Study of the Enzymatic Function of Myosin Cross Reactive Antigen from <em>Bifidobacterium animalis</em>

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China Biotechnology ›› 2012, Vol. 32 ›› Issue (12) : 30-36.

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

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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 PichiaPink^TM expression vector pPinkα-HC, and transformed into PichiaPink^TM wild strain. The recombinant MCRA protein was confirmed through SDS-PAGE and Western blot. The lipid profile of the recombinant PichiaPink^TM 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 PichiPink^TM, with a molecular weight of 82 kDa. With adding LA in the media, the recombinant PichiaPink^TM 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.

Key words

Myosin cross reactive antigen / Linoleic acid / Pichiapink^TM expression system / Recombinant expression

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YANG Bo, CHEN Hai-qin, SONG Yuan-da, ZHANG Hao, CHEN Wei. Study of the Enzymatic Function of Myosin Cross Reactive Antigen from Bifidobacterium animalis[J]. China Biotechnology, 2012, 32(12): 30-36

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