Modified gram positive bacteria and uses thereof
The present invention relates to a gram positive bacterium, preferably a lactic acid bacterium (LAB) or Bifidobacterium , with increased stress resistance and/or improved manufacturing, processing and/or storage characteristics. In particular, the invention relates to a gram positive bacterium which accumulate intracellular trehalose. The gram positive bacterium according to the invention lack trehalose 6-phosphate phosphorylase (TrePP) activity. The gram positive bacterium may further lack cellobiose-specific PTS system IIC component (ptcC) activity. The gram positive bacterium may further overexpress trehalose transporters. The invention further relates to compositions comprising such gram positive bacterium as well as methods and uses thereof.
1. A composition comprising a gram positive bacterium selected from a lactic acid bacterium (LAB) and a Bifidobacterium , the gram positive bacterium lacking trehalose 6-phosphate phosphorylase (TrePP) activity and cellobiose-specific PTS system IIc component (“ptcC”) activity, and pharmaceutically acceptable carrier.
2. The composition according to claim 1 , wherein the gram positive bacterium does not contain functional heterologous trehalose 6-phosphate phosphatase.
3. The composition according to claim 1 , wherein the gram positive bacterium overexpresses one or more genes encoding a trehalose transporter.
4. The composition according to claim 1 , wherein the gram positive bacterium contains one or more heterologous gene product.
5. The composition according to claim 1 , wherein the gram positive bacterium is dried, spray-dried, frozen or freeze-dried.
6. A food additive, a probiotic composition or a starter culture comprising a gram positive bacterium selected from a lactic acid bacterium (LAB) and a Bifidobacterium , the gram positive bacterium lacking TrePP activity and ptcC activity.
7. The starter culture according to claim 6 , wherein the starter culture is a starter culture for the preparation of a food product.
8. The food additive, probiotic composition or starter culture according to claim 6 , wherein the gram positive bacterium does not contain functional heterologous trehalose 6-phosphate phosphatase.
9. The food additive, probiotic composition or starter culture according to claim 6 , wherein the gram positive bacterium overexpresses one or more genes encoding a trehalose transporter.
10. The food additive, probiotic composition or starter culture according to claim 6 , wherein the gram positive bacterium expresses one or more heterologous gene products.
11. The food additive, probiotic composition or starter culture according to claim 6 , wherein the food additive or starter culture is frozen, dried, spray-dried or freeze-dried.
12. A method for preparing a food product, comprising admixing the food additive, the probiotic composition or the starter culture according to claim 6 with a substrate material capable of being fermented by the gram positive bacterium, optionally further comprising fermenting said substrate material.
13. A food product comprising the food additive according to claim 6 , and a substrate material capable of being fermented by the gram positive bacterium.
14. The composition according to claim 1 , wherein a gene encoding endogenous TrePP has been partially or completely deleted, disrupted or inactivated such as being incapable of producing functional TrePP gene product.
15. The composition according to claim 1 , wherein a gene encoding endogenous ptcC has been partially or completely deleted, disrupted or inactivated such as being incapable of producing functional ptcC gene product.
16. The composition according to claim 3 , wherein overexpression of the one or more genes encoding a trehalose transporter is constitutive.
17. The composition according to claim 3 , wherein the one or more genes encoding a trehalose transporter is endogenous to the gram positive bacterium.
18. The composition according to claim 4 , wherein the one or more heterologous gene product is one or more prophylactic or therapeutic gene product or antigen.
19. The food additive, the probiotic composition or the starter culture according to claim 6 , wherein a gene encoding endogenous TrePP has been partially or completely deleted, disrupted or inactivated such as being incapable of producing functional TrePP gene product.
20. The food additive, the probiotic composition or the starter culture according to claim 6 , wherein a gene encoding endogenous ptcC has been partially or completely deleted, disrupted or inactivated such as being incapable of producing functional ptcC gene product.
21. The food additive, the probiotic composition or the starter culture according to claim 9 , wherein overexpression of the one or more genes encoding a trehalose transporter is constitutive.
22. The food additive, the probiotic composition or the starter culture according to claim 9 , wherein the one or more genes encoding a trehalose transporter is endogenous to the gram positive bacterium.
23. The food additive, the probiotic composition or the starter culture according to claim 10 , wherein the one or more heterologous gene product is one or more prophylactic or therapeutic gene product or antigen.
24. The composition according to claim 1 , wherein the lactic acid bacterium is Lactococcus lactis.
25. The food additive, probiotic composition or starter culture according to claim 6 , wherein the lactic acid bacterium is Lactococcus lactis.