Mining for new β-galactosidases form metagenome


Wei Wang;


β-galactosidases can be used to produce low-lactose milk and dairy products for lactose intolerant people.Because of the great economic value,identification of novelβ-galactosidases with excellent lactose hydrolysis from unculturable microorganisms accounting for over 99%of microorganisms in the environment via metagenomic strategy is still urgently in demand.In my study,an E.coli strain△lacZ(DE3)for effectively screeningβ-Galactosidases was constructed,which knocked out theβ-galactosidase gene lacZ in E.coli expression strain BL21(DE3)by CRISPR-Cas9 system.From the data of metagenome sequencing of cotton field in Shandong,China,175β-galactosidase homologs were identified,of which thirty-five candidates were successfully expressed in E.coli△lacZ(DE3).Thirteen candidates showedβ-galactosidase activities.They were all monomeric enzymes,which were conducive to industrial production.Five novelβ-D-galactosidases of their own features such as thermostable,alkaline and acidic were purifying and further investigated the lactose hydrolysis.This study will enrich the source ofβ-galactosidases,and attract some attentions toβ-galactosidases from metagenomic library,which is important for milk products industry.


galactosidase;hydrolysis;lacZ;sequencing;accounting;Shandong;Mining for new;alkaline;library;enrich;


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