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Chapter 30 : Genetics of Lactococci

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Abstract:

This chapter presents current information in different areas of lactococcal genetics, keeping in mind (where possible) pertinence of findings to related pathogens. It highlights major recent work in lactococci, including surprising metabolic capacities, physiology, stress response, and studies leading to novel successful uses of lactococci for protein delivery. Lactococcal metabolism has been intensively studied for its industrial importance in fermentation processes, with a focus on metabolic pathways and their engineering. However, basic metabolic functions may have far-reaching effects, metabolic shifts can result in dramatic changes in growth characteristics and survival. Researchers confirmed and developed a 1970 study showing that lactococci not only ferment sugars, but are also capable of forming an active electron transport chain to generate respiration metabolism. Laboratory results demonstrate that respiration metabolism in lactococci is an efficient means of eliminating oxygen, compared to fermentation, leading to good survival in stationary phase. Some of the most spectacular applications of lactococci concern their use in "bioprotein" delivery. Some tools developed in are adaptable to other gram-positive bacteria. The development of surface display systems in lactic acid bacteria (LAB) will be potentially useful in the development of oral vaccines based on the nontoxic LAB. As an organism present on plants, in milk, in dairy products, and in the gut, may be the organism of choice for studies on the influence of environmental stress on evolution.

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30

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Lactic Acid Fermentation
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Proteins
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Integrative and Conjugative Elements
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Figures

Image of FIGURE 1
FIGURE 1

Phylogenetic tree reveals similarities between lactococci and streptococcal pathogens. Asterisks indicate species commonly found in dairy foods. The tree is based on alignments using the conserved gene (http://prodes.toulouse.inra.fr/multalin).

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30
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Image of FIGURE 2
FIGURE 2

Carbon metabolism in Fermentation of sugar results in ATP production, which in turn is used for anabolism. During anaerobic conditions and rapid sugar flux, all sugar is converted to lactate (homolactic fermentation). When the sugar flux is slower, or the oxygen concentration high, mixed acid fermentation is observed. The latter two conditions are characterized by lower NADH/NAD ratios than those found during homolactic fermentations. Note that the major part of the carbon for anabolism is derived from amino acids (or casein) supplied in the medium.

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30
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Image of FIGURE 3
FIGURE 3

Increases in GAPDH activity do not increase glycolytic flux in steadily growing MG1363. Levels of GAPDH activity were modulated by driving expression using different strength promoters. At the wild-type enzyme level (set to 1), GAPDH has zero control on the flux. gdw, grams dry weight. (Reproduced with permission from reference .)

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30
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Image of FIGURE 4
FIGURE 4

Activation of respiration metabolism in . Lactococci were found capable of respiration metabolism. An active respiration chain requires three components: an electron donor (NADH dehydrogenase, potentially encoded by and genes; it expulses H and transfers e); quinone electron transfer molecules (synthesized by enzymes encoded by the genes; it further transfers e); and an oxidoreductase (-encoded cytochrome bd quinol oxidase; it transfers e to its final acceptor, oxygen, which then reacts with H to produce water). An ATP synthase presumably recovers expulsed H to produce ATP during its entry.

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30
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Image of FIGURE 5
FIGURE 5

Respiration metabolism increases survival capacity of lactococci. When supplemented with hemin, aerobically grown lactococci can undergo respiration metabolism. As a result, cells stored at 4°C show a markedly better survival, as compared to cells grown aerobically in the absence of hemin or in static conditions. Improved survival was also observed when cells are maintained at 30°C. Experiment shown was performed by Karin Vido (URLGA, INRA, France).

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30
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Image of FIGURE 6
FIGURE 6

Bacterial root formation in semiliquid medium. Bacterial chains (here, an mutant of ; “parental strain”) sediment slowly in a semiliquid (0.035% agar) medium. A bacterial “dechained” mutant sediments more quickly to form a “root.” In such experiments, all the roots corresponded to independent mutants in the same gene, , encoding PBP1A. (Photograph kindly provided by S. Kulakauskas, URLGA, INRA, France.)

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30
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Tables

Generic image for table
TABLE 1

Characteristics of

Requires plasmid-encoded factors.

Citation: Gaudu P, Yamamoto Y, Jensen P, Hammer K, Gruss A. 2006. Genetics of Lactococci, p 356-368. In Fischetti V, Novick R, Ferretti J, Portnoy D, Rood J (ed), Gram-Positive Pathogens, Second Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816513.ch30

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