Expression of lactonohydrolase Zhd101 for detoxifying zearalenone by Lactobacillus reuteri Pg4
Date Issued
2014
Date
2014
Author(s)
Yang, Wen-Chun
Abstract
Mycotoxins are fungal secondary metabolites and contaminate agricultural products during crops growing, harvest, transportation, processing, or storage. Zearalenone （ZEN） is a nonsteriod estrogenic mycotoxin, produced by Fusarium species, and has been frequently implicated in reproductive disorders of farm animals and occasionally in hyperoestrogenic syndromes in humans. Biological, chemical, and physical detoxification procedures are trying to figure out mycotoxin contaminations by absorption or biotransformation, and enzymatic detoxification could be an efficient method for ZEN detoxification. ZEN-detoxifying gene, zhd101, is isolated from Clonostachys rosea IFO 7063. Zhd101 converts ZEN to the nonestrogenic product. Maximal activity of Zhd101 toward ZEN was measured at pH 10.5 and 37 to 45°C. Lactobacillus reuteri Pg4 is a lactic acid bacteria isolated from the gastrointestinal tract of broilers and possesses probiotic characteristics, so it has great potential for application. This study is aimed to construct expression vector with zhd101 and electroporation into L. reuteri Pg4 to express Zhd101. The zhd101 gene was cloned into Escherichia coli-Lactobacillus shuttle vector, pNZ3004, and the plasmid pNZ-zhd101 was electoporotion into L. reuteri Pg4. Zhd101 was successfully expressed by recomninant L. reuteri pNZ-zhd101 at DNA, mRNA, and protein levels. L. reuteri pNZ-zhd101 transformants showed ZEN degradation activity at pH 7.0 and highly plasmid stability.
Subjects
玉米烯酮
內酯水解酶
乳酸桿菌
Type
thesis
File(s)![Thumbnail Image]()
Loading...
Name
ntu-103-R01641017-1.pdf
Size
23.54 KB
Format
Adobe PDF
Checksum
(MD5):ee7fdab1ec8e84b6fcb9637ef58291e6