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On-line Access: 2010-04-28

Received: 2009-08-08

Revision Accepted: 2010-01-28

Crosschecked: 2010-03-17

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Journal of Zhejiang University SCIENCE B 2010 Vol.11 No.5 P.342-349

http://doi.org/10.1631/jzus.B0900254


Expression of a bee venom phospholipase A2 from Apis cerana cerana in the baculovirus-insect cell


Author(s):  Li-rong Shen, Mei-hui Ding, Li-wen Zhang, Wei-guang Zhang, Liang Liu, Duo Li

Affiliation(s):  Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China, Department of Agriculture and Natural Resources, Delaware State University, Dover, DE 19901, USA

Corresponding email(s):   shenlirong@zju.edu.cn

Key Words:  Apis cerana cerana, Bee venom phospholipase A2 (BvPLA2), Insect cell, Expression


Li-rong Shen, Mei-hui Ding, Li-wen Zhang, Wei-guang Zhang, Liang Liu, Duo Li. Expression of a bee venom phospholipase A2 from Apis cerana cerana in the baculovirus-insect cell[J]. Journal of Zhejiang University Science B, 2010, 11(5): 342-349.

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author="Li-rong Shen, Mei-hui Ding, Li-wen Zhang, Wei-guang Zhang, Liang Liu, Duo Li",
journal="Journal of Zhejiang University Science B",
volume="11",
number="5",
pages="342-349",
year="2010",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B0900254"
}

%0 Journal Article
%T Expression of a bee venom phospholipase A2 from Apis cerana cerana in the baculovirus-insect cell
%A Li-rong Shen
%A Mei-hui Ding
%A Li-wen Zhang
%A Wei-guang Zhang
%A Liang Liu
%A Duo Li
%J Journal of Zhejiang University SCIENCE B
%V 11
%N 5
%P 342-349
%@ 1673-1581
%D 2010
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0900254

TY - JOUR
T1 - Expression of a bee venom phospholipase A2 from Apis cerana cerana in the baculovirus-insect cell
A1 - Li-rong Shen
A1 - Mei-hui Ding
A1 - Li-wen Zhang
A1 - Wei-guang Zhang
A1 - Liang Liu
A1 - Duo Li
J0 - Journal of Zhejiang University Science B
VL - 11
IS - 5
SP - 342
EP - 349
%@ 1673-1581
Y1 - 2010
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0900254


Abstract: 
bee venom phospholipase A2 (BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids. In this work, a new BvPLA2 (AccPLA2) gene from the Chinese honeybee (Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector. Tn-5B-4 (Tn) cells were transfected with the recombinant bacmid DNA for expression. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa. Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins. The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2 (BvPLA2 of the European honeybee, Apis mellifera) polyclonal serum. The reaction resulted in a double glycosylation band, which agrees with the band generated by the native AmPLA2 in Western blot analysis. The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg). In summary, the recombinant AccPLA2 protein, a native BvPLA2-like structure with corresponding biological activities, can be glycosylated in Tn cells. These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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