Expression of Bile Salt Hydrolase Genes from Two Different Origins in Escherichia coli Rosetta

doi:10.7506/spkx1002-6630-201307030

Abstract

Abstract: Bile salt hydrolase (BSH) gene of Bifidobacterium bifidum ATCC 29521 and Lactobacillus acidophilus NCFM was amplified by gradient PCR using genome DNA as template and cloned into pET28a (+) expression vector. The expression vector was then transfected into E. coli Rosetta and expression of BSH was induced by IPTG. SDS-PAGE analysis showed the presence of three target proteins estimated to be 40, 43 kD and 45 kD, respectively. Ninhydrin analysis revealed that the recombinant BSH hydrolyzed sodium glycocholate to glycine, showing its hydrolysis activity on conjugated bile salts.

Key words: bile salt hydrolase, cloning and expression, activity analysis

CLC Number:

TS252

YANG Shi-qin1，MAN Chao-xin2，QU Xing-guang1，LI Bin1，XIE Kun-hao1，JIANG Yu-jun1,2,*. Expression of Bile Salt Hydrolase Genes from Two Different Origins in Escherichia coli Rosetta[J]. FOOD SCIENCE, 2013, 34(7): 144-147.

References

(1. Oh, Hae-Keun, Ji Yoon Lee and Soo Jin Lim et al. Molecular Cloning and Characterization of a Bile Salt Hydrolase from Lactobacillus acidophilus PF01[J]. J. Microbiol. Biotechnol,2008,18(3):449-456.
(2. G.CORZO and S.E.GILLILAND. Measurement of Bile Salt Hydrolase Activity from Lactobacillus acidophilus Based on disappearance of Conjugated Bile Salts[J]. journal of dairy science,1999,82(3):466-471.
(3. G.CORZO and S.E.GILLILAND. Bile Salt Hydrolase Activity of Three Strains of Lactobacillus acidophilus[J]. journal of dairy science,1999,82(3):472-480.
(4. 黄茜,黄璐,潘道东,等.植物乳杆菌Lactobacillu plantarum Y1菌株胆盐水解酶基因（bsh）的克隆及重组表达[J].南京师大学报,2010,33(3):91-96.
(5. 战媛媛,王长远,于长青.植物乳杆菌中胆盐水解酶基因的原核表达及纯化[J].中国生物制品学杂志2009,22(9):876-879.
(6. Olivia McAuliffe, Raul J. Cano, and Todd R. Klaenhammer et al. Genetic Analysis of Two Bile Salt Hydrolase Activities in Lactobacillus acidophilus NCFM[J]. Applied and enveronmental microbiology, 2005,71(8):4925-4929.
(7. Geun-Bae Kim,Mathien Brochet，Byong H.Lee. Cloning and characterization of a bile salt hydrolase (bsh) from Bifibobacterium adolescentis. Biotechnology Letters,2005,27:817-822.
(8. Geun-Bae Kim, Carol M.Miyamoto, Edward A. Meighen et al. Cloning and Characterization of the Bile Salt Hydrolase Genes (bsh) from Bifidobacterium bifidum Strains[J]. Applied and enveronmental microbiology, 2005,70(9):5603-5612.
(9. JEAN-PIERRE GRILL, FRANC?OIS SCHNEIDER and JOEL CROCIANI, Purification and Characterization of Conjugated Bile Salt Hydrolase from Bifidobacterium longum BB536, Applied and Environmental Microbiology[J].1995,61(7):2577-2582.
(10. Hiroshi Tanaka,Honoo Hashiba,Jan Kok et al. Bile Salt Hydrolase of Bifidobacterium longum-Biochemical and Genetic Characterization[J].Applied and environmental Microbiology,2000,66(6):2502-2511.
(11. 居述林，王雪奇，马向前，等.两歧双岐杆菌培养基成分的改进[J].复旦学报，1998,37(2):186-190.
(12. 王昂,王丽丽,仪宏,等.茚三酮比色法测定谷氨酸含量的研究[J].中国调味品,2005,8:50-52.
(13. 刘慧,杜薇,张红星.乳酸乳球菌乳酸亚种高产胆盐水解酶发酵条件的优化研究[J].食品科学,2006,27(11):322-326.
(14. 王玉文，刘慧，李平兰，等.产胆盐水解酶乳酸菌的分离、鉴定及讲解胆固醇机理的初步研究[J].食品科学,2006,27(10):215-218.
(15. 王莜兰，孙兵兵，杨志平.D-氨基酸氧化酶基因在大肠杆菌中的诱导表达.南昌大学学报[J].2008,32(4):401-404.
(16. 魏杰，易忠，魏玉荣，等.IPTG诱导浓度、时间对Asia I型口蹄疫病毒VP1蛋白表达的影响[J],2009,36(5):63-66.

[1]	GUO Shuangshuang, ZHENG Fangfang, KE Congxue, WANG Zilong, WEI Yutuo, HUANG Ribo, DU Liqin. Enzymatic Characterization of Recombinant α-L-Rhamnosidase from Pediococcus acidilactici AS1.2696 [J]. FOOD SCIENCE, 2019, 40(4): 160-169.
[2]	REN Dayong, QU Tianming, YANG Liu, AN Bin, WANG Guochao, FENG Shirong. Screening of Lactic Acid Bacterial Isolates from Traditional Fermented Foods in Northeast China for Cholesterol-Lowering Property and Mechanism of Action Analysis [J]. FOOD SCIENCE, 2019, 40(22): 199-206.
[3]	TANG Leli, WANG Xiaomeng, WU Xiuling, HUANG Qing, LI He. Screening, Cloning and Characterization of a Novel β-Glucosidase from Soil Metagenomic Library [J]. FOOD SCIENCE, 2018, 39(4): 118-124.
[4]	Lü Xiaoqian, WANG Jingxue, LIN Hong. Characteristics and Expression of Endolysin from Staphylococcus aureus Bacteriophage qdsa001 [J]. FOOD SCIENCE, 2018, 39(18): 133-138.
[5]	LIAO Lan, ZHANG Fengli, LI Zhangfa, CHEN Linping, LIN Weijie, YANG Yanhong, WEN Xiaoyan, NI Li. In Vitro Expression of Cloned α-Gliadin and Effect of Deamidated α-Gliadin on the Mechanic Properties of Noodles [J]. FOOD SCIENCE, 2018, 39(17): 8-13.
[6]	HOU Jinhui, ZHANG Xiang, QIAO Gaoxiang. Cloning, Expression and Activity of an Endo-1,4-β-D-Glucanase from Pantoea ananatis [J]. FOOD SCIENCE, 2016, 37(23): 211-215.
[7]	WANG Jilian, LI Mingyuan, CHEN Yun. Progress in Cloning and Expression of Protease Genes [J]. FOOD SCIENCE, 2015, 36(23): 377-381.
[8]	ZHAO Rui-xiang, LI Gang, NIU Sheng-yang, YANG Da-guang, LU Si-hai, DUAN Gai-li, LI Yin-na. Purification and Enzymatic Properties of Bile Salt Hydrolase Produced by Lactobacillus acidophilus [J]. FOOD SCIENCE, 2014, 35(5): 165-168.
[9]	REN Jun, MIN Wei-hong*, ZHAN Dong-ling, FANG Li, LI Hui-ying, ZHU Yun-ming. Gene Cloning, Expression and Enzymatic Characterization of Aspartokinase Mutant G277K [J]. FOOD SCIENCE, 2014, 35(11): 149-154.
[10]	LIU Fei-fei，ZHANG Liang，GU Zheng-hua，DING Chong-yang，SHI Gui-yang. Cloning and Heterologous Expression in E. coli of Phospholipase C Gene from Bacillus cereus [J]. FOOD SCIENCE, 2013, 34(11): 182-187.
[11]	. Hypolipidemic Effect of Lactobacillus with Bile Salt Hydrolase Activity in Hyperlipidemic Rats [J]. FOOD SCIENCE, 2013, 34(1): 257-262.
[12]	HUANGQian，HUANGLu，PANDao-dong，YANGYao. Expression in Escherichia coli, Purification and Functional Characterization of Recombination Fusion Protein MBP-BSH [J]. FOOD SCIENCE, 2012, 33(7): 198-203.
[13]	. Expression and Activity Analysis of Bile Salt Hydrolases from Lactobacillus plantarum ST-Ⅲ [J]. FOOD SCIENCE, 2012, 33(17): 165-168.
[14]	LIUYue，LIShu-qian，ZHANGBo，LIUChang-jiang，XINGuang. Optimization of Extraction Conditions for Polygalacturonase from Actinidia arguta [J]. FOOD SCIENCE, 2012, 33(10): 115-118.
[15]	ZHANG Hong-xing1，WANG Nian1，LIU Yong2，LIU Li1，TENG Guo-xin3，LIU Hui1,*. Research Progress in Physiological Characteristics and Gene Sequencing of Bile Salt Hydrolase from Lactic Acid Bacteria [J]. FOOD SCIENCE, 2011, 32(1 ): 302-305.