IP Library › Patent Application 17434529
Patent Application
App. No. 17/434,529

METHODS, APPARATUSES, AND SYSTEMS FOR IMPROVING MICROBIAL PRESERVATION YIELD THROUGH RESCUE AND SERIAL PASSAGE OF PRESERVED CELLS

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Patent No.
US None
App. No.
17/434,529
Abstract

The present disclosure provides methods of improving microbe viability after preservation comprising subjecting a population of target microbial cells to one or more preservation challenges and preparing a product using the population of preserved viability-enhanced microbial cells produced from said methods. The present disclosure further provides products comprising preserved viability-enhanced microbial cells produced by the methods described herein.

Claims (43)

1 . A method of improving microbe viability after preservation comprising:

a. subjecting a population of target microbial cells to a first preservation challenge to provide a population of challenged microbial cells;

b. harvesting viable challenged microbial cells from the population of challenged microbial cells;

c. preserving the viable challenged microbial cells to provide a population of preserved viability-enhanced microbial cells; and

d. preparing a product using the population of preserved viability-enhanced microbial cells.

2 . The method of claim 1 , wherein the first preservation challenge includes one of freeze drying, lyophilization, cryopreservation, preservation by evaporation, preservation by foam formation, vitrification, stabilization by glass formation, preservation by vaporization, spray drying, adsorptive drying, extrusion, or fluid bed drying.

3 . The method of claim 1 , wherein preserving the viable challenged cells includes freeze drying, lyophilization, cryopreservation, preservation by evaporation, preservation by foam formation, vitrification, stabilization by glass formation, preservation by vaporization, spray drying, adsorptive drying, extrusion drying, or fluid bed drying.

4 . The method of claim 1 , further comprising subjecting the population of challenged cells to at least one additional preservation challenge

5 . A method for microbe viability enhancement and preservation, the method comprising:

a. subjecting a population of target microbial cells to a first preservation challenge to provide a first population of challenged microbial cells;

b. harvesting viable challenged microbial cells from the first population of challenged microbial cells to provide a first population of viable challenged microbial cells;

c. subjecting the first population of viable challenged microbial cells to a second preservation challenge to provide a second population of challenged microbial cells;

d. harvesting viable challenged microbial cells from the second population of challenged microbial cells to provide a second population of viable challenged microbial cells;

e. preserving the second population of viable challenged microbial cells to provide a population of preserved viability-enhanced microbial cells; and

f. preparing a product using the population of preserved viability-enhanced microbial cells.

6 . The method of claim 5 , wherein the first preservation challenge and the second preservation challenge are of the same challenge type.

7 . The method of claim 5 , wherein the first preservation challenge and the second preservation challenge are of different challenge types.

8 . The method of claim 5 , wherein the first preservation challenge and the second preservation challenge are selected from a combination described in Table 1.

9 . The method of claim 5 , further comprising subjecting the second population of challenged cells to at least one additional preservation challenge.

10 . The method of claim 5 , wherein preserving the second viable challenged cell population includes freeze drying, lyophilization, cryopreservation, preservation by evaporation, preservation by foam formation, vitrification, stabilization by glass formation, preservation by vaporization, spray drying, adsorptive drying, extrusion drying, or fluid bed drying.

11 . The method of claim 1 , wherein the population of target microbial cells comprises a Clostridium spp. bacterium, a Succinivibrio spp. bacterium, a Butyrivibio spp. bacterium, a Bacillus spp. bacterium, a Lactobacillus spp. bacterium, a Prevotella spp. bacterium, a Syntrophococcus spp. bacterium, a Ruminococcus spp. bacterium, a Caecomyces spp. fungus, a Pichia spp. fungus, an Orpinomyces spp. fungus, a Piromyces spp. fungus, or a species of the Lachnospiraceae family.

12 . (canceled)

13 . (canceled)

14 . The method of claim 11 , wherein:

a. the Clostridium spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 6;

b. the Succinivibrio spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 11;

c. the Pichia spp. comprises an ITS sequence comprising at least 97% sequence identity to SEQ ID NO: 2;

d. the Bacillus spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 4;

e. the Lactobacillus spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9;

f. the Prevotella spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 10; or

g. the species of the Lachnospiraceae family comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 12.

15 . The method of claim 1 , wherein the population of target microbial cells comprises a Ruminococcus bovis bacterium, a Succinivibrio dextrinosolvens bacterium, or a Caecomyces spp. fungus.

16 . The method of claim 1 , wherein the population of target microbial cells comprises a Clostridium butyricum bacterium, a Pichia kudriazevii fungus, a Butyrivibio fibrosolvens bacterium, a Ruminococcus bovis bacterium, or a Succinivibrio dextrinosolvens bacterium.

17 . A product prepared by the methods of claim 1 , comprising a population of preserved viability-enhanced microbial cells.

18 . The product of claim 17 , wherein the population of preserved viability-enhanced microbial cells comprises a Clostridium spp. bacterium, a Succinivibrio spp. bacterium, a Caecomyces spp. bacterium, a Pichia spp. fungus, a Butyrivibio spp. bacterium, an Orpinomyces spp. fungus, a Piromyces spp. fungus, a Bacillus spp. bacterium, a Lactobacillus spp. bacterium, a Prevotella spp. bacterium, a Syntrophococcus spp. bacterium, a Ruminococcus spp bacterium, or a species of the Lachnospiraceae family.

19 . The product of claim 18 , wherein:

a. the Clostridium spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 6;

b. the Succinivibrio spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 11;

c. the Pichia spp. comprises an ITS sequence comprising at least 97% sequence identity to SEQ ID NO: 2;

d. the Bacillus spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 4;

e. the Lactobacillus spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9; or

f. the Prevotella spp. comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 10; or

g. the species of the Lachnospiraceae family comprises a 16S rRNA sequence comprising at least 97% sequence identity to SEQ ID NO: 12.

Assignments (2)
SECURITY INTEREST Recorded Jul 30, 2025
From: NATIVE MICROBIALS, INC.
To: WESTERN ALLIANCE BANK
Reel/Frame 072324/0001 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 27, 2021
From: GILMORE, SEAN; DODGE, COREY
To: NATIVE MICROBIALS, INC.
Reel/Frame 057313/0791 →