Display of Candida antarctica lipase B on Pichia pastoris and its application to flavor ester synthesis.
Two alternative cell-surface display systems were developed in Pichia pastoris using the alpha-agglutinin and Flo1p (FS) anchor systems, respectively. Both the anchor cell wall proteins were obtained originally from Saccharomyces cerevisiae. Candida antarctica lipase B (CALB) was displayed functionally on the cell surface of P. pastoris using the anchor proteins alpha-agglutinin and FS. The activity of CALB displayed on P. pastoris was tenfold higher than that of S. cerevisiae. The hydrolytic and synthetic activities of CALB fused with alpha-agglutinin and FS anchored on P. pastoris were investigated. The hydrolytic activities of both lipases displayed on yeast cells surface were more than 200 U/g dry cell after 120 h of culture (200 and 270 U/g dry cell, respectively). However, the synthetic activity of CALB fused with alpha-agglutinin on P. pastoris was threefold higher than that of the FS fusion protein when applied to the synthesis of ethyl caproate. Similarly, the CALB displayed on P. pastoris using alpha-agglutinin had a higher catalytic efficiency with respect to the synthesis of other short-chain flavor esters than that displayed using the FS anchor. Interestingly, for some short-chain esters, the synthetic activity of displaying CALB fused with alpha-agglutinin on P. pastoris was even higher than that of the commercial CALB Novozyme 435.
Su
Guo-Dong
GD
Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, People's Republic of China.
Huang
Deng-Feng
DF
Han
Shuang-Yan
SY
Zheng
Sui-Ping
SP
Lin
Ying
Y
D001709:Biotechnology / Q000379:methods*; D002175:Candida / Q000201:enzymology*; D002384:Catalysis; D003001:Cloning, Molecular; D004952:Esters / Q000378:metabolism; D005421:Flavoring Agents / Q000378:metabolism*; D005656:Fungal Proteins; D008049:Lipase / Q000235:genetics* / Q000378:metabolism; D037241:Mannose-Binding Lectins / Q000235:genetics / Q000378:metabolism; D000072235:Mating Factor; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
D004952:Esters; C087578:FLO1 protein, S cerevisiae; D005421:Flavoring Agents; D005656:Fungal Proteins; D037241:Mannose-Binding Lectins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D000072235:Mating Factor; D008049:Lipase; C431349:lipase B, Candida antarctica