Development of a Potential Protein Display Platform in Corynebacterium glutamicum Using Mycolic Acid Layer Protein, NCgl1337, as an Anchoring Motif. Issue 2 (15th November 2017) - Choi, Jae Woong
Abstract : In the cell surface display, the choice of host cell and anchoring motif are the most crucial for the efficient display of passenger proteins. Corynebacterium glutamicum has mycolic acid layer in outer membrane and the use of protein in the mycolic acid layer as an anchoring motif can provide a potential platform for surface display in C. glutamicum . All 19 mycolic acid layer proteins of C. glutamicum are analyzed, and two proteins, NCgl0535 and NCgl1337, which have a signal peptide and predicted O ‐mycoloylation site, are selected as anchoring motifs candidates. Among them, NCgl1337, which shows better expression with higher display efficiency, is chosen as a potential anchoring motif. Two forms of the NCgl1337 anchoring motif, a full‐length (1–324 amino acids) and a short‐length (1–50 amino acids) containing only signal peptide and O ‐mycoloylation site, are constructed and their abilities for surface display are examined using two protein models, endoxylanase from Streptomyces coelicolor and α‐amylase from Streptococcus bovis . For both model proteins, the short‐length NCgl1337 anchoring motif exhibits higher yield of protein display on the surface of C. glutamicum than the full‐length NCgl1337. Finally, with C. glutamicum displaying α‐amylase, a batch fermentation is performed for the production ofl ‐lysine from starch degradation, and a production ofl ‐lysine as high as 10.8 ± 0.92 g L −1 was achieved after 18 h of culture. Abstract : Using the mycolic acid layer proteins as an anchoring proteins, the target proteins can be displayed on the most exterior layer of Corynebacterium glutamicum . This work is the potential basis for the use of C. glutamicum as a whole cell catalyst.