Dsm IP Assets B.v.

Dsm N.v.

Gist-brocades B.v.

Dsm Anti-infectives B.v.

Hendrik Louis Bijl

Johannes Martinus Jacobus Visser

Albert Schaap

Johannes Hendrik Wolf

Daniel Verkoeijen

Johannes Martinus Visser

Mieke Sibeijn

Daniel Veroeijen

Fenna Johanna Catharina Jolink

Johannes Hendrick Wolf

An improved pasteurisation protocol for pasteurising microbial cells is disclosed. The protocol has three stages, a first heating stage, a second plateau stage at which the cells are held at a (maximum and) constant temperature, and a third cooling stage. Both the heating and the cooling stages are rapid, with the temperature of the cells passing through 40 to 80&#xb0; C. in no more than 30 minutes in the heating stage. The heating rate is at least 0.5&#xb0; C./minute and during cooling is at least &#x2212;0.5&#xb0; C./minute. The plateau maximum temperature is from 70 to 85&#xb0; C. By plotting the pasteurisation protocol on a time (t, minutes) versus temperature (T, &#xb0; C.) graph, one obtains a trapezium having an area less than 13,000&#xb0; C. minute. Not only does this result in a smaller energy input (and so a reduction in costs), but a better quality (and less oxidised) oil results having a peroxide value (POV) of less than 1.5 and an anisidine value (AnV) of less than 1.0.

Pasteurisation process for microbial cells and microbial oil

An improved pasteurisation protocol for pasteurising microbial cells is disclosed. The protocol has three stages, a first heating stage, a second plateau stage at which the cells are held at a (maximum and) constant temperature, and a third cooling stage. Both the heating and the cooling stages are rapid, with the temperature of the cells passing through 40 to 80&#xb0; C. in no more than 30 minutes in the heating stage. The heating rate is at least 0.5&#xb0; C./minute and during cooling is at least &#x2212;0.5&#xb0; C./minute. The plateau maximum temperature is from 70 to 85&#xb0; C. By plotting the pasteurisation protocol on a time (t, minutes) versus temperature (T, &#xb0; C.) graph, one obtains a trapezium having an area less than 13,000&#xb0; C. minute. Not only does this result in a smaller energy input (and so a reduction in costs), but a better quality (and less oxidized) oil results having a peroxide value (POV) of less than 1.5 and an anisidine value (AnV) of less than 1.0.

A stable aqueous liquid lactase formulation is provided, comprising lactase and further comprising sodium, calcium or potassium--lactate or a combination thereof and optionally a sugar, and/or optionally comprising sodium or potassium chloride or a combination thereof, preferably wherein the concentration of each of the components is such that the water activity Ais at most 0.82. The formulation is particularly suitable when using invertase-free lactase, allowing the use of sucrose as stabilizer. Also provided is a process to produce the liquid lactase formulation, an infant formula (e.g. as powder of granulate) comprising the liquid lactase formulation, a method to produce said infant formula, and the use of the formulation in the production of infant formula.

Stable liquid lactase compositions

A process for producing an oil, or a polyunsaturated fatty acid (PUFA), is described where an aqueous liquid comprising cells is deaerated, and the oil or PUFA is obtained from the cells. Deaeration can be performed by a wide variety of techniques, including the application of a vacuum (or reduced pressure), mechanical deaeration or degassing by reduced stirring or subjecting the broth to centrifugal forces, reducing viscosity (by dilution or heating), reduction in the supply of oxygen or air during fermentation or a reduction in stirring rate, lowering the pH (to lower the solubility of CO), filtration using PTFE capillaries, gas displacement (by bubbling in nitrogen or helium) or chemical deaeration (using oxygen scavengers).

Deaeration process

A pasteurization protocol for pasteurizing microbial cells is disclosed. The protocol has a heating stage, a plateau stage at which the cells are held at a (maximum and) constant temperature, and a cooling stage. The heating and cooling stages are rapid, the temperature of the cells passing through 40 to 80&#xb0; C. in no more than 30 minutes in the heating stage. The heating rate is at least 0.5&#xb0; C./minute and during cooling is at least &#x2212;0.5&#xb0; C./minute. The plateau maximum temperature is from 70 to 85&#xb0; C. By plotting the pasteurization protocol on a time (t, minutes) versus temperature (T, &#xb0; C.), one obtains a trapezium having an area less than 13,000&#xb0; C. minute. This results in a smaller energy input (so a reduction in costs) and a better quality oil having a peroxide value (POV) of less than 1.5 and an anisidine value (AnV) of less than 1.0.

A protocol for pasteurizing microbial cells is disclosed. The protocol has three stages, a first heating stage, a second plateau stage at which the cells are held at a (maximum and) constant temperature, and a third cooling stage. The heating and cooling stages are rapid, with the temperature of the cells passing through 40 to 80&#xb0; C. in no more than 30 minutes in the heating stage. The heating rate is at least 0.5&#xb0; C./minute and during cooling is at least &#x2212;0.5&#xb0; C./minute. The plateau maximum temperature is from 70 to 85&#xb0; C. By plotting the pasteurization protocol on a time (t, minutes) versus temperature (T, &#xb0; C.) graph, a trapezium is obtained having an area less than 13,000&#xb0; C. minute. This results in a smaller energy input and a better quality oil results having a peroxide value (POV) of less than 1.5 and an anisidine value (AnV) of less than 1.0.

An improved pasteurization protocol for pasteurizing microbial cells is disclosed. The protocol has three stages, a first heating stage, a second plateau stage at which the cells are held at a (maximum and) constant temperature, and a third cooling stage. Both the heating and the cooling stages are rapid, with the temperature of the cells passing through 40 to 80&#xb0; C. in no more than 30 minutes in the heating stage. The heating rate is at least 0.5&#xb0; C./minute and during cooling is at least &#x2212;0.5&#xb0; C./minute. The plateau maximum temperature is from 70 to 85&#xb0; C. By plotting the pasteurization protocol on a time (t, minutes) versus temperature (T, &#xb0; C.) graph, one obtains a trapezium having an area less than 13,000&#xb0; C. minute. Not only does this result in a smaller energy input (and so a reduction in costs), but a better quality (and less oxidized) oil results having a peroxide value (POV) of less than 1.5 and an anisidine value (AnV) of less than 1.0.

The extraction of a microbial or single cell oil, for example comprising one or more polyunsaturated fatty acids (PUFAs), directly from microbial cells is disclosed which avoids the need for solvents. After fermentation, the microbial cells are pasteurised, washed and the cell walls lysed or disrupted by a mechanical (e.g. homogenisation), physical (boiling or drying), chemical (solvents) or enzymatic (cell wall degrading enzymes) technique. The oil (containing the PUFA) is then separated from the resulting cell wall debris. This is achieved by centrifugation, which results in an oily phase (top layer) that contains the oil which that can be separated from an aqueous phase (containing the cell wall debris). The oil can then be extracted and if necessary the PUFA can be purified or isolated from the oil.

Isolation of microbial oils

The present invention relates to a crystalline carotenoid compound, such as &#x3b2;-carotene, with a purity of at least 95% and with substantially no solvent enclosed in the crystal lattice. The present invention further describes a process to prepare such a highly pure crystalline carotenoid compound from microbial biomass, without the use of a solvent extraction and/or an anti-solvent crystallization process.

Isolation of carotenoid crystals

The present invention discloses a microbial polyunsaturated fatty acid(PUFA)-containing oil with a high triglyceride content and a high oxidative stability. In addition, a method is described for the recovery of such oil from a microbial biomass deriverd from a pasteurized fermentation broth, wherein the microbial biomass is subjected to extrusion to form granular particles, dried and the oil then extracted from the dried granules using an appropriate solvent.

Preparation of microbial polyunsaturated fatty acid containing oil from pasteurised biomass

The present invention discloses a microbial polyunsaturated fatty acid(PUFA)-containing oil with a high triglyceride content and a high oxidative stability. In addition, a method is described for the recovery of such oil from a microbial biomass derived from a pasteurized fermentation broth, wherein the microbial biomass is subjected to extrusion to form granular particles, dried and the oil then extracted from the dried granules using an appropriate solvent.

Preparation of microbial polyunsaturated fatty acid containing oil from pasteurized biomass

The present invention discloses a microbial polyunsaturated fatty acid(PUFA)-containing oil with a high triglyceride content and a high oxidative stability. In addition, a method is described for the recovery of such oil from a microbial biomass derived from a pasteurised fermentation broth, wherein the microbial biomass is subjected to extrusion to form granular particles, dried and the oil then extracted from the dried granules using an appropriate solvent.

Growing community of inventors

Barendrecht, Netherlands

Albert Schaap