2011 33 3 0597-0601 http / /xuebao. jxau. edu. cn Acta Agriculturae Universitatis Jiangxiensis E - mail ndxb7775@ sina. com * 311400 Alcalase 2. 4 L 90. 48% Alcalase 2. 4 L 91. 40% Alcalase 2. 4 L TS225. 1 + 6 A 1000-2286 2011 03-0597 - 05 A Preliminary Study on the Extraction Techology of Oil-tea Camellia Seed Oil by Aqueous Enzymatic Method SUN Hong FEI Xue-qian * FANG Xue-zhi Research Institute of Subtropical Forestry CAF Fuyang 311400 China Abstract An aqueous enzymatic extraction technology was adopted to extract oil from Oil - tea camellia seed. The influences of different enzymes and enzyme complex formulas on oil yield were investigated. The result showed that Alcalase 2. 4 L was the best single enzyme with the highest oil yield of 90. 48%. The best enzyme complex formula was composed by pectinase cellulose and Alcalase 2. 4 L which was gained by multilevel optimization procedure. The oil yield efficiency could reach 91. 40% when this enzyme complex formula was used in sequence. By comprehensive consideration Alcalase 2. 4 L is the best hydrolase. Key words oil-tea camellia seed oil aqueous enzymatic method enzymatic selection oil yield 1-4 5-7 8 8-10 2010-12 - 08 2011-03 - 03 2009BADB1B03 948 2009-4 - 56 1982 E - mail sunh206@ 163. com * E - mail fxq6565@ 163. com
598 33 1 1. 1 5. 81% 48. 12% 60 7 5 sigma C2730 Trichoderma reesei ATCC 26921 50 ~ 60 ph 4. 5 ~ 6. 0 sigma H2125 Aspergillus niger 40 ph 4. 5 sigma P2736 Aspergillus niger 50 ph 8. 0 sigma 10069 Bacillus subtilis 25 ph 6. 9 Alcalase 2. 4 L Novozymes Alcalase 2. 4 L Bacillus licheniformis 50 ~ 60 ph 8 ~ 9 Buchi B - 811 Thermo Fisher ph 1. 2 1. 2. 1 5 g 50 ml 25 ml 90 10 min ph 90 10 min 8 000 r /min 30 min 1. 2. 2 GB /T 5009. 6-2003 11 = - / 100% 1. 2. 3 / 1 5 g 5 50 ml 2% 1 5 60 4 h 8 000 r /min 30 min 2 2% 1 5 2 h 1 Tab. 1 1 Enzymatic hydrolysis conditions of simultaneous enzyme supplementation /% ph / /h Emzyme combination Enzyme dosage Reaction ph Reaction temperature Reaction time + Alcalase Cellulose + Alcalase 2 7. 0 55 4 + Alcalase Hemicellulase + Alcalase 2 8. 0 50 4 + Alcalase Pectinase + Alcalase 2 7. 0 45 4 + Alcalase Amylase + Alcalase 2 7. 5 40 4 3 2% 1 5 2 h
3 599 2 2. 1 1 Alcalase Alcalase 85. 38% 90. 48% Alcalase 4 1 14. 20% 13. 39% 11. 22% 5. 97% Fig. 1 The influences of different enzyme type on oil yield 12-13 7 Alcalase 2. 2 2. 2. 1 1 + Alcalase 2 + Alcalase 3 + 4 + Alcalase 1 Cellulase + Alcalase 2 Pectinase + Alcalase 3 Hemicellulase + Alcalase 4 Amylase + Alcalase. 2 Fig. 2 The influences of different two - enzymes complex formulas on oil yield 14-15 2 Alcalase Alcalase 90. 65% 90. 53% 1. 73% Alcalase 1 + Alcalase 2 /Alcalase 3 Alcalase / 4 + Alcalase 5 /Alcalase 6 Alcalase / Alcalase + / 1 Cellulase + Alcalase 2 Cellulase /Alcalase 3 Alcalase /Cellulase 4 Pectinase + Alcalase 5 Pectinase /Alcalase 6 Alcalase /Pectinase. + means simultaneous enzyme supplementation / means different adding sequence. Alcalase 3 Fig. 3 The influences of different adding sequence on oil yield 2. 2. 2 3
600 33 1 /Alcalase 2 3 /Alcalase 4 1 / /Alcalase 2 / /Alcalase 3 / /Alcalase 4 / /Alcalase 5 / /Alcalase 1 Cellulase/Pectinase/Alcalase 1 Cellulase /Alcalase 2 Pectinase /Alcalase 2 Cellulase/Hemicellulase/Alcalase 3 Cellulase/Amylase/Alcalase 3 /Alcalase 4 Amylase + Alcalase. 4 Pectinase/Hemicellulase/Alcalase 5 Pectinase /Amylase /Alcalase. 4 5 Fig. 4 The influences of different enzymes Fig. 5 The influences of different tri enzymes complex formulas on oil yield complex formulas on oil yield Alcalase 90. 53% Alcalase Alcalase 90. 81% 0. 31% Alcalase 2 ph Alcalase Alcalase 2. 2. 3 4 Alcalase Alcalase 90. 92% 90. 81% 0. 39% Alcalase Alcalase 2. 3 2. 3. 1 5 1 / /Alcalase 2 / /Alcalase 3 Alcalase + /Alcalase 4 + + Alcalase 5 Alcalase / / 6 Alcalase / / 91. 31% 0. 44% 1 Cellulase /Pectinase /Alcalase 2 Pectinase /Cellulase /Alcalase 3 91% Cellulase + Pectinase /Alcalase 4 Cellulase + Pectinase + Alcalase 5 Alcalase /Cellulase /Pectinase 6 Alcalase /Pectinase /Cellulase. 6 Alcalase Fig. 6 The influences of different adding sequence on oil yield
3 601 2. 3. 2 6 Alcalase 91. 40% Alcalase 6 Alcalase 2. 3. 3 91. 40% 90. 48% 90. 92% 91. 40% 0. 44% 0. 92% Alcalase 2. 4 L 90. 48% Alcalase2. 4 L 3 1 Alcalase 91. 40% 2 Alcalase 2. 4 L 90. 48% Alcalase 2. 4 L 3 1. J. 1994 13 3 179-191. 2. J. 2004 23 5 58-62. 3 Zhang S B Wang Z Xu S Y. Optimization of the aqueous enzymatic extraction of rapeseed oil and protein hydrolysates J. Journal of the American Oil Chemists Society 2007 84 1 97-105. 4 Moreau R A Johnston D B Powell M J et al. A comparison of commercial enzymes for the aqueous enzymatic extraction of corn oil from corn germ J. Journal of the American Oil Chemists Society 2004 81 11 1071-1075. 5. J. 2009 34 1 23-26. 6. J. 2010 23 2 165-169. 7. J. 2007 33 1 83-86. 8. J. 2006 14 6 41-42. 9 Dominguez H Nunez M J Lema J M. Enzymatic pretreatment to enhance oil extraction from fruits and oilseeds A review J. Food Chemistry 1994 49 3 271-286. 10 Rosenthal A Pyle D L Niranjan K. Aqueous and enzymatic processes for edible oil extraction J. Enzyme and Microbial Technology 1996 19 6 402-420. 11 GB /T 5009. 6-2003 S. 2003. 12 Huang A. Structure of plant seed oil bodies J. Current Opinion in Structural Biology 1994 4 4 493-498. 13 Yoon SH Kim IH Kwon TW. Effects of enzyme treatments and ultrasonification on extraction yields of lipids and protein from soybean by aqueous process J. Korean J 1991 23 673-676. 14. J. 2005 6 10-12. 15. J. 2007 32 2 27-29.