Novel minor lipase from Rhizopus chinensis during solid-state fermentation: Biochemical characterization and its esterification potential for ester synthesis |
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| Authors: | Shu Yang Sun Yan Xu Dong Wang |
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| Affiliation: | 1. Joint Graduate School for Energy and Environment (JGSEE), King Mongkut''s University of Technology Thonburi, Bangmod, Bangkok 10140, Thailand;2. Enzyme Technology Laboratory, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand;1. Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, ZC 28049 Madrid, Spain;2. Escuela de Química, GIBIM, Edificio Camilo Torres 210, Universidad Industrial de Santander, Bucaramanga, Colombia;3. Escuela de Bacteriología y Laboratorio Clínico, Universidad Industrial de Santander, Bucaramanga, Colombia;4. Biocatalyst and Enzyme Technology Lab., Institute of Food Science and Technology, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, 9500, P.O. Box 15090, ZC 91501-970 Porto Alegre, RS, Brazil;1. Bioengineering College, Chongqing University, Chongqing 400044, China;2. Research Center for Tobacco Bioengineering and Technology, Chongqing Science and Technology Commission, Yubei District, Chongqing 401147, China;1. CEB—Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal;2. Biochemical Engineering Department, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil |
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| Abstract: | Rhizopus chinensis produces two lipases that catalyze ester synthesis when cultured under solid-state fermentation. The Lip2 was purified to homogeneity by ammonium sulphate precipitation, hydrophobic interaction chromatography and gel filtration chromatography. It has an apparent molecular weight of 33 kDa estimated from SDS–PAGE and 32 kDa calculated from analytical gel permeation, with synthetic activity and purification fold of 96.8 U/mg and 138.3, respectively. Maximum hydrolytic activity was obtained at pH 8.0–8.5 and 40 °C using pNPP as substrate. Slight activation of the enzyme was observed when Mn2+ is present. The enzyme was most active on p-nitrophenyl laurate (C12). The purified lipase exhibited maximum synthetic activity at pH memory of 6.0 and 30 oC. Most of ethyl esters synthesized by lyophilized enzyme achieved good yields (>90%), and caprylic acid served as the best acyl donor. The enzyme presented a particular affinity for ethanol, n-propanol and n-hexanol, with conversion of 92%, 93% and 92%, respectively, after 20 h incubation. |
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