การแยกและเพิ่มความเข้มข้นของเอนไซม์แอซิคโปรโปรติเอสและอะไมเลส จาก Aspergillus niger

Name: เอกสิทธิ์ โฆษิตวงษา

Organization : มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

keyword: Solid State Fermentation

; Aspergillus niger

; Acid Proteases

; Amylases

; Preparation

Abstract: In this research, the production of acid proteases and amylases from Aspergillus niger in solid statefermentation were studied. The appropriate ratio of tapioca pulp and soybean pulp in the substratewere determined. The results showed that 60:40 ratio of tapioca pulp and soybean pulp gave thehighest growth and enzyme production. Cultivation was performed at moisture content of 60percent, 30?C and initial pH 5.0 for 48 hours. The maximum acid protease and amylase activitieswere 1349 Unit/g. dry substrate and 107141 unit/g. dry substrate, respectively.The activities of acid proteases and amylases when extracted from substrate by 0.5 percent sodiumchloride in phosphate citrate buffer pH 4 was higher than when extracted by only phosphate citratebuffer by 6.89 and 4.69 percent, respectively. The crude enzyme solution can be stored at–20?C and 4?C for at least 120 days without loosing any activity. But the activity dropped if thecrude enzyme solution was stored at 30?C for 15 days.The acid protease and amylase were precipitated by addition of acetone, ethanol and ammoniumsulphate. It was found that the optimum ratio of crude enzyme solution and solvent were 1:2 and 1:3for acetone and ethanol, respectively. In case of ammonium sulphate, maximum activity wasobtained at 70 percent salt saturation. The acetone precipitation gave recovery yields of 74.18 and73.81 percent and purity 4.98 folds and 5.00 folds for acid protease and amylase respectively, andthere were the bast results when recovery, purity and economic benefit are concerned.The activity of enzyme powder had optimum condition of pH 3.0, 60?C and pH 4.0, 60?C for acidproteases and amylases respectively. In the solution form, acid proteases and amylases were stableat pH range of 4.0 to 5.0 and 5.0 to 6.0 respectively. Acid proteases and amylases were quite stablewhen incubated for 30 minutes at 60?C and 65?C, respectively. However, the enzymes were quitestable when incubated at 40?C for 3 hours. Ki (inactivation cofficients) of both enzymes show thatamylases were more stable than acid proteases at the same temperature. The enzyme powder can bestored at –20?C, 4?C and 30?C for at least 120 days without worse of their activity.

มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

Address: กรุงเทพมหานคร (Bangkok)

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

Name: ผศ.ดร.อนันต์ ทองทา

Role: อาจารย์ที่ปรึกษา

Created: 2546

Issued: 2005-11-10

Modified: 2006-05-10

วิทยานิพนธ์/Thesis

ISBN: 9741833857

CallNumber: FDE217

tha

DegreeName: วิศวกรรมศาสตรมหาบัณฑิต

Level: ปริญญาโท

Descipline: วิศวกรรมอาหาร

Grantor: มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

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3	FDE217ab.txt	2.56 KB	79	2018-06-20 16:10:29

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