Novel minor lipase from Rhizopus chinensis during solid-state fermentation: biochemical characterization and its esterification potential for ester synthesis.
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 degrees C using pNPP as substrate. Slight activation of the enzyme was observed when Mn(2+) 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 degrees C. 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.
Sun
Shu Yang
SY
Key Laboratory of Industrial Biotechnology of Ministry of Education and School of Biotechnology, Jiangnan University, Wuxi 214122, PR China.
Xu
Yan
Y
Wang
Dong
D
D000438:Alcohols / Q000378:metabolism; D002210:Caprylates / Q000378:metabolism; D004789:Enzyme Activation; D004795:Enzyme Stability; D004951:Esterification; D004952:Esters / Q000378:metabolism*; D000431:Ethanol / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007848:Laurates / Q000737:chemistry; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008345:Manganese / Q000737:chemistry; D019216:Metals, Heavy / Q000737:chemistry; D009930:Organic Chemicals / Q000737:chemistry; D012233:Rhizopus / Q000201:enzymology* / Q000378:metabolism; D012997:Solvents / Q000737:chemistry; D013379:Substrate Specificity; D013696:Temperature
D000438:Alcohols; D002210:Caprylates; D004952:Esters; D005227:Fatty Acids; D007848:Laurates; D019216:Metals, Heavy; D009930:Organic Chemicals; D012997:Solvents; C050014:4-nitrophenyl laurate; D000431:Ethanol; D008345:Manganese; D008049:Lipase; C031492:octanoic acid