Construction and optimization of trans‐4‐hydroxy‐L‐proline production recombinant E. coli strain taking the glycerol as carbon source. Issue 9 (2nd June 2016)

• Record Type: Journal Article
• Authors: Wang, Jinxia; Zhang, Zhenyu; Liu, Hedong; Sun, Fubao Fuelbiol; Yue, Chun; Hu, Jinguang; Wang, Chundi
• Is Part Of: Journal of chemical technology & biotechnology. Volume 91:Issue 9(2016:Sep.)
• Page Start: 2389
• Page End: 2398
• Publication Date: 2016-06-02
• Subjects: glycerol -- trans‐4‐hydroxy‐L‐proline -- proline 4‐hydroxylase -- Escherichia coli recombinant -- fermentation optimization; Biotechnology -- Periodicals; Chemistry, Technical -- Periodicals; Chemical engineering -- Periodicals; Industries -- Environmental aspects -- Periodicals; 660
• DOI: 10.1002/jctb.5024
• Languages: English
• ISSNs: 0268-2575

Abstract

Abstract: BACKGROUND: Trans ‐4‐hydroxy‐L‐proline (Hyp) is a value‐added amino acid that is an applicable chiral building block in some industries, such as the pharmaceutical, cosmetic and healthcare industries. Given that glycerol is a good carbon source for microbial fermentation and a large amount of glycerol is produced by the biodiesel industry, this study investigated a value‐added Hyp fermentation from glycerol using an engineered Escherichia coli strain. RESULTS: By optimizing the proline‐4‐hydroxylase ( P4H ) gene codons and vectors, a constructed recombinant Escherichia coli BL21 strain successfully converted L‐proline to Hyp. With a chemo‐physical combination mutagenesis, a high Hyp production strain that can grow with glycerol as a sole carbon source, NA45 was isolated, yielding 1.24 g L −1 from 20 g L −1 glycerol at 12 h in a 30 mL/250 mL conical flask fermentation. With further systemic optimization of nutritional elements, the output was enhanced to 1.62 g L −1 . In a 5 L fermentator, the strain achieved a surprisingly high output of 25.4 g L −1 at 48 h in fed batch mode. CONCLUSION: A high Hyp production strain growing well on glycerol as carbon source was successfully engineered, which is a promising candidate for the industrial Hyp production. © 2016 Society of Chemical Industry