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Extracting of Urease from Soybean and Its Influencing Factors(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2008年04期
Page:
704-707
Research Field:
Publishing date:

Info

Title:
Extracting of Urease from Soybean and Its Influencing Factors
Author(s):
ZHOU Dong-kai; LIU Ying; MA Xue-liang; LIU Dong-mei; WANG Zhan-yong; MA Juan
Liaoning Shihua University，Fushun 113001，Liaoning,China
Keywords:
Soybean(Glycine max); Soybean urease; Km; Enzyme activity
PACS:
Q550.3
DOI:
10.11861/j.issn.1000-9841.2008.04.0704
Abstract:
The problem of lacking soybean urease in Chinese has been becoming more and more serious thanks to the increasing market demand although soybean（Glycine max)was planted in large area all across China,the experiment was designed to extract soybean urease by ethanol-water solution in laboratory and realized the predetermined target.The Km value of the soybean urease was 3.576×10-2，the enzyme activity of 1.0 mL extracting liquid was 26.58 U.The optimum conditions of extracting soybean urease were investigated by the concentration of ethanol in water，the ratio of soybean fine powder weight to extract liquid volume(W/V)and pH value of the extract solution.Urea could be decomposed to（NH4)2O3by soybean urease,and NH+4could be quantitatively determinated by Nessler’s reagent colorimetry.The absorption value at 480 nm（A-480）was proportional to NH+4concentration in the reaction solution，so it had linear correlation with the soybean urease activity.When the concentration of ethanol in water was 30%，A-480 showed the best value 0.885 than those were 18%、24%、42% and 36%.W/V equal to 1:10 was the best ratio for A-480 than those were 1:6，1:8，1:12 and 1:14.Kjeldahl method was used to determine the NH+4concentration at different pH values，the HCl dosage in titration operation was proportional to NH+4concentration thus to manifest the soybean urease activity，the experiment result showed that 7.0 was the optimum pH value.

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Last Update: 2014-10-07