Abstract
Lysine is an essential amino acid for both humans and animals; and it is usually the first or second limiting amino acid in most formulated diets. In order to estimate the lysine content in feeds and feed sources, rapid amino acid bioassays have been developed. The objective of this work is to assess a rapid assay for lysine supplementation in chicken feeds, using a luminescent Escherichia coli lysine-auxotrophic strain, to avoid prior thermal sterilization. An E. coli lysine auxotroph carrying a plasmid with lux genes was used as the test organism. The lysine assay was conducted using depleted auxotrophic cells in lysine samples. Luminescence was measured with a Dynex MLX luminometer after addition of the aldehyde substrate. Growth response (monitored as optical density at 600 nm) and light emission response of the assay E. coli strain were monitored to generate standard curves. Bioluminescent analysis of feed samples indicated that the method works well in the presence of a complex feed matrix. Comparison of both optical density and luminescent-based methods indicated that, when the assay takes place under optimal conditions, both methodologies correlated well (r 2=0.99). Except for the 0.64% lysine-supplemented feed, estimates for lysine based on the bacterial assay were over 80% (82–97%) of the theoretical values. Animal data showed that the bacterial bioluminescent method correlated well with the chick bioassay when diets with different levels of lysine supplementation were assayed for lysine bioavailability (r 2=0.97). Luminescent methodology coupled with a bacterial growth assay is a promising technique to assess lysine availability in supplemented animal feeds.
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References
Anantharaman K, Parard C, Decarli B, Reinhardt P, Graf E, Finot P-A (1983) The microbiological estimation of available lysine in industrial milk products using a lysine- Escherichia coli, M 26-26. In: McLoughlin JV, McKenna BM (eds) Basic studies in food science, research in food science and nutrition, vol 2. Boole Press, Dublin, pp 81–82
Baker DH (1996) Advances in amino acid nutrition and metabolism of swine and poultry. In: Kornegay ET (ed) Nutrient management of food animals to enhance and protect the environment. CRC Press, Boca Raton, Fla., pp 41–53
Baker JM, Griffiths MW, Collins-Thompson DL (1992) Bacterial bioluminescence: applications in food microbiology. J Food Prot 55:62–70
Baldwin TO, Devine JH, Heckel RC, Lin J-N, Shadel GS (1989) The complete nucleotide sequence of the lux regulon of Vibrio fischeri and the luxABN region of Photobacterium leiognathi and the mechanism of control of bacterial bioluminescence. J Biolumin Chemilumin 4:326–341
Bjarnason J, Carpenter KJ (1970) Mechanism of heat damage in proteins. 2. Chemical changes in pure proteins. Br J Nutr 24:313–329
Boctor AM, Harper AE (1968) Measurement of available lysine in heated and unheated feedstuffs by chemical and biological methods. J Nutr 94:289–296
Bronstein I, Fortin J, Stanley PE, Stewart GSAB, Kricka LJ (1994) Chemiluminescent and bioluminescent reporter gene assays. Anal Biochem 219:169–181
Carpenter KJ, Booth VH (1973) Damage to lysine in foods processing: its measurement and significance. Nutr Abstr Rev 43:423–451
Cork LC, Clarkson TB, Jacoby RO, Gaertner DJ, Leary SL, Linn JM, Pakes SP, Ringler DH, Strandberg JD, Swindle MM (1997) The cost of animal research: origins and options. Science 276:758–759
Erickson AM, Li X, Woodward CL, Ricke SC (1999) Optimisation of enzyme treatment for the degradation of feed proteins for an Escherichia coli auxotroph lysine availability assay. J Sci Food Agric 79:1929–1935
Erickson AM, Zabala Díaz IB, Kwon YM, Ricke SC. (2000) A bioluminescent Escherichia coli auxotroph for use in an in vitro lysine availability assay. J Microbiol Methods 40:207–212
Han Y, Baker DH (1994) Digestible lysine requirement of male and female broiler chicks during the period three to six weeks posthatching. Poult Sci 73:1739–1745
Heitzer A, Malachowsky K, Thonnard J, Bienkowski PR, White DC, Sayler GS (1994) Optical biosensor for environmental on-line monitoring of naphthalene and salicylate bioavailability with an immobilized bioluminescent catabolic reporter bacterium. Appl Environ Microbiol 60:1487–1494
Hill PJ, Stewart GSAB (1994) Use of lux genes in applied biochemistry. J Biolumin Chemilumin 9:211–215
Hitchins AD, McDonough FR, Wells PA (1989) The use of Escherichia coli mutants to measure the bioavailability of essential amino acids in foods. Plant Food Hum Nutr 39:109–120
Labadan MC Jr, Hsu K-N, Austic RE (2001) Lysine and arginine requirements of broiler chickens at two- to three-week intervals of eight weeks of age. Poult Sci 80:599–606
Meighen EA (1994) Genetics of bacterial bioluminescence. Annu Rev Genet 28:117–139
Nordheim JP, Coon CN (1984) A comparison of four methods for determining available lysine in animal protein meals. Poult Sci 63:1040–1051
Payne JW, Bell G, Higgins CF (1977) The use of an Escherichia coli lys − auxotroph to assay nutritionally available lysine in biological materials. J Appl Bacteriol 42:165–177
Rinehart KE (1996) Environmental challenges as related to animal agriculture—poultry. In: Kornegay ET (ed) Nutrient management of food animals to enhance and protect the environment. CRC Press, Boca Raton, Fla., pp 21–28
Ripp S, Applegate BM, Simpson ML, Sayler GS (2001) Whole-cell bioluminescent bioreporter sensing of foodborne intoxicants. In: Chen Y-R, Tu S-I (eds) Photonic detection and intervention technologies for safe food. Proc Soc Photo-Opt Instr Eng 4206:13–21
Salanitro JP, Blake LG, Muirhead PA, Maglio M, Goodman JR (1978) Bacteria isolated from the duodenum, ileum, and cecum of young chicks. Appl Environ Microbiol 35:782–790
SAS Institute (1988) SAS user's guide. SAS Institute, Cary, N.C.
Shockman GD (1963) Amino acids. In: Kavanagh F (ed) Analytical microbiology. Academic Press, New York, pp 567–673
Stewart GSAB, Williams P (1992) lux genes and the application of bacterial bioluminescence. J Gen Microbiol 138:1289–1300
Tesseraud S, Le Bihan-Duval E, Peresson R, Michel J, Chagneau AM (1999) Response of chick lines selected on carcass quality to dietary lysine supply: live performance and muscle development. Poult Sci 78:80–84
Tuffnell JM, Payne JW (1985) A colorimetric enzyme assay using Escherichia coli to determine nutritionally available lysine in biological materials. J Appl Bacteriol 58:333–341
Acknowledgements
This research was supported by Hatch grant H8311 (administered by the Texas A&M Agricultural Experiment Station) and by the Texas Advanced Technology Program, grant 000517-0220-2001 (Texas Higher Education Board, Austin, Tex.). The LUZ-CONICIT Graduate Fellowship (Maracaibo, Venezuela) supported I.B.Z.D. We thank C. Weiser and D. Jackson for conducting the chick growth assay.
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Zabala Díaz, I.B., Ricke, S.C. Quantitative detection of crystalline lysine supplementation in poultry feeds using a rapid bacterial bioluminescence assay. Appl Microbiol Biotechnol 62, 268–273 (2003). https://doi.org/10.1007/s00253-003-1271-1
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DOI: https://doi.org/10.1007/s00253-003-1271-1