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利用基于全细胞绿色荧光蛋白生长的大肠杆菌生物传感器对饲料蛋白质来源中的总赖氨酸和可利用赖氨酸进行定量分析。

Quantification of total and bioavailable lysine in feed protein sources by a whole-cell green fluorescent protein growth-based Escherichia coli biosensor.

作者信息

Chalova V I, Kim W K, Woodward C L, Ricke S C

机构信息

Department of Poultry Science, Texas A&M University, College Station, TX 77843, USA.

出版信息

Appl Microbiol Biotechnol. 2007 Aug;76(1):91-9. doi: 10.1007/s00253-007-0989-6. Epub 2007 May 4.

Abstract

Using a fluorescent whole-cell Escherichia coli biosensor previously developed in our laboratory, we determined total and bioavailable lysine in four feed ingredients (soybean, cottonseed, meat and bone meal, and sorghum) and three complete feeds (chick starter and finisher, and swine starter). The same feed sources were analyzed for total lysine by high performance liquid chromatography (HPLC) and bioavailable lysine by chick bioassay. No significant differences were found between bioavailable lysine estimates for soybean, cottonseed, meat and bone meal, chick starter and finisher, and swine starter obtained by the fluorescent E. coli biosensor and chick bioassay. Except for sorghum, the E. coli biosensor estimates for total lysine were highly comparable to those obtained by HPLC. Comparisons were also conducted between conventionally performed optical density-based and the newly developed fluorescence-based lysine assay. The lack of significant differences in data obtained for total and bioavailable lysine by both detection modes indicated reliance and accuracy of the fluorescent E. coli biosensor. Overall results suggest that the microbial assay based on green fluorescent protein fluorescence represents a promising alternative method for lysine quantification.

摘要

我们使用先前在本实验室开发的荧光全细胞大肠杆菌生物传感器,测定了四种饲料原料(大豆、棉籽、肉骨粉和高粱)以及三种全价饲料(雏鸡育雏料和育成料,以及仔猪开食料)中的总赖氨酸和可利用赖氨酸。通过高效液相色谱法(HPLC)分析相同饲料来源的总赖氨酸,并通过雏鸡生物测定法分析可利用赖氨酸。通过荧光大肠杆菌生物传感器和雏鸡生物测定法获得的大豆、棉籽、肉骨粉、雏鸡育雏料和育成料以及仔猪开食料的可利用赖氨酸估计值之间没有显著差异。除高粱外,大肠杆菌生物传感器对总赖氨酸的估计值与通过HPLC获得的估计值高度可比。还对传统的基于光密度的赖氨酸测定法和新开发的基于荧光的赖氨酸测定法进行了比较。两种检测模式获得的总赖氨酸和可利用赖氨酸数据缺乏显著差异,表明荧光大肠杆菌生物传感器的可靠性和准确性。总体结果表明,基于绿色荧光蛋白荧光的微生物测定法是一种有前景的赖氨酸定量替代方法。

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