Degeest B, Mozzi F, De Vuyst L
Department of Applied Biological Sciences, Vrije Universiteit Brussel, Brussels, Belgium.
Int J Food Microbiol. 2002 Dec 15;79(3):161-74. doi: 10.1016/s0168-1605(02)00116-2.
To increase the exopolysaccharide (EPS) yields from Streptococcus thermophilus LY03 and to unravel the nature of the EPS degradation process, fermentation experiments were carried out with this strain in a customized MRS medium, using different additional carbohydrates or amino acids possibly related to growth and EPS production. No significant increase of the EPS yields or activities of the enzymes alpha-phosphoglucomutase, UDP-glucose pyrophosphorylase and UDP-galactose 4-epimerase that are correlated with EPS production, or of the activity of dTDP-glucose pyrophosphorylase involved in the rhamnose synthetic branch of EPS biosynthesis, was observed. The EPS monomer composition remained unchanged for all experiments. Fermentations with a sudden temperature increase or lowered pH were carried out as well to try to avoid EPS degradation upon prolonged fermentation. It was demonstrated that EPS degradation took place enzymatically. Incubations of purified high-molecular-mass EPS with cell-free culture supernatant or cell extracts showed its degradation by enzymes with an endo-activity. This glycohydrolytic activity probably encompasses several enzymes having a molecular mass lower than 50,000 and 10,000 Da, and seems to be rather stable at high temperature and low pH. These results contribute to a better understanding of the physiological and chemical factors influencing EPS production and degradation.
为提高嗜热链球菌LY03的胞外多糖(EPS)产量并揭示EPS降解过程的本质,使用定制的MRS培养基对该菌株进行发酵实验,添加了可能与生长和EPS产生相关的不同碳水化合物或氨基酸。与EPS产生相关的α-磷酸葡萄糖变位酶、UDP-葡萄糖焦磷酸化酶和UDP-半乳糖4-表异构酶的活性,以及参与EPS生物合成鼠李糖合成分支的dTDP-葡萄糖焦磷酸化酶的活性,均未显著增加。所有实验中EPS单体组成均保持不变。还进行了温度突然升高或pH降低的发酵,以避免长时间发酵导致EPS降解。结果表明EPS降解是由酶催化的。将纯化的高分子量EPS与无细胞培养上清液或细胞提取物一起孵育,结果显示其被具有内切活性的酶降解。这种糖水解活性可能包含几种分子量低于50,000和10,000 Da的酶,并且在高温和低pH下似乎相当稳定。这些结果有助于更好地理解影响EPS产生和降解的生理和化学因素。