เอกสารการประชุมวิชาการและเสนอผลงานวิจัย มหาวิทยาลัยทักษิณ ครั้งที่ 19 2552 - page 893

1
„µ¦ž¦³»¤ª·µ„µ¦Â¨³Áœ°Ÿ¨Šµœª·‹´¥¤®µª· š¥µ¨´¥š´„¬·– ‡¦´ÊŠš¸É 19 ž¦³‹Îµže 2552
„µ¦„´—¨³„µ¦Â¥„Á°œÅŽ¤r ŨÁž‹µ„ÁœºÊ°¤³¡¦oµª—oª¥¦³µ¦¨³¨µ¥œÊ尊ª´’£µ‡
Extraction and Partition of cocoanut lipase in aqueous two-phase systems (ATPS)
曦 ˜´œ˜¸¤´œ˜r1 „»¨›·—µ ‡¦´˜œr 2 žd¥µ£¦–r £µ¬·˜„»¨3*
Sarothorn Tantasiman1, Kulthida Kotchrat 2 Piyaporn Pasitkul3*
š‡´—¥n°
„µ¦„´—Â¥„Á°œÅŽ¤rŨÁž‹µ„ÁœºÊ°¤³¡¦o µª¡ªnµ µ¦„´ —Äœ´¢Á¢°¦rš¸É ¡¸ Á°Áž}œÁ‹³¤¸‡nµÂ°‡˜·ª· ˜¸Ê
°ŠÁ°œÅŽ¤r ¼Š„ªnµµ¦„´—Äœ¡¸Á°š¸É Áž}œ„¦— ¨³Á¤ºÉ°œÎµ¤µšÎ µ„µ¦Â¥„Á°œÅŽ¤rŨÁž—oª¥¦³ ATPS ץčo
PEG 1500 „´Á„¨º° Tri-potassium citrate ¡ªnµ Á¤ºÉ°Á¡·É ¤‡ªµ¤Á o¤ oœ °Š PEG ‹³šÎµÄ®oÁ°œÅŽ¤rŨÁž ¹ÊœÅž°¥¼n Ĝ
´Êœ PEG ¤µ„ ¹Êœ ×¥š¸É‹³¤¸‡ªµ¤¦· »š›·Í°ŠÃž¦˜¸ œÄœ´Êœ PEG ™¹Š 4.18 ÁšnµÁ¤ºÉ °Áš¸¥„´µ¦„´—®¥µ ץĮoŸ¨
°—‡¨o °Š„´„µ¦Â¥„Þ¦˜¸œ—oª¥Áš‡œ·‡ SDS-PAGE ´Êœ PEG ‹³¡Â™Ãž¦˜¸œ 3 ™š¸É´—Á‹œ ¤¸œµ—äÁ¨„» ¨
44.38 19.72 ¨³ 9.64 kDa Á¤ºÉ °œÎµ´ÊœÁ„¨º°¤µŸnµœ Sephadex-G 100 ¤¸ 5 ¡¸‡Ãž¦˜¸œ ¡Â°‡˜·ª·˜¸Ê°ŠÁ°œÅŽ¤r ŨÁž¡¸
‡š¸É 2 Ášnµ„´ 0.121±0.007 units/ml nªœ´Êœ PEG ‹³¤¸ 8 ¡¸‡Ãž¦˜¸œ ¡Â°‡˜·ª·˜¸Ê°ŠÁ°œÅŽ¤rŨÁž¡¸‡š¸É 7 Ášnµ„´
0.098±0.003 units/ml °‡˜·ª·˜¸ÊÁ°œÅŽ¤rŨÁžš¸É ¡š´ÊŠ 2 ¡¸‡ ´œœ·¬“µœÅ—oªnµ°µ‹Áž}œ 2 isozyme š¸É ¤¸ÄœÁœºÊ°¤³¡¦oµª
Ž¹É Šµ¤µ¦™Â¥„Å—o—oª¥Áš‡œ· ‡ ATPS Ÿ¨‡ªµ¤šœ˜n °°»–®£¼¤·°ŠÅ¨ÁžÄœ´Êœ PEG µ¤µ¦™šÎ µŠµœÅ—o—¸ÄœnªŠ
°»–®£¼¤· 50-80 oC ¨³¤¸šœ‡ªµ¤°»–®£¼¤·š¸É 80 oC š¸É Áª¨µ 1 ´ÉªÃ¤Š ¤¸Â°‡˜·ª·˜¸Ê°¥¼n™¹Š 85.14 % Ĝ –³š¸É´ÊœÁ„¨º°‹³
Á®¨º °°¥¼nÁ¡¸¥Š 67.06 % ‹µ„„µ¦œÎ µÁ°œÅŽ¤rŨÁžž¦·¤µ– 15.34 unit/ml ¤µÁž}œ˜´ªÁ¦nŠž’·„·¦·¥µš¦µœÁ°Áš°¦·¢dÁ‡´œ
š¸É °»–®£¼¤· 40 oC ¡ªn µ‹³Å—oÁ¤š·¨Á°Áš°¦r(ŝð—¸ÁŽ¨) ¼Š™¹Š 75% Ž¹É ŠÅÃ°—¸ÁŽ¨š¸É ŗo¤¸‡»–¤´˜·šµŠ„µ¥£µ¡Â¨³
šµŠÁ‡¤¸ š¸ÉĄ¨oÁ‡¸¥Š„´œ„´ „µ¦Á¦nŠž‘·„·¦·¥µ—oª¥ÁÃ—¥ÅÃ°—¸ÁŽ¨š¸Éŗo‹µ„„µ¦Á¦nŠ—oª¥Á ¨³Á°œÅŽ¤rŨÁž¤¸ ‡nµ
Rf Ášnµ„´ 0.71 ¨³ 0.78 ¤¸‡nµ‡ªµ¤®œº—´¤¡´š›rÁš¸¥„´œÊεÁšnµ„´ 1.6 ¨³2.6 ˜µ¤¨Îµ—´
‡ÎµÎµ‡´ : Á°œÅŽ¤rŨÁž¦³µ¦¨³¨µ¥œÊ尊ª´’£µ‡ š¦µœÁ°Áš°¦·¢d Á‡´œÅÃ°—¸ÁŽ¨ ‡ªµ¤®œº—´¤¡´ š›r
Abstract
Extraction enzyme lipase from cocoanut was found that base pH condition of extraction will promote
high lipase activity better than acid pH condition. And then, the extracted was partition by aqueous two-phase
systems (ATPS) with polyethylene glycol 1500 and tripotassium citrate. From result, when increasing of PEG
concentration, the lipase from aqueous phase was moved to PEG phase and found that 4.18-fold purification
compare with crude extraction, related with the result by SDS-PAGE, sample from PEG phase showed 3 bands
1 œ· ·˜ž¦· µÃš £µ‡ª·µÁ‡¤¸ ‡–³ª·š¥µ«µ˜¦r ¤®µª· š¥µ¨´¥š´„¬·– Š ¨µ 90000
2 œ· ·˜„µ¦«¹„¬µ´–š·˜ µ µª·µª·š¥µ«µ˜¦r-Á‡¤¸ ‡–³«¹„¬µ«µ˜¦r ¤®µª·š¥µ¨´¥š´„¬·– Š ¨µ 90000
3 °µ‹µ¦¥rž¦³‹Îµ £µ‡ª·µÁ‡¤¸ ‡–³ª·š¥µ«µ˜¦r ¤®µª· š¥µ¨´¥š´„¬·– Š ¨µ 90000
* Corresponding author :Ú¦«´¡šr/Ú¦µ¦ : 074-443955 e-mail : ppasitkul@hotmail.com