Immobilization of Lactococcus lactis to cellulosic material by cellulose-binding domain of Cellvibrio japonicus

• Published in: Journal of applied microbiology 2010-10, Vol.109 (4), p.1274-1283
• Main Authors: Kylä-Nikkilä, K, Alakuijala, U, Saris, P.E.J
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - genetics; biofilm(s); Biological and medical sciences; Bioreactors; biotechnology; Cells, Immobilized; Cellulose - metabolism; Cellvibrio - enzymology; Endo-1,4-beta Xylanases - chemistry; Endo-1,4-beta Xylanases - genetics; Fermentation; Fundamental and applied biological sciences. Psychology; Industrial Microbiology; lactic acid bacteria; Lactococcus; Lactococcus lactis; Lactococcus lactis - genetics; Lactococcus lactis - metabolism; membrane; Microbiology; Muramidase - genetics; Protein Structure, Tertiary; Recombinant Fusion Proteins - analysis; Recombinant Fusion Proteins - chemistry
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2010.04757.x

Immobilization of whole cells can be used to accumulate cells in a bioreactor and thus increase the cell density and potentially productivity, also. Cellulose is an excellent matrix for immobilization purposes because it does not require chemical modifications and is commercially available in many different forms at low price. The aim of this study was to construct a Lactococcus lactis strain capable of immobilizing to a cellulosic matrix. In this study, the Usp45 signal sequence fused with the cellulose-binding domain (CBD) (112 amino acids) of XylA enzyme from Cellvibrio japonicus was fused with PrtP or AcmA anchors derived from L. lactis. A successful surface display of L. lactis cells expressing these fusion proteins under the P45 promoter was achieved and detected by whole-cell ELISA. A rapid filter paper assay was developed to study the cellulose-binding capability of these recombinant strains. As a result, an efficient immobilization to filter paper was demonstrated for the L. lactis cells expressing the CBD-fusion protein. The highest immobilization (92%) was measured for the strain expressing the CBD in fusion with the 344 amino acid PrtP anchor. The result from the binding tests indicated that a new phenotype for L. lactis with cellulose-binding capability was achieved with both PrtP (LPXTG type anchor) and AcmA (LysM type anchor) fusions with CBD. We demonstrated that an efficient immobilization of recombinant L. lactis cells to cellulosic matrix is possible. This is a step forward in developing efficient immobilization systems for lactococcal strains for industrial-scale fermentations.