การพัฒนาการผลิตโบรโมเปอร์ออกซิเดสจากสาหร่ายสีแดง polycarvernosa sp.

The Development of Bromoperoxidase Production from Red Algae Polycarvernosa sp.

Name: ศยามพงษ์ พงษ์ดำ

Organization : มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

keyword: Bromoperoxidase

; polycarvernosa sp

; Red algae

Abstract: Effect of various cell disintegration techniques on enzyme extraction were studied for thepurification. These techniques were ; grinding with detergent (Triton X-100 and sodiumdeoxycholate), grinding with ultrasonication, freeze and thaw, grinding with freeze and thaw,grinding with agitation. Grinding with agitation (agitation time of 6 hours) provided a good result.Specific activity of bromoperoxidase was increased up to 57.54% when compared with grindingalone. Effects of various precipitants on fractionation by precipitation were also investigated. Theseprecipitants were ; ammonium sulphate, methanol, ethanol and acetone. Fractionation by acetonegave the highest degree of purification (degree of purification is 4.21). At the purification stage,DEAE-Toyopearl was shown to be a greater anion-exchanger than that of DEAE-Sephadex, with7.24 degree of purification. Comparison of the effect of separation by gel filtration using SephadexG-75 and Biogel A-O.5 M were also studied. The degree of purification from Sephadex G-75 was14.42, while from Biogel A-O.5 M was lower. The protease concentration in crude enzyme was 267pU/ml, but in partial purified enzyme was only 0.002 pU/ml. Molecular weight of the extractedbromoperoxidase was approximately 6 15,000 dalton. Bromoperoxidase was deactivated by dialysisagainst EDTA, then reactivated specifically by vanadium. It was found that vanadium 0.6 mM wassuitable for partial purified enzyme reactivation. Effect of temperature on stability was studied,partial purified enzyme was more stable than crude enzyme. The partial purified enzyme was stableat acidic pHs down to 4 and at alkaline pHs up to 9, The optimum pH and temperature of the partialpurified enzyme activity were 6 and 55ฐC respectively. Crude enzyme was stable at acidic pHsdown to 4 and at alkaline pHs up to 9. The optimum pH and temperature of crude activity were 6and 55OC respectively. After maintaining at 4’C for 40 days, crude enzyme activity was decreasedto 47.50%, then partial purified enzyme activity was decreased to 26.07 %. Crude enzyme activityand partial purified enzyme activity were decreased to 34.48 % and 30.63 “/q respectively, whenkept at -2O’C for 40 days.

มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

Address: กรุงเทพมหานคร (Bangkok)

Email: info.lib@mail.kmutt.ac.th

Name: ดร. จิรศักดิ์ คงเกียรติขจร

Role: อาจารย์ที่ปรึกษา

Created: 2541

Issued: 2005-10-17

Modified: 2006-05-30

วิทยานิพนธ์/Thesis

ISBN: 9746245678

CallNumber: BIT266

tha

DegreeName: วิทยาศาสตรมหาบัณฑิต

Level: ปริญญาโท

Descipline: เทคโนโลยีชีวภาพ

Grantor: มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

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