A combined cell-consortium approach for lignocellulose degradation by specialized Lactobacillus plantarum cells

Biotechnology for Biofuels

Sarah Moraïs²  Naama Shterzer¹  Raphael Lamed³  Edward A Bayer²  Itzhak Mizrahi¹ 

[1] Department of Ruminant Science, Institute of Animal Sciences, Agricultural Research Organization, P.O.B. 6, Bet-Dagan 50250, Israel

[2] Department of Biological Chemistry, The Weizmann Institute of Science, 234 Herzl St., Rehovot 7610001, Israel

[3] Department of Molecular Microbiology and Biotechnology, Tel Aviv University, P.O. Box 39040, Ramat Aviv 69978, Israel

关键词: Biomimicry;    Spatial differentiation;    Enzymatic paradigm;    Glycoside hydrolase;    Wheat straw;    Bioprocessing;    Biomass;    Designer cellulosome;   

Others  :  1084797
DOI  :  10.1186/1754-6834-7-112

 received in 2014-04-27, accepted in 2014-07-09,  发布年份 2014

【 摘 要 】

Background

Lactobacillus plantarum is an attractive candidate for metabolic engineering towards bioprocessing of lignocellulosic biomass to ethanol or polylactic acid, as its natural characteristics include high ethanol and acid tolerance and the ability to metabolize the two major polysaccharide constituents of lignocellulolytic biomass (pentoses and hexoses). We recently engineered L. plantarum via separate introduction of a potent cellulase and xylanase, thereby creating two different L. plantarum strains. We used these strains as a combined cell-consortium for synergistic degradation of cellulosic biomass.

Results

To optimize enzymatic degradation, we applied the cell-consortium approach to assess the significance of enzyme localization by comparing three enzymatic paradigms prevalent in nature: (i) a secreted enzymes system, (ii) enzymes anchored to the bacterial cell surface and (iii) enzymes integrated into cellulosome complexes. The construction of the three paradigmatic systems involved the division of the production and organization of the enzymes and scaffold proteins into different strains of L. plantarum. The spatial differentiation of the components of the enzymatic systems alleviated the load on the cell machinery of the different bacterial strains. Active designer cellulosomes containing a xylanase and a cellulase were thus assembled on L. plantarum cells by co-culturing three distinct engineered strains of the bacterium: two helper strains for enzyme secretion and one producing only the anchored scaffoldin. Alternatively, the two enzymes were anchored separately to the cell wall. The secreted enzyme consortium appeared to have a slight advantage over the designer cellulosome system in degrading the hypochlorite pretreated wheat straw substrate, and both exhibited significantly higher levels of activity compared to the anchored enzyme consortium. However, the secreted enzymes appeared to be less stable than the enzymes integrated into designer cellulosomes, suggesting an advantage of the latter over longer time periods.

Conclusions

By developing the potential of L. plantarum to express lignocellulolytic enzymes and to control their functional combination and stoichiometry on the cell wall, this study provides a step forward towards optimal biomass bioprocessing and soluble fermentable sugar production. Future expansion of the preferred secreted-enzyme and designer-cellulosome systems to include additional types of enzymes will promote enhanced deconstruction of cellulosic feedstocks.

【 授权许可】

   
2014 Moraïs et al.; licensee BioMed Central Ltd.

【 图 表 】

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