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BR-112019021396-B1 - PROCESS FOR MANUFACTURING PULULANO

BR112019021396B1BR 112019021396 B1BR112019021396 B1BR 112019021396B1BR-112019021396-B1

Abstract

The present disclosure provides a process for manufacturing pullulan.

Inventors

  • JEFFREY F. BREIT
  • BRANDON J. DOWNEY
  • Justin BELLER

Assignees

  • CAPSUGEL BELGIUM NV

Dates

Publication Date
20260310
Application Date
20180409
Priority Date
20170414

Claims (15)

  1. 1. Process for manufacturing pullulan characterized by comprising: a. preparing a fermentation medium comprising water, a nitrogen source, a carbon source, a phosphate source and a magnesium source; b. inoculating said fermentation medium with a pullulan-producing microorganism; c. cultivating said microorganism in said fermentation medium so as to produce pullulan in said fermentation medium; wherein said phosphate source comprises calcium phosphate and the pullulan has a (Mg+2 + Ca+2)/K+ ratio greater than 5.
  2. 2. Process according to claim 1, characterized in that said phosphate source consists of calcium phosphate.
  3. 3. Process according to claim 1, characterized in that said fermentation medium has an initial calcium phosphate concentration of 0.25 g/L to 4 g/L.
  4. 4. Process according to claim 1, characterized in that the magnesium source is magnesium chloride.
  5. 5. Process according to claim 1, characterized in that said fermentation medium has an initial magnesium chloride concentration of 0.01 g/L to 1 g/L.
  6. 6. Process according to claim 1, characterized in that the nitrogen source is yeast extract, NH4OH, L-glutamine, NaNO3, NH4Cl and L-arginine.
  7. 7. Process according to claim 1, characterized in that the nitrogen source is yeast extract.
  8. 8. Process according to claim 1, characterized in that the carbon source is glucose, maltose, lactose, sucrose, fruit and vegetable syrups, and fruit and vegetable molasses.
  9. 9. Process according to claim 1, characterized in that the carbon source is sucrose.
  10. 10. Process according to claim 1, characterized in that the microorganism in question is Aureobasidium pullulans.
  11. 11. Process according to claim 1, characterized in that said microorganism is Aureobasidium pullulans ATCC No. 42023.
  12. 12. Process, according to claim 1, characterized in that the cultivation of said microorganism is stopped by pasteurization.
  13. 13. Process according to claim 1, characterized in that the molecular weight of said pullulan is reduced by pullulanase produced by said microorganism, and said process is free of a step involving the addition of an acid or heat to reduce the molecular weight of said pullulan.
  14. 14. Process according to claim 1, characterized in that said fermentation medium has an initial pH of 2.5 to 8.
  15. 15. Process, according to claim 1, characterized in that said microorganism is cultivated for at least 3 days.

Description

[001] This application claims the benefit of U.S. Provisional Application No. 62/485,855, filed April 14, 2017, which is incorporated herein by reference in its entirety. SUMMARY [002] Processes for the production of pullulan by microorganisms in a fermentation medium containing calcium phosphate as a main phosphate source are disclosed. [003] Pullulan is a glucose polymer, more particularly, a polysaccharide consisting of α-1,4-linked maltotriose units that are connected by α-1,6 linkages between the terminal glycosidic residues of the trisaccharide. [004] Pullulan is obtained from an extracellular Aureobasidium pullulans strain when the strain is grown aerobically in a fermentation medium. The degrees of polymerization of the pullulan thus obtained are reported as falling in the range of a few hundred to several thousand kilodaltons (kDa). High-quality consistency of the product can be obtained by controlling the fermentation process. [005] Films formed with pullulan exhibit several properties that make them suitable for forming products such as capsules. These resulting films have excellent homogeneity and transparency. Furthermore, these films have very low oxygen permeability. Consequently, capsules made from pullulan are particularly useful for filling oxygen-sensitive products such as fish and vegetable oils. These films and resulting capsules made from pullulan also have a relatively low water content and exhibit high stability during storage, both in terms of mechanical properties and dissolution. [006] The natural origin of pullulan and its superior properties as a film-forming material have made pullulan a desirable polymer for the manufacture of capsules containing entirely natural ingredients. This would allow capsules manufactured with pullulan to meet the requirements for "organic" labeling. Notwithstanding the commercial need for a capsule meeting organic labeling requirements, to date no satisfactory pullulan capsules have been developed that meet organic labeling requirements. This is primarily due to the absence of pullulan as a raw material that has been manufactured to meet organic labeling requirements. [007] In one embodiment, a process for manufacturing pullulan is provided, comprising: a. preparing a fermentation medium comprising water, a nitrogen source, a carbon source, a phosphate source, and a magnesium source; b. inoculating said fermentation medium with a pullulan-producing microorganism; c. cultivating said microorganism in said fermentation medium so as to produce pullulan in said fermentation medium; wherein said phosphate source comprises calcium phosphate. [008] In one embodiment, the phosphate source comprises, consists essentially of, or consists of calcium phosphate. [009] In one embodiment, the fermentation medium has an initial calcium phosphate concentration of about 0.25 g/L to about 4 g/L. [0010] In one embodiment, the source of magnesium is magnesium chloride. [0011] In one embodiment, the fermentation medium has an initial magnesium chloride concentration of 0.01 g/L to 1 g/L. [0012] In one embodiment, the nitrogen source is yeast extract, NH4OH, L-glutamine, NaNO3, NH4Cl, arginine, or a mixture thereof. [0013] In one embodiment, the nitrogen source is yeast extract. [0014] In one embodiment, the carbon source is glucose, maltose, lactose, sucrose, fruit and vegetable syrups, fruit and vegetable molasses, or a mixture thereof. [0015] In one embodiment, the carbon source is sucrose. [0016] In one embodiment, the microorganism is Aureobasidium pullulans. [0017] In one embodiment, the microorganism is Aureobasidium pullulans ATCC No. 42023. [0018] In one embodiment, the cultivation of the microorganism is stopped by pasteurization. [0019] In one embodiment, the molecular weight of pullulan is reduced by pullulanase produced by the microorganism, and the process is free of a step involving the addition of an acid or heat to reduce the molecular weight of pullulan. [0020] In one embodiment, the fermentation medium has an initial pH of 2.5 to 8. [0021] In one embodiment, the microorganism is cultivated for at least 3 days. [0022] In one embodiment, pullulan is produced from the above processes. [0023] In one embodiment, pullulan has a molecular weight of 200 to 500 kDa. [0024] In one embodiment, pullulan has a Ca2+ content of at least 50 mg/kg, or at least 100 mg/kg, or at least 200 mg/kg, or at least 300 mg/kg, or even at least 500 mg/kg. [0025] In one embodiment, pullulan has a Mg2+ content of at least 10 mg/kg, or 20 mg/kg, or 30 mg/kg, or at least 40 mg/kg, or even at least 50 mg/kg. [0026] In one embodiment, pullulan has a ratio of (Mg2++ Ca2+)/K+ that is greater than 3, or greater than 5, or greater than 10, or even greater than 15. [0027] It should be understood that both the general description above and the detailed description below are exemplary and explanatory and are not limited to the subject matter claimed. BRIEF DESCRIPTION OF THE DRAWING [0028] Figur