A visual method for direct selection of high-producing Pichia pastorisclones

BMC Biotechnology

Methodology Article

Tao Ke¹  Ben Rao²  Xiang Dong Ma²  Li Xin Ma²  Yu Lin Zhou²  Fan Hu²  Peng Zheng²  Jie Lü³  Pei Hong Mao³  Xiang Jin³  Xin Li⁴  Sheng Yi Liu⁴ 

[1] College of Life Science and Technology, Nanyang Normal University, 473061, Nanyang, PR China;Hubei Key Laboratory of Industrial Biotechnology, College of Life Science, Hubei University, 430062, Wuhan, PR China;Institute of Ion Beam Biotechnology, College of Physics Science and Technology, Xinjiang University, 830008, Urumqi, PR China;Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, 430062, Wuhan, PR China;

关键词: Pichia Pastoris;    Methylotrophic Yeast;    Glucomannan;    Pastoris GS115;    Mannanase Activity;   

DOI  :  10.1186/1472-6750-11-23

 received in 2010-08-09, accepted in 2011-03-21,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThe methylotrophic yeast, Pichia pastoris, offers the possibility to generate a high amount of recombinant proteins in a fast and easy way to use expression system. Being a single-celled microorganism, P. pastoris is easy to manipulate and grows rapidly on inexpensive media at high cell densities. A simple and direct method for the selection of high-producing clones can dramatically enhance the whole production process along with significant decrease in production costs.ResultsA visual method for rapid selection of high-producing clones based on mannanase reporter system was developed. The study explained that it was possible to use mannanase activity as a measure of the expression level of the protein of interest. High-producing target protein clones were directly selected based on the size of hydrolysis holes in the selected plate. As an example, the target gene (9elp-hal18) was expressed and purified in Pichia pastoris using this technology.ConclusionsA novel methodology is proposed for obtaining the high-producing clones of proteins of interest, based on the mannanase reporter system. This system may be adapted to other microorganisms, such as Saccharomyces cerevisiae for the selection of clones.

【 授权许可】

Unknown   
© Hu et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.