Modified gram positive bacteria and uses thereof
AssigneeIntrexon Actobiotics NV
InventorsLothar Steidler, Karolien Van Huynegem, Klaas Vandenbroucke
Life of this patent
Estimating the term…
Abstract
The present invention relates to gram positive bacteria with increased stress resistance and/or improved storage characteristics. In particular, the invention relates to gram positive bacterium which accumulate intracellular trehalose. The gram positive bacterium according to the invention lack cellobiose-specific PTS system IIC component (PtcC)activity. The gram positive bacterium may further lack trehalose 6-phosphate phosphorylase (TrePP)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.
Claims30
1-18. (canceled)
1-28. (canceled)
A method for improving stress resistance or storage characteristics of a gram positive bacterium, preferably a lactic acid bacterium (LAB) or Bifidobacterium, preferably one or more stress resistance or storage characteristics selected from the group comprising resistance to acid conditions, resistance to bile salts, resistance to drying, freezing or freeze-drying, and osmotic resistance, comprising modifying the gram positive bacterium such as to lack PtcC activity, preferably wherein the gene encoding endogenous PtcC has been partially or completely deleted, disrupted or inactivated such as being incapable of producing functional ptcC gene product.
20-26. (canceled)
A method for preparing a food product, comprising admixing a food additive or starter culture comprising a non-pathogenic gram positive bacterium with a substrate material that is capable of being fermented by the non-pathogenic gram positive bacterium, optionally further comprising the step of fermenting said substrate material, wherein the non-pathogenic gram positive bacterium lacks cellobiose-specific PTS system IIC component (PtcC) activity, preferably wherein the gene encoding endogenous PtcC has been partially or completely deleted, disrupted or inactivated such as being incapable of producing functional ptcC gene product.
(canceled)
A method for improving a stress resistance or storage characteristic of a Gram-positive bacterium, comprising partially or completely deleting, disrupting, or inactivating an endogenous ptcC in the Gram-positive bacterium such that the Gram-positive bacterium is incapable of producing a functional PtcC.
A method for preparing a composition comprising a Gram-positive bacterium, the method comprising:
- i) culturing the Gram-positive bacterium in a medium comprising a substrate material capable of being fermented by the Gram-positive bacterium, thereby forming propagated Gram-positive bacterium;
- and
- ii) formulating the propagated Gram-positive bacterium into the composition, wherein the Gram-positive bacterium lacks PtcC activity, and wherein the Gram-positive bacterium further possesses at least one feature selected from the group consisting of:
- (a) constitutively overexpressing a gene encoding a trehalose transporter;
- (b) lacking trehalose 6-phosphate phosphorylase (TrePP) activity;
- and
- (c) containing a functional heterologous trehalose 6-phosphatase gene.
Dependencies are inferred from the claim text.