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Coenzyme B12 Can Be Produced by Engineered Escherichia Coli Under Both Anaerobic and Aerobic Conditions

Coenzyme B12 Can Be Produced by Engineered Escherichia Coli Under Both Anaerobic and Aerobic Conditions

Yeounjoo Ko, Somasundar Ashok, Satish Kumar Ainala, Mugesh Sankaranarayanan, Ah Yeong Chun, Gyoo Yeol Jung, Sunghoon Park

Biotechnol J. 2014 Dec;9(12):1526-35.

PMID: 25146562

Abstract:

Coenzyme B12 (Vitamin B12 ) is one of the most complex biomolecules and an essential cofactor required for the catalytic activity of many enzymes. Pseudomonas denitrificans synthesizes coenzyme B12 in an oxygen-dependent manner using a pathway encoded by more than 25 genes that are located in six different operons. Escherichia coli, a robust and suitable host for metabolic engineering was used to produce coenzyme B12 . These genes were cloned into three compatible plasmids and expressed heterologously in E. coli BL21 (DE3). Real-time PCR, SDS-PAGE analysis and bioassay showed that the recombinant E. coli expressed the coenzyme B12 synthetic genes and successfully produced coenzyme B12 . However, according to the quantitative determination by inductively coupled plasma-mass spectrometry, the amount of coenzyme B12 produced by the recombinant E. coli (0.21 ± 0.02 μg/g cdw) was approximately 13-fold lower than that by P. denitrificans (2.75 ± 0.22 μg/g cdw). Optimization of the culture conditions to improve the production of coenzyme B12 by the recombinant E. coli was successful, and the highest titer (0.65 ± 0.03 μg/g cdw) of coenzyme B12 was obtained. Interestingly, although the synthesis of coenzyme B12 in P. denitrificans is strictly oxygen-dependent, the recombinant E. coli could produce coenzyme B12 under anaerobic conditions.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP13870901	Coenzyme B12		13870-90-1	Price

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