Score: 11.00 |
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| Title: Metabolism of zearalenone by genetically modified organisms expressing the detoxification gene from Clonostachys rosea .
| | Author: Takahashi-Ando N Ohsato S Shibata T Hamamoto H Yamaguchi I Kimura M | | Journal: Appl . Environ . Microbiol . Citation: V : 70 ( 6 ) P : 3239-45 Year: 2004 Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub15184117 Accession (PMID): 15184117 | | Abstract: Zearalenone ( ZEN ) is converted to a nontoxic product by a lactonohydololase encoded by zhd101 .
An enhanced green fluorescent protein ( EGFP ) gene was fused to zhd101 ( ie , egfp : : zhd101 ) and expressed in Escherichia coli .
Both recombinant ZHD101 and EGFP : : ZHD101 were purified to homogeneity and characterized .
Maximal activity of ZHD101 toward ZEN was measured at approximately 37 to 45 degrees C and pH 10 . 5 ( k ( cat ) at 30 degrees C , 0 . 51 s ( -1 ) ) .
The enzyme was irreversibly inactivated at pH values below 4 . 5 or by treatment with serine protease inhibitors .
ZHD101 was also active against five ZEN cognates , although the efficiencies were generally low ; eg , the k ( cat ) was highest with zearalanone ( 1 . 5 s ( -1 ) ) and lowest with beta-zearalenol ( 0 . 075 s ( -1 ) ) .
EGFP : : ZHD101 had properties similar to those of the individual proteins with regard to the EGFP fluorescence and lactonohydrolase activity .
Fortuitously , EGFP : : ZHD101 exhibited a good correlation between the fluorescence intensity and reaction velocity under various pH conditions .
We therefore used egfp : : zhd101 to visually monitor the lactonohydrolase activity in genetically modified organisms and evaluated the usefulness of zhd101 for in vivo detoxification of ZEN .
While recombinant E coli and transgenic rice calluses exhibited strong EGFP fluorescence and completely degraded ZEN in liquid media , recombinant Saccharomyces cerevisiae gave poor fluorescence and did not eliminate all the toxicity of the mycotoxin in the medium ; ie , the rest of ZEN was transformed into an unfavorable substrate , beta-zearalenol , by an as-yet-unidentified reductase and remained in the medium .
Even so , as much as 75% of ZEN was detoxified by the yeast transformant , which is better than the detoxification system in which food-grade Lactobacillus strains are used ( H El-Nezami , N Polychronaki , S Salminen , and H Mykkune , Appl . Environ . Microbiol . 68 : 3545-3549 , 2002 ) .
An appropriate combination of a candidate host microbe and the codon-optimized synthetic gene may contribute significantly to establishing a mycotoxin detoxification system for food and feed . | Matching Sentences:
[ Sen. 2, subscore: 2.00 ]: An enhanced green fluorescent protein ( EGFP ) gene was fused to zhd101 ( ie , egfp : : zhd101 ) and expressed in Escherichia coli .
[ Sen. 3, subscore: 2.00 ]: Both recombinant ZHD101 and EGFP : : ZHD101 were purified to homogeneity and characterized .
[ Sen. 9, subscore: 2.00 ]: We therefore used egfp : : zhd101 to visually monitor the lactonohydrolase activity in genetically modified organisms and evaluated the usefulness of zhd101 for in vivo detoxification of ZEN .
[ Sen. 1, subscore: 1.00 ]: Zearalenone ( ZEN ) is converted to a nontoxic product by a lactonohydololase encoded by zhd101 .
[ Sen. 4, subscore: 1.00 ]: Maximal activity of ZHD101 toward ZEN was measured at approximately 37 to 45 degrees C and pH 10 . 5 ( k ( cat ) at 30 degrees C , 0 . 51 s ( -1 ) ) .
[ Sen. 6, subscore: 1.00 ]: ZHD101 was also active against five ZEN cognates , although the efficiencies were generally low ; eg , the k ( cat ) was highest with zearalanone ( 1 . 5 s ( -1 ) ) and lowest with beta-zearalenol ( 0 . 075 s ( -1 ) ) .
[ Sen. 7, subscore: 1.00 ]: EGFP : : ZHD101 had properties similar to those of the individual proteins with regard to the EGFP fluorescence and lactonohydrolase activity .
[ Sen. 8, subscore: 1.00 ]: Fortuitously , EGFP : : ZHD101 exhibited a good correlation between the fluorescence intensity and reaction velocity under various pH conditions .
| | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
Score: 4.00 |
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| Title: A model transgenic cereal plant with detoxification activity for the estrogenic mycotoxin zearalenone .
| | Author: Higa-Nishiyama A Takahashi-Ando N Shimizu T Kudo T Yamaguchi I Kimura M | | Journal: Transgenic Res .
Citation: V : 14 ( 5 ) P : 713-7 Year: 2005 Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub16245162 Accession (PMID): 16245162 | | Abstract: Zearalenone ( ZEN ) is an estrogenic mycotoxin produced by the necrotrophic cereal pathogen Fusarium graminearum .
This mycotoxin is detoxified by ZHD101 , a lactonohydrolase from Clonostachys rosea , or EGFP : ZHD101 , its fusion to the C-terminus of an enhanced green fluorescence protein .
We previously showed that egfp : zhd101 is efficiently expressed in T ( 0 ) leaves of rice .
In this study , we assessed the feasibility of in planta detoxification of the mycotoxin using progeny .
When protein extract from T ( 1 ) leaves was incubated with ZEN , the amount of the toxin decreased significantly as measured by HPLC .
ZEN degradation activity was also detected in vivo in transgenic T ( 2 ) seeds .
These results suggest that zhd101 can be exploited as an efficient and cost-effective system for protection of important cereals that are more susceptible to the pathogen ( eg , wheat and maize ) from contamination with the estrogenic mycotoxin . | Matching Sentences:
[ Sen. 2, subscore: 2.00 ]: This mycotoxin is detoxified by ZHD101 , a lactonohydrolase from Clonostachys rosea , or EGFP : ZHD101 , its fusion to the C-terminus of an enhanced green fluorescence protein .
[ Sen. 3, subscore: 1.00 ]: We previously showed that egfp : zhd101 is efficiently expressed in T ( 0 ) leaves of rice .
[ Sen. 7, subscore: 1.00 ]: These results suggest that zhd101 can be exploited as an efficient and cost-effective system for protection of important cereals that are more susceptible to the pathogen ( eg , wheat and maize ) from contamination with the estrogenic mycotoxin .
| | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
Score: 2.00 |
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| Title: Expression in cereal plants of genes that inactivate Fusarium mycotoxins .
| | Author: Higa A Kimura M Mimori K Ochiai-Fukuda T Tokai T Takahashi-Ando N Nishiuchi T Igawa T Fujimura M Hamamoto H Usami R Yamaguchi I | | Journal: Biosci . Biotechnol . Biochem . Citation: V : 67 ( 4 ) P : 914-8 Year: 2003 Type: ARTICLE | | Literature: oryza Field: abstract Doc ID: pub12784641 Accession (PMID): 12784641 | | Abstract: Trichothecene 3-O-acetyltransferase ( encoded by Tri101 ) inactivates the virulence factor of the cereal pathogen Fusarium graminearum .
Zearalenone hydrolase ( encoded by zhd101 ) detoxifies the oestrogenic mycotoxin produced by the same pathogen .
These genes were introduced into a model monocotyledon rice plant to evaluate their usefulness for decontamination of mycotoxins .
The strong and constitutive rice Act1 promoter did not cause accumulation of TRI101 protein in transgenic rice plants .
In contrast , the same promoter was suitable for transgenic production of ZHD101 protein ; so far , five promising T0 plants have been generated .
Low transgenic expression of Tri101 was suggested to be increased by addition of an omega enhancer sequence upstream of the start codon .
| Matching Sentences:
[ Sen. 2, subscore: 1.00 ]: Zearalenone hydrolase ( encoded by zhd101 ) detoxifies the oestrogenic mycotoxin produced by the same pathogen .
[ Sen. 5, subscore: 1.00 ]: In contrast , the same promoter was suitable for transgenic production of ZHD101 protein ; so far , five promising T0 plants have been generated .
| | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
