FP	16.14±0.40	22.70±0.17	6.97±0.06
PP	12.55±0.11	1.32±0.03	0.66±00.05
CSL	11.59±0.31	5.32±0.06	2.48±0.02

Nitrogen sources （2.0%,W/V）	OD₆₀₀	Enzyme activity （U/mL）	Specific activity （U/mg）
Tryptone	12.69±0.32	19.41±0.20	12.15±0.09
SM	15.16±0.37	9.50±0.07	4.22±0.06
BBP	12.88±0.25	20.99±0.19	6.66±0.07
FP	16.14±0.40	22.70±0.17	6.97±0.06
PP	12.55±0.11	1.32±0.03	0.66±00.05
CSL	11.59±0.31	5.32±0.06	2.48±0.02