|Table of Contents|

Cloning of Douchi Fibrinolytic Enzyme Gene and Its Expression in Bacillus subtilis(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2012年01期
Page:
159-161
Research Field:
Publishing date:

Info

Title:
Cloning of Douchi Fibrinolytic Enzyme Gene and Its Expression in Bacillus subtilis
Author(s):
QI Hai-ping; GANG jie; HU Wen-zhong; JIANG Ai-li; TIAN Mi-xia
College of Life Science,Dalian Nationalities University,Education Department and National Nationality Key Lab，Dalian 116600,Liaoning,China
Keywords:
Douchi fibrinolytic enzyme; Gene cloning; Highly expression; Bacillus subtilis
PACS:
TS201.1
DOI:
10.3969/j.issn.1000-9841.2012.01.037
Abstract:
With the total DNA extracted from Bacillaceae UD8 as template,one pair of primers were designed according to DNA sequence of Douchi fibrinolytic enzyme(DFE)gene(AY720895.2)on Genbank website,to amplify DFE gene by PCR.The PCR prodcuct was inserted into expression plasmid EC-BS8,and the plasmid was transformed into the host strain Bacillus subtilis. DFE gene was successfully expressed in Bacillus subtilis and the maximum Douchi fibrinolytic enzyme level reached 9 830 IU·mL-1,which was 2.9 fold higher of the initial strain.The cloned Douchi fibrinolytic enzyme purified molecular weight was 31.0 kDa with a single band from SDS-PAGE.

References:

[1]Sumi H,Banba T,Kishimoto N.Strong pro-urokinase activators proved in Japanese soybean cheese natto[J].Journal of Japanese Society of Food Science and Technology,1996,43(10):1124-1127.

[2]Urano T,Ihara H,Umemura K,et al.The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis?Cleaves and inactivates plasminogen activator inhibitor type I[J].The Journal of Biological Chemistry,2001,276:24690-24696.

[3]刘宇峰，王金英，孙岸弢，等.豆豉纤溶酶保健功能食品的研制[J].大豆通报，2000，（2）：22-23.(Liu Y F,Wang J Y,Sun A T,et al.Development of Douchi fibrinolytic enzyme functional food[J].Soybean Bulletin,2000，（2）：22-23.

[4]段智变，江汉湖，张书霞，等.纳豆激酶粗制液对家兔溶血栓作用及其机制研究[J].营养学报，2003，25（1）：46-51.(Duan Z B,Jiang H H,Zhang S X,et al.Studies on fibrinolytic function of Nattokinase and its mechanism[J].Acta Nutrimenta Sinica,2003,25（1）：46-51.)

[5]吕美云，张新，陆云华.豆豉纤溶酶基因的克隆及其在毕赤酵母中的表达[J].安徽农业科学，2008，36(6)：2270-2271.(Lu M Y,Zhang X,Lu Y H.Cloning of Douchi Fibrinolytic enzyme gene and its expression in Pichia pastoris[J].Journal of Anhui Agricultural Sciences，2008，36(6)：2270-2271.)

[6]刘成君,黄庆,赵莲,等.大肠杆菌-枯草杆菌穿梭质粒载体pSUGV4的构建[J].四川大学学报(自然科学版),2001,38(2):243-246.(Liu C J,Huang Q,Zhao L,et al.Construction of a Escherichia coli-bacillus subtilis shuttle vector PSUGV4[J].Journal of Sichuan University(Natural Science Edition),,2001,38(2):243-246.)

[7]Astrup T,Mullertz S.The fibrin plate method for estimating fibrinolytic activity[J].Archives of biochemistry and biophysics,1952，40：346-351.

[8]Sambrook J,Fritsch E F,Maniatis T..Molecular cloning:A laboratory manual[M].Cold Spring Harbor Laboratory Press,1989.齐海萍,钱和,王璋,等，淡豆豉纤溶酶的分离纯化[J].中国调味品,2005(11):21-27.

[9]Qi H P,Qian H,Wang Z,et al.The purification of Douchi fibrinolytic enzyme from Dochi[J].China Condiment,2005(11):21-27.


Memo

Memo:
-
Last Update: 2014-08-15