เอกสารการประชุมวิชาการและเสนอผลงานวิจัย มหาวิทยาลัยทักษิณ ครั้งที่ 19 2552 - page 896

4
„µ¦ž¦³»¤ª·µ„µ¦Â¨³Áœ°Ÿ¨Šµœª·‹´¥¤®µª· š¥µ¨´¥š´„¬·– ‡¦´ÊŠš¸É 19 ž¦³‹Îµže 2552
„µ¦Á¦n Šž’·„·¦·¥µ—oª¥Á°œÅŽ¤r œÎ µÁ°œÅŽ¤rŨÁž¤µŸ¤„´œÊε¤´œ¡º Â¨³Á¤šµœ°¨ Ĝ°´˜¦µnªœœÊε¤´œ¡º:
Á¤šµœ°¨:Á°œÅŽ¤rŨÁž 10:2:1 (v/v) ‡œÄ®oÁ oµ„´œš¸É°»–®£¼¤·®o °Š œÎµÅž˜´ÊŠš¸É°»–®£¼¤· 40 oC Áž}œÁª¨µ 3 ´ÉªÃ¤Š
®¨´Š‹µ„šÎµž’· „·¦·¥µš¦µœÁ°Áš°¦·¢dÁ‡´ÉœÄœ„¦³ªœ„µ¦Ÿ¨· ˜ÅÃ°—¸ÁŽ¨ ‹³Á„· —„µ¦Â¥„Áž}œ 2 ´Êœ ×¥
Á¤š· ¨Á°Áš°¦rš¸É °¥¼n´Êœœ ¨³„¨¸ ÁŽ°¦°¨°¥¼n´Êœ¨nµŠ šÎµ„µ¦™nµ¥„¨¸ÁŽ°¦°¨°°„ œÎµn ªœœ¤µšÎµ„µ¦¨oµŠ ץčo ¼˜¦
„µ¦¨oµŠ ‡º ° Á¤š·¨Á°Áš°¦r:œÊεÁ„¨º°:Heptane Ĝ°´˜¦µnªœ 3:1.5:0.5 (v/v) Á ¥nµÄ®oÁ oµ„´œÂ¨o ªªµŠš·ÊŠÄ®o Â¥„´Êœ œÎµ
´Êœ Heptane Ž¹ÉŠÁž}œ´ÊœœÅžšÎµ„µ¦ª·Á‡¦µ³®r ‡»–¤´˜·šµŠ„µ¥£µ¡Â¨³šµŠÁ‡¤¸
„µ¦ª·Á‡¦µ³®r ‡ªµ¤®œº—¨³„µ¦Â¥„ŝð—¸ÁŽ¨—oª¥Áš‡œ·‡ Thin layer chromatography(TLC)
„µ¦ª·Á‡¦µ³®r ‡ªµ¤®œº— šÎµÃ—¥ždÁž˜µ¦š¸É˜o°Š„µ¦š—°ž¦·¤µ˜¦ 1 ml ‹µ„œ´ÊœšÎµ„µ¦ž¨n°¥ ‹´Áª¨µÁ¤ºÉ°
µ¦Á¦·É ¤®¥—°°„‹œ„¦³š´ÉŠ™¹Š ¸—°„ž¦·¤µ˜¦ 0.5ml ´œš¹„Áª¨µÂ¨oª´ ŠÁ„˜ÂœªÃœo¤ °ŠÁª¨µš¸Éŗo ¦³®ªnµŠœÊε¤´œ
Ä®¤n œÊÎ µ¤´ œÄo¨oª Ž¹É ŠÄoÁž}œµ¦˜´Ê Š˜oœÄœ„µ¦Ÿ¨·˜ÅÃ°—¸ ÁŽ¨Â¨³¦³®ªnµŠÅÃ°—¸ÁŽ¨š¸É Á¦n Š—oª¥Á°œÅŽ¤r ŨÁžÂ¨³ÅÃ°—¸
ÁŽ¨š¸ÉÁ¦nŠ—oª¥Á Áž¦¸ ¥Áš¸¥Áž}œ‡ªµ¤®œº—´¤¡´š›r—oª¥œÊε
„µ¦Â¥„ŝð—¸ÁŽ¨—oª¥Áš‡œ·‡ Thin layer chromatography (TLC) ×¥œÎµÂŸnœ TLC Á‡¨º °—oª¥ silica gel
Áž}œ stationary phase ¨³Äo˜´ªšÎ µ¨³¨µ¥Ÿ¤¦³®ªnµŠ Hexane : Acetic acid : Diethly ether Ĝ°´˜¦µnªœ 2:7:3
˜µ¤¨Îµ—´ Áž}œ mobile phase ®¥—ŝð—¸ÁŽ¨š¸É Á¦nŠ—oª¥Á°œÅŽ¤rŨÁž ŝ𗸠ÁŽ¨š¸ÉÁ¦nŠ—oª¥ÁœÊε¤´œ¡º œÂŸnœ
TLC œÎµ¤µªµŠÄœ™´ŠÂ„o ªš¸É¤¸˜´ªšÎ µ¨³¨µ¥Ÿ¤°·É¤˜´ª ‹µ„œ´ÊœœÎµÂŸnœ TLC ¤µ¥o °¤ Iodine ¨oªªµŠš·ÊŠÅªo Ä®o®oŠ
´ŠÁ„˜Ã‡¦¤µÃš¦Â„¦¤š¸É ŗo
Ÿ¨„µ¦ª·‹´¥Â¨³°£·ž¦µ¥Ÿ¨
„µ¦„´ —Á°œÅŽ¤rŨÁž‹µ„ÁœºÊ °¤³¡¦oµª
„µ¦„´ —Á°œÅŽ¤rŨÁž—oª¥µ¦¨³¨µ¥´¢Á¢°¦r 50 mM Sodium acetate ¡¸Á° 5 - 6 50 mM Tris-base
¡¸ Á° 7 - 9 ×¥„´—Äœ°´˜¦µn ªœ °ŠÁœºÊ°¤³¡¦oµª˜n°µ¦¨³¨µ¥´¢Á¢°¦rÁž}œ 1:1(w/v) ¡ªn µÁž}œÅž—´Š˜µ¦µŠš¸É 1
˜µ¦µŠš¸É 1 —Šž¦·¤µ–Á°œÅŽ¤r ŨÁž‹µ„„µ¦„´—Äœµ¦¨³¨µ¥´ ¢Á¢°¦r¡¸ Á°˜nµŠ Ç
Treatment Protein
(mg/ml)
Lipase Activity
(Units/ml)
Total protein
(mg)
Total activity
(Units)
Specific
activity
œÊÎ µ„¨´Éœ 1.65 0.087 248.17 13.01 5.24
pH5.0 1.12 0.066 168.62 9.96 5.90
pH6.0 1.70 0.067 254.95 10.05 3.94
pH7.0 2.79 0.080 418.99 11.96 2.85
pH8.0 3.06 0.107 459.31 16.06 3.50
pH9.0 3.24 0.108 485.52 16.25 3.35
‹µ„Ÿ¨š¸Éŗo‹³¡ªnµ„µ¦„´—Á°œÅŽ¤r ŨÁžÄœ¡¸Á°š¸É ¼Š‹³¤¸„µ¦šÎµŠµœ °ŠÁ°œÅŽ¤r š¸É‡n°œ oµŠ¼ŠÁ¤ºÉ °
Áž¦¸¥Áš¸¥„´„µ¦„´—Äœ¡¸Á°˜Éε Ç —o ª¥Ž¹ÉŠšÎµÄ®oš¦µªnµÁ°œÅŽ¤rŨÁžš¸Éŗo ‹³¤¸„µ¦šÎµŠµœÅ—o—¸Äœ£µª³š¸É Áž}œ
Á‡º°¡¸ Á° 8 – 9 Ž¹É ŠÅ¤n˜„˜nµŠ„´ œ¤µ„œ´„ ˜nš¸É ¡¸Á° 8 Ä®o ‡nµ Specific activity š¸É ¼Š„ªnµÁ¨È„œo °¥ —´Šœ´Êœ£µª³š¸É
Á®¤µ³¤Äœ„µ¦„´ —Á°œÅŽ¤rŨÁžÄœÁœºÊ°¤³¡¦oµª ‡º°š¸É¡¸Á° 8 ˜n µ¦„´—š¸É¡¸ Á°˜Éε‹³¤¸‡nµ Specific activity ¤µ„