156 ◾ Fundamental Food Microbiology
4. Define and list the advantages and disadvantages of the following methods used in gene transfer: transduction, transfection, transformation, and protoplast fusion in lactic acid bacteria.
5. Describe the process involved in conjugal transfer of DNA in lactic acid bacteria. What are
the advantages and disadvantages of this method?
6. Describe the technique involved in electrotransformation of DNA in lactic acid bacteria.
Discuss the advantages of transferring DNA by this method over the other methods in lactic
acid bacteria.
7. Define the following terms and briefly discuss their importance in lactic acid bacteria: cryptic plasmid, plasmid replication, electrotransformation, IS-elements, and transposons.
8. Define the term “metabolic engineering.” With a proper example, explain how metabolic
engineering in lactic acid bacteria can be used to produce diacetyl, l(+)-lactic acid, and folic
acid in strains that have a low ability to produce them.
9. Explain with one example the importance of (a) protein targeting and (b) protein engineering research in lactic acid bacteria.
10. Explain how information on the genome sequence of lactic acid bacteria is helpful in their
application in strain development.
11. List three important applications that can be developed from the genome sequences of
phages of lactic acid bacteria.
12. Briefly discuss the functions of (a) lac genes and (b) las genes.
References
1. McKay, L.L., Functional properties of plasmids in lactic streptococci, Antonie Van Leeuwenhoek J.
Microbiol., 49, 259–274, 1983.
2. Gasson, M.J., Progress and potential in the biotechnology of lactic acid bacteria, FEMS Microbiol. Rev.,
12, 3–20, 1993.
3. Peterbauer, C., Maischberger, T., and Haltrich, D., Food-grade gene expression in lactic acid bacteria,
Biotechnol. J., 6, 1147–1161, 2011.
4. Douglas, G., Goh, Y., and Klaenhammer, T., Integrative food grade expression system for lactic acid
bacteria. In Strain Engineering, Williams, J.A., Ed., Humana Press, 2011, pp. 373–387.
5. Lee, S.Y., Mattanovich, D., and Villaverde, A., Systems metabolic engineering, industrial biotechnology and microbial cell factories, Microb. Cell. Fact., 11, 2012.
6. de Vos, W.M., Systems solutions by lactic acid bacteria: From paradigms to practice, Microb. Cell
Factories, 10, 2011.
7. Hugenholtz, J., The lactic acid bacterium as a cell factory for food ingredient production, Int. Dairy J.,
18, 466–475, 2008.
8. Daniel, C., Roussel, Y., Kleerebezem, M., and Pot, B., Recombinant lactic acid bacteria as mucosal
biotherapeutic agents, Trends Biotechnol., 29, 499–508, 2011.
9. Bron, P.A. and Kleerebezem, M., Engineering lactic acid bacteria for increased industrial functionality,
Bioengineered, 2, 80–87, 2011.
10. Villatoro-Hernández, J., Kuipers, O., Saucedo-Cárdenas, O., and Montes-de-Oca-Luna, R., Heterologous
protein expression by Lactococcus lactis. In Recombinant Gene Expression, Lorence, A., Ed., Springer,
New York, 2012, pp. 155–165.
11. Steidler, L. and Neirynck, S., In situ delivery of therapeutic proteins by recombinant Lactococcus lactis,
J. Microbiol., 41, 63–72, 2003.
12. Amalaradjou, M.A.R. and Bhunia, A.K., Bioengineered probiotics, a strategic approach to control
enteric infections, Bioengineered, 4, 2013.
13. Ray, B. and Miller, K.W., Pediocins (Pediococcus species). Naidu, A.S., Ed., CRC Press, Boca Raton, FL,
2000.
4. Define and list the advantages and disadvantages of the following methods used in gene transfer: transduction, transfection, transformation, and protoplast fusion in lactic acid bacteria.
5. Describe the process involved in conjugal transfer of DNA in lactic acid bacteria. What are
the advantages and disadvantages of this method?
6. Describe the technique involved in electrotransformation of DNA in lactic acid bacteria.
Discuss the advantages of transferring DNA by this method over the other methods in lactic
acid bacteria.
7. Define the following terms and briefly discuss their importance in lactic acid bacteria: cryptic plasmid, plasmid replication, electrotransformation, IS-elements, and transposons.
8. Define the term “metabolic engineering.” With a proper example, explain how metabolic
engineering in lactic acid bacteria can be used to produce diacetyl, l(+)-lactic acid, and folic
acid in strains that have a low ability to produce them.
9. Explain with one example the importance of (a) protein targeting and (b) protein engineering research in lactic acid bacteria.
10. Explain how information on the genome sequence of lactic acid bacteria is helpful in their
application in strain development.
11. List three important applications that can be developed from the genome sequences of
phages of lactic acid bacteria.
12. Briefly discuss the functions of (a) lac genes and (b) las genes.
References
1. McKay, L.L., Functional properties of plasmids in lactic streptococci, Antonie Van Leeuwenhoek J.
Microbiol., 49, 259–274, 1983.
2. Gasson, M.J., Progress and potential in the biotechnology of lactic acid bacteria, FEMS Microbiol. Rev.,
12, 3–20, 1993.
3. Peterbauer, C., Maischberger, T., and Haltrich, D., Food-grade gene expression in lactic acid bacteria,
Biotechnol. J., 6, 1147–1161, 2011.
4. Douglas, G., Goh, Y., and Klaenhammer, T., Integrative food grade expression system for lactic acid
bacteria. In Strain Engineering, Williams, J.A., Ed., Humana Press, 2011, pp. 373–387.
5. Lee, S.Y., Mattanovich, D., and Villaverde, A., Systems metabolic engineering, industrial biotechnology and microbial cell factories, Microb. Cell. Fact., 11, 2012.
6. de Vos, W.M., Systems solutions by lactic acid bacteria: From paradigms to practice, Microb. Cell
Factories, 10, 2011.
7. Hugenholtz, J., The lactic acid bacterium as a cell factory for food ingredient production, Int. Dairy J.,
18, 466–475, 2008.
8. Daniel, C., Roussel, Y., Kleerebezem, M., and Pot, B., Recombinant lactic acid bacteria as mucosal
biotherapeutic agents, Trends Biotechnol., 29, 499–508, 2011.
9. Bron, P.A. and Kleerebezem, M., Engineering lactic acid bacteria for increased industrial functionality,
Bioengineered, 2, 80–87, 2011.
10. Villatoro-Hernández, J., Kuipers, O., Saucedo-Cárdenas, O., and Montes-de-Oca-Luna, R., Heterologous
protein expression by Lactococcus lactis. In Recombinant Gene Expression, Lorence, A., Ed., Springer,
New York, 2012, pp. 155–165.
11. Steidler, L. and Neirynck, S., In situ delivery of therapeutic proteins by recombinant Lactococcus lactis,
J. Microbiol., 41, 63–72, 2003.
12. Amalaradjou, M.A.R. and Bhunia, A.K., Bioengineered probiotics, a strategic approach to control
enteric infections, Bioengineered, 4, 2013.
13. Ray, B. and Miller, K.W., Pediocins (Pediococcus species). Naidu, A.S., Ed., CRC Press, Boca Raton, FL,
2000.