Perturbation of the peptidoglycan network and utilization of the signal recognition particle-dependent pathway enhances the extracellular production of a truncational mutant of CelA in Escherichia coliopen access
- Authors
- Kang, Tae-Gu; Hong, Seok-Hyun; Jeon, Gi-Beom; Yang, Yung-Hun; Kim, Sun-Ki
- Issue Date
- Jun-2021
- Publisher
- OXFORD UNIV PRESS
- Keywords
- Signal recognition particle-dependent pathway; Extracellular secretion; CelA
- Citation
- JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, v.48, no.5-6
- Journal Title
- JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
- Volume
- 48
- Number
- 5-6
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/49655
- DOI
- 10.1093/jimb/kuab032
- ISSN
- 1367-5435
1476-5535
- Abstract
- Caldicellulosiruptor bescii is the most thermophilic, cellulolytic bacterium known and has the native ability to utilize unpretreated plant biomass. Cellulase A (CelA) is the most abundant enzyme in the exoproteome of C. bescii and is primarily responsible for its cellulolytic ability. CelA contains a family 9 glycoside hydrolase and a family 48 glycoside hydrolase connected by linker regions and three carbohydrate-binding domains. A truncated version of the enzyme (TM1) containing only the endoglucanase domain is thermostable and actively degrades crystalline cellulose. A catalytically active TM1 was successfully produced via the attachment of the PelB signal peptide (P-TM1), which mediates post-translational secretion via the SecB-dependent translocation pathway. We sought to enhance the extracellular secretion of TM1 using an alternative pathway, the signal recognition particle (SRP)-dependent translocation pathway. The co-translational extracellular secretion of TM1 via the SRP pathway (D-TM1) resulted in a specific activity that was 4.9 times higher than that associated with P-TM1 overexpression. In batch fermentations, the recombinant Escherichia coli overexpressing D-TM1 produced 1.86 +/- 0.06 U/ml of TM1 in the culture medium, showing a specific activity of 1.25 +/- 0.05 U/mg cell, 2.7- and 3.7-fold higher than the corresponding values of the strain overexpressing P-TM1. We suggest that the TM1 secretion system developed in this study can be applied to enhance the capacity of E. coli as a microbial cell factory for the extracellular secretion of this as well as a variety proteins important for commercial production.
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