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| Study of the Enzymatic Function of Myosin Cross Reactive Antigen from Bifidobacterium animalis |
| YANG Bo, CHEN Hai-qin, SONG Yuan-da, ZHANG Hao, CHEN Wei |
| State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China |
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Abstract Objective: To express the myosin cross reactive antigen (MCRA) from Bifidobacterium animalis subsp. lactis BB-12 and to determine its enzymatic function. Methods: MCRA gene was amplified by PCR from chromosome of B. animalis subsp. lactis BB-12, and then the gene was cloned into PichiaPinkTM expression vector pPinkα-HC, and transformed into PichiaPinkTM wild strain. The recombinant MCRA protein was confirmed through SDS-PAGE and Western blot. The lipid profile of the recombinant PichiaPinkTM was analyzed via GC-MS. Results: It was indicated from the results of SDS-PAGE and Western blot that the MCRA protein was expressed successfully and targeted into the cell membrane in the recombinant PichiPinkTM, with a molecular weight of 82 kDa. With adding LA in the media, the recombinant PichiaPinkTM converted LA into 10-hydroxy-cis12-octadecenoic acid (10-HOE). Conclusion: This is the first report of expressing the MCRA gene with successful target and function. The MCRA of B. animalis subsp. lactis BB-12 was successfully and actively expressed in Pichia pastoris, and the product is 10-HOE.
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Received: 10 September 2012
Published: 25 December 2012
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