A bioluminescent Escherichia coli auxotroph for use in an in vitro lysine availability assay

Erickson, A.M.; Díaz, Irene; Kwon, Yong-Kook; Ricke, Steven C.

doi:10.1016/s0167-7012(00)00121-4

Cited by 23 publications

(13 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This bioassay allows for rapid determination of amino acid concentrations within 60 min, a remarkably shorter time than necessary for any other conventional bioassays (Bertels et al 2012;Cardinal and Hedrick 1948;Chalova et al 2006;Erickson et al 2000;Kim et al 2010;Li and Ricke 2003;Palanza et al 2016;Steele et al 1949;Tamura et al 1952). The present results also demonstrate the high versatility of this bioassay in terms of both Fig.…”

Section: Discussionsupporting

confidence: 66%

“…Some recent studies have substituted auxotrophic Escherichia coli mutants for lactic acid bacteria (Li and Ricke 2003), since the former exhibits a lower doubling time. In addition, marker proteins-such as green fluorescent protein (GFP) and β-galactosidase-can be introduced to increase the detection sensitivity for cell growth (Bertels et al 2012;Chalova et al 2006;Chalova et al 2007;Erickson et al 2000;Kim et al 2010). Although these modifications have succeeded in shortening the incubation time when compared to conventional methods, they still require no less than a 4-6-h incubation period due to the above restriction intrinsic to growth-dependent bioassays.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Kameya

Asano

2016

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Bioassay for amino acid quantification is an important technology for a variety of fields, which allows for easy, inexpensive, and high-throughput analyses. Here, we describe a novel translation-dependent bioassay for the quantification of amino acids. For this, the gene encoding firefly luciferase was introduced into Lactococcus lactis auxotrophic to Glu, His, Ile, Leu, Pro, Val, and Arg. After a preculture where luciferase expression was repressed, the cells were mixed with analytes, synthetic medium, and an inducer for luciferase expression. Luminescence response to the target amino acid appeared just after mixing, and linear standard curves for these amino acids were obtained during 15-60-min incubation periods. The rapid quantification of amino acids has neither been reported in previous works on bioassays nor is it theoretically feasible with conventional methods, which require incubation times of more than 4 h to allow for the growth of the microbe used. In contrast, our assay was shown to depend on protein translation, rather than on cell growth. Furthermore, replacement of the luciferase gene with that of the green fluorescent protein (GFP) or β-galactosidase allowed for fluorescent and colorimetric detection of the amino acids, respectively. Significantly, when a Gln-auxotrophic Escherichia coli mutant was created and transformed by a luciferase expression plasmid, a linear standard curve for Gln was observed in 15 min. These results demonstrate that this methodology can provide versatile bioassays by adopting various combinations of marker genes and host strains according to the analytes and experimental circumstances.

show abstract

Section: Discussionsupporting

confidence: 66%

Section: Introductionmentioning

confidence: 99%

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Kameya

Asano

2016

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…In previous studies from our laboratory, bacterial growth response to lysine has been measured turbidimetrically (Zabala- Erickson et al 1999;Li and Ricke 2003) or by bioluminescence (Erickson et al 2000, Zabala-Díaz and. Both methods have drawbacks as discussed in the introduction.…”

Section: Construction Of E Coli Biosensor and Growth Characteristicsmentioning

confidence: 99%

Development of a whole cell green fluorescent sensor for lysine quantification

Chalova

Zabala-Díaz

Woodward

et al. 2007

World J Microbiol Biotechnol

View full text Add to dashboard Cite

As an essential amino acid, lysine is an important component of animal and human diets and its bioavailability can depend on a variety of factors. Therefore, an accurate pre-determination of bioavailable lysine in foods and feeds is important. In this study a whole cell fluorescent biosensor for the quantification of lysine in protein sources was constructed. A gene encoding for green fluorescent protein (GFPmut3) was introduced into an E. coli lysine auxotroph genome as a part of a mini-Tn5-Km transposon. The location of the transposon was determined and the growth kinetics of the newly constructed biosensor were examined. The transposon disrupted the ybhM gene, which encodes for the synthesis of a protein with an unknown function. No effect of the transposon's location in the genome or the expression of gfp on bacterial growth rates was observed. Based on the fluorescence emitted by GFPmut3, a standard curve after 6-h growth of the strain was generated. A correlation coefficient of 0.95 was observed when the fluorescence method was compared to the conventional optical density (OD) growth-based lysine assay. Using the newly developed lysine fluorescent whole cell sensor we determined the total lysine in casein acid hydrolyzate (7.13 ± 0.34%). When lysine added to 12 lg/ml and 30 lg/ml of casein acid hydrolyzate was quantified, recoveries of 97 ± 1.65% and 103 ± 4.66% respectively were detected. The results suggest that the microbial assay using GFP fluorescence represents a promising alternative method for the potential estimation of lysine in protein sources.

show abstract

“…Previously, bioluminescence‐based approaches have been shown to be quantitatively comparable to OD‐based assays for microbial growth‐based assessment of methionine and lysine estimations (Erickson et al. 2000; Froelich et al.…”

Section: Resultsmentioning

confidence: 99%

“…E. coli strains genetically constructed as auxotrophs for various amino acids have been utilized as potential bioassay organisms (Tuffnell and Payne 1985; Hitchins et al. 1989; Erickson et al. 1999a, b, 2000).…”

Section: Introductionmentioning

confidence: 99%

QUANTIFYING METHIONINE WITH A GREEN FLUORESCENT ESCHERICHIA COLI METHIONINE AUXOTROPH

Froelich¹,

Díaz²,

Chalova³

et al. 2005

Rapid Methods Automation Mic

View full text Add to dashboard Cite

Methionine is one of the essential and primary limiting amino acids in animal nutrition. In this study, an Escherichia coli methionine auxotroph bacterial strain that exhibited a linear growth response to methionine concentrations was transformed with a plasmid containing genes encoding for a green fluorescent protein, in order to develop a fluorescent bioassay technique for methionine quantitation. Transformants were grown on arabinose‐supplemental media and kept at 27C. The strain was also tested in supplemented media with increasing methionine concentrations, and growth response (measured as optical density [OD]) was compared with fluorescence. The transformed E. coli methionine auxotroph exhibited low OD values, but the OD and fluorescent standard curves used for estimating methionine concentrations were linear with increasing increments of methionine concentration (OD, R2 = 0.97 and 0.89; fluorescent, R2 = 0.90 and 0.89) as were the corresponding fluorescent responses (OD versus fluorescent, R2 = 0.85 and 0.91). The transformed fluorescent strain also responded to enzymatic digests of selected animal feed proteins, yielding differences in available methionine among the different feed protein sources. Using the fluorescence‐based bacterial growth assay to quantify methionine in microtiter plates has potential for assessment of methionine levels in animal feed samples.

show abstract

A bioluminescent Escherichia coli auxotroph for use in an in vitro lysine availability assay

Cited by 23 publications

References 23 publications

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Translation-dependent bioassay for amino acid quantification using auxotrophic microbes as biocatalysts of protein synthesis

Development of a whole cell green fluorescent sensor for lysine quantification

QUANTIFYING METHIONINE WITH A GREEN FLUORESCENT ESCHERICHIA COLI METHIONINE AUXOTROPH

Contact Info

Product

Resources

About