เอกสารการประชุมวิชาการและเสนอผลงานวิจัย มหาวิทยาลัยทักษิณ ครั้งที่ 19 2552 - page 924

1
„µ¦ž¦³»¤ª·µ„µ¦Â¨³Áœ°Ÿ¨Šµœª·‹´¥¤®µª·š¥µ¨´¥š´„¬·– ‡¦´Ê Šš¸É 19 ž¦³‹Îµže 2552
„µ¦Â¥„Á°œÅŽ¤rÁŽ¨¨¼ Á¨Â¨³Ãž¦˜· Á°ÄœœÊ室´„Áž¨º°„´ž³¦——oª¥Áš‡œ·‡¦³µ¦¨³¨µ¥œÊÎ µ
°Šª´’£µ‡
Isolation of cellulase and protease in fermented pine apple rinds juice by using aqueous two
phase system
¥»ªµ„¦ «·¦·¡·š¥µ1 ¨³žd¥µ£¦–r £µ¬·˜„»¨2*
Yuwakorn Siripithaya1 and Piyaporn Pasitkul2*
š‡´—¥n°
«¹„¬µ„µ¦Â¥„Á°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Á°œÅŽ¤rÞ¦˜·Á°ÄœœÊ室´„Áž¨º°„´ž³¦——oª¥Áš‡œ·‡ Aqueous
two-phase system ץčo Polyethylene glycol 1500 (PEG 1500) š¸É‡ªµ¤Á o¤ oœ 5%, 10%, 15% ¨³ 20% w/v ¨³
čoÁ„¨º° 3 œ·— ŗo „n Tripotassium citrate, Ammonium sulphate ¨³ Dipotassium hydrogen phosphate Ž¹ÉŠ®µ
ž¦·¤µ–Á°œÅŽ¤r ÁŽ¨¨¼Á¨Â¨³Ãž¦˜· Á°Äœ´Êœµ¦¨³¨µ¥ PEG ¨³´Êœµ¦¨³¨µ¥Á„¨º°¡ªn µ „µ¦Äo PEG 1500 15%
¨³Á„¨º° Ammonium sulphate 19.2% ‹³šÎµÄ®o„µ¦Â¥„Á°œÅŽ¤rÁŽ¨¨¼Á¨°°„‹µ„Á°œÅŽ¤rÞ¦˜·Á°ÄœœÊ室´„
Áž¨º °„´ž³¦—‹³Å—o Ÿ¨—¸š¸É »— ×¥Á°œÅŽ¤r Þ¦˜·Á°‹³Â¥„Åž°¥¼nĜ´Êœ PEG ¤¸ %Yield Ášnµ„´ 90.26% n ªœ
Á°œÅŽ¤rÁŽ¨¨¼Á¨‹³Â¥„Åž°¥¼nĜ´Ê œÁ„¨º° ¤¸ %Yield Ášnµ„´ 78.86% ¨³¤¸‡nµ Kprotease ¨³‡n µ Kcellulase Ášnµ„´ 6.50
¨³ 0.48 ˜µ¤¨Îµ—´ ‹µ„œ´Êœ šÎ µÁ°œÅŽ¤rÄ®o¦· »š›·Í—oª¥„µ¦„ε‹´—¡°¨·Á¤°¦r¨³Á„¨º°°°„—oª¥Áš‡œ·‡ Gel filtration
ץčo Sepharose 6B œµ—‡°¨´¤œr 30 cm × 2 cm ´¢Á¢°¦rš¸É čo‡º° 50 mM Tris-HCl pH 7.0 ¨³Äo °´˜¦µ„µ¦Å®¨
Ášnµ„´ 0.22 ml/min ¡ªnµ Á°œÅŽ¤rÞ¦˜·Á°‹³¤¸‡ªµ¤¦· »š›·ÍÁ¡·É¤ ¹ÊœÁž}œ 3.83 Ášnµ ¨³¤¸ %Yield Ášn µ„´ 48.45%
n ªœÁ°œÅŽ¤rÁŽ¨¨¼Á¨‹³¤¸‡ªµ¤¦· » š›·ÍÁ¡·É¤ ¹ÊœÁž} œ 2.71 Ášnµ ¨³¤¸ %Yield Ášnµ„´ 64.04% ‹µ„„µ¦®µœÊ宜´ „
äÁ¨„» ¨ °ŠÁ°œÅŽ¤rš´Ê Š°Šœ·——oª¥Áš‡œ·‡ Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-
PAGE) ¡ªn µ Á°œÅŽ¤rÞ¦˜·Á°‹³¤¸ 2 Sub unit ×¥¤¸ œÊ宜´„äÁ¨„»¨Ášn µ„´ 20.0 ¨³ 19.3 kDa nªœÁ°œÅŽ¤r
ÁŽ¨¨¼Á¨¤¸ 1 Sub unit Ž¹É Š¤¸œÊ宜´„äÁ¨„»¨Ášnµ„´ 7.3 kDa
‡ÎµÎµ‡´ : ¦³µ¦¨³¨µ¥œÊ尊ª´’£µ‡
Abstract
In this study, the cellulose and protease in fermented pine apple rind juice were isolated by using
aqueous two-phase system (ATPS) which varied polyethylene glycol 1500 (PEG 1500) concentration 5%, 10%,
15% and 20% w/v and used tripotassium citrate, ammonium sulphate and dipotassium hydrogen phosphate salts.
Cellulase and protease activity were measured in both PEG and salt phase after partition. The results found that
____________________
1 œ· ·˜´ –”·˜«¹„¬µ £µ‡ª·µÁ‡¤¸ ‡–³ª·š¥µ«µ˜¦r ¤®µª·š¥µ¨´¥š´„¬·– Š ¨µ 90000
2 °µ‹µ¦¥rž¦³‹Îµ£µ‡ª·µÁ‡¤¸ ‡–³ª·š¥µ«µ˜¦r ¤®µª· š¥µ¨´¥š´„¬·– Š ¨µ 90000
* Ú¦«´¡šr/Ú¦µ¦ : 074-443955 e-mail : ppasitkul@hotmail.com