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| De Novo Design and High-throughput Screening Strategy Achieved Over Expression of Yarrowia lipolytica Lipase YLL in Pichia patoris |
| YANG Jiang-ke, MAO Ling, ZHOU Wen-jing, CHEN Jiang-shan, HU Chen |
| School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China |
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Abstract Lipase(EC 3.1.1.3) is a important biocatalyzer used in a variety of industrial fields. High-level lipase producing strains is the prerequisite for industrial bioapplication of lipase. De novo design, gene synthesis and high-throughput screening strategies were used to realize the overexpression of Yarrowia lipolytica lipase YLL in Pichia pastoris. By de novo design and gene synthesis of YLL lipase gene, the expression level of synthesized lipase gene was improved 1.26 fold than the original YLL gene. After that, a high throughput screening strategy including olive oil Rhodamine B plates phenotype check, 96 well plates and flask fermentation was used to acquire the over expression Pichia recombinant SILVER. In the 14 L fermentor, Pichia recombinant strain SILVER achieved the highest activity of 40 500 U/ml broths, and the protein content reached 2.52 g/L broths, which set the foundation for industrial application of YLL lipase.
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Received: 26 May 2014
Published: 25 August 2014
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