The Genome Mining and Expression for β-Glucosidase of Thermomyces lanuginosus
Genome mining is an efficient tool for discovering biocatalysts,and developing thermal stability β-glucosi-dase has important application value.Seven genes,annotated as β-glucosidase gene or β-glucosidase predicts pro-tein,are excavated using genome mining technology from thermophilic fungus Thermomyces lanuginosus genomic da-ta.Among them,TLG-1,TLG-2,TLG-4,TLG-5,and TLG-6 belong to the GH3 family,while TLG-3 and TLG-7 be-long to the GH1 family and GH17 family,respectively.TLG-7 has low homology(48.43％)with the existing se-quences in the GenBank database.Four β-glucosidase(TLG-2,TLG-5,TLG-6,and TLG-7),with low homology and limited research on homologous sequences,are selected for gene cloning and heterologous expression in Pichia pas-toris.After the recombinant Pichia pastoris is induced by methanol for expression,the supernatant of the fermentation broth is taken for SDS-PAGE analysis and β-glucosidase activity testing.The results show that TLG-2,TLG-5,TLG-6,and TLG-7 are successfully secretory expressed.TLG-2 has the highest activity,reaching 166.6 U·mL-1,fol-lowed by TLG-5 with 79.8 U·mL-1,while TLG-6 and TLG-7 have lower activity,only 9.9 U·mL-1 and 9.5 U·mL-1,respectively.The above results provid some references for the application ofβ-glucosidase from T.lanuginosus.
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