title: C-terminal region of Candida rugosa lipases affects enzyme activity and interfacial activation.
abstract: Candida rugosa contains several lipase (CRLs) genes, and CRLs show diverse enzyme activity despite being highly homologous across their entire protein family. Previous studies found that LIP4 has a high esterase activity and a low lipolytic activity and lacks interfacial activation. To investigate whether the C-terminal region of the CRLs mediates enzymatic activity, chimeras were generated in which the C-terminus of LIP4 from either residue 374, 396, 417, or 444 to residue 534 was swapped with the corresponding peptide from the isoform LIP1. A chimeric lipase containing the C-terminus from 396 to 534 of LIP1 on a LIP4 scaffold showed activity similar to that of commercial CRL on triolein, and lipolytic activity increased 2-6-fold over that of LIP4. Moreover, interfacial activation was also observed in the chimeric lipase. To improve its enzymatic properties, a novel glycosylation site was added at residue 314. The new glycosylated lipase showed improved thermostability and enhancement in enzymatic activity, indicating its potential for use in further application.
authors.affiliation: Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan.
authors.forename: Kuo-Sheng
authors.initials: KS
authors.lastname: Hung
authors.forename: Shiow-Yi
authors.initials: SY
authors.lastname: Chen
authors.forename: Hsu-Feng
authors.initials: HF
authors.lastname: Liu
authors.forename: Bing-Reui
authors.initials: BR
authors.lastname: Tsai
authors.forename: Hung-Wei
authors.initials: HW
authors.lastname: Chen
authors.forename: Chin-Yen
authors.initials: CY
authors.lastname: Huang
authors.forename: Ji-Long
authors.initials: JL
authors.lastname: Liao
authors.forename: Kuang-Hui
authors.initials: KH
authors.lastname: Sun
authors.forename: Shye-Jye
authors.initials: SJ
authors.lastname: Tang
mesh_terms: D000595:Amino Acid Sequence; D002175:Candida / Q000201:enzymology*; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008066:Lipolysis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D005656:Fungal Proteins; D010446:Peptide Fragments; D020033:Protein Isoforms; C428110:Lip4 protein, Candida rugosa; D008049:Lipase