Overexpressing <i>CrePAPS</i> Polyadenylate Activity Enhances Protein Translation and Accumulation in <i>Chlamydomonas reinhardtii</i>

The alga <i>Chlamydomonas reinhardtii</i> is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of <i>C. reinhardtii</i> strains producing genetically engineered recombinant therapeutic protein have been reported. However,...

Full description

Bibliographic Details
Main Authors: Quan Wang, Jieyi Zhuang, Shuai Ni, Haolin Luo, Kaijie Zheng, Xinyi Li, Chengxiang Lan, Di Zhao, Yongsheng Bai, Bin Jia, Zhangli Hu
Format: Article
Language:English
Published: MDPI AG 2022-04-01
Series:Marine Drugs
Subjects:
Online Access:https://www.mdpi.com/1660-3397/20/5/276
Description
Summary:The alga <i>Chlamydomonas reinhardtii</i> is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of <i>C. reinhardtii</i> strains producing genetically engineered recombinant therapeutic protein have been reported. However, owing to extremely low protein expression efficiency, none have been applied for industrial purposes. Improving protein expression efficiency at the molecular level is, therefore, a priority. The 3′-end poly(A) tail of mRNAs is strongly correlated with mRNA transcription and protein translation efficiency. In this study, we identified a canonical <i>C. reinhardtii</i> poly(A) polymerase (CrePAPS), verified its polyadenylate activity, generated a series of overexpressing transformants, and performed proteomic analysis. Proteomic results demonstrated that overexpressing CrePAPS promoted ribosomal assembly and enhanced protein accumulation. The accelerated translation was further verified by increased crude and dissolved protein content detected by Kjeldahl and bicinchoninic acid (BCA) assay approaches. The findings provide a novel direction in which to exploit photosynthetic green algae as a recombinant protein expression platform.
ISSN:1660-3397