Enhanced expression of heterologous inulinase in Kluyveromyces lactis by disruption of hap1 gene.
Inulinase gene (Kcinu) derived from Kluyveromyces cicerisporus was expressed extracellularly in Kluyveromyces lactis using an episomal vector directed by Kcinu promoter. The influence of hap1 gene disruption on the expression of inulinase was studied. Inulinase activity in the supernatant of the recombinant Klhap1Delta strain was 391 U ml(-1) after cultured 120 h, which was 2.2-fold that of the wild type host. The relative inulinase mRNA level of the Klhap1Delta strain was 11.3-fold that of the wild type strain, and the expression plasmid was more stable in the mutant host. Based on these results, the disruption of hap1 facilitated the high and stable expression of inulinase controlled by Kcinu promoter in K. lactis.
Yu
Jing
J
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China.
Jiang
Jiaxi
J
Fang
Zian
Z
Li
Yuyang
Y
Lv
Hong
H
Liu
Jianping
J
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics*
D005656:Fungal Proteins; D014157:Transcription Factors; D006026:Glycoside Hydrolases; C023086:inulinase