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Abstract
Lipase (E.C.3.1.1.3) was found from the lower liquid solution by centrifugation of intense fresh coconut
milk. It was a waste product from virgin coconut oil processing. Lipase was partitioned on aqueous two-phase
systems (ATPS), between PEG 1500 or PEG 6000 and trisodium citrate. The PEG 1500(10% w/v) and trisodium
citrate(30% w/v) was the best condition, 30.81% lipase was partitioned to the top of PEG phase, it had 0.0718
Units/mg, 5.03 of patition coefficient( Klipase) and 231 purification factor when compare with the lower liquid
solution. The Coconut lipase – PEG phase (CLP1500) showed 3 bands of protein on SDS-PAGE and molecular
weight were estimated to be 50, 24 and 15 kDa. The CLP1500 and CLP6000 were stable in broad ranges of
temperature (40-80 oC), the thermostability was done at 70 oC for 1 h and the activity were 106.26 and 136.69 %,
respectively. The effect of various divalent metal ions indicated that the Ag+ Ca2+ Mg2+ Cu2+ and Co 2+could
inhibited enzyme activity whereas 10 mM of Cu2+ and Co2+ could inhibited 53.37 and 35.58 %, respectively.
However, the PMSF, EDTA and DTT were promoted the lipase activity at 1 and 10 mM. Finally. this lipase was
simple to preparation and stable at high temperature, so we propose to apply it as a biocatalyst in food industry.
Keywords: Lipase; Virgin coconut oil; Aqueous Two- Phase Systems, ATPS; Coconut lipase PEG phase, CLP
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