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Production of Bioactive Recombinant Bovine Chymosin in Tobacco Plants.


ABSTRACT: Chymosin (also known as rennin) plays an essential role in the coagulation of milk in the cheese industry. Chymosin is traditionally extracted from the rumen of calves and is of high cost. Here, we present an alternative method to producing bovine chymosin from transgenic tobacco plants. The CYM gene, which encodes a preprochymosin from bovine, was introduced into the tobacco nuclear genome under control of the viral 35S cauliflower mosaic promoter. The integration and transcription of the foreign gene were confirmed with Southern blotting and reverse transcription PCR (RT-PCR) analyses, respectively. Immunoblotting analyses were performed to demonstrate expression of chymosin, and the expression level was quantified by enzyme-linked immunosorbent assay (ELISA). The results indicated recombinant bovine chymosin was successfully expressed at an average level of 83.5 ng/g fresh weight, which is 0.52% of the total soluble protein. The tobacco-derived chymosin exhibited similar native milk coagulation bioactivity as the commercial product extracted from bovine rumen.

SUBMITTER: Wei ZY 

PROVIDER: S-EPMC4881450 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature

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Production of Bioactive Recombinant Bovine Chymosin in Tobacco Plants.

Wei Zheng-Yi ZY   Zhang Yu-Ying YY   Wang Yun-Peng YP   Fan Ming-Xia MX   Zhong Xiao-Fang XF   Xu Nuo N   Lin Feng F   Xing Shao-Chen SC  

International journal of molecular sciences 20160428 5


Chymosin (also known as rennin) plays an essential role in the coagulation of milk in the cheese industry. Chymosin is traditionally extracted from the rumen of calves and is of high cost. Here, we present an alternative method to producing bovine chymosin from transgenic tobacco plants. The CYM gene, which encodes a preprochymosin from bovine, was introduced into the tobacco nuclear genome under control of the viral 35S cauliflower mosaic promoter. The integration and transcription of the forei  ...[more]

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