Cell Surface-Displaying the Lactose Hydrolase on <i>Yarrowia lipolytica</i>: a New Approach to Lactose Hydrolysis

doi:10.13523/j.cb.20180806

China Biotechnology

2018, Vol. 38

Issue (8): 41-49 DOI: 10.13523/j.cb.20180806

Cell Surface-Displaying the Lactose Hydrolase on Yarrowia lipolytica: a New Approach to Lactose Hydrolysis

Yi-ying WANG,Hai-rong CHENG(

)

School of Life Science and Biotechnology, State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, China

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Abstract

Lactose is one of the important carbon sources for infants to obtain energy, but lactose needs to be hydrolyzed by enzyme to galactose and glucose before being absorbed. Infants and young children lacking lactose hydrolase can fall in lactose intolerance when consuming lactose-containing food. Lactose intolerance is a symptom of high incidence in China. Therefore, lactose hydrolysis with high efficiency in vitro is of great importance to alleviate the symptoms. The lactose hydrolase (also known as β-galactosidase) was displayed on the cell surface of the food-satefy-grade yeast Yarrowia lipolytica, and the engineered yeast cells are used directly to hydrolyze lactose to produce galactose and glucose. The engineered yeast cell (HCY10) can completely hydrolyze 50g/L lactose in 24 hours to produce galactose and glucose. The method has the advantages of high efficiency and simplicity, and it can provide an alternative solution for the efficient lactose hydrolysis in vitro.

Key words： Lactose β-galactosidase; Lactose hydrolase Yarrowia lipolytica Yeast surface display technology

Received: 08 March 2018 Published: 11 September 2018

ZTFLH:

Q819

Corresponding Authors: Hai-rong CHENG E-mail: chrqrq@sjtu.edu.cn

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Cite this article:

Yi-ying WANG,Hai-rong CHENG. Cell Surface-Displaying the Lactose Hydrolase on Yarrowia lipolytica: a New Approach to Lactose Hydrolysis. China Biotechnology, 2018, 38(8): 41-49.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20180806 OR https://manu60.magtech.com.cn/biotech/Y2018/V38/I8/41

Fig.1 Surface display plasmid containing lactose hydrolase gene from Aspergillus oryzae 112 (a) Plasmid pHyg-Pir1-P.dSIase (b) Plasmid pHyg-Pir1-Gal

Fig.2 Yeast transformants grown on YNLHX plate (a) Transformants grown on YNLHX plate (b) The yeast positive transformants grown on YNLHX plate (c) Color variations of yeast positive transformants by successive seven purification on YNLHX plate (d) Color variations of yeast positive transformants by successive twenty purification on YNLHX plate

Fig.3 Molecular identification of Y.lipolytica transformants M: DNA molecular standard; 1~4: The PCR results of lactose hydrolase gene of four transformants that remained dark blue; 5~8: The PCR results of lactose hydrolase gene of four light blue transformants; 9~10: The PCR results of lactose hydrolase gene of two white transformants

Fig.4 HPLC results of transformation of 5% lactose by different yeast transformants

Fig.5 Comparison of lactose hydrolase activity of 20 dark blue transformants

Fig.6 The hydrolysis efficiency of the HCY10 strain in different lactose concentrations

Fig.7 Comparison of erythritol synthesis by HCY10 transformant and control Y.lipolytica CGMCC7326 strain


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