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„µ¦Â¥„Á°œÅŽ¤rŨÁž‹µ„nªœš¸ÉÁ®¨º °š·ÊŠ °Š„¦³ªœ„µ¦„´—œÊε¤´œ¤³¡¦o µªÃ—¥Äo
Aqueous Two- Phase Systems (ATPS)
Isolation of Lipase from Waste of Coconut Oil Processing by using Aqueous Two- Phase Systems
(ATPS)
曦 ˜´œ˜¸¤´œ˜r1 ¨³žd¥µ£¦–r £µ¬·˜„»¨2
Sarothorn Tantasiman1 and Piyaporn Pasitkul2
š‡´—¥n°
Á°œÅŽ¤rŨÁžŽ¹ÉŠµ¤µ¦™˜¦ª‹¡Å—o‹µ„ °ŠÁ®¨ª´Êœ¨n µŠš¸Éŗo‹µ„„µ¦ÁŽœ˜¦· ¢dª‹r œÊε„³š· Á o¤ oœ Áž}œ
n ªœš¸ÉÁ®¨º°š·ÊŠ °Š„¦³ªœ„µ¦„µ¦Ÿ¨·˜œÊε¤´ œ¤³¡¦oµª¦· »š›·Í ¨³Á¤ºÉ°Â¥„Á°œÅŽ¤rŨÁž—oª¥Áš‡œ· ‡ aqueous
two- phase systems (ATPS) ¦³®ªnµŠ PEG 1,500 ®¦º° PEG 6,000 „´Á„¨º° trisodium citrate ¡ªnµš¸É 10% PEG
¨³ 30% trisodium citrate Áž}œ£µª³š¸É—¸š¸É »—š¸ÉšÎ µÄ®o 30.81 % °ŠÅ¨Áž™¼„Â¥„Åž°¥¼nĜ´Êœ PEG —o µœœ ¤¸‡nµ
°‡˜·ª·˜¸Ê 0.0718 Units/mg ‡n µ Partition coeffient(K lipase) Ášnµ„´ 5.03 ¨³¤¸‡ªµ¤¦· »š›·ÍÁ¡·É¤ ¹Êœ 231 Ášnµ Á¤ºÉ°Áš¸¥
„´ °ŠÁ®¨ª´Êœ¨nµŠ„n°œÂ¥„ Á¤ºÉ°œÎµ´Êœ PEG 1500 ¤µšÎµ SDS-PAGE ‹³ž¦µ„’™Ãž¦˜¸œ 3 ™ š¸É¤¸œµ— 50
24 ¨³15 kDa ‹µ„„µ¦«¹„¬µ‡»–¤´˜·‡ªµ¤šœ˜n°°»–®£¼¤·¡ªnµÁ°œÅŽ¤rŨÁžÄœ´Êœ PEG 1500 ¨³ PEG 6000
µ¤µ¦™šœ°» –®£¼¤·Å—o—¸Äœn ªŠ„ªoµŠ(40-80 oC) ¨³¤¸‡ªµ¤šœ˜n°°»–®£¼¤·š¸É 70 oC Áž}œÁª¨µ 1 ¤. ×¥¤¸Â°‡˜· ª·˜¸Ê
°¥¼n ™¹Š 106.26 % ¨³136.69% ˜µ¤¨Îµ—´ œ°„‹µ„œ¸ÊÅ°°°œ Ag+ Ca2+ Mg2+ Cu2+ ¨³ Co2+ µ¤µ¦™¥´¥´ÊŠÁ°œÅŽ¤r
ŨÁžÄœ PEG 1500 ŗo ×¥ÁŒ¡µ³Á¤ºÉ°Äo Cu2+ ¨³ Co2+ š¸É ‡ªµ¤Á o¤ oœ 10 mM ‹³¤¸„µ¦¥´ ¥´ÊŠ™¹Š 53.37 ¨³ 35.58
% ˜µ¤¨Î µ—´ Ĝ –³š¸É PMSF EDTA ¨³ DTT ‹³µ¤µ¦™Á¡·É¤Â°‡˜· ª·˜¸Êŗo š´ÊŠ‡ªµ¤Á o¤ oœ 1 ¨³ 10 mM —oª¥
ª·›¸ „µ¦Â¥„Á°œÅŽ¤rš¸É Šnµ¥ ¨³Á°œÅŽ¤r¤¸‡ªµ¤šœ°»–®£¼¤· ¼Š —´Šœ´Êœ‹¹ŠœÄ‹œÎµÁ°œÅŽ¤rŨÁžÄœ´Êœ PEG 1500 Ş
ž¦³¥»„˜rčoÁž}œ˜´ªÁ¦nŠž’·„·¦·¥µšµŠ¸ª£µ¡Äœ°»˜µ®„¦¦¤°µ®µ¦˜n°Åž
‡ÎµÎµ‡´ : Á°œÅŽ¤rŨÁžœÊε¤´œ¤³¡¦oµª¦· »š›·Í ˜´ªÁ¦nŠž’·„·¦·¥µšµŠ¸ª£µ¡
1 œ· ·˜ž¦· µÃš µ µª·µÁ‡¤¸ ‡–³ª·š¥µ«µ˜¦r ¤®µª· š¥µ¨´¥š´„¬·– Š ¨µ 90000
Master's Degree Student, Department of Chemistry, Faculty of Science, Thaksin University, Songkhla, 90000
Corresponding author : Ú¦«´¡šr 0872872058 E-mail:tantasi@hotmail.com
2 °µ‹µ¦¥r —¦. µ µª·µÁ‡¤¸ ‡–³ª·š¥µ«µ˜¦r ¤®µª· š¥µ¨´¥š´„¬·– Š ¨µ 90000
Dr., Department of Chemistry, Faculty of Science, Thaksin University, Songkhla, 90000