เอกสารการประชุมวิชาการและเสนอผลงานวิจัย มหาวิทยาลัยทักษิณ ครั้งที่ 19 2552 - page 930

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„µ¦ž¦³»¤ª·µ„µ¦Â¨³Áœ°Ÿ¨Šµœª·‹´¥¤®µª·š¥µ¨´¥š´„¬·– ‡¦´Ê Šš¸É 19 ž¦³‹Îµže 2552
‡·—Áž}œœÊ宜´ „äÁ¨„»¨Ášnµ„´ 7.3 kDa Ž¹ÉŠ¤¸ œµ—Á¨È„„ªn µÁ°œÅŽ¤r Þ¦˜·Á° ×¥‹³¡š´Ê ŠÄœœÊ室´„Áž¨º °„
´ž³¦— µ¦¨³¨µ¥´Ê œÁ„¨º° Ammonium sulphate ¨³ Fraction µ¦¨³¨µ¥´ÊœÁ„¨º°š¸Éŗo‹µ„„µ¦šÎ µ Gel filtration
˜nŤn¡Â™Ãž¦˜¸œ °ŠÁ°œÅŽ¤rÁŽ¨¨¼Á¨Äœ´Êœ °Šµ¦¨³¨µ¥ PEG ÁœºÉ°Š‹µ„Á°œÅŽ¤r ÁŽ¨¨¼Á¨nªœÄ®n‹³™¼„
Â¥„Åž°¥¼nĜ´Ê œ °ŠÁ„¨º°®¨´Š‹µ„„µ¦Â¥„—o ª¥¦³µ¦¨³¨µ¥°Šª´’£µ‡ (ATPS)
£µ¡š¸É 4 —ŠŸ¨„µ¦šÎµ SDS-PAGE ×¥ Lane 1 : Marker, Lane 2 : œÊ室´ „Áž¨º°„´ ž³¦—, Lane 3 : PEG phase,
Lane 4 : Salt phase, Lane 5 ¨³ 6 : Purified protease ¨³ Lane 7 ¨³ 8 : Purified cellulase
¦»žŸ¨„µ¦ª·‹´¥
‹µ„„µ¦®¤´„Áž¨º°„´ž³¦——o ª¥°´˜¦µnªœ °ŠÁž¨º°„´ž³¦— (g) ˜n°œÊ嘵¨ (g) ‡º ° 3:1 Áž}œÁª¨µ 12
ª´œ ×¥«¹„¬µž¦·¤µ–Þ¦˜¸œ ¨³ž¦·¤µ– °ŠÁ°œÅŽ¤rÁŽ¨¨¼Á¨Â¨³Ãž¦˜·Á°¡ªnµ ¤¸ž¦· ¤µ–Þ¦˜¸œÁšnµ„´ 13.19
mg/ml ¤¸ ž¦·¤µ–Á°œÅŽ¤rÁŽ¨¨¼Á¨Ášnµ„´ 124.24 Units/ml ¨³¤¸ ž¦·¤µ–Á°œÅŽ¤rÞ¦˜·Á°Ášnµ„´ 11.25 Units/ml
Á¤ºÉ °œÎµœÊ室´ „Áž¨º°„´ž³¦— ¤µšÎµ„µ¦Â¥„Á°œÅŽ¤r ÁŽ¨¨¼Á¨Â¨³Ãž¦˜· Á°—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 ¨³ÄoÁ„¨º° 3
œ·— ŗo „n Tripotassium citrate, Ammonium sulphate ¨³ Dipotassium hydrogen phosphate ¡ªn µ „µ¦Äo PEG
15% ¨³Á„¨º ° Ammonium sulphate 19.2% ‹³µ¤µ¦™Â¥„Á°œÅŽ¤rÞ¦˜·Á°Â¨³Á°œÅŽ¤rÁŽ¨¨¼Á¨°°„‹µ„„´œÅ—o—¸
š¸É »— ×¥Á°œÅŽ¤rÞ¦˜·Á°‹³Â¥„Åž°¥¼nĜ´Ê œ PEG ¤¸ %Yield Ášnµ„´ 90.26% nªœÁ°œÅŽ¤rÁŽ¨¨¼Á¨‹³Â¥„Åž°¥¼n
Ĝ´Ê œÁ„¨º° ¤¸ %Yield Ášnµ„´ 78.86% ¨³¤¸‡nµ Kprotease ¨³‡nµ Kcellulase Ášnµ„´ 6.50 ¨³ 0.48 ˜µ¤¨Îµ—´ ‹µ„œ´Êœ šÎµ
Á°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 Á„ȝ
Fraction ¨³ 2 ml ¡ªnµ Sepharose 6B ‹³µ¤µ¦™Â¥„Á°œÅŽ¤r š´ÊŠ°Šœ·—°°„‹µ„„´œÅ—o ¨³šÎµÄ®oÁ°œÅŽ¤rš¸É ŗo¤¸
‡ªµ¤¦· »š›·ÍÁ¡·É¤¤µ„ ¹Êœ ×¥Á°œÅŽ¤rÞ¦˜· Á°‹³¤¸‡ªµ¤¦· » š›·ÍÁ¡·É¤ ¹ÊœÁž}œ 3.83 Ášnµ ¨³¤¸ %Yield Ášnµ„´
48.45% nªœÁ°œÅŽ¤rÁŽ¨¨¼Á¨‹³¤¸‡ªµ¤¦· »š›·ÍÁ¡·É¤ ¹ÊœÁž}œ 2.71 Ášnµ ¨³¤¸ %Yield Ášnµ„´ 64.04% »—šoµ¥‹¹Š®µ
œÊÎ µ®œ´„äÁ¨„»¨ °ŠÁ°œÅŽ¤rš´Ê Š°Šœ·——oª¥Áš‡œ·‡ Sodium dodecyl sulphate polyacrylamide gel electrophoresis
Lane 1 Lane 2 Lane 3 Lane 4 Lane 7 Lane 8 Lane 6 Lane 5
20.0 kDa
19.3 kDa
7.3 kDa
7.3 kDa