title: Characterization of an acidic cold-adapted cutinase from Thielavia terrestris and its application in flavor ester synthesis.
abstract: An acidic cutinase (TtcutB) from Thielavia terrestris CAU709 was purified to apparent homogeneity with 983 Um g(-1) specific activity. The molecular mass of the enzyme was estimated to be 27.3 and 27.9 kDa by SDS-PAGE and gel filtration, respectively. A peptide sequence homology search revealed no homologous cutinases from T. terrestris, except for one putative cutinase gene (XP003656017.1), indicating that TtcutB is a novel enzyme. TtcutB exhibited an acidic pH optimum of 4.0, and stability at pH 2.5-10.5. Optimal activity was at 55 °C, it was stable up to 65 °C, and retained over 30% activity at 0 °C. Km values toward p-nitrophenyl (pNP) acetate, pNP-butyrate and pNP-caproate were 8.3, 1.1 and 0.88 mM, respectively. The cutinase exhibited strong synthetic activity on flavor ester butyl butyrate under non-aqueous environment, and the highest esterification efficiency of 95% was observed under the optimized reaction conditions. The enzyme's unique biochemical properties suggest great potential in flavor esters-producing industries.
authors.affiliation: Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Haibo
authors.initials: H
authors.lastname: Xu
authors.affiliation: Bioresource Utilization Laboratory, College of Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Qiaojuan
authors.initials: Q
authors.lastname: Yan
authors.affiliation: Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Duan
authors.affiliation: Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. Electronic address: ysq9926@163.com.
authors.forename: Shaoqing
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. Electronic address: zhqjiang@cau.edu.cn.
authors.forename: Zhengqiang
authors.initials: Z
authors.lastname: Jiang
mesh_terms: D002087:Butyrates / Q000737:chemistry; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D003080:Cold Temperature; D004591:Electrophoresis, Polyacrylamide Gel; D004951:Esterification; D004952:Esters / Q000737:chemistry*; D005421:Flavoring Agents / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D020037:Sordariales / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D002087:Butyrates; D004952:Esters; D005421:Flavoring Agents; C022793:n-butyl n-butyrate; C033592:4-nitrophenyl butyrate; D002265:Carboxylic Ester Hydrolases; C019952:cutinase
keywords: Characterization; Cold adaption; Cutinase; Flavor ester synthesis; Thielavia terrestris