title: Displaying Candida antarctica lipase B on the cell surface of Aspergillus niger as a potential food-grade whole-cell catalyst.
abstract: Aspergillus niger is a recognized workhorse used to produce food processing enzymes because of its extraordinarily high protein-producing capacity. We have developed a new cell surface display system de novo in A. niger using expression elements from generally recognized as safe certified microorganisms. Candida antarctica lipase B (CALB), a widely used hydrolase, was fused to an endogenous cell wall mannoprotein, CwpA, and functionally displayed on the cell surface. Localization of CALB was confirmed by enzymatic assay and immunofluorescence analysis using laser scanning confocal microscopy. After induction by maltose for 45 h, the hydrolytic activity and synthesis activity of A. niger mycelium-surface displayed CALB (AN-CALB) reached 400 and 240 U/g dry cell, respectively. AN-CALB was successfully used as a whole-cell catalyst for the enzymatic production of ethyl esters from a series of fatty acids of different chain lengths and ethanol. In a solvent-free system, AN-CALB showed great synthetic activity and afforded high substrate mole conversions, which amounted to 87 % for ethyl hexanoate after 2 h, 89 % for ethyl laurate after 2 h, and 84 % for ethyl stearate after 3 h. These results suggested that CwpA can act as an efficient anchoring motif for displaying enzyme on A. niger, and AN-CALB is a robust, green, and cost-effective alternative food-grade whole-cell catalyst to commercial lipase.
authors: Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, People's Republic of China.
authors: Zhi-You
authors: ZY
authors: Pan
authors: Zhi-Ming
authors: ZM
authors: Yang
authors: Li
authors: L
authors: Pan
authors: Sui-Ping
authors: SP
authors: Zheng
authors: Shuang-Yan
authors: SY
authors: Han
authors: Ying
authors: Y
authors: Lin
mesh_terms: D001234:Aspergillus niger / Q000235:genetics* / Q000378:metabolism; D055162:Biocatalysis; D061505:Cell Surface Display Techniques / Q000379:methods*; D004951:Esterification; D005525:Food-Processing Industry; D005656:Fungal Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D008049:Lipase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000235:genetics; D025282:Mycelium / Q000737:chemistry
chemical_list: D005656:Fungal Proteins; D008562:Membrane Glycoproteins; C025363:mannoproteins; D008049:Lipase; C431349:lipase B, Candida antarctica