African Journal of Biotechnology Vol. 5 (4), pp. 378-383, 16 February 2006   

ISSN 1684–5315 © 2006 Academic Journals        

Full Length Research Paper

A new manometric method for measuring carbon dioxide production by dairy starter culture: a case of Leuconostoc mesenteroides

Mebrouk KIHAL¹*, Jamal Eddine Henni¹, Hérvé Prevost² and Charles Diviès²

¹Laboratoire de Microbiologie-Appliquée, Département de Biologie, Faculté des Sciences, Université d’Oran, Bp 16 Es-sénia, Oran, 31100 Algérie.

²Laboratoire de Microbiologie- Biotechnologie, ENSBANA,1 esplanade Erasme, Université de Bourgogne, 21000 Dijon, France.

*Corresponding authors E-mail: Kihalm@hotmail.com.

Accepted 26 August, 2005

This paper describe a  technique  for carbon dioxide (CO₂) measurement  based on the displacement of acidifying liquid in burette at constant pressure. It was adapted to allow the growth of Leuconostoc sp. on semi-synthetic media and milk. A highly precise measurement was obtained using the chemical reaction of sodium carbonate with hydrochloric acid. This method has been proven to present an excellent linear response from 0 to 0.895 mM of CO₂ with a high sensitivity. With cultures in milk medium, kinetics of CO₂ evolved by Leuconostoc mesenteroides subsp. dextranicum L4 were higher than L. mesenteroides subsp. mesenteroides 19D and L. mesenteroides subsp. cremoris Ec195 grown on milk.. The use of L. mesenteroides subsp. dextranicum L4 and their  variants Lac(-)Cit(+), Lac(+)Cit(-) and Lac(-)Cit(-) showed that the CO₂ production correlated to the substrate utilization. The curve of CO₂ measured was close to the amount of CO₂ calculated from lactose and citrate used. The CO₂ production rate was higher with the parental strain (6.5 mM/ h) than with the variants. The amount of CO₂ produced was always close to the theoritical value, and was also correlated to the growth rate and substrates utilization by L. mesenteroides. The volume of CO₂ produced by Leuconostoc in milk was highly correlated with the D-lactate production (r = 0.995). This method could be used routinely for the evaluation and the selection of bacteria having potential ability for CO₂ production.

Key words: Leuconostoc sp., CO₂ production, lactic acid, milk.