CN118076232A

CN118076232A - Compositions and methods for producing fermented plant-based compositions with creamy flavor

Info

Publication number: CN118076232A
Application number: CN202280050888.2A
Authority: CN
Inventors: E·马努里
Original assignee: International Nutrition And Health Denmark Private Ltd
Current assignee: International Nutrition And Health Denmark Private Ltd
Priority date: 2021-07-29
Filing date: 2022-07-28
Publication date: 2024-05-24

Abstract

Provided herein are compositions and methods for producing fermented plant-based compositions, such as fermented milk alternative foods, having a creamy flavor. The compositions and methods provided herein include bacteria, such as lactobacillus rhamnosus, that are useful for producing an acetaldehyde content to diacetyl content ratio that produces a creamy flavor in a fermented plant-based composition. In some embodiments, a bacterium, such as lactobacillus rhamnosus, reduces the acetaldehyde content and/or increases the diacetyl content in the fermented plant-based composition. Provided are fermented plant-based compositions, including stored fermented plant-based compositions, produced according to the methods described herein. Methods for identifying bacteria useful for producing fermented plant-based compositions having a creamy flavor are also provided.

Description

Compositions and methods for producing fermented plant-based compositions with creamy flavor

Technical Field

Provided herein are compositions and methods for producing fermented plant-based compositions (e.g., fermented milk alternative foods) having a creamy flavor. The compositions and methods provided herein include bacteria (e.g., lactobacillus rhamnosus (Lacticaseibacillus rhamnosus)) useful for producing an acetaldehyde content to diacetyl content ratio that results in a creamy flavor in a fermented plant-based composition. In some embodiments, the bacteria (e.g., lactobacillus rhamnosus) reduce the acetaldehyde content and/or increase the diacetyl content in the fermented plant-based composition. Provided are fermented plant-based compositions, including stored fermented plant-based compositions, produced according to the methods described herein. Methods for identifying bacteria useful for producing fermented plant-based compositions having a creamy flavor are also provided.

Background

Flavor perception is a complex sensory phenomenon that contributes to consumer acceptance and desirability of food. In fermented plant-based milk alternative products (such as plant-based yoghurt alternatives), it is preferred to achieve organoleptic properties similar to those of the fermented milk product and to minimize the plant-based notes that are often identified as off-flavours. In particular, cream flavor can be an important flavor feature for consumer acceptance of fermented plant-based milk alternatives.

Achieving milk-like organoleptic properties of fermented plant-based milk alternatives remains a challenge in the industry, and as consumer interest in plant-based diets and natural ingredients continues to grow, consumer demand has created an urgent need for identification solutions that do not rely on manual means.

Accordingly, there is a need for compositions and methods for naturally generating and improving cream flavor in fermented plant-based compositions (e.g., fermented plant-based milk replacers). The compositions and methods provided herein address such a need.

Disclosure of Invention

In aspects, methods for producing a fermented plant-based composition having a creamy flavor are provided, the methods comprising reducing the acetaldehyde content in the fermented plant-based composition. Also provided are methods for producing a stored fermented plant-based composition having a creamy flavor, the methods comprising: (a) Storing the fermented plant-based composition to produce a stored fermented plant-based composition; and (b) reducing the acetaldehyde content in the fermented plant-based composition during storage. In some embodiments, the methods comprise producing a fermented plant-based composition comprising fermenting a plant-based substrate inoculated with a starter culture to produce a fermented plant-based composition.

In various aspects, methods for producing a fermented plant-based composition having a creamy flavor are provided, the methods comprising: (a) Fermenting the plant base inoculated with the starter culture to produce a fermented plant-based composition; and (b) reducing the acetaldehyde content in the fermented plant-based composition.

In some embodiments, the methods provided herein further comprise increasing the diacetyl content in the fermented plant-based composition. In some embodiments, the vegetable base comprises soybean plant matter. In some embodiments, the soybean plant matter is contained in an aqueous suspension. In some embodiments, the methods comprise storing the fermented plant-based composition to produce a stored fermented plant-based composition. In some embodiments, the acetaldehyde content in the fermented plant-based composition is reduced during storage. In some embodiments, the methods comprise increasing the diacetyl content of the fermented plant-based composition during storage. In some embodiments, the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

In some embodiments, the starter culture contains one or more lactic acid bacteria. In some embodiments, the starter culture contains at least two lactic acid bacteria. In some embodiments, the starter culture contains bacteria from the following genera: lactococcus (Lactobacillus), lactobacillus (Streptococcus), lactobacillus (Lacticaseibacillus), leuconostoc (Leuconostoc), pediococcus (Pediococcus), or Bifidobacterium), or any combination of the foregoing. In some embodiments, the starter culture contains one or more strains of lactobacillus rhamnosus. In some embodiments, one or more of the lactobacillus rhamnosus strains are strains of vegetable origin. In some embodiments, the one or more lactobacillus rhamnosus strains are or comprise: (a) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM33650 or a mutant thereof; (b) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM22876 or a mutant thereof; or (c) the strain of Lactobacillus rhamnosus deposited with the DSMZ under accession number DSM33850 or a mutant thereof.

In some embodiments, the starter culture comprises one or more strains of streptococcus thermophilus (Streptococcus thermophilus). In some embodiments, the one or more streptococcus thermophilus strains is or includes a streptococcus thermophilus strain deposited at the DSMZ under accession No. DSM33651 or a mutant thereof.

In some embodiments, the starter culture is or comprises: (a) Lactobacillus rhamnosus strain or a mutant thereof deposited at the DSMZ under accession No. DSM33650, and streptococcus thermophilus strain or a mutant thereof deposited at the DSMZ under accession No. DSM 33651; (b) Lactobacillus rhamnosus strain or a mutant thereof deposited at the DSMZ under accession No. DSM22876, and streptococcus thermophilus strain or a mutant thereof deposited at the DSMZ under accession No. DSM 33651; or (c) the strain of Lactobacillus rhamnosus or a mutant thereof deposited with DSMZ under accession number DSM33850, and the strain of Streptococcus thermophilus or a mutant thereof deposited with DSMZ under accession number DSM 33651.

In some embodiments, the starter culture contains one or more of the following: lactobacillus delbrueckii subsp bulgaricus (Lactobacillus delbrueckii subsp bulgaricus) strain, lactobacillus acidophilus (Lactobacillus acidophilus) strain, bifidobacterium lactis (Bifidobacterium lactis) strain, lactobacillus mucilaginosus (Limosilactobacillus fermentum) strain, lactobacillus paracasei (Lacticaseibacillus paracasei) strain, lactobacillus plantarum (Lactiplantibacillus plantarum) strain, propionibacterium freudenreichii (Propionibacteriafreudenreichii) strain, pediococcus acidilactici (Pediococcus acidilactici) strain, lactobacillus lactici subsp lactici (Lactococcus lactis subsp lactis) strain, or Lactobacillus milk subsp lactis (Lactococcus cremoris subsp cremoris) strain.

In some embodiments, the inoculated plant substrate is fermented at a temperature above about 33 ℃, 35 ℃, 36 ℃,37 ℃, 38 ℃, 39 ℃, 40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, or 50 ℃. In some embodiments, the fermented plant-based composition is stored at about 1 ℃,2 ℃,3 ℃,4 ℃,5 ℃,6 ℃, 7 ℃,8 ℃,9 ℃, 10 ℃,11 ℃,12 ℃, 13 ℃,14 ℃, 15 ℃,16 ℃,17 ℃, 18 ℃, 19 ℃,20 ℃,21 ℃,22 ℃, 23 ℃, 24 ℃,25 ℃, or about 26 ℃. In some embodiments, the fermented plant-based composition is stored: a) At least about 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days; or b) up to about 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days. In some embodiments, the fermented plant-based composition is stored at about 6 ℃ for about 28 days. In some embodiments, the fermented plant-based composition is stored at about 6 ℃ for about 14 days. In some embodiments, the fermented plant-based composition is stored at about 6 ℃ for about 7 days.

In some embodiments, the cream flavor includes a cream aroma and/or a cream scent. In some embodiments, the cream flavor is an increased cream flavor as compared to the cream flavor of a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of equal to or greater than 0.7. In some embodiments, the increased cream flavor comprises an increased cream aroma and/or cream odor as compared to a cream aroma and/or cream odor of a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio equal to or greater than 0.7.

In various aspects, compositions are provided that include a lactobacillus rhamnosus strain, wherein the lactobacillus rhamnosus strain reduces acetaldehyde in a fermented plant-based composition to produce a creamy flavor. In some embodiments, the lactobacillus rhamnosus strain is a strain of vegetable origin. In some embodiments, the lactobacillus rhamnosus strain is: (a) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM33650 or a mutant thereof; (b) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM22876 or a mutant thereof; or (c) the strain of Lactobacillus rhamnosus deposited with the DSMZ under accession number DSM33850 or a mutant thereof. In some embodiments, the composition further comprises a streptococcus thermophilus strain. In some embodiments, the streptococcus thermophilus strain is the streptococcus thermophilus strain deposited at the DSMZ under accession No. DSM33651 or a mutant thereof. In some embodiments, the composition is or comprises: (a) Lactobacillus rhamnosus strain or a mutant thereof deposited at the DSMZ under accession No. DSM33650, and streptococcus thermophilus strain or a mutant thereof deposited at the DSMZ under accession No. DSM 33651; (b) Lactobacillus rhamnosus strain or a mutant thereof deposited at the DSMZ under accession No. DSM22876, and streptococcus thermophilus strain or a mutant thereof deposited at the DSMZ under accession No. DSM 33651; or (c) the strain of Lactobacillus rhamnosus or a mutant thereof deposited with DSMZ under accession number DSM33850, and the strain of Streptococcus thermophilus or a mutant thereof deposited with DSMZ under accession number DSM 33651. In some embodiments, the composition further comprises one or more of the following: lactobacillus delbrueckii subsp bulgaricus strain, lactobacillus acidophilus strain, bifidobacterium lactis strain, lactobacillus mucilaginosus strain, lactobacillus paracasei strain, lactobacillus plantarum strain, propionibacterium freudenreichii strain, pediococcus acidilactici strain, lactococcus lactici subsp lactici strain, or lactococcus cremoris subsp. In some embodiments, the composition is a starter culture. In some embodiments, the composition is a fermented plant-based food.

In various aspects, there is provided the use of lactobacillus rhamnosus to reduce the acetaldehyde content in a fermented plant-based composition to produce a creamy flavour. In some embodiments, the lactobacillus rhamnosus increases the diacetyl content in the fermented plant-based composition. In some embodiments, the lactobacillus rhamnosus strain is: (a) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM33650 or a mutant thereof; (b) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM22876 or a mutant thereof; or (c) the strain of Lactobacillus rhamnosus deposited with the DSMZ under accession number DSM33850 or a mutant thereof. In some embodiments, the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0. In some embodiments, the cream flavor includes a cream aroma and/or a cream scent. In some embodiments, the cream flavor is an increased cream flavor as compared to the cream flavor of a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of equal to or greater than 0.7. In some embodiments, the increased cream flavor comprises an increased cream aroma and/or cream odor as compared to a cream aroma and/or cream odor of a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio equal to or greater than 0.7.

In various aspects, provided are fermented plant-based compositions obtained according to the methods or uses provided herein. In some embodiments, the fermented plant-based composition is a fermented plant-based food. In some embodiments, the fermented plant-based composition is a fermented plant-based milk-substitute food product. In some embodiments, wherein the fermented plant-based composition is a plant-based yogurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese (from milk beverages) alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir (kefir) alternative.

In various aspects, a fermented plant-based food product having a creamy flavor is provided that contains an acetaldehyde content to diacetyl content ratio of less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0. In some embodiments, the fermented plant-based food product contains a strain of lactobacillus rhamnosus that reduces acetaldehyde in the fermented plant-based food product. In some embodiments, the lactobacillus rhamnosus strain increases diacetyl in the fermented plant-based food product.

In some embodiments, the fermented plant-based food product contains: (a) The lactobacillus rhamnosus strain deposited under accession number DSM33650 or a mutant thereof; (b) The lactobacillus rhamnosus strain deposited under accession number DSM22876 or a mutant thereof; and/or (c) the lactobacillus rhamnosus strain deposited under accession number DSM33850 or a mutant thereof. In some embodiments, the fermented plant-based food product further comprises a streptococcus thermophilus strain. In some embodiments, the streptococcus thermophilus strain is the streptococcus thermophilus strain deposited under accession number DSM33651 or a mutant thereof. In some embodiments, the fermented plant-based food product is a fermented plant-based milk-alternative food product. In some embodiments, the fermented plant-based food product is a plant-based yogurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative.

In various aspects, a method of identifying bacteria capable of producing a fermented plant-based composition having a creamy flavor is provided, the method comprising quantifying the acetaldehyde content and the diacetyl content in a fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the fermented plant-based composition is stored and the acetaldehyde content and diacetyl content are quantified during storage. In some embodiments, one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the acetaldehyde content and diacetyl content are quantified such that the ratio of acetaldehyde content to diacetyl content is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0. In some embodiments, the one or more bacteria include a strain of lactobacillus rhamnosus. In some embodiments, the lactobacillus rhamnosus is of vegetable origin. In some embodiments, the one or more bacteria include a streptococcus thermophilus strain.

In some embodiments, the ratios of acetaldehyde to diacetyl described herein in relation to cream flavor are achieved after 28 days or less of storage. In some embodiments, the ratios of acetaldehyde to diacetyl described herein in relation to cream flavor are achieved after 21 days or less of storage. In some embodiments, the ratios of acetaldehyde to diacetyl described herein in relation to cream flavor are achieved after 14 days or less of storage. In some embodiments, the ratios of acetaldehyde to diacetyl described herein in relation to cream flavor are achieved after 7 days or less of storage.

In various aspects, a composition is provided that comprises one or more bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor according to the methods provided herein. In some embodiments, the composition is a starter culture or a fermented plant-based food. In some embodiments, the fermented plant-based food product is a fermented plant-based milk-alternative food product.

Kits comprising the compositions described herein and instructions for use are also provided herein.

In various aspects, a lactobacillus rhamnosus strain deposited under accession number DSM33850 or a mutant thereof is provided. In various aspects, a lactobacillus rhamnosus strain deposited under accession number DSM22876 or a mutant thereof is provided. In various aspects, a streptococcus thermophilus strain deposited at the DSMZ under accession No. DSM33651, or a mutant thereof, is provided.

Each of the aspects and embodiments described herein can be used together unless expressly or clearly excluded from the context of the embodiments or aspects.

Drawings

Figure 1 shows a scatter plot of 2, 3-butanedione (diacetyl) and acetaldehyde content (in ppm) of five soy-based yoghurt substitutes produced by fermentation with the exemplary streptococcus thermophilus strain (DSM 33651) alone or in combination with the exemplary lactobacillus rhamnosus strains shown (DSM 24616, DSM33650, DSM22876 or DSM 33850) after storage for 1, 7, 14, 21 and 28 days at 6 ℃. The 2, 3-butanedione and acetaldehyde content (in ppm) of the soy substrate prior to fermentation is also shown.

Figure 2 shows a scatter plot of the 2, 3-butanedione (diacetyl) and acetaldehyde content (in ppm) of soy-based yoghurt substitutes produced by fermentation with one of two industrial starter cultures (industrial starter 1 or industrial starter 2) alone or in combination with an exemplary lactobacillus rhamnosus strain DSM33650 after storage for 7 days at 6 ℃.

Figure 3 shows the acetaldehyde and 2, 3-butanedione content (in ppm, axis 1) and sensory cream/milk odor assessment (axis 2) of the soy yoghurt alternative. 1: yoghurt substitutes fermented with a separate industrial starter culture. 2: yogurt alternative fermented with a combination of an industrial starter culture and a lactobacillus rhamnosus strain DSM33650

Detailed Description

Provided herein are compositions and methods for producing fermented plant-based compositions (e.g., fermented plant-based foods) having a creamy flavor. As described herein, it has surprisingly been found that a plant based substrate (e.g., soybean) fermented with the bacteria described herein can produce levels of acetaldehyde and diacetyl (also referred to as 2, 3-butanedione) that result in an increased creamy flavor in a fermented plant-based composition.

While it has been recognized that diacetyl provides a characteristic flavor of butter and can contribute to the perception of cream flavor, it has not previously been considered that a diacetyl content greater than acetaldehyde content will be related to (e.g., correlated with) the cream flavor in a fermented plant-based food, nor that such cream flavor will increase as compared to the cream flavor in a fermented plant-based food having similar levels of acetaldehyde and diacetyl or having an acetaldehyde content greater than diacetyl content. Thus, the methods and compositions provided herein take advantage of the surprising discovery described herein (see, e.g., examples) that fermented plant-based compositions having a level of diacetyl that is greater than acetaldehyde have an increased cream flavor as compared to the cream flavor in fermented plant-based compositions having levels of acetaldehyde and diacetyl that are about the same, or having an acetaldehyde content that is greater than the diacetyl content.

The use of the bacterial strains described herein for achieving the diacetyl and acetaldehyde levels described herein in connection with cream flavor provides a natural solution to increase cream flavor in fermented plant-based compositions. The demonstrated ability of the compositions and methods provided herein to increase cream flavor without the use of additives or artificial flavors may be desirable to certain consumers.

In some aspects, the methods and compositions described herein allow the relative amounts (e.g., ratios) of acetaldehyde to diacetyl associated with creamer flavor in fermented plant-based compositions to be achieved by adjusting the levels of acetaldehyde and diacetyl in the fermented plant-based compositions. In some embodiments, the methods and compositions described herein help reduce the level of acetaldehyde and/or increase the level of diacetyl in a fermented plant-based composition to achieve a relative content (e.g., ratio) of acetaldehyde to diacetyl associated with a creamer flavor. In some embodiments, the cream flavor produced by the methods and compositions provided herein is increased as compared to methods and compositions for producing a cream flavor in a fermented plant-based composition that does not reach the relative content (e.g., ratio) of acetaldehyde to diacetyl as described herein.

In some embodiments, the methods provided herein include using a bacterium or bacterial composition (e.g., starter culture) that contains lactobacillus rhamnosus. In some embodiments, the lactobacillus rhamnosus modulates (e.g., reduces) the acetaldehyde content in the fermented plant-based composition. In some embodiments, the lactobacillus rhamnosus independently modulates (e.g., independently increases or decreases) the acetaldehyde and diacetyl content in the fermented plant-based composition. In some embodiments, the lactobacillus rhamnosus reduces the acetaldehyde content in the fermented plant-based composition. In some embodiments, the lactobacillus rhamnosus increases the diacetyl content in the fermented plant-based composition. In some embodiments, the lactobacillus rhamnosus reduces the acetaldehyde content and increases the diacetyl content in the fermented plant-based composition. In some embodiments, the bacterial composition (e.g., starter culture) further comprises streptococcus thermophilus.

In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein produce a diacetyl content greater than an acetaldehyde content in the fermented plant-based composition. In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein produce increased diacetyl content as compared to acetaldehyde content, the relative amounts (e.g., ratios) of acetaldehyde to diacetyl being related to cream flavor (e.g., increased cream flavor). In some cases, achieving the relative content (e.g., ratio) of acetaldehyde to diacetyl associated with a cream flavor (e.g., increased cream flavor) is accomplished by fermenting a plant substrate using bacteria or bacterial compositions (e.g., starter cultures) that modulate (e.g., increase, decrease) the levels of acetaldehyde and/or diacetyl as described herein. In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein reduce acetaldehyde levels in a fermented plant-based composition. In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein increase the level of diacetyl in a fermented plant-based composition. In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein increase the level of diacetyl or decrease the level of acetaldehyde in a fermented plant-based composition. In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein increase the level of diacetyl and decrease the level of acetaldehyde in a fermented plant-based composition.

In some embodiments, the acetaldehyde to diacetyl content (e.g., relative content) is expressed as a ratio of acetaldehyde content to diacetyl content (e.g., as a quotient). In some embodiments, the bacteria and bacterial compositions (e.g., starter cultures) described herein independently regulate (e.g., decrease or increase) the levels of acetaldehyde and diacetyl such that the ratio of acetaldehyde to diacetyl is less than 1, e.g., less than about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.075, 0.05, 0.01, or 0. In some cases, the content (e.g., relative content) of acetaldehyde and diacetyl is expressed as a percentage. For example, in some cases, the bacteria and bacterial compositions (e.g., starter cultures) described herein independently regulate (e.g., increase or decrease) the levels of acetaldehyde and diacetyl such that the percentage of acetaldehyde to diacetyl is less than 100%, e.g., less than about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 1O, 7.5%, 5%, 1%, or 0%. In some cases, the content (e.g., relative content) of acetaldehyde and diacetyl is expressed as a multiple, e.g., 2-fold, 3-fold. Any means suitable for digitally representing the relationship between the acetaldehyde content and the diacetyl content may be used.

In some cases, the creamer flavor in the fermented plant-based compositions produced according to the methods and compositions provided herein is not present in these fermented plant-based compositions immediately after fermentation, e.g., according to sensory analysis. For example, in some cases, the acetaldehyde to diacetyl content (e.g., acetaldehyde to diacetyl ratio of less than 1) associated with the creamer flavor (e.g., increased creamer flavor) in the fermented plant-based composition occurs over time (e.g., a certain length of time after fermentation has ceased). As such, in some embodiments, a creamy flavor in the fermented plant-based composition occurs over time due to variations in acetaldehyde content and/or diacetyl content release. Thus, in some embodiments, methods provided herein include storing a fermented plant-based composition, e.g., as described in sections I-D below. In some embodiments, the fermented plant-based composition is stored under refrigeration, e.g., as described in sections I-D below. In some embodiments, the level of acetaldehyde decreases over time of storage. In some embodiments, the level of diacetyl increases over time. In some embodiments, the level of acetaldehyde decreases or the level of diacetyl increases over time. In some embodiments, the level of acetaldehyde decreases and the level of diacetyl increases over time.

In some embodiments, the creamy flavor in the fermented plant-based composition develops during storage. In some embodiments, the creamy flavor in the fermented plant-based composition increases during storage. For example, in some cases, the sensory characteristics indicative of the cream flavor change such that an increase in cream aroma and/or cream smell occurs with additional storage time. In some embodiments, the creamy flavor in the fermented plant-based composition develops during storage and then reaches a stable creamy flavor level. For example, in some cases, the sensory properties indicative of the cream flavor remain stable, e.g., no perceptible (e.g., statistically significant, organoleptically detectable) increase or decrease in cream aroma and/or cream odor occurs over additional storage time. Methods for generating and evaluating organoleptic properties include those described in sections I-E, e.g., as described in example 3.

In some embodiments, the fermented plant-based composition having a creamy flavor is a fermented plant-based composition that has been stored for a duration of time. In some embodiments, the fermented plant-based composition having a creamy flavor is a stored fermented plant-based composition. In some embodiments, the storage duration of the fermented plant-based composition is at or about the length of time required for the acetaldehyde to decrease, and optionally for the diacetyl level to increase, to a level (e.g., relative amount, ratio) associated with the creamer flavor in the fermented plant-based composition (e.g., stored fermented plant-based composition).

In some embodiments, the creamer flavor in a fermented plant-based composition produced according to the methods and compositions provided herein is related (e.g., correlated) to a particular level of diacetyl and/or acetaldehyde in the fermented plant-based composition, and/or relative amounts and ratios thereof. See, for example, sections I-E below. For example, in some embodiments, the creamy flavor is associated with a level of diacetyl that is greater than the level of acetaldehyde. In some embodiments, the cream flavor is related to (e.g., correlated to) the acetaldehyde content level and the diacetyl level in the fermented plant-based composition, which results in a ratio of acetaldehyde to diacetyl of less than 1, such as less than about 0.9, 0.85, 0.8, 0.75, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.075, 0.05, 0.025, or 0.01, or 0. In some embodiments, the level of diacetyl in the fermented plant-based composition is at least 1ppm, e.g., at least about 1.5ppm, 2ppm, 3ppm, 4ppm, 5ppm, 10ppm, 15ppm, or 20ppm. In some embodiments, the acetaldehyde level in the fermented plant-based composition is less than 1ppm, e.g., less than about 0.9ppm, 0.8ppm, 0.7ppm, 0.6ppm, 0.5ppm, 0.4ppm, 0.3ppm, 0.2ppm, 0.1ppm, 0.05ppm, 0.01ppm, or below the lower instrumentation and/or quantification limit. In some embodiments, the lower limit of quantitation is 0.125ppm. In some embodiments, the level of diacetyl in the fermented plant-based composition is at least 1ppm, e.g., at least about 1.5ppm, 2ppm, 3ppm, 4ppm, 5ppm, 10ppm, 5ppm, or 20ppm, and the level of acetaldehyde is such that the fermented plant-based composition has an acetaldehyde to diacetyl ratio of less than 1, e.g., less than about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.075, 0.05, 0.025, or 0.01, or is 0 (e.g., when the level of acetaldehyde is below a quantitative or detection level). In some embodiments, only the level of acetaldehyde is reduced to produce a ratio of acetaldehyde to diacetyl related to creamy flavor. In some embodiments, the fermented plant-based composition has an increased creamy flavor as compared to a fermented plant-based composition having an acetaldehyde to diacetyl ratio of 1, about 1, or greater than 1. The terms level, content and concentration are used interchangeably herein to describe the amount of acetaldehyde, diacetyl, or other volatile organic compounds.

In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is a creamy aroma. The creamy aroma is perceived via the post-nasal pathway (e.g., once the food product (e.g., fermented plant-based composition) is in the intake port). In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is a creamy smell. Cream odors are perceived (smelled) through the nose using a positive nasal passage. In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is a creamy aroma or creamy smell. In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is creamy aroma and creamy smell.

In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein is an increased creamy flavor. For example, the creamy flavor of a fermented plant-based composition produced according to the methods and compositions provided herein may be increased as compared to a fermented plant-based composition that does not have the acetaldehyde and diacetyl levels, or relative amounts (e.g., ratios) thereof, associated with the creamy flavor according to the compositions and methods provided herein, or that does not achieve the acetaldehyde and diacetyl levels, or relative amounts (e.g., ratios) thereof, associated with the creamy flavor according to the compositions and methods provided herein. Thus, in some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is an increased creamy aroma. In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is an increased creamy smell. In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is an increased creamy aroma or an increased creamy smell. In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods and compositions provided herein includes or is increased creamy aroma and increased creamy smell. In some embodiments, the increase in cream flavor (e.g., increased cream aroma and/or increased cream odor) is determined as compared to a fermented plant-based composition that does not have, or produces, or does not achieve, the relative levels of acetaldehyde and diacetyl (e.g., ratios) associated with the cream flavor according to the compositions and methods provided herein.

In some embodiments, the cream flavor (e.g., increased cream flavor), including its cream aroma and/or cream smell, is determined by a trained sensory evaluator panel.

Also provided herein are fermented plant-based compositions that are food products (e.g., fermented plant-based food products) produced according to the methods provided herein (including embodiments thereof). In some embodiments, the fermented plant-based composition (e.g., a fermented plant-based food) is a fermented milk-alternative food or feed product. In some embodiments, the fermented plant-based composition (e.g., fermented plant-based food) is a plant-based yogurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative. In some embodiments, the term "alternative" is used herein to indicate that the food product is similar to a dairy food product, but is not produced using a dairy substrate (e.g., dairy milk). In some embodiments, the fermented plant-based composition is a stored fermented plant-based composition (e.g., a stored fermented plant-based food).

Also provided herein are methods of identifying bacteria capable of producing a fermented plant-based composition with a creamy flavor, optionally stored fermented plant-based composition. In some embodiments, the identification method comprises quantifying the acetaldehyde and diacetyl content, and optionally the variation thereof, in a fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the identification method comprises determining the relative content (e.g., ratio) of acetaldehyde to diacetyl in a fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the identification method comprises quantifying the acetaldehyde content and the diacetyl content, and optionally the changes thereof, in a fermented plant-based composition fermented in the presence of one or more bacteria and stored after fermentation. For example, quantifying the acetaldehyde content and the diacetyl content, and optionally the changes thereof, occurs during storage of the fermented plant-based composition. In some embodiments, the identification method comprises determining the relative content (e.g., ratio) of acetaldehyde to diacetyl in a fermented plant-based composition fermented in the presence of one or more bacteria and stored after fermentation. For example, determining the relative amounts (e.g., ratios) of acetaldehyde to diacetyl occurs during storage of the fermented plant-based composition. In some embodiments, the identification method comprises quantifying the acetaldehyde content and the diacetyl content, and optionally the variation thereof, in a stored fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the identification method comprises determining the relative content (e.g., ratio) of acetaldehyde to diacetyl in a stored fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, quantifying the amount and/or determining the relative amount (e.g., ratio) as described herein is performed at different points in time over the duration of storage. In some embodiments, quantifying the amount and/or determining the relative amount (e.g., ratio) as described herein is performed continuously for the duration of the storage. In some embodiments, the storing is performed as described in sections I-D below. In some embodiments, one or more bacteria useful for achieving diacetyl and/or acetaldehyde levels, as well as relative amounts and ratios thereof, associated with creamy flavor in a fermented plant-based composition as described herein (see, e.g., sections I-E) are identified as being capable of producing a fermented plant-based composition (optionally, a stored fermented plant-based composition) having creamy flavor. In some embodiments, the composition (e.g., starter culture) contains one or more identified bacteria.

The compositions and methods provided herein can be used to naturally produce fermented plant-based compositions (e.g., fermented plant-based foods) having a creamy flavor (e.g., increased creamy flavor) without the need for potentially undesirable and/or expensive additives. By using bacteria and bacterial compositions for fermentation as described herein, the methods provided herein do not require additional processing steps (e.g., introducing additives into the composition) such that the fermented plant-based composition is produced without additional time delay. The methods and compositions provided herein for producing fermented plant-based compositions having a creamy flavor can help to increase consumption of plant-based products that can promote nutritional and health benefits and can have a positive impact on the environment, such as when consumed as an alternative to dairy products. Thus, the methods and compositions described herein provide numerous advantages while also satisfying consumer cravings for fermented plant-based foods with naturally increased cream flavor.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

The present disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the embodiments of the present disclosure.

The headings provided herein are not limitations of the various aspects or embodiments of the disclosure which can be had by reference to the specification as a whole. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Any terminology that is defined is more fully defined when the description is given herein as a whole.

Definitions of terms may appear throughout this specification. It is to be understood that this disclosure is not limited to the particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

It must be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "a" includes "at least one" or "one or more".

As used herein, the terms "comprises" (comprising, comprises) and "consisting of" … … are synonymous with "comprising" (including, includes) or "containing" (containing, contains), and grammatical variants thereof, are inclusive or open-ended and do not exclude additional, unrecited members, elements or method steps. The terms "comprising" (comprising, comprises) and "consisting of (… …)", "including" (including, includes) or "containing (containing, contains)" and grammatical variants thereof also include the term "consisting of … … (consisting of)".

Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range is also specifically disclosed. Every smaller range between any stated or intermediate value in the stated range and any other stated or intermediate value in the stated range is encompassed within this disclosure. The upper and lower limits of these smaller ranges may independently be included in the range or excluded from the range, and each range (either, neither, or both limits are included in the smaller ranges) is also encompassed within the present disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

Values and ranges may be expressed herein as the numerical value preceded by the term "about". The term "about" is used herein to provide literal support for the exact number following it and numbers near or approximating the number following the term. In determining whether a number is close or approximate to a particular recited number, the close or approximate non-recited number may be a number that provides a substantial equivalent of the particular recited number in the context in which it is presented. For example, with respect to a numerical value, the term "about" refers to a range of-10% to +10% of the numerical value, unless the term is specifically defined in the context. All values and ranges can implicitly include the term "about" unless the context dictates otherwise.

All publications (including patent documents, scientific articles, and databases) mentioned in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication was individually incorporated by reference. Nothing herein is to be construed as an admission that such publication constitutes prior art. If the definition set forth herein is contrary to or otherwise inconsistent with the definition set forth in the patents, applications, published applications and other publications, which are incorporated by reference, the definition set forth herein takes precedence over the definition set forth herein by reference.

I. method for producing fermented plant-based compositions with creamy flavor

The methods and compositions provided herein allow for the production of a fermented plant-based composition having a creamy flavor, wherein the creamy flavor can be increased compared to a fermented plant-based composition produced according to a different method. In some embodiments, the creamy flavor of the fermented plant-based composition produced according to the methods provided herein is increased compared to the creamy flavor of the fermented plant-based composition produced according to other methods, such as not controlling (e.g., adjusting) the relative amounts of acetaldehyde and diacetyl in the fermented plant-based composition to levels related (e.g., correlated) to creamy flavor as described herein (see, e.g., sections I-E).

In some aspects, the methods and compositions provided herein achieve a fermented plant-based composition having a relative amount of acetaldehyde and diacetyl that is related (e.g., correlated) to increased cream flavor. In some embodiments, the increased cream flavor has increased cream aroma. In some embodiments, the increased cream flavor has an increased cream odor. In some embodiments, the increased cream flavor has an increased cream aroma or an increased cream odor. In some embodiments, the increased cream flavor has an increased cream aroma and an increased cream odor.

In some aspects, a method for producing a fermented plant-based composition having a creamy flavor is provided, the method comprising reducing the acetaldehyde content in the fermented plant-based composition. In some aspects, a method for producing a fermented plant-based composition having a creamy flavor is provided, the method comprising reducing the acetaldehyde content in the fermented plant-based composition and increasing the diacetyl content therein.

In some aspects, a method for producing a fermented plant-based composition having a creamy flavor is provided, the method comprising fermenting a plant base inoculated with a starter culture to produce a fermented plant-based composition, and reducing the acetaldehyde content in the fermented plant-based composition. In some aspects, a method for producing a fermented plant-based composition having a creamy flavor is provided, the method comprising fermenting a plant base inoculated with a starter culture to produce a fermented plant-based composition, and reducing the acetaldehyde content and increasing the diacetyl content in the fermented plant-based composition.

In some aspects, a method for producing a stored fermented plant-based composition having a creamy flavor is provided, the method comprising storing a fermented plant-based composition to produce a stored fermented plant-based composition, and reducing the acetaldehyde content in the fermented plant-based composition during storage. In some aspects, a method for producing a stored fermented plant-based composition having a creamy flavor is provided, the method comprising storing the fermented plant-based composition to produce a stored fermented plant-based composition, and reducing the acetaldehyde content and increasing the diacetyl content in the fermented plant-based composition during storage.

In some aspects, a method of producing a creamy flavor in a fermented plant-based composition during storage of the fermented plant-based composition is provided, the method comprising reducing the acetaldehyde content in the fermented plant-based composition during storage. In some aspects, a method of producing a creamy flavor in a fermented plant-based composition during storage of the fermented plant-based composition is provided, the method comprising reducing the acetaldehyde content and increasing the diacetyl content in the fermented plant-based composition during storage.

In some embodiments, the methods provided herein comprise producing a fermented plant-based composition. In some embodiments, methods for producing a fermented plant-based composition include inoculating a plant-based substrate (e.g., as described in section I-a below) with bacteria or a composition thereof (e.g., as described in section I-B below), and fermenting the inoculated plant-based substrate (e.g., as described in section I-C below).

As described above, in some embodiments, the fermented plant-based composition is stored. The plant-based binders that have been subjected to thermophilic fermentation and subsequently stored may be referred to herein as stored fermented plant-based compositions. In some embodiments, the storing is performed under refrigeration. In some embodiments, the storage of the fermented plant-based composition is performed according to the methods described in sections I-D to produce a stored fermented plant-based composition.

In some embodiments, the relative amounts of acetaldehyde and diacetyl in the fermented plant-based composition that are related (e.g., correlated) to the cream flavor (e.g., increased cream flavor) are achieved during storage of the fermented plant-based composition. In some embodiments, the levels, relative amounts, and ratios of acetaldehyde and diacetyl associated with cream flavor are described in sections I-E.

The methods and compositions provided herein can be used to produce a fermented plant-based composition that is a fermented plant-based food having a creamy flavor (e.g., increased creamy flavor). Exemplary fermented plant-based compositions as food products are described in section I-F.

A. vegetable base material

In some embodiments, the fermented plant-based composition having a creamy flavor is produced by, for example, inoculating with bacteria or a bacterial composition as described herein, and fermenting the plant-based base. The plant-based substrate contemplated herein for use in producing a fermented plant-based composition may be any raw and/or processed plant-based material (e.g., plant matter) capable of being fermented according to the methods provided herein.

In some embodiments, the plant based binder is or includes a completely or partially hydrolyzed plant material (e.g., cereal). In some embodiments, the cereal is wheat, rye, spelt, barley, oats, millet, sorghum, rice, bran, or a combination thereof.

In some embodiments, the plant based binder is or includes plant matter from leguminous plants. In some embodiments, the vegetable base is or includes beans, such as split peas, purple peas, dried peas, lentils, chickpeas (chickpea, garbanzo beans), konda, kidney beans, white peas, cowpeas, broad beans, lentils, spot beans, red beans, mexican red beans, kidney beans, black tortoise beans, cranberry beans, luo Madou, spot sugar beans, lima beans, pigtails, cotton beans, mung beans (GREEN GRAM, mung bean), kidney beans, black gei beans, indian black beans, soybeans, and/or lupins. In some embodiments, the vegetable base is or includes soybean (e.g., soybean plant matter).

In some embodiments, the vegetable base is or includes vegetable matter from nuts. Non-limiting examples of nuts contemplated for use herein include almonds, cashews, pecans, macadamia nuts, hazelnuts, pistachios, walnuts, or combinations thereof.

In some embodiments, the vegetable base is or includes seeds. Non-limiting examples of seeds contemplated for use herein include pumpkin, quinoa, sesame, tiger nuts, flax, chia seeds (chia), sunflower, coconut, or combinations thereof.

In some embodiments, the vegetable base is milk of vegetable origin. For example, milk is not a milk secretion obtained by milking any mammal (e.g., cattle, sheep, goats, buffalo, zebra, horses, donkeys, camels, etc.). In some embodiments, the vegetable base is or includes vegetable-based milk. In some embodiments, the plant-based milk is or includes milk of any plant matter described herein. For example, in some cases, the plant-based milk is milk of grains, legumes, nuts, seeds, or a combination thereof. In some embodiments, the plant-based milk is nut milk. In some embodiments, the plant-based milk is cereal milk. In some embodiments, the plant-based milk is legume milk. In some embodiments, the plant-based milk is seed milk. In some embodiments, the plant-based milk is soy milk. In some embodiments, the vegetable base is soy milk.

In some embodiments, the vegetable base is a suspension. In some embodiments, the plant-based binder is an aqueous suspension comprising water and plant matter as described herein. Methods of preparing plant matter suspensions suitable for carrying out the fermentation of a plant based material as described herein are known in the art. Exemplary methods for preparing the plant matter include mechanically and/or enzymatically disrupting the plant matter, optionally in combination with a solution or other hydrated form, to produce an aqueous fraction that can be separated (e.g., decanted, centrifuged, and/or filtered) from the remaining plant matter (e.g., starch matter, fiber matter).

In some embodiments, the vegetable base is a butter alternative or beverage, such as a nut, seed, or soy butter alternative or beverage. Methods for preparing butter alternatives for use as described herein are known in the art. Exemplary methods of preparing butter alternatives include wet or dry grinding roasted or unroasted, e.g., nuts, seeds, beans, into a paste. In some embodiments, the particle size of the paste is suitable for use in preparing a beverage.

In some embodiments, the vegetable base does not include added sugar or protein. For example, in some cases, the carbohydrates of the vegetable base are provided solely by the vegetable matter.

In some embodiments, the vegetable base comprises a protein. In some embodiments, the protein is provided solely by plant matter. In some embodiments, the amount of protein is in the range of 1% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the amount of protein is in the range of 2% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the amount of protein is in the range of 3% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the amount of protein is in the range of 4% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the amount of protein is in the range of 1% to 4% (wt.%) of the total plant base or about this range. In some embodiments, the amount of protein is in the range of 1% to 3% (wt.%) of the total plant base or about this range. In some embodiments, the amount of protein is in the range of 1% to 2% (wt.%) of the total plant base or about this range.

In some embodiments, sugar may be added to the vegetable base. In some embodiments, the added sugar is in the range of 0.5% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the added sugar is in the range of 1% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the added sugar is in the range of 2% to 5% (wt.%) of the total plant base or about this range. In some embodiments, the added sugar is in the range of 2% to 4% (wt.%) of the total plant base or about this range. In some embodiments, the added sugar is in the range of 2% to 3% (wt.%) of the total plant base or about this range. In some embodiments, the added sugar is 2% (wt%) or about 2% of the total vegetable base. In some embodiments, the sugar is dextrose or sucrose. In some embodiments, the sugar is dextrose.

In some embodiments, the plant based binder has a low, undetectable, or unquantifiable level of acetaldehyde. In some embodiments, the vegetable base has a low, undetectable, or unquantifiable level of diacetyl. In some embodiments, the vegetable base has low, undetectable or unquantifiable levels of acetaldehyde and diacetyl. Methods for detecting and/or quantifying the levels of volatile organic compounds such as acetaldehyde and diacetyl include, but are not limited to, mass spectrometry such as gas chromatography-mass spectrometry (GC-MS), selective ion flow tube mass spectrometry (SIFT-MS), proton transfer reaction mass spectrometry (PTR-MS), ion-molecule reaction mass spectrometry (IMR-MS). In some embodiments, the low acetaldehyde or diacetyl level is less than 0.5ppm, such as less than 0.4, 0.3, 0.2, or 0.1ppm. In some embodiments, the low acetaldehyde or diacetyl level is from about 0.5 to about 0.1ppm, from about 0.4 to about 0.1ppm, from about 0.3 to about 0.1ppm, or from about 0.2 to about 0.1ppm. In some embodiments, undetectable levels of acetaldehyde and diacetyl refer to amounts of volatile organic compounds below the detection and/or quantification lower limit of the instrument used to detect the compounds. In some embodiments, acetaldehyde and diacetyl are detected and quantified using GC-MS. In some embodiments, the lower limit of quantitation is 0.125ppm. In some embodiments, fermentation of a plant substrate according to the methods and compositions described herein results in an increase in the amount of acetaldehyde and/or diacetyl as compared to the amount of acetaldehyde and/or diacetyl in the plant substrate prior to fermentation.

In some embodiments, the plant substrate is inoculated with a bacterium or bacterial composition (e.g., starter culture) as described in section I-B to facilitate fermentation.

B. Bacteria and bacterial compositions

One or more bacterial strains may be used to inoculate a plant substrate as described in section 1-a to facilitate fermentation. In some embodiments, the bacterial composition (e.g., starter culture) contains one or more bacteria. In some embodiments, the starter culture contains one or more bacterial strains. In some embodiments, the starter culture is a formulation of living bacteria capable of assisting fermentation. In some embodiments, the plant substrate is inoculated with a starter culture. In some embodiments, the plant substrate is inoculated with a starter culture and a protective culture. In some embodiments, the protective cultures are cultures that are capable of reducing or preventing the growth of biological contaminants such as yeast and mold. In some embodiments, the protective cultures may include one or more bacterial strains described herein, such as the lactobacillus rhamnosus strains described herein. The terms inoculating and adding may be used interchangeably to refer to contacting the plant substrate with one or more bacteria (e.g., as contained in a bacterial composition such as a starter culture or a protective culture).

In some embodiments, the plant substrate is inoculated with one or more bacterial strains, respectively. For example, the bacterial strains are not mixed together prior to addition to the plant base. In some embodiments, the bacterial strains are mixed together prior to addition to the plant base. The strain or strain mixture used for inoculation may be referred to as starter culture or protective culture or contained therein, regardless of how the bacterial strain is added to the plant base. In some embodiments, the starter culture is a thermophilic starter culture, and the composition of the starter culture is suitable for use in thermophilic fermentation as described in sections 1-C. In some embodiments, the starter culture comprises a combination of bacteria suitable for mesophilic and/or thermophilic fermentation. In some embodiments, the starter culture comprises mesophilic and thermophilic bacteria, and the composition of the starter culture is suitable for use in mesophilic and/or thermophilic fermentation as described, for example, in sections I-C.

In some embodiments, thermophilic microorganisms (e.g., thermophilic bacteria) refer to microorganisms that preferentially function at temperatures above 37 ℃ (e.g., 37 ℃ to 50 ℃). In some embodiments, mesophilic microorganisms (e.g., mesophilic bacteria) refer to microorganisms that preferentially function at 37 ℃ or at temperatures below 37 ℃ (e.g., 25 ℃ to 37 ℃).

In some embodiments, the starter culture is a pure culture, i.e., comprises or consists of a single bacterial strain. In some embodiments, the starter culture is a mixed culture, i.e., comprising or consisting of at least one bacterial strain of the invention as described herein and at least one other bacterial strain. For example, the starter culture comprises at least 1 or more, and in particular 1,2, 3, 4 or 5 other bacterial strains.

In some embodiments, the starter culture contains one or more lactic acid bacteria. In some embodiments, the starter culture contains at least two lactic acid bacteria. Since it is normal to use a mixed culture as starter culture during fermentation of lactic acid bacteria, in some embodiments, the composition may comprise multiple strains belonging to the same species or belonging to different species. For example, in some cases, the lactic acid bacteria in the starter culture are or comprise a mixture of lactobacillus delbrueckii subsp bulgaricus strains, streptococcus thermophilus strains, and lactobacillus rhamnosus strains.

In some embodiments, the starter culture comprises at least bacteria from the genera streptococcus and lactobacillus. In some embodiments, the starter culture comprises or consists of bacteria from the following genera: lactococcus, lactobacillus, streptococcus, lactobacillus, leuconostoc, pediococcus, enterococcus, bifidobacterium, lactobacillus paracasei (Paralactobacillus), lactobacillus aceti (Acetilactobacillus), lactobacillus in farmland (Agrilactobacillus), lactobacillus amyloliquefaciens (Amylolactobacillus), lactobacillus bee (Apilactobacillus), lactobacillus in a culture medium, Lactobacilli (Bombilactobacillus), associated Lactobacillus (Companilactobacillus), de-Lagerita (Dellaglioa), lactobacillus (Fructilactobacillus), lactobacillus furgus (Furfurilactobacillus), hozapofil (Holzapfelia), lactobacillus (Lactiplantibacillus), lactobacillus stone wall (Lapidilactobacillus), lactobacillus, Lactobacillus (Latilactobacillus), lactobacillus lentus (Lentilactobacillus), lactobacillus helveticus (Levilactobacillus), lactobacillus bifidus (Ligilactobacillus), lactobacillus mucilaginosus (Limosilactobacillus), lactobacillus liquidus (Liquorilactobacillus), lactobacillus putrefaciens (Loigolactobacilus), lactobacillus oligovorus (Paucilactobacillus), Shi Laifu lactobacillus (Schleiferilactobacillus), lactobacillus paracasei (Secundilactobacillus), or any combination thereof. In some embodiments, the starter culture comprises or consists of bacteria from the following genera: lactococcus, lactobacillus, streptococcus, lactobacillus, leuconostoc, pediococcus, enterococcus or Bifidobacterium, or any combination thereof. In some embodiments, the starter culture comprises or consists of bacteria from the following genera: lactococcus, lactobacillus, streptococcus, lactobacillus, leuconostoc, pediococcus, or Bifidobacterium, or any combination thereof. In some embodiments, the starter culture comprises or consists of bacteria from the following genera: lactococcus, lactobacillus, streptococcus, lactobacillus, or any combination thereof. In some embodiments, the starter culture comprises or consists of bacteria from the following genera: lactobacillus, streptococcus, lactobacillus, or any combination thereof. In some embodiments, the starter culture comprises or consists of bacteria from the genera streptococcus and lactobacillus.

In some embodiments, the starter culture comprises at least a streptococcus thermophilus strain and a lactobacillus rhamnosus strain. In some embodiments, the starter culture comprises one or more of the following: streptococcus thermophilus strains, lactobacillus rhamnosus strains, lactobacillus acidophilus strains, bifidobacterium lactis strains, lactobacillus mucilaginosus strains, lactobacillus paracasei strains, lactobacillus plantarum strains, lactobacillus delbrueckii subsp. Bulgaricus strains, propionibacterium freudenreichii strains, pediococcus acidilactici strains, enterococcus faecium (Enterococcus faecium) strains, lactococcus lactici (Lactococcus lactis) strains, lactococcus cremoris (Lactococcus cremoris) strains, or any combination of the foregoing. In some embodiments, the starter culture comprises one or more of the following: streptococcus thermophilus strains, lactobacillus acidophilus strains, lactobacillus rhamnosus strains, bifidobacterium lactis strains, lactobacillus mucilaginosus strains, lactobacillus paracasei strains, lactobacillus plantarum strains, lactobacillus delbrueckii subsp. Bulgaricus strains, propionibacterium freudenreichii strains, pediococcus acidilactici strains, lactococcus lactici strains, lactococcus cremoris strains, or any combination of the foregoing. In some embodiments, the starter culture comprises one or more of the following: streptococcus thermophilus strains, lactobacillus rhamnosus strains, lactobacillus delbrueckii subsp bulgaricus strains, or any combination of the foregoing. In some embodiments, the starter culture comprises a streptococcus thermophilus strain and a lactobacillus rhamnosus strain. In some embodiments, the starter culture comprises more than one species strain.

In some embodiments, the starter culture comprises at least one strain of lactococcus. For example, the starter culture may include one or more strains of the genus lactococcus known in the art, such as a strain of lactococcus cremoris (previously known as lactococcus lactis subsp. Cremoris (Lactococcus lactis subsp. Cremoris)), lactococcus lactis subsp. Holori (Lactococcus lactis subsp hordniae), or lactococcus lactis subsp. In some embodiments, the starter culture comprises lactococcus cremoris subspecies and/or lactococcus lactis subspecies lactis strains.

As described in the examples, the fermented plant-based composition inoculated with starter cultures comprising streptococcus thermophilus and lactobacillus rhamnosus strains has a diacetyl content greater than the acetaldehyde content associated with increased cream flavor. Thus, in some embodiments, the starter culture is or comprises a strain of the species streptococcus thermophilus and lactobacillus rhamnosus.

In some embodiments, the starter culture comprises one or more lactobacillus rhamnosus strains. In some embodiments, the starter culture comprises one or more lactobacillus rhamnosus strains of vegetable origin. For example, in some cases, the lactobacillus rhamnosus strain is derived, selected or isolated from plant material (e.g., fermented plant material). In some embodiments, the starter culture comprises lactobacillus rhamnosus strain DGCC1179 or a mutant strain thereof deposited with the german collection of microorganisms and cell culture (DSMZ) under accession No. DSM 33650. In some embodiments, the mutant strain is obtained by using the deposited strain as a starting material. In some embodiments, the mutant strain is a strain having all of the identifying characteristics of the strain deposited at DSMZ under number DSM 33650. In some embodiments, the starter culture comprises lactobacillus rhamnosus strain DGCC1460 or a mutant strain thereof deposited at the DSMZ under accession No. DSM 22876. In some embodiments, the mutant strain is obtained by using the deposited strain as a starting material. In some embodiments, the mutant strain is a strain having all of the identifying characteristics of the strain deposited at DSMZ under number DSM 22876. In some embodiments, the starter culture comprises lactobacillus rhamnosus strain DGCC13617 or a mutant strain thereof deposited at the DSMZ under accession No. DSM 33850. In some embodiments, the mutant strain is obtained by using the deposited strain as a starting material. In some embodiments, the mutant strain is a strain having all of the identifying characteristics of the strain deposited at the DSMZ under the number DSM 33850.

In some embodiments, the starter culture comprises one or more streptococcus thermophilus strains. In some embodiments, the starter culture comprises streptococcus thermophilus strain DGCC11042, or a mutant strain thereof, deposited at the DSMZ under accession No. DSM 33651. In some embodiments, the mutant strain is obtained by using the deposited strain as a starting material. In some embodiments, the mutant strain is a strain having all of the identifying characteristics of the strain deposited at DSMZ under number DSM 33651.

In some embodiments, the starter culture comprises one or more lactobacillus rhamnosus strains and/or one or more streptococcus thermophilus strains. In some embodiments, the starter culture comprises one or more lactobacillus rhamnosus strains disclosed herein and/or one or more streptococcus thermophilus strains disclosed herein.

In some embodiments, the starter culture comprises or consists of strain DSM33650 or a mutant thereof and strain DSM33651 or a mutant thereof. In some embodiments, the starter culture comprises or consists of strain DSM22876 or a mutant thereof and strain DSM33651 or a mutant thereof. In some embodiments, the starter culture comprises or consists of strain DSM33850 or a mutant thereof and strain DSM33651 or a mutant thereof. In some embodiments, the mutant strain is obtained by using the deposited strain as a starting material. In some embodiments, the mutant is a strain having all of the identifying characteristics of the strain deposited at the DSMZ.

As used herein, a mutant may be understood as a strain derived from a strain described herein and treated by, for example, genetic engineering, radiation, UV light and/or chemical treatment and/or methods of inducing genomic changes. The mutant may be a functionally equivalent mutant, e.g., a mutant having substantially the same or improved properties as the parent strain (e.g., with respect to the modulation and/or production of diacetyl and acetaldehyde content in a fermented plant-based composition; flavor; post-acidification; acidification rate; and/or phage robustness). In some embodiments, the mutant reduces acetaldehyde content in the fermented plant-based composition. In some embodiments, the mutant increases the diacetyl content in the fermented plant-based composition. In some embodiments, the mutant reduces acetaldehyde content and/or increases diacetyl content in the fermented plant-based composition. In some embodiments, the mutant reduces the acetaldehyde content and increases the diacetyl content in the fermented plant-based composition when used in combination with other strains or mutants. In some embodiments, a single mutant or combination of mutants and another strain or mutant can be used to achieve acetaldehyde and diacetyl levels, as well as relative amounts and ratios thereof, associated with a creamy flavor (e.g., increased creamy flavor) in a fermented plant-based composition. In some embodiments, the mutant may exhibit the same or different phage resistance and/or reduced phage sensitivity as the phage resistance and/or reduced phage sensitivity of the parent strain. In some embodiments, phage resistance and/or reduced phage sensitivity may be caused by differences in one or more CRISPR loci compared to the parent strain. In some embodiments, the difference may be the addition of one or more spacers in one or more CRISPR loci. In some embodiments, phage resistance and/or reduced phage sensitivity may be caused by a different non-CRISPR-mediated phage resistance mechanism than the parent strain. Thus, in some embodiments, the CRISPR locus of the mutant strain is the same as the CRISPR locus of the parent strain, but the mutant strain exhibits different phage resistance and/or reduced phage sensitivity compared to the parent strain. Methods for inducing or generating phage resistance or reduced sensitivity and detecting such features are known in the art. Non-limiting methods for inducing phage resistance or reduced phage sensitivity can be found, for example, in published international applications WO 2007/025097, WO 2007/136815 and WO 2008/108989, which are incorporated by reference in their entirety. Such mutants are part of the present invention.

Mutants as referred to herein (e.g., mutant strains) may be strains obtained by subjecting the strains described herein to any conventionally used mutagenesis treatment, including treatment with a chemical mutagen such as Ethyl Methanesulfonate (EMS) or N-methyl-N' -nitro-N-Nitroguanidine (NTG), UV light, or spontaneously occurring mutants. Mutants may have undergone several mutagenesis treatments (a single treatment is understood to include one mutagenesis step followed by a screening/selection step). In some embodiments, no more than 20, or no more than 10, or no more than 5 treatments (or screening/selection steps) are performed to generate mutants. In some embodiments, less than 5%, or less than 1% or less than 0.1% of the nucleotides in the bacterial genome are shifted, deleted, substituted or added by another nucleotide as compared to the parent strain. In some embodiments, the mutant contains no more than 20, particularly no more than 10, particularly no more than 5, particularly no more than 4, particularly no more than 3, particularly no more than 2, and particularly no more than 1 bacterial genomic nucleotide mutations compared to the parent strain, wherein the mutation is a substitution, insertion, or deletion of a nucleotide. In some embodiments, the mutants are obtained by subjecting the strains described herein to a selective pressure or stress source.

In some embodiments, the starter culture as a pure culture or mixed culture as defined above is in frozen, dried, lyophilized, liquid or solid form, in pellet or frozen pellet form, or in powder or dry powder. In some embodiments, the starter culture is in frozen form or in the form of pellets or frozen pellets, in particular contained in one or more boxes or pouches. In some embodiments, the starter culture as defined herein is in powder form, such as a dry or lyophilized powder, particularly contained in one or more cartridges or pouches. In some embodiments, the liquid form is a bulk starter, such as a bacterial strain or a mixture of bacterial strains that have been previously propagated in a growth medium to obtain a desired inoculation concentration.

Commercial non-concentrated bacterial cultures are referred to in the industry as "parent cultures" and these cultures are propagated at the production site prior to fermentation by addition to a starting material (e.g., a plant base).

In some embodiments, the starter culture, as a pure culture or a mixed culture as defined above and in whatever form (frozen, dried, lyophilized, liquid or solid, in pellet or frozen pellet form or in powder or dry powder) comprises one or more bacterial strains described herein at a concentration in the range of 10 ⁵ to 10 ¹³ cfu (colony forming units)/gram starter culture or about 10 ⁵ to 10 ¹³ cfu/gram starter culture. In some embodiments, the concentration of one or more bacterial strains described herein within the starter culture of the present invention is in the range of 10 ⁵ to 10 ¹³ CFU/gram of starter culture or about 10 ⁵ to 10 ¹³ CFU/gram of starter culture, and in particular at least 10 ⁶, at least 10 ⁷, at least 10 ⁸, at least 10 ⁹, at least 10 ¹⁰ or at least 10 ¹¹ CFU/gram of starter culture. In some embodiments, the concentration of one or more bacterial strains described herein (as pure cultures or as mixed cultures) in the starter culture is in the range of 10 ⁶ to 10 ¹³ cfu/g frozen or dried concentrate or about 10 ⁶ to 10 ¹³ cfu/g frozen or dried concentrate, when in the form of a frozen or dried concentrate, and more preferably at least 10 ⁷, at least 10 ⁸, at least 10 ⁹, at least 10 ¹⁰, At least 10 ¹¹, at least 10 ¹², or at least 10 ¹³ cfu/g frozen concentrate or dry concentrate. In some embodiments, the concentration of one or more bacterial strains described herein within the starter culture of the present invention is in the range of 10 ⁵ to 10 ⁹ CFU/g starter culture or about 10 ⁵ to 10 ⁹ CFU/g starter culture. In some embodiments, the concentration of one or more bacterial strains described herein within the starter culture of the present invention is in the range of 10 ⁶ to 10 ⁹ CFU/g starter culture or about 10 ⁶ to 10 ⁹ CFU/g starter culture. In some embodiments, the concentration of one or more bacterial strains described herein within the starter culture of the present invention is in the range of 10 ⁶ to 10 ⁸ CFU/g starter culture or about 10 ⁶ to 10 ⁸ CFU/g starter culture. In some embodiments, the concentration of one or more bacterial strains described herein within the starter culture of the present invention is in the range of 10 ⁶ to 10 ⁷ CFU/g starter culture or about 10 ⁶ to 10 ⁷ CFU/g starter culture. In some embodiments, the concentration of one or more bacterial strains described herein within a starter culture of the present invention is 10 ⁶ CFU/g starter culture, at least 10 ⁶ CFU/g starter culture, or about 10 ⁶ CFU/g starter culture. In some embodiments, the concentration of one or more bacterial strains described herein within a starter culture of the present invention is 10 ⁷ CFU/g starter culture, at least 10 ⁷ CFU/g starter culture, or about 10 ⁷ CFU/g starter culture.

Starter cultures may be prepared by techniques well known in the art, such as those disclosed in US 4,621,058. For example, a starter culture may be prepared by: an inoculum (e.g., bacteria) is introduced into the growth medium to produce an inoculated medium, and the inoculated medium is matured to produce a starter culture. Dry starter cultures may be prepared by techniques well known in the art, such as those discussed in US 4,423,079 and US 4,140,800. In some embodiments, the starter culture is in the form of concentrated frozen pellets. In some embodiments, the addition of starter culture to the plant base is direct, e.g., as a direct barrel inoculant, direct barrel starter, direct barrel group culture.

In some embodiments, the plant substrate may be inoculated with the starter culture prior to starting the fermentation. In some embodiments, the plant substrate is inoculated with the starter culture after the fermentation is started (e.g., at some point in time after the fermentation is started). In some embodiments, the plant substrate is inoculated with a starter culture during fermentation. For example, in some cases where the strains of the starter culture are added separately (e.g., not in a mixture), the strains may be added at different times prior to or during fermentation.

Additional microorganisms and/or additives may be added to the vegetable base before, during or after fermentation of the vegetable base. For example, in addition to starter cultures, enhancers (e.g., yeast extracts and/or amino acid-containing compositions) may be added to the plant base to support or promote microbial function and promote successful production of the fermented plant-based composition. Microorganisms that may be added to the plant base include those that will contribute in an advantageous manner to the characteristics of the fermented plant-based composition (e.g., fermented plant-based food). For example, the added microorganism, yeast extract, or amino acid composition may improve or support diacetyl production, acetaldehyde conversion and decomposition, viscosity, gel firmness, paste mouthfeel, flavor, post-acidification, and/or acidification rate of the fermented plant-based composition. Optionally, other ingredients may be added to the vegetable base, such as colorants, stabilizers (e.g., pectin, starch, modified starch, CMC, etc.); or polyunsaturated fatty acids, such as omega-3 fatty acids. Such ingredients may be added at any point during the production process (e.g., before or after fermentation).

Components such as cryoprotectants and/or conventional additives (including nutrients such as yeast extract, sugars and vitamins (e.g., vitamins A, C, D, K or vitamins of the vitamin B group)) may also be added to the vegetable base either together with or separately from the starter culture or other additives. Suitable cryoprotectants that may be added include components that improve cold tolerance of the microorganism, such as mannitol, sorbitol, sodium tripolyphosphate, xylitol, glycerol, raffinose, maltodextrin, erythritol, threitol, trehalose, glucose, and fructose. Other additives may include carbohydrates, fragrances, minerals, enzymes (e.g., chymosin, lactase, and/or phospholipase).

In some embodiments, the additive, microorganism, or other component as described in the preceding paragraphs is added to the plant base at the same time or nearly the same time as the starter culture (e.g., at the same time as the plant base is inoculated with the starter culture). In some embodiments, the additives, microorganisms, or other components as described in the preceding paragraphs are added to the plant-based substrate prior to (e.g., shortly before) inoculating the plant-based substrate with the starter culture. In some embodiments, the additives, microorganisms, or other components as described in the preceding paragraphs are added after (e.g., shortly after) inoculating the plant substrate with the starter culture. In some embodiments, additives, microorganisms or other components as described in the preceding paragraphs may be added to the vegetable base before, during or after fermentation, e.g. independently of inoculation with starter cultures.

In some embodiments, the plant substrate is inoculated with the starter culture and optionally additives, microorganisms or other components as described in the preceding paragraphs by any suitable method. For example, the plant base may be received by direct inoculation into a fermentation vessel.

C. Fermentation

Methods for producing the fermented plant-based compositions provided herein having a creamy flavor include fermentation (e.g., thermophilic fermentation) of a plant-based substrate (e.g., as described in sections 1-a) inoculated with starter cultures and optionally other additives, microorganisms, and components (as described in sections I-B).

Fermentation according to the methods provided herein facilitates the conversion of carbohydrates to alcohols or acids by the action of microorganisms (e.g., bacteria present in, for example, starter cultures). In some embodiments, fermentation according to the methods provided herein comprises converting lactose to lactic acid.

The fermentation process may be selected to optimize parameters (e.g., temperature, oxygen, processing time) to achieve a fermented plant-based composition having a creamy flavor as described herein. In some embodiments, the fermentation is mesophilic fermentation. In some embodiments, the fermentation is thermophilic fermentation.

In some embodiments, the inoculated plant substrate is fermented at 25 ℃, 30 ℃, 35 ℃,40 ℃, or 50 ℃, or at about these temperatures. In some embodiments, the inoculated plant base is fermented at or about 20 ℃ to 50 ℃, 30 ℃ to 50 ℃, or 40 ℃ to 50 ℃. In some embodiments, the inoculated plant substrate is fermented at a temperature above or about 30 ℃, 33 ℃, 35 ℃,36 ℃, 37 ℃, 38 ℃, 39 ℃,40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, or 50 ℃. In some embodiments, the inoculated plant based matrix is fermented at or about 35 ℃ to 47 ℃, 35 ℃ to 46 ℃, 35 ℃ to 45 ℃, 35 ℃ to 44 ℃, 35 ℃ to 43 ℃,36 ℃ to 43 ℃, 37 ℃ to 43 ℃, 38 ℃ to 43 ℃, 39 ℃ to 43 ℃,40 ℃ to 43 ℃, 41 ℃ to 43 ℃, or 42 ℃ to 43 ℃. In some embodiments, the inoculated plant substrate is fermented at 36 ℃, 37 ℃, 38 ℃, 39 ℃,40 ℃, 41 ℃, 42 ℃, 43 ℃, or 44 ℃, or at temperatures about these temperatures. In some embodiments, the inoculated vegetable substrate is fermented at 37 ℃ or a temperature of about 37 ℃ (e.g., 37 ℃ ±1 ℃). In some embodiments, the inoculated vegetable substrate is fermented at a temperature of 40 ℃ or about 40 ℃ (e.g., 40 ℃ ±1 ℃). In some embodiments, the inoculated vegetable substrate is fermented at a temperature of 43 ℃ or about 43 ℃ (e.g., 43 ℃ ±1 ℃). In some embodiments, more than one temperature or temperature range may be used for fermentation.

In some embodiments, the inoculated plant substrate is fermented for at least 4 hours, e.g., 5, 6, 7, 8, 9, 1O, 15, 20, or 24 hours. In some embodiments, the inoculated plant substrate is fermented for 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours, or about these times. In some embodiments, the inoculated vegetable substrate is fermented for up to 24 hours. In some embodiments, the inoculated plant substrate is fermented for 4 to 24, 4 to 23, 4 to 22, 4 to 21, 4 to 20, 4 to 19, 4 to 18, 4 to 17, 4 to 16, 4 to 15, 4 to 14, 4 to 13, 4 to 12, 4 to 11, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, or 4 to 5 hours, or about these times. In some embodiments, the inoculated vegetable substrate is fermented for 10 hours or about 10 hours. In some embodiments, the inoculated vegetable substrate is fermented for 9 hours or about 9 hours. In some embodiments, the inoculated vegetable substrate is fermented for 8 hours or about 8 hours. In some embodiments, the inoculated vegetable substrate is fermented for 7 hours or about 7 hours. In some embodiments, the inoculated vegetable substrate is fermented for 6 hours or about 6 hours. In some embodiments, the inoculated vegetable substrate is fermented for 5 hours or about 5 hours. In some embodiments, the inoculated vegetable substrate is fermented for 4 hours or about 4 hours.

In some embodiments, the inoculated vegetable base stock is fermented for a length of time required to bring the vegetable base stock to the target pH. In some embodiments, the target pH is a pH suitable for producing a fermented plant-based composition as a food product, e.g., as described in sections I-F below. In some embodiments, the target pH is in the range of 3.4 to 5, or about 3.4 to 5. In some embodiments, the target pH is in the range of 3.8 to 5, or about 3.8 to 5. In some embodiments, the target pH is in the range of 4 to 5, or about 4 to 5. In some embodiments, the target pH is 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, or 4.7, or about these values. In some embodiments, the target pH is in the range of 4.2 to 4.7, or about 4.2 to 4.7. In some embodiments, the target pH is 4.6 or about 4.6.

In some embodiments, at the end of fermentation, the pH of the fermented plant-based composition is in the range of 3.4 to 5, or about 3.4 to 5. In some embodiments, at the end of fermentation, the pH of the fermented plant-based composition is in the range of 3.8 to 5, or about 3.8 to 5. In some embodiments, at the end of fermentation, the pH of the fermented plant-based composition is in the range of 4 to 5, or about 4 to 5. In some embodiments, at the end of fermentation, the pH of the fermented plant-based composition is 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, or 4.7, or about these values. In some embodiments, at the end of fermentation, the pH of the fermented plant-based composition is in the range of 4.2 to 4.7, or about 4.2 to 4.7. In some embodiments, at the end of fermentation, the pH of the fermented plant-based composition is 4.6, or about 4.6.

In some embodiments, the pH of the inoculated plant substrate is measured (e.g., monitored) during fermentation. In some embodiments, the pH of the inoculated vegetable substrate is measured (e.g., monitored) at a particular point in time during fermentation. For example, pH measurements may be made every 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 minutes, or every hour, every 2 hours, every 3 hours, every 4 hours, or every 5 hours. In some embodiments, the pH of the inoculated vegetable substrate is measured (e.g., monitored) continuously during fermentation.

In some embodiments, a fermented plant-based composition produced by fermenting a plant base with a starter culture as described herein contains increased levels of acetaldehyde and/or diacetyl as compared to the levels of acetaldehyde and/or diacetyl in the plant base prior to fermentation.

D. Storage of

In some aspects, the fermented plant-based composition produced according to the methods provided herein is stored. For example, in some embodiments, after fermentation of the inoculated plant base material as described herein (see sections I-A, I-B and I-C), the fermented plant-based composition is stored to produce a stored fermented plant-based composition.

In some embodiments, the fermented plant-based composition is stored in one or more containers. Suitable containers for storage include, but are not limited to, containers capable of preserving and protecting the fermented plant-based composition (e.g., protecting the fermented plant-based composition from contamination). In some embodiments, the fermented plant-based composition is packaged in a sealed container. In some embodiments, after fermentation occurs, the fermented plant-based composition is packaged in a container. In some embodiments, fermentation occurs in a sealable container. For example, in some cases, a vegetable base (e.g., an inoculated vegetable base) may be fermented in a container such that at the end of the fermentation, the container contains a fermented vegetable-based composition, and the container is then sealed. In some embodiments, the nutrient substrate is inoculated prior to placing the nutrient substrate in the vessel for fermentation. In some embodiments, the nutrient substrate is inoculated after the nutrient substrate is placed in the vessel for fermentation.

In some embodiments, one or more containers may be used to store the fermented plant-based composition at a cooled temperature, e.g., a temperature colder than the temperature used for fermentation, such as 1 ℃,2 ℃,3 ℃,4 ℃,5 ℃,6 ℃,7 ℃,8 ℃,9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃, 14 ℃, 15 ℃,16 ℃, 17 ℃, 18 ℃,19 ℃, 20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, or 26 ℃, or about these temperatures. In some embodiments, the fermented plant-based composition is stored at a temperature in the following range or about the range: 1 ℃ to 26 ℃,1 ℃ to 24 ℃,1 ℃ to 22 ℃,1 ℃ to 20 ℃,1 ℃ to 18 ℃,1 ℃ to 16 ℃,1 ℃ to 14 ℃,1 ℃ to 12 ℃,1 ℃ to 10 ℃,1 ℃ to 8 ℃,1 ℃ to 6 ℃,1 ℃ to 4 ℃, or 1 ℃ to 2 ℃. In some embodiments, the fermented plant-based composition is stored at a temperature in the following range or about the range: 1 ℃ to 14 ℃,1 ℃ to 12 ℃,1 ℃ to 10 ℃,1 ℃ to 8 ℃,1 ℃ to 6 ℃, or 1 ℃ to 4 ℃. In some embodiments, the fermented plant-based composition is stored at a temperature in the range of 4 ℃ to 8 ℃, or in the range of about 4 ℃ to 8 ℃. In some embodiments, the fermented plant-based composition is stored at or about the temperature: 1 ℃,2 ℃,3 ℃,4 ℃,5 ℃,6 ℃,7 ℃,8 ℃,9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃, 14 ℃, 15 ℃,16 ℃, 17 ℃, 18 ℃,19 ℃, 20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, or 26 ℃. In some embodiments, the fermented plant-based composition is stored at or about the temperature: 1 ℃,2 ℃,3 ℃,4 ℃,5 ℃,6 ℃,7 ℃,8 ℃,9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃ or 14 ℃. In some embodiments, the fermented plant-based composition is stored at or about the temperature: 2 ℃,3 ℃,4 ℃,5 ℃,6 ℃,7 ℃ or 8 ℃. In some embodiments, the fermented plant-based composition is stored at a temperature of 6 ℃ or about 6 ℃ (e.g., 6 ℃ ± 1 ℃).

In some embodiments, the stored fermented plant-based composition maintains a target pH reached during fermentation. In some embodiments, the target pH is a pH suitable for producing a fermented plant-based composition as a food product, e.g., as described in sections I-F below. In some embodiments, the target pH is in the range of 3.8 to 5, or about 3.8 to 5. In some embodiments, the target pH is in the range of to 5, or about to 5. In some embodiments, the target pH is 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, or 4.7, or about these values. In some embodiments, the target pH is in the range of 4.2 to 4.7, or about 4.2 to 4.7. In some embodiments, the target pH is 4.6 or about 4.6.

In some embodiments, the pH of the stored fermented plant-based composition is in the range of 3.4 to 4.5, or about 3.4 to 4.5. In some embodiments, the pH of the stored fermented plant-based composition is in the range of 3.8 to 4.2, or about 3.8 to 4.2. In some embodiments, the pH of the stored fermented plant-based composition is 3.8, 3.9, 4.0, 4.1, 4.2, or 4.3, or about these values. In some embodiments, the pH of the stored fermented plant-based composition is in the range of 3.9 to 4.1, or about 3.9 to 4.1. In some embodiments, the pH of the stored fermented plant-based composition is 4.0 or about 4.0.

In some embodiments, the pH of the stored fermented plant-based composition is measured (e.g., monitored) during storage. In some embodiments, the pH of the stored fermented plant-based composition is measured (e.g., monitored) at a specific point in time during storage. For example, pH measurements may be made every day of storage, or every other day, every third day, every fourth day, every fifth day, every sixth day, every seventh day, every eighth day, every ninth day, every tenth day, or every twelve days, every thirteenth day, or every fourteen days during the duration of storage. In some embodiments, the pH of the stored fermented plant-based composition is measured (e.g., monitored) about every 5 hours, 10 hours, 15 hours, 20 hours, 24 hours, 36 hours, 48 hours, 60 hours, or every 72 hours for the duration of storage. In some embodiments, the pH of the stored fermented plant-based composition is measured (e.g., monitored) continuously during storage. In some embodiments, the pH of the stored fermented plant-based composition is measured at a temperature of 20 ℃ or about 20 ℃. For example, the stored fermented plant-based composition may be removed from storage for a period of time to increase the temperature of the stored fermented plant-based composition prior to measuring the pH.

In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days, or at least about these times. In some embodiments, the fermented plant-based composition (e.g., a stored fermented plant-based composition) is stored for at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100, 110, or 120 days, or at least about these times. In some embodiments, the fermented plant-based composition (e.g., a stored fermented plant-based composition) is stored for at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, or 90 days, or at least about these times. In some embodiments, the fermented plant-based composition (e.g., a stored fermented plant-based composition) is stored for at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, or 60 days, or at least about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 days, or at least about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 4, 3, 2, or 1 month, or at least about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 120 days or at least about 120 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 90 days or at least about 90 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 60 days or at least about 60 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 30 days or at least about 30 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 28 days or at least about 28 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 21 days or at least about 21 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 14 days or at least about 14 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for at least 7 days or at least about 7 days.

In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days, or up to about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 30, 40, 50, 60, 70, 80, 90, 100, 110, or 120 days, or up to about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29, 30, 40, 50, 60, 70, 80, or 90 days, or up to about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, or 60 days, or up to about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 days, or up to about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 4, 3, 2, or 1 month, or up to about these times. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 90 days or up to about 90 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 60 days or up to about 60 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 30 days or up to about 30 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 28 days or up to about 28 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 21 days or up to about 21 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 14 days or up to about 14 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for up to 7 days or up to about 7 days.

In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 1 to 120, 1 to 90, 1 to 60, 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, or 1 to 5 days, or about these durations. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 120, 7 to 90, 7 to 60, or 7 to 30 days, or about these durations. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 28, 7 to 21, or 7 to 14 days, or about these durations. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 120 days, or about this duration. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 90 days, or about this duration. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 60 days, or about this duration. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 30 days, or about this duration. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 to 21, or 7 to 14 days, or about these durations. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 120 days or about 120 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 90 days or about 90 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 60 days or about 60 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 30 days or about 30 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for 28 days or a duration of about 28 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 21 days or about 21 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 14 days or about 14 days. In some embodiments, the fermented plant-based composition (e.g., stored fermented plant-based composition) is stored for a duration of 7 days or about 7 days.

In some embodiments, the fermented plant-based composition (e.g., a stored fermented plant-based composition) is stored for a length of time or at least for a length of time suitable for the acetaldehyde and diacetyl levels in the fermented plant-based composition (e.g., a stored fermented plant-based composition) to reach a relative amount or ratio associated with a creamy flavor (e.g., an increased creamy flavor). In some embodiments, the fermented plant-based composition (e.g., a stored fermented plant-based composition) is stored until the relative amount or ratio of acetaldehyde to diacetyl as described in sections 1-E is reached. In some embodiments, the fermented plant-based composition (e.g., a stored fermented plant-based composition) is stored until a creamy flavor (e.g., increased creamy flavor) as described in sections I-E is achieved.

E. Evaluation of acetaldehyde, diacetyl and creamer flavor

In some aspects, the fermented plant-based composition produced according to the methods provided herein has a creamy flavor (e.g., increased creamy flavor). In some embodiments, the cream flavor imparts a cream aroma and/or cream smell. Without being bound by theory, the cream flavor may be related to the relative amounts of diacetyl and acetaldehyde present in the fermented plant-based composition. Thus, in some embodiments, the methods provided herein include reducing the acetaldehyde content in a fermented plant-based composition and increasing the diacetyl content therein to reach a level perceptually relevant to the creamer flavor (e.g., increased creamer aroma and/or creamer odor level) of the fermented plant-based composition. In some embodiments, the acetaldehyde and diacetyl levels achieved according to the methods provided herein produce increased cream flavor (e.g., cream aroma and/or cream smell) as compared to other methods of producing a fermented plant-based composition that does not result in the relative amounts of acetaldehyde and diacetyl as described herein. In some embodiments, the increased cream flavor is an increased cream aroma and/or an increased cream odor, e.g., as compared to a fermented plant-based composition produced via an alternative method.

Methods for detecting and optionally quantifying the levels of volatile organic compounds such as acetaldehyde and diacetyl include, but are not limited to, mass spectrometry such as gas chromatography-mass spectrometry (GC-MS), selective ion flow tube mass spectrometry (SIFT-MS), proton transfer reaction mass spectrometry (PTR-MS), ion-molecule reaction mass spectrometry (IMR-MS). In some embodiments, acetaldehyde and diacetyl in the fermented plant-based composition are detected and quantified using GC-MS.

Methods for determining sensory characteristics (including cream flavor, e.g., cream aroma, cream smell) are known in the art and include, for example, quantitative Descriptive Analysis (QDA) using a trained sensory panel. In some embodiments, the trained sensory evaluator evaluates the descriptive index using a linear scale (e.g., a 7-wire linear scale). In some embodiments, the descriptive indicators of the evaluation include cream aroma and cream scent. In some embodiments, QDA is performed in a blind manner, e.g., the evaluator is unaware of the identity of the fermented plant-based composition being evaluated or the method of producing it.

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the acetaldehyde content in the fermented plant-based composition. In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the acetaldehyde content and increasing the diacetyl content in the fermented plant-based composition. In some embodiments, the reduction in acetaldehyde results in a ratio of acetaldehyde to diacetyl that is related to cream flavor (e.g., increased cream flavor). In some embodiments, the reduction in acetaldehyde and the increase in diacetyl results in a ratio of acetaldehyde to diacetyl that is related to cream flavor (e.g., increased cream flavor).

The acetaldehyde content may be considered to be reduced relative to the initial acetaldehyde content. In some embodiments, the initial acetaldehyde content is the acetaldehyde content of a fermented plant-based composition. In some embodiments, the initial acetaldehyde content is the acetaldehyde content of the fermented plant-based composition at the end of the fermentation process. In some embodiments, the initial acetaldehyde content is the acetaldehyde content of the fermented plant-based composition at the beginning of storage (e.g., the first day of storage). Similarly, the diacetyl content may be considered to increase relative to the initial diacetyl content. In some embodiments, the initial diacetyl content is the diacetyl content of the fermented plant-based composition at the end of the fermentation process. In some embodiments, the initial diacetyl content is the diacetyl content of the fermented plant-based composition at the beginning of storage (e.g., the first day of storage).

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the level of acetaldehyde in the fermented plant-based composition (e.g., initial acetaldehyde content) as compared to the level of acetaldehyde present in a fermented plant-based composition produced by fermenting an inoculated plant base. In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes increasing the level of diacetyl in the fermented plant-based composition as compared to the level of diacetyl (e.g., initial diacetyl content) present in the fermented plant-based composition produced by fermenting an inoculated vegetable base. In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the level of acetaldehyde in the fermented plant-based composition (e.g., initial acetaldehyde content) as compared to the level of acetaldehyde present in the fermented plant-based composition produced by fermenting an inoculated plant base; and increasing the level of diacetyl in the fermented plant-based composition as compared to the level of diacetyl (e.g., the initial diacetyl content) present in the fermented plant-based composition produced by fermenting the inoculated plant-based material. In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the level of acetaldehyde in the fermented plant-based composition (e.g., initial acetaldehyde content) as compared to the level of acetaldehyde present in the fermented plant-based composition produced by fermenting an inoculated plant base; and/or increasing the level of diacetyl in the fermented plant-based composition as compared to the level of diacetyl (e.g., the initial diacetyl content) present in a fermented plant-based composition produced by fermenting the inoculated plant-based material to produce an acetaldehyde to diacetyl ratio associated with a creamer flavor (e.g., increased creamer flavor) as described herein.

In some embodiments, the acetaldehyde content is reduced by at least a factor of 1.5, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 30, 40, or 50. In some embodiments, the acetaldehyde content is reduced by at least a factor of 2. In some embodiments, the fold reduction in acetaldehyde content is in the range of 1.5 to 15, 1.5 to 10, 1.5 to 9, 1.5 to 8, 1.5 to 7, 1.5 to 6, 1.5 to 5, 1.5 to 4, 1.5 to 3, or 1.5 to 2, or about these ranges. In some embodiments, the fold reduction in acetaldehyde content is in the range of 1.5 to 10, 1.5 to 9, 1.5 to 8, 1.5 to 7, or 1.5 to 6, or about these ranges. In some embodiments, the fold reduction in acetaldehyde content is in the range of 2 to 10, or about this range. In some embodiments, the acetaldehyde content is reduced by a factor of 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 20. In some embodiments, the acetaldehyde content is reduced by a factor of 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15, or about these factors. In some embodiments, the acetaldehyde content is reduced by a factor of 1.5, 2, 3, 4, or 5, or about these factors. In some embodiments, the acetaldehyde content is reduced by a factor of 1.5 or about 1.5. In some embodiments, the acetaldehyde content is reduced by a factor of 2 or about 2.

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor comprises reducing the acetaldehyde content by a factor described herein.

In some embodiments, the acetaldehyde content is reduced to 1ppm、0.5ppm、0.4ppm、0.3ppm、0.2ppm、0.1ppm、0.09ppm、0.08ppm、0.07ppm、0.06ppm、0.05ppm、0.03ppm、0.02ppm、0.01ppm or less, or less than these values. In some embodiments, the acetaldehyde content is reduced to the following range or about the following range: 0.8ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.7ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.6ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.5ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.4ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.3ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.2ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), or 0.1ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit). In some embodiments, the acetaldehyde content is reduced to a specified amount and/or a lower limit of examination, or below. In some embodiments, the acetaldehyde content is reduced to a range of 0.6ppm to about 0ppm or about this range, e.g., below the lower quantification and/or lower examination limit. In some embodiments, the acetaldehyde content is reduced to a range of 0.2ppm to about 0ppm or about this range, e.g., below the lower quantification and/or inspection limits. In some embodiments, the acetaldehyde content in the fermented plant-based composition is negligible. In some embodiments, the lower detection and/or quantification limit is specific to the instrument used to detect the compound (e.g., acetaldehyde). In some embodiments, acetaldehyde is detected and quantified using GC-MS. In some embodiments, the lower limit of quantitation is 0.125ppm.

In some embodiments, the method of producing a fermented plant-based composition having a creamy flavor produces a fermented plant-based composition having an acetaldehyde content of 1ppm、0.9ppm、0.8ppm、0.7ppm、0.6ppm、0.5ppm、0.4ppm、0.3ppm、0.2ppm、0.1ppm、0.09ppm、0.08ppm、0.07ppm、0.06ppm、0.05ppm、0.03ppm、0.02ppm、0.01ppm, or less. In some embodiments, the acetaldehyde content produced is in the following range or about the range: 0.8ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.7ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.6ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.5ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.4ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.3ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), 0.2ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit), or 0.1ppm to about 0ppm (e.g., below a lower quantification and/or inspection limit). In some embodiments, the amount of acetaldehyde produced is a quantitative and/or check lower limit, about the lower limit, or below the lower limit. In some embodiments, the acetaldehyde content is reduced to a range of 0.6ppm to about 0ppm or about this range, e.g., below the lower quantification and/or lower examination limit. In some embodiments, the acetaldehyde content produced is in the range of 0.2ppm to about 0ppm or about this range, e.g., below the lower quantification and/or inspection limits. In some embodiments, the amount of acetaldehyde produced in the fermented plant-based composition is negligible. In some embodiments, the lower detection and/or quantification limit is specific to the instrument used to detect the compound (e.g., acetaldehyde). In some embodiments, acetaldehyde is detected and quantified using GC-MS. In some embodiments, the lower limit of quantitation is 0.125ppm.

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the acetaldehyde content to the amounts described herein.

In some embodiments, the diacetyl content is increased by at least a factor of 1.5, e.g., 2,5, 6,7,8,9, 10, 11, 12, 13, 14, or 15. In some embodiments, the diacetyl content is increased by at least a factor of 2. In some embodiments, the diacetyl content is increased by a factor in the range of 1.5 to 20, 1.5 to 15, 1.5 to 10, 1.5 to 9, 1.5 to 8, 1.5 to 7, 1.5 to 6, 1.5 to 5, 1.5 to 4, 1.5 to 3, or 1.5 to 2, or about these ranges. In some embodiments, the diacetyl content is increased by a factor in the range of 1.5 to 15, 1.5 to 10, 1.5 to 9, 1.5 to 8, 1.5 to 7, or 1.5 to 6, or about these ranges. In some embodiments, the diacetyl content is increased by a factor in the range of 1.5 to 10, or about this range. In some embodiments, the diacetyl content is increased 1.5, 2,3, 4,5, 6,7,8,9, 10, 11, 12, 13, 14, 15, or 20 times. In some embodiments, the diacetyl content is increased by a factor of 1.5, 2,3, 4,5, 6,7,8,9, 10, 11, 12, 13, 14, or 15, or about these factors. In some embodiments, the diacetyl content is increased by a factor of 1.5, 2,3, 4, or 5, or about these factors. In some embodiments, the diacetyl content is increased by a factor of 1.5 or about 1.5. In some embodiments, the diacetyl content is increased by a factor of 2 or about 2. In some embodiments, the diacetyl content is increased 3-fold or about 3-fold. In some embodiments, the diacetyl content is increased by a factor of 4 or about 4. In some embodiments, the diacetyl content is increased by a factor of 5 or about 5. In some embodiments, the diacetyl content is increased by a factor of 6 or about 6. In some embodiments, the diacetyl content is increased by a factor of 7 or about 7. In some embodiments, the diacetyl content is increased by a factor of 8 or about 8. In some embodiments, the diacetyl content is increased by a factor of 9 or about 9. In some embodiments, the diacetyl content is increased by a factor of 10 or about 10.

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor comprises increasing the diacetyl content by a factor described herein.

In some embodiments, the diacetyl content is increased to 1ppm, 2ppm, 3ppm, 4ppm, 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm, 11ppm, 12ppm, 13ppm, 14ppm, 15ppm, 20ppm, or more, or at least these values. In some embodiments, the diacetyl content is increased to 1.5ppm or at least 1.5ppm. In some embodiments, the diacetyl content is increased to 2ppm or at least 2ppm. In some embodiments, the diacetyl content is increased to 3ppm or at least 3ppm. In some embodiments, the diacetyl content is increased to 4ppm or at least 4ppm. In some embodiments, the diacetyl content is increased to 5ppm or at least 5ppm. In some embodiments, the diacetyl content is increased to 6ppm or at least 6ppm. In some embodiments, the diacetyl content is increased to 7ppm or at least 7ppm. In some embodiments, the diacetyl content is increased to 8ppm or at least 8ppm. In some embodiments, the diacetyl content is increased to 9ppm or at least 9ppm. In some embodiments, the diacetyl content is increased to 10ppm or at least 10ppm. In some embodiments, the diacetyl content is increased to 15ppm or at least 15ppm. In some embodiments, the diacetyl content is increased to 20ppm or at least 20ppm.

In some embodiments, the diacetyl content is increased to 1ppm to 20ppm, 1ppm to 15ppm, 1ppm to 14ppm, 1ppm to 13ppm, 1ppm to 12ppm, 1ppm to 11ppm, 1ppm to 10ppm, 1ppm to 9ppm, 1ppm to 8ppm, 1ppm to 7ppm, 1ppm to 6ppm, 1ppm to 5ppm, 1ppm to 4ppm, 1ppm to 3ppm, 1ppm to 2ppm, or 1ppm to 1.5ppm, or about these ranges. In some embodiments, the diacetyl content is increased to 1ppm to 7ppm, 1ppm to 6ppm, 1ppm to 5ppm, 1ppm to 4ppm, 1ppm to 3ppm, 1ppm to 2ppm, or 1ppm to 1.5ppm, or about these ranges. In some embodiments, the diacetyl content is increased to 1ppm to 7ppm, or about 1ppm to 7ppm. In some embodiments, the diacetyl content is increased to 1.5ppm to 7ppm, or about 1.5ppm to 7ppm. In some embodiments, the diacetyl content is increased to 1ppm, 2ppm, 3ppm, 4ppm, 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm, 11ppm, 12ppm, 13ppm, 14ppm, or 15ppm, or about these values.

In some embodiments, the method of producing a fermented plant-based composition having a creamy flavor produces a diacetyl content of 1ppm, 2ppm, 3ppm, 4ppm, 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm, 11ppm, 12ppm, 13ppm, 14ppm, 15ppm, 20ppm, or more, or at least these values. In some embodiments, the diacetyl content produced is 1.5ppm or at least 1.5ppm. In some embodiments, the diacetyl content produced is 2ppm or at least 2ppm. In some embodiments, the diacetyl content produced is 3ppm or at least 3ppm. In some embodiments, the diacetyl content produced is 4ppm or at least 4ppm. In some embodiments, the diacetyl content produced is 5ppm or at least 5ppm. In some embodiments, the diacetyl content produced is 6ppm or at least 6ppm. In some embodiments, the diacetyl content produced is 7ppm or at least 7ppm. In some embodiments, the diacetyl content produced is 8ppm or at least 8ppm. In some embodiments, the diacetyl content produced is 9ppm or at least 9ppm. In some embodiments, the diacetyl content produced is 10ppm or at least 10ppm. In some embodiments, the diacetyl content produced is 15ppm or at least 15ppm. In some embodiments, the diacetyl content produced is 20ppm or at least 20ppm. In some embodiments, the diacetyl produced is in the range of 1ppm to 20ppm, 1ppm to 15ppm, 1ppm to 14ppm, 1ppm to 13ppm, 1ppm to 12ppm, 1ppm to 11ppm, 1ppm to 10ppm, 1ppm to 9ppm, 1ppm to 8ppm, 1ppm to 7ppm, 1ppm to 6ppm, 1ppm to 5ppm, 1ppm to 4ppm, 1ppm to 3ppm, 1ppm to 2ppm, or 1ppm to 1.5ppm, or about these ranges. In some embodiments, the diacetyl produced is in the range of 1ppm to 7ppm, 1ppm to 6ppm, 1ppm to 5ppm, 1ppm to 4ppm, 1ppm to 3ppm, 1ppm to 2ppm, or 1ppm to 1.5ppm, or about these ranges. In some embodiments, the diacetyl content produced is in the range of 1ppm to 7ppm or about this range. In some embodiments, the diacetyl content produced is in the range of 1.5ppm to 7ppm or about this range. In some embodiments, the diacetyl content produced is 1ppm, 2ppm, 3ppm, 4ppm, 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm, 11ppm, 12ppm, 13ppm, 14ppm, 15ppm, or 20ppm, or about these values.

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes increasing the diacetyl content to the amounts described herein.

In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the acetaldehyde content by a factor described herein, and increasing the diacetyl content by a factor described herein. In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the acetaldehyde content by an amount described herein, and increasing the diacetyl content by an amount described herein. In some embodiments, a method of producing a fermented plant-based composition having a creamy flavor includes reducing the acetaldehyde content to an amount described herein, and increasing the diacetyl content to an amount described herein.

In some embodiments, the method of producing a fermented plant-based composition having a cream flavor (e.g., increased cream flavor) results in a ratio of acetaldehyde to diacetyl that is related to the cream flavor (e.g., increased cream flavor). In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamer flavor (e.g., increased creamer flavor) is less than 1, such as about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.3, about 0.2, about 0.1, about 0.08, about 0.07, about 0.06, about 0.05, about 0.04, about 0.03, about 0.02, or about 0.01, or less. In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamer flavor (e.g., increased creamer flavor) is less than 0.85, 0.75, 0.5, 0.25, 0.1, 0.075, 0.05, 0.025, or 0.01, or less than about these values. In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamy flavor (e.g., increased creamy flavor) is less than 0.7, 0.5, 0.4, 0.3, 0.2, or 0.1, or less than about these values. In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamy flavor (e.g., increased creamy flavor) is 0.6 or about 0.6. In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamy flavor (e.g., increased creamy flavor) is less than 0.25, 0.1, 0.075, 0.05, 0.025, or 0.01, or less than about these values. In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamer flavor (e.g., increased creamer flavor) is in the range of 0.8 to 0, 0.7 to 0, 0.6 to 0, 0.5 to 0, 0.4 to 0, 0.3 to 0, 0.2 to 0, or 0.1 to 0, or about these ranges. In some embodiments, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition that is associated with a creamy flavor (e.g., increased creamy flavor) is in the range of 0.6 to 0, or about this range. In some embodiments (ratio embodiments), for example, when the acetaldehyde content is at or below a lower detection and/or quantification limit, the ratio of acetaldehyde to diacetyl in the fermented plant-based composition in relation to cream flavor (e.g., increased cream flavor) is 0 or about 0.

In some embodiments, the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is less than 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0.01, or 0.

In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0.01, or less than these values, or 0. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0.01, or less than these values, or 0, and the diacetyl content is increased to at least 1ppm or at least about 1ppm. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less than these values, or 0 or about 0, and the diacetyl content is increased to at least 1ppm or at least about 1ppm. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0.01, or less than these values, or 0, and the diacetyl content is increased to at least 1.5ppm or at least about 1.5ppm. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less than these values, or 0 or about 0, and the diacetyl content is increased to at least 1.5ppm or at least about 1.5ppm. In some embodiments, the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is 0.6, 0.5, 0.25, 0.1, or less than these values, or 0 or about v, and the diacetyl content is increased to the level disclosed in any of the embodiments herein.

In some embodiments, the diacetyl content is increased to about 1ppm, about 2ppm, about 3ppm, about 4ppm, about 5ppm, about 6ppm, about 7ppm, about 8ppm, about 9ppm, about 10ppm, about 11ppm, about 12ppm, about 13ppm, about 14ppm, about 15ppm, or about 20ppm, and the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, or less than these values, or 0. In some embodiments, the diacetyl content is increased to about 1ppm, about 2ppm, about 3ppm, about 4ppm, about 5ppm, about 6ppm, about 7ppm, and the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less than these values, or 0. In some embodiments, the diacetyl content is increased to about 1ppm, about 2ppm, about 3ppm, about 4ppm, about 5ppm, about 6ppm, about 7ppm, and the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less than these values, or 0. In some embodiments, the acetaldehyde content is reduced to the levels disclosed herein to achieve the ratios disclosed herein in relation to cream flavor (e.g., increased cream flavor).

In some embodiments, the acetaldehyde content is reduced to less than about 1ppm and the diacetyl content is increased to greater than 1ppm, or greater than about 1ppm. In some embodiments, the acetaldehyde content is reduced to less than about 0.7ppm and the diacetyl content is increased to greater than about 1ppm. In some embodiments, the acetaldehyde content is reduced to about 0.6ppm and the diacetyl content is increased to greater than 1.5ppm, or greater than about 1.5ppm. In some embodiments, the acetaldehyde content is reduced to less than about 1ppm and the diacetyl content is increased to greater than 1.5ppm, or greater than about 1.5ppm. In some embodiments, the acetaldehyde content is reduced to less than about 0.5ppm and the diacetyl content is increased to greater than about 1.5ppm. In some embodiments, the acetaldehyde content is reduced to about 0ppm and the diacetyl content is increased to greater than 1.5ppm, or greater than about 1.5ppm. In some embodiments, the acetaldehyde content is reduced to any level disclosed herein and the diacetyl content is increased to any level disclosed herein to obtain the acetaldehyde to diacetyl ratio disclosed herein, e.g., a ratio related to cream flavor (e.g., increased cream flavor).

In some embodiments, the method of producing a fermented plant-based composition having a creamy flavor (e.g., increased creamy flavor) produces an acetaldehyde content of less than about 1ppm, and a diacetyl content of greater than 1ppm, or greater than about 1ppm. In some embodiments, the acetaldehyde content produced is less than about 0.6ppm and the diacetyl content produced is greater than about 1ppm. In some embodiments, the acetaldehyde content produced is about 0.6ppm and the diacetyl content produced is greater than 1.5ppm, or greater than about 1.5ppm. In some embodiments, the acetaldehyde content produced is at any level disclosed herein and the diacetyl content produced is at any level disclosed herein to obtain the acetaldehyde to diacetyl ratio disclosed herein, e.g., a ratio related to cream flavor (e.g., increased cream flavor).

In some embodiments, the creamer flavor of the fermented plant-based composition having the acetaldehyde and diacetyl levels and/or ratios thereof described herein has increased creamer aroma and/or increased creamer odor as compared to a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of about 0.7 or greater. In some embodiments, the creamer flavor of the fermented plant-based composition having the acetaldehyde and diacetyl levels or ratios thereof described herein has an increased creamer aroma and/or an increased creamer smell as compared to a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of about 0.8 or greater. In some embodiments, the creamer flavor of the fermented plant-based composition having the acetaldehyde and diacetyl levels or ratios thereof described herein has an increased creamer aroma and/or an increased creamer smell as compared to a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of about 0.9 or greater. In some embodiments, the creamer flavor of the fermented plant-based composition having the acetaldehyde and diacetyl levels or ratios thereof described herein has an increased creamer aroma and/or an increased creamer odor as compared to a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of about 1 or greater.

In some embodiments, the content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is reached at the end of fermentation. In some embodiments, the content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is reached at some point in time after fermentation is complete. For example, the levels or ratios described in this section in relation to cream flavor (e.g., increased cream flavor) are achieved during storage of the fermented plant-based composition. In some embodiments, the content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved in a stored fermented plant-based composition. For example, in some cases, the content or ratio described in this section in relation to a creamer flavor (e.g., increased creamer flavor) is achieved during storage of a fermented plant-based composition (e.g., a stored fermented plant-based composition according to any of the storage methods described in sections 1-D above).

In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved after more than 1 day of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved in the range of 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, or 1 to 5 days, or about the range of these days, of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved over a period of days or about a range of days in which the fermented plant-based composition is stored of 1 to 28, 1 to 21, or 1 to 14, or 1 to 7.

In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 28 days or less of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 21 days or less of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 14 days or less of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 7 days or less of storage of the fermented plant-based composition.

In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 28 days or about 28 days of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 21 days or about 21 days of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 14 days or about 14 days of storage of the fermented plant-based composition. In some embodiments, the acetaldehyde to diacetyl content or ratio described in this section in relation to cream flavor (e.g., increased cream flavor) is achieved within 7 days or about 7 days of storage of the fermented plant-based composition.

F. Fermented plant-based food

Also provided herein are fermented plant-based foods obtained by any of the methods described herein and having any of the characteristics (e.g., ratios) described herein that produce a creamy flavor (e.g., increased creamy flavor). In some aspects, the fermented plant-based composition is a fermented plant-based food.

In some embodiments, the methods provided herein for producing a fermented plant-based composition having a creamy flavor produce a fermented plant-based food. In some embodiments, the fermented plant-based food product contains an acetaldehyde content to diacetyl content ratio of less than 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, or 0.01, or 0. In some embodiments, the fermented plant-based food product contains an acetaldehyde content to diacetyl content ratio of less than 0.7, 0.6, 0.5, 0.25, 0.1, 0.075, 0.05, 0.025, or 0.01, or 0. In some embodiments, the fermented plant-based food product contains an acetaldehyde content to diacetyl content ratio of less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0. In some embodiments, the fermented plant-based food product contains an acetaldehyde content to diacetyl content ratio of 0.7 or less than 0.7. In some embodiments, the fermented plant-based food product contains an acetaldehyde content to diacetyl content ratio of 0.6 or less than 0.6. In some embodiments, the amount or ratio of acetaldehyde to diacetyl in the fermented plant-based food is any amount or ratio associated with a creamy flavor (e.g., increased creamy flavor) as described in sections I-E.

In some embodiments, the fermented plant-based food product contains a strain of lactobacillus rhamnosus that reduces acetaldehyde in the fermented plant-based food product. In some embodiments, the fermented plant-based food product contains a strain of lactobacillus rhamnosus that increases diacetyl in the fermented plant-based food product. In some embodiments, the acetaldehyde-reducing lactobacillus rhamnosus strain and the diacetyl-increasing lactobacillus rhamnosus strain are the same strain. Alternatively, different strains of lactobacillus rhamnosus may reduce acetaldehyde and increase diacetyl.

In some embodiments, the fermented plant-based food product contains lactobacillus rhamnosus strain DSM33650 or a mutant thereof as described herein. In some embodiments, the fermented plant-based food product contains lactobacillus rhamnosus strain DSM22876 or a mutant thereof as described herein. In some embodiments, the fermented plant-based food product contains lactobacillus rhamnosus strain DSM33850 or a mutant thereof as described herein. In some embodiments, the fermented plant-based food product contains streptococcus thermophilus.

In some embodiments, the fermented plant-based food product contains streptococcus thermophilus strain DSM33651, or a mutant thereof, as described herein, and lactobacillus rhamnosus strain DSM33650, or a mutant thereof, as described herein. In some embodiments, the fermented plant-based food product contains streptococcus thermophilus strain DSM33651, or a mutant thereof, as described herein, and lactobacillus rhamnosus strain DSM22876, or a mutant thereof, as described herein. In some embodiments, the fermented plant-based food product contains streptococcus thermophilus strain DSM33651, or a mutant thereof, as described herein, and lactobacillus rhamnosus strain DSM33850, or a mutant thereof, as described herein.

In some embodiments, the fermented plant-based food product is a soy-based composition. In some embodiments, the fermented plant-based food is a fermented soybean-based food.

Fermented plant-based food products contemplated herein include, but are not limited to, food products such as, but not limited to, fermented milk alternative food products. In some embodiments, the fermented plant-based food is a fermented milk-substitute food.

The term "food" is used in a broad sense and includes feed, foodstuff, food ingredients, food supplements and functional foods. Herein, the term "food" is used in a broad sense-and covers both human food as well as food for animals (i.e. feed). In a preferred aspect, the food is for human consumption.

In some embodiments, the fermented plant-based composition produced according to the methods described herein produces a fermented plant-based food product that is a functional food. As used herein, the term "functional food" means a food that is not only capable of providing a nutritional effect and/or a taste satisfaction but also capable of delivering additional beneficial effects to the consumer. Although there is no legal definition for functional foods, most parties interested in this field agree on foods that exist for sale with specific health effects.

In some embodiments, the bacterial strains or bacterial compositions described herein (e.g., bacterial strains or bacterial compositions in starter cultures) can be food ingredients, food supplements, or functional foods, or can be added thereto.

In some embodiments, the food may be in the form of a solution or as a solid, depending on the use and/or mode of application and/or mode of administration.

In some embodiments, the bacterial strains or bacterial compositions (e.g., starter cultures) described herein can be used to prepare a food product, such as one or more fermented plant-based food products.

For example, a bacterial strain or bacterial composition (e.g., starter culture) may be used to prepare the following ingredients: milk beverages, lactobacillus beverages, yogurt substitutes, and/or drinking yogurt substitutes made from plant matter (e.g., plant based binders) and referred to as milk-substitute foods (e.g., plant based yogurt substitutes).

In some embodiments, the fermented plant-based food product is a stirred milk-substitute food product. In some embodiments, the fermented plant-based food product is a set-type (set-type) milk-substitute food product. In some embodiments, the fermented plant-based food product is a plant-based yogurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative.

Method for identifying bacteria capable of producing fermented plant-based compositions with creamy flavour

Provided herein are methods for identifying bacteria for producing a fermented plant-based composition (e.g., a fermented plant-based food product) having a creamy flavor (e.g., an increased creamy flavor). In some embodiments, methods for identifying bacteria and bacterial compositions capable of producing a fermented plant-based composition having a creamy flavor (e.g., an increased creamy flavor) include fermenting a plant base inoculated with one or more bacteria as described herein, and determining whether the one or more bacteria cause an increase in diacetyl content and/or a decrease in acetaldehyde content in the fermented plant-based composition that produces a ratio and/or level of acetaldehyde and diacetyl that is related (e.g., correlated) to the creamy flavor (e.g., an increased creamy flavor). In some cases, one or more bacteria capable of producing such a result are identified as being capable of producing a fermented plant-based composition having a creamy flavor (e.g., increased creamy flavor). In some aspects, methods of identifying such bacteria include quantifying the acetaldehyde and diacetyl content in a fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the fermented plant-based composition is produced (e.g., fermented and stored) according to the methods provided herein, wherein the ability of the bacteria or bacterial composition to produce levels and ratios of acetaldehyde and diacetyl related to creamer flavor is evaluated. In some embodiments, one or more bacteria capable of producing a fermented plant-based composition having a creamy flavor are identified.

In some embodiments, a method of identifying bacteria capable of producing a fermented plant-based composition having a creamy flavor as described herein comprises determining the ratio of acetaldehyde to diacetyl in a fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the identification method comprises quantifying the acetaldehyde content and the diacetyl content in a fermented plant-based composition (e.g., as described herein) fermented in the presence of one or more bacteria and stored after fermentation. In some embodiments, the identification method comprises determining the ratio of acetaldehyde to diacetyl in a fermented plant-based composition fermented in the presence of one or more bacteria and stored after fermentation as described herein. In some embodiments, the identification method comprises quantifying the acetaldehyde content and the diacetyl content in a stored fermented plant-based composition fermented in the presence of one or more bacteria. In some embodiments, the identification method comprises determining the ratio of acetaldehyde to diacetyl in a stored fermented plant-based composition fermented in the presence of one or more bacteria as described herein. In some embodiments, quantifying the amount and/or determining the ratio as described herein is performed at different points in time over the duration of the storage. Thus, in some cases, the change in diacetyl and/or acetaldehyde content may be determined over time. In some embodiments, quantifying the amount and/or determining the ratio as described herein is performed continuously for the duration of the storage.

In some embodiments, one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the one or more bacteria cause a reduction in the level of acetaldehyde as described in sections I-E. In some embodiments, one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the one or more bacteria cause a decrease in acetaldehyde levels and an increase in diacetyl levels as described in sections I-E. In some embodiments, one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the one or more bacteria produce a ratio of acetaldehyde to diacetyl related to the creamy flavor as described in section I-E. In some embodiments, one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the one or more bacteria produce levels and/or ratios of acetaldehyde to diacetyl related to the creamy flavor as described in sections I-E.

In some embodiments, bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor result in reduced levels of acetaldehyde as described in sections I-E. In some embodiments, bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor result in reduced levels of acetaldehyde and increased levels of diacetyl as described in sections I-E. In some embodiments, the bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor produce a ratio of acetaldehyde to diacetyl related to the creamy flavor as described in sections I-E. In some embodiments, the bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor produce levels and/or ratios of acetaldehyde to diacetyl related to the creamy flavor as described in sections I-E.

In some embodiments, the bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor are or include lactobacillus rhamnosus strains. In some embodiments, the lactobacillus rhamnosus strain is a strain of vegetable origin. In some embodiments, the bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor are or include streptococcus thermophilus strains. In some embodiments, the bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor are or include lactobacillus rhamnosus strain and streptococcus thermophilus strain.

In some aspects, the composition contains one or more bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor. In some embodiments, the composition is a starter culture. In some embodiments, the starter culture further comprises one or more bacteria described in section I-B. In some embodiments, the composition is a fermented plant-based food. In some embodiments, the fermented plant-based food product is any one of those described in section 1-F.

III kit

Kits are also provided, including compositions (e.g., starter cultures) containing bacteria and bacterial compositions described herein, which may further include instructions for using the compositions (as described herein). The kits described herein may also include other materials desirable from a commercial and user perspective, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for performing any of the methods described herein. In some embodiments, the kits provided herein include bacteria and/or bacterial compositions capable of producing a fermented plant-based composition having a creamy flavor (e.g., increased creamy flavor).

Preservation and professional solutions

The following depositions are made in accordance with the budapest treaty on the international recognition of the deposit of microorganisms for the purposes of the patent procedure.

The Lactobacillus rhamnosus strain (DGCC 1179) deposited with DSMZ [ German collection of microorganisms and cell cultures, britain Hopfen street 7B, post code D-38124] under accession number DSM33650, 9/2020.

The Lactobacillus rhamnosus strain (DGCC 1460) deposited with DSMZ [ German collection of microorganisms and cell cultures, briorexin Hopofungin street 7B, post code D-38124] under accession number DSM22876, month 7, 2021.

The Lactobacillus rhamnosus strain (DGCC 13617) deposited with DSMZ [ German collection of microorganisms and cell cultures, briorexin Hopfen street 7B, post code D-38124] under accession number DSM33850 at 20 of 2021.

Streptococcus thermophilus strain (DGCC 11042) deposited with DSMZ [ German collection of microorganisms and cell cultures, briorexin Hopfen street 7B, post code D-38124] under accession number DSM 33651, month 9, 2020.

It is required that the biological material is obtained only by dispensing a sample to an expert designated by the applicant. For those designations seeking European patent, a sample of the deposited microorganism may be obtained before mention of an announcement granting European patent or before the application is refused or withdrawn, or is considered withdrawn, and such sample is only issued to the specialist designated by the person requesting the sample and is required to be informed of i) the applicant and/or ii) the European patent office (as the case may be) consent (European patent convention rule No. 32)

Exemplary embodiment

Examples provided are:

1. a method for producing a fermented plant-based composition having a creamy flavor, the method comprising reducing the acetaldehyde content in the fermented plant-based composition.

2. A method for producing a stored fermented plant-based composition having a creamy flavor, the method comprising:

(a) Storing the fermented plant-based composition to produce a stored fermented plant-based composition; and

(B) The acetaldehyde content in the fermented plant-based composition is reduced during storage.

3. The method of embodiment 1 or embodiment 2, wherein the method further comprises producing the fermented plant-based composition, the producing comprising fermenting a plant-based substrate inoculated with a starter culture.

4. A method for producing a fermented plant-based composition having a creamy flavor, the method comprising:

(a) Fermenting the plant base inoculated with the starter culture to produce a fermented plant-based composition; and

(B) Reducing the acetaldehyde content of the fermented plant-based composition.

5. The method of any one of embodiments 1-4, further comprising increasing the diacetyl content in the fermented plant-based composition.

6. The method of any of embodiments 3-5, wherein the vegetable base comprises soybean plant matter, optionally in an aqueous suspension.

7. The method of any one of embodiments 1 and 3-6, comprising storing the fermented plant-based composition to produce a stored fermented plant-based composition.

8. The method of embodiment 7, wherein the acetaldehyde content in the fermented plant-based composition is reduced during storage.

9. The method of embodiment 7 or embodiment 8, comprising increasing the diacetyl content of the fermented plant-based composition during storage.

10. The method of any of embodiments 1-9, wherein the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

11. The method of any of embodiments 5-10, wherein the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

12. The method of any of embodiments 3-11, wherein the starter culture comprises one or more lactic acid bacteria, optionally at least two lactic acid bacteria.

13. The method of any one of embodiments 3-12, wherein the starter culture comprises bacteria from the following genera: lactococcus, lactobacillus, streptococcus, lactobacillus, leuconostoc, pediococcus, or Bifidobacterium, or any combination of the foregoing.

14. The method of any one of embodiments 3-13, wherein the starter culture comprises one or more lactobacillus rhamnosus strains.

15. The method of embodiment 14, wherein the one or more lactobacillus rhamnosus strains comprises or consists of:

(a) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM33650 or a mutant thereof;

(b) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM22876 or a mutant thereof; and/or

(C) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM33850 or a mutant thereof.

16. The method of any one of embodiments 3-15, wherein the starter culture comprises one or more streptococcus thermophilus strains.

17. The method of embodiment 16, wherein the one or more streptococcus thermophilus strains comprises or consists of: the Streptococcus thermophilus strain deposited under DSMZ accession number DSM33651 or a mutant thereof.

18. The method of any one of embodiments 3-17, wherein the starter culture comprises or consists of:

(a) Lactobacillus rhamnosus strain or a mutant thereof deposited at the DSMZ under accession No. DSM33650, and streptococcus thermophilus strain or a mutant thereof deposited at the DSMZ under accession No. DSM 33651;

(b) Lactobacillus rhamnosus strain or a mutant thereof deposited at the DSMZ under accession No. DSM22876, and streptococcus thermophilus strain or a mutant thereof deposited at the DSMZ under accession No. DSM 33651; or (b)

(C) The strain of Lactobacillus rhamnosus or a mutant thereof deposited with DSMZ under accession number DSM33850, and the strain of Streptococcus thermophilus or a mutant thereof deposited with DSMZ under accession number DSM 33651.

19. The method of any one of embodiments 3-18, wherein the starter culture comprises one or more of: lactobacillus delbrueckii subsp bulgaricus strain, lactobacillus acidophilus strain, bifidobacterium lactis strain, lactobacillus mucilaginosus strain, lactobacillus paracasei strain, lactobacillus plantarum strain, propionibacterium freudenreichii strain, pediococcus acidilactici strain, lactococcus cremoris strain, lactococcus lactici subsp lactici strain, or lactococcus cremoris subsp.

20. The method of any one of embodiments 3-19, wherein the inoculated plant substrate is fermented at a temperature above about 33 ℃, 35 ℃, 36 ℃, 37 ℃, 38 ℃, 39 ℃, 40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃, or 50 ℃.

21. The method of any one of embodiments 2, 3 and 7-20, wherein the fermented plant-based composition is stored at about 1 ℃,2 ℃, 3 ℃,4 ℃,5 ℃, 6 ℃,7 ℃,8 ℃,9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃, 14 ℃,15 ℃, 16 ℃, 17 ℃, 18 ℃, 19 ℃, 20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, or about 26 ℃.

22. The method of any one of embodiments 2, 3, and 7-21, wherein the fermented plant-based composition is stored:

(a) At least about 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days; or (b)

(B) Up to about 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days.

23. The method of any one of embodiments 2,3 and 7-22, wherein the fermented plant-based composition is stored at about 6 ℃ for about 28 days.

24. The method of any one of embodiments 2, 3 and 7-22, wherein the fermented plant-based composition is stored at about 6 ℃ for about 14 days.

25. The method of any one of embodiments 2,3 and 7-22, wherein the fermented plant-based composition is stored at about 6 ℃ for about 7 days.

26. The method of any of embodiments 1-25, wherein the cream flavor comprises a cream aroma and/or a cream scent.

27. The method of any of embodiments 1-26, wherein the cream flavor is an increased cream flavor as compared to the cream flavor of a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of equal to or greater than 0.7.

28. The method of embodiment 27, wherein the increased cream flavor comprises an increased cream aroma and/or cream odor as compared to the cream aroma and/or cream odor of a fermented plant-based composition having an acetaldehyde content and a diacetyl content resulting in an acetaldehyde to diacetyl ratio of equal to or greater than 0.7.

29. A composition comprising a lactobacillus rhamnosus strain, wherein the lactobacillus rhamnosus strain reduces acetaldehyde in a fermented plant-based composition to produce a creamy flavour.

30. The composition of embodiment 29 wherein the lactobacillus rhamnosus strain is a strain of vegetable origin.

31. The composition of example 29 or example 30, wherein the lactobacillus rhamnosus strain is:

(b) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM22876 or a mutant thereof; or (b)

32. The composition of any one of embodiments 29-31, further comprising a streptococcus thermophilus strain.

33. The composition of example 32, wherein the streptococcus thermophilus strain is the streptococcus thermophilus strain deposited at the DSMZ under accession No. DSM33651 or a mutant thereof.

34. The composition of example 32 or example 33, comprising or consisting of:

35. The composition of any one of embodiments 29-34, further comprising one or more of the following: lactobacillus delbrueckii subsp bulgaricus strain, lactobacillus acidophilus strain, bifidobacterium lactis strain, lactobacillus mucilaginosus strain, lactobacillus paracasei strain, lactobacillus plantarum strain, propionibacterium freudenreichii strain, pediococcus acidilactici strain, lactococcus cremoris subsp lactici strain, or lactococcus lactici subsp lactici strain.

36. The composition of any one of embodiments 29-35, wherein the composition is a starter culture or a fermented plant-based food.

37. Use of lactobacillus rhamnosus to reduce the acetaldehyde content in a fermented plant-based composition to produce a creamy flavour.

38. The use of example 37, wherein the lactobacillus rhamnosus strain is:

39. The composition of any one of embodiments 29-36 or the use of embodiment 37 or embodiment 38, wherein the lactobacillus rhamnosus increases the diacetyl content in the fermented plant-based composition.

40. The composition of any of embodiments 29-36 and 39 or the use of any of embodiments 37-39, wherein the acetaldehyde content is reduced such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

41. The composition of embodiment 39 or embodiment 40 or the use of embodiment 39 or embodiment 40, wherein the acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

42. The composition of any one of embodiments 29-36 and 39-41 or the use of any one of embodiments 37-41, wherein the cream flavor comprises a cream aroma and/or a cream scent.

43. The composition of any one of examples 29-36 and 39-42 or the use of any one of examples 37-42, wherein the cream flavor is an increased cream flavor as compared to a cream flavor of a fermented plant-based composition having an acetaldehyde to diacetyl ratio of equal to or greater than 0.7 due to acetaldehyde content and diacetyl content.

44. The composition of example 43 or the use of example 43, wherein the increased cream flavor comprises an increased cream aroma and/or cream odor compared to a cream aroma and/or cream odor of a fermented plant-based composition having an acetaldehyde to diacetyl content resulting in an acetaldehyde to diacetyl ratio of equal to or greater than 0.7.

45. A fermented plant-based composition obtained by the method according to any one of embodiments 1-28 or the use according to any one of embodiments 37-44.

46. The fermented plant-based composition of embodiment 45, wherein the fermented plant-based composition is a fermented plant-based food, optionally a fermented plant-based milk-replacable food.

47. The fermented plant-based composition of embodiment 45 or embodiment 46, wherein the fermented plant-based composition is a plant-based yogurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative.

48. A fermented plant-based food product having a creamy flavor, the fermented plant-based food product comprising an acetaldehyde content to diacetyl content ratio of less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

49. The fermented plant-based food with cream flavor of example 48, wherein the fermented plant-based food comprises a lactobacillus rhamnosus strain that reduces acetaldehyde in the fermented plant-based food.

50. The fermented plant-based product with creamy flavor of example 48 or example 49, wherein the lactobacillus rhamnosus strain increases diacetyl in the fermented plant-based food.

51. The fermented plant-based product with creamy flavor of any one of embodiments 48-50, wherein the lactobacillus rhamnosus strain is:

(a) The lactobacillus rhamnosus strain deposited under accession number DSM33650 or a mutant thereof;

(b) The lactobacillus rhamnosus strain deposited under accession number DSM22876 or a mutant thereof; and/or

(C) The lactobacillus rhamnosus strain deposited under accession number DSM33850 or a mutant thereof.

52. The fermented plant-based food product with cream flavor of any one of embodiments 48-51, further comprising a streptococcus thermophilus strain.

53. The fermented plant-based food product with cream flavor of example 52, wherein the streptococcus thermophilus strain is the streptococcus thermophilus strain deposited under accession No. DSM33651 or a mutant thereof.

54. The fermented plant-based food with cream flavor of any one of embodiments 48-53, wherein the fermented plant-based food is a fermented plant-based milk-replaceable food.

55. The fermented plant-based food product of any one of embodiments 48-54, wherein the fermented plant-based food product is a plant-based yoghurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative.

56. A method of identifying bacteria capable of producing a fermented plant-based composition having a creamy flavor, the method comprising quantifying the acetaldehyde content and the diacetyl content in a fermented plant-based composition fermented in the presence of one or more bacteria.

57. The method of example 56, wherein the fermented plant-based composition is stored and the acetaldehyde content and diacetyl content are quantified during storage.

58. The method of example 56 or example 57, wherein the one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the acetaldehyde content and diacetyl content are quantified such that the ratio of acetaldehyde content to diacetyl content is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

59. The method of any one of embodiments 56-58, wherein the one or more bacteria comprises lactic acid bacteria.

60. The method of any one of embodiments 56-59, wherein the one or more bacteria comprises a lactobacillus rhamnosus strain, optionally wherein the lactobacillus rhamnosus strain is a plant-derived strain.

61. The method of any one of embodiments 56-60, wherein the one or more bacteria comprises a streptococcus thermophilus strain.

62. The method of any one of embodiments 10-28 and 56-61, the composition of any one of embodiments 40-44, the use of any one of embodiments 40-44, the fermented plant-based composition of any one of embodiments 45-47, or the fermented plant-based food of any one of embodiments 48-55, wherein the acetaldehyde to diacetyl ratio is achieved after 28 days or less of storage.

63. The method of any one of embodiments 10-28 and 56-62, the composition of any one of embodiments 40-44 and 62, the use of any one of embodiments 40-44 and 62, the fermented plant-based composition of any one of embodiments 45-47 and 62, or the fermented plant-based food of any one of embodiments 48-55 and 62, wherein the acetaldehyde to diacetyl ratio is achieved after 21 days or less of storage.

64. The method of any one of embodiments 10-28 and 56-63, the composition of any one of embodiments 40-44, 62 and 63, the use of any one of embodiments 40-44, 62 and 63, the fermented plant-based composition of any one of embodiments 45-47, 62 and 63, or the fermented plant-based food of any one of embodiments 48-55, 62 and 63, wherein the acetaldehyde to diacetyl ratio is achieved after 14 days or less of storage.

65. The method of any one of embodiments 10-28 and 56-64, the composition of any one of embodiments 40-44 and 62-64, the use of any one of embodiments 40-44 and 62-64, the fermented plant-based composition of any one of embodiments 45-47 and 62-64, or the fermented plant-based food of any one of embodiments 48-55 and 62-64, wherein the acetaldehyde to diacetyl ratio is achieved after storage for 7 days or less.

66. A composition comprising one or more bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor according to the method of any one of examples 56-65.

67. The composition of embodiment 66, wherein the composition is a starter culture or a fermented plant-based food, optionally a fermented plant-based milk-replacer.

68. A kit, the kit comprising:

(a) The composition of any one of embodiments 29-36, 40-44, and 62-67; and

(B) Instructions for use.

69. A strain of lactobacillus rhamnosus deposited with the DSMZ under accession No. DSM33850 or a mutant thereof.

70. A strain of lactobacillus rhamnosus deposited under accession No. DSM22876 or a mutant thereof.

71. A streptococcus thermophilus strain deposited under the accession number DSMZ 33651 or a mutant thereof at the DSMZ.

V. examples

The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.

Example 1: production of volatile organic compounds related to creamy flavor in fermented plant-based yoghurt alternatives

In a fermented plant-based yoghurt alternative product, the ability to produce the levels of the volatile organic compounds acetaldehyde and diacetyl associated with the flavour of cream was assessed using a bacterial composition.

A. Materials and methods

Substrate(s)

Exemplary soy substrates for producing thermophilic fermented plant-based yoghurt alternatives were evaluated. Table E1 describes the raw materials and plant substrates used for fermentation.

Table E1: characteristics of exemplary soy raw materials and plant-based substrates for fermentation.

Thermophilic fermentation

Soy plant substrate (1L) as described in table E1 was inoculated with streptococcus thermophilus DSM33651 alone or in combination with one of the following four exemplary lactobacillus rhamnosus strains: DSM33650, DSM33850, DSM22876, or DSM24616. Streptococcus thermophilus was inoculated at 2U/100L and Lactobacillus rhamnosus strain was inoculated at 1X10 ⁷ CFU/mL (frozen form).

The inoculated substrate was gently stirred and dispensed into 8x125mL yoghurt cups. The cups were placed in a 43℃water bath for fermentation. Once the plant substrate reached a pH of 4.6, the yoghurt cup was stored at 6 ℃ for 28 days.

Volatile organic compounds

After storage at 6 ℃ for 1, 7, 14, 21 and 28 days, volatile organic compound analysis was performed on the fermented plant substrate.

Volatile organic compounds were analyzed by gas chromatography-mass spectrometry (GC-MS) (Sesameimer's technologies Co., ltd. (ThermoFisher Scientific); TRACE1300GC coupled to ISQ mass detector).

Samples were prepared in 10mL headspace vials by combining 1g NaCl, 2g fermentation substrate, 1mL MilliQ cooling water, and 100 μl internal standard (25 ppm butyl acetate solution). The sample was vortexed and then placed on a refrigerated autosampler of gas chromatography.

After heating (80 ℃ for 15 min) and shaking, volatile organic compounds were extracted from the samples by ITEX (dynamic headspace). The ITEX traps were then desorbed in a GC-MS syringe at 200 ℃. Molecules were separated on a ZB-Wax (60m x0.25mm x0.25 μm) column from Feinomex (Phenomenex) at 1mL/min with helium as carrier gas.

The volatile organic compounds were ionized by electron impact (70 eV) and detected by quadrupole mass spectrometry in a full scan mode of 30 to 200 amu.

The key molecules of interest (including diacetyl and acetaldehyde) were quantified by external calibration and expressed in ppm, with calibration using internal standards. Samples were injected in triplicate.

B. results

At various time points during storage of the fermented soy-based thermophilic yoghurt alternative, diacetyl (2, 3-butanedione) and acetaldehyde content were detected using GC-MS as described above. Figure 1 shows a scatter plot of the 2, 3-butanedione and acetaldehyde content (in ppm) after 1, 7, 14, 21 and 28 days at 6 c for five soy-based yoghurt alternatives. The 2, 3-butanedione and acetaldehyde content of the soybean substrate prior to inoculation and fermentation are also shown.

As shown in fig. 1, after one day of storage, the acetaldehyde levels of all yoghurt substitutes were similar to the 2, 3-butanedione levels measured in the corresponding yoghurt substitutes. However, the acetaldehyde content is generally higher in yogurt alternatives fermented with streptococcus thermophilus alone or in combination with DSM24616 than in other yogurt alternatives.

Between day 1 and day 7 of storage, a decrease in acetaldehyde content was observed in the yoghurt alternative fermented with a combination of streptococcus thermophilus with one of the lactobacillus rhamnosus strains (DSM 33650 or DSM 22876). For these yogurts, after 7 days, the level of acetaldehyde was generally maintained, and an increase in 2, 3-butanedione was observed.

On days 14, 21 and 28 of storage, yogurt alternatives fermented with streptococcus thermophilus alone or in combination with one of the lactobacillus rhamnosus strains (DSM 24616 or DSM 33850) show similar 2, 3-butanedione levels, which are higher compared to other soy yogurt alternatives. However, the acetaldehyde level of the yogurt alternative fermented with a combination of streptococcus thermophilus and DSM33850 is lower than that of yogurt fermented with streptococcus thermophilus alone or in combination with DSM 24616. See fig. 1.

These results support that compositions comprising streptococcus thermophilus and lactobacillus rhamnosus (including exemplary strains DSM33650, DSM33850 or DSM 22876) are capable of reducing the acetaldehyde content to produce acetaldehyde and diacetyl levels associated with cream flavor (e.g., cream odor and cream aroma) in fermented soy-based thermophilic yogurt alternatives. These results also demonstrate that inclusion of lactobacillus rhamnosus strains during fermentation can promote a change in these volatile compounds, thereby imparting a creamy flavor to the fermented soy-based thermophilic yogurt alternative.

Example 2: production of volatile organic compounds related to creamy flavor in fermented plant-based yoghurt substitutes produced using industrial starter cultures

In a fermented plant-based yoghurt alternative product, the ability to produce levels of the volatile organic compounds acetaldehyde and diacetyl associated with cream flavour using a combination of an industrial starter culture with an exemplary strain of lactobacillus rhamnosus was evaluated.

A. Materials and methods

Substrate(s)

Exemplary soy substrates for use in producing a thermophilic fermented yoghurt alternative were evaluated. Table E2 describes the raw materials and plant substrates used for fermentation.

Table E2: characteristics of exemplary Soybean raw materials and plant-based substrates for fermentation

Thermophilic fermentation

The plant substrate was inoculated with one of two industrial starter cultures each containing streptococcus thermophilus and lactobacillus delbrueckii subsp bulgaricus, alone or in combination with the exemplary lactobacillus rhamnosus strain DSM 33650. Industrial starter cultures were inoculated at 20DCU/100L and Lactobacillus rhamnosus strains were inoculated at 1X10 ⁷ CFU/mL.

The inoculated substrate was gently stirred and placed at 40℃for fermentation. Once the plant substrate reached a pH of 4.6, the yoghurt cup was stored for 7 days at 6 ℃.

After 7 days of storage at 6 ℃, the volatile organic compound analysis was performed as described in example 1.

B. results

For each fermented soy-based thermophilic yoghurt alternative, after 7 days of storage, diacetyl (2, 3-butanedione) and acetaldehyde content were detected using GC-MS as described above. FIG. 2 shows a scatter plot of 2, 3-butanedione and acetaldehyde content (in ppm) after storage at 6℃for 7 days.

As shown in fig. 2, after 7 days of storage, the acetaldehyde and 2, 3-butanedione content of the yoghurt alternative fermented with the combination of industrial starter culture and lactobacillus rhamnosus strain DSM33650 was similar (about 3.0ppm of 2, 3-butanedione and less than 1.0ppm of acetaldehyde). In a yoghurt alternative fermented with a separate industrial starter culture, the acetaldehyde content is higher than the diacetyl content. The diacetyl level of the yogurt alternatives fermented with the industrial starter culture alone is lower than the yogurt alternatives fermented with the combination of the industrial starter culture with the exemplary lactobacillus rhamnosus strain.

These results support the ability of lactobacillus rhamnosus strains to reduce the acetaldehyde content and increase the diacetyl content in fermented soy-based yoghurt substitutes even when used in combination with industrial starter cultures.

Example 3: evaluation of creamy flavor of fermented plant-based yoghurt

The ability to naturally produce and improve cream flavor in fermented plant based yogurt alternatives will be assessed by using bacteria including industrial starter cultures (containing streptococcus thermophilus and lactobacillus delbrueckii subsp. Bulgaricus) and exemplary lactobacillus rhamnosus strains (including DSM 33650).

Plant-based soybean substrates were inoculated, fermented and stored as generally described in example 2 above. Sensory analysis will be performed by a panel of trained sensory evaluators who will evaluate fermented plant-based yoghurt substitutes via Quantitative Descriptive Analysis (QDA) using a linear point scale. Statistical tests will be performed.

Sensory analysis of the fermented plant-based yoghurt alternative will be determined at different time points during storage.

These results will support the correlation between the reduced level of acetaldehyde relative to diacetyl levels in the plant-based fermented yoghurt and the cream flavour. These results will further indicate that inclusion of exemplary bacteria, and in particular exemplary lactobacillus rhamnosus strains, during fermentation may promote a change in volatile compounds, thereby improving the creamy flavor of the fermented plant-based yoghurt alternative.

The scope of the invention is not limited to the specific embodiments disclosed, which are provided, for example, to illustrate various aspects of the invention. Various modifications to the compositions and methods will be apparent from the description and teachings herein. Such changes may be made without departing from the true scope and spirit of the disclosure, and such changes are intended to fall within the scope of the disclosure. Although the invention may be described in connection with certain preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims.

As shown in fig. 3, after 14 days of storage, the acetaldehyde content of the soy yoghurt alternative fermented with the industrial starter culture alone was higher than the soy yoghurt alternative fermented with the combination of the industrial starter culture with lactobacillus rhamnosus strain DSM 33650. After 14 days of storage, the 2, 3-butanedione content of the soy yoghurt alternative fermented with the industrial starter culture alone is lower than that of the soy yoghurt alternative fermented with the combination of the industrial starter culture with lactobacillus rhamnosus strain DSM 33650. In soy yoghurt alternatives where fermentation is performed with a separate industrial starter culture, the acetaldehyde content is higher than the diacetyl content. In soy yoghurt substitutes fermented with industrial starter cultures with exemplary lactobacillus rhamnosus strains, the acetaldehyde content is lower than the diacetyl content.

The sensory panel perceived a higher cream/milk odor in the soy yoghurt alternative fermented with the combination of industrial starter culture and lactobacillus rhamnosus strain DSM33650 than in the soy yoghurt alternative fermented with the industrial starter culture alone.

These results support the ability of the lactobacillus rhamnosus strain to reduce the acetaldehyde content and increase the diacetyl content in the fermented soy-based yoghurt alternative, as well as to increase the perception of cream/milk smell.

These results demonstrate that inclusion of exemplary bacteria, and in particular exemplary lactobacillus rhamnosus strains, during fermentation promotes a change in volatile compounds, thereby improving the creamy flavor of the fermented plant-based yoghurt alternative.

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Claims

1. A method for producing a fermented plant-based composition having a creamy flavor, the method comprising:

(B) Reducing the acetaldehyde content in the fermented plant-based composition.

2. The method of claim 1, comprising storing the fermented plant-based composition.

3. The method of claim 2, wherein

(A) Reducing the acetaldehyde content in the fermented plant-based composition during storage such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0; or (b)

(B) The acetaldehyde content is reduced and the diacetyl content is increased during storage such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

4. A method according to any one of claims 1-3, wherein the starter culture comprises one or more strains of lactobacillus rhamnosus (Lacticaseibacillus rhamnosus); optionally wherein the one or more lactobacillus rhamnosus strains comprise or consist of:

5. The method of any one of claims 1-4, wherein the starter culture comprises one or more streptococcus thermophilus (Streptococcus thermophilus) strains, optionally wherein the one or more streptococcus thermophilus strains comprises or consists of: the Streptococcus thermophilus strain deposited under DSMZ accession number DSM33651 or a mutant thereof.

6. The method of any one of claims 1-5, wherein the starter culture comprises or consists of:

7. The method of any one of claims 2-6, wherein the fermented plant-based composition is:

(a) Stored at about 1 ℃,2 ℃,3 ℃, 4 ℃, 5 ℃,6 ℃,7 ℃,8 ℃, 9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃, 14 ℃,15 ℃, 16 ℃, 17 ℃, 18 ℃, 19 ℃,20 ℃, 21 ℃, 22 ℃, 23 ℃, 24 ℃, 25 ℃, or about 26 ℃; and

(B) Storing for at least about 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days; or (b)

(C) Stored for up to about 1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、40、50、60、70、80、90、100、110 or 120 days.

8. A composition comprising a lactobacillus rhamnosus strain, wherein the lactobacillus rhamnosus strain reduces acetaldehyde in a fermented plant-based composition to produce a creamy flavor; optionally, wherein

(A) Reducing the acetaldehyde content such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0; or (b)

(B) Decreasing the acetaldehyde content and increasing the diacetyl content such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0; and

Optionally wherein the composition is a starter culture or a fermented plant-based food.

9. Use of lactobacillus rhamnosus to reduce the acetaldehyde content in a fermented plant-based composition to produce a creamy flavour; optionally, wherein

(B) The acetaldehyde content is reduced and the diacetyl content is increased such that the ratio of acetaldehyde to diacetyl is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

10. The composition of claim 8 or the use of claim 9, wherein the lactobacillus rhamnosus strain is:

11. A fermented plant-based composition obtained by the method of any one of claims 1-7 or the use of claim 9 or claim 10, optionally wherein the fermented plant-based composition is a fermented plant-based food product, optionally a plant-based yoghurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative.

12. A fermented plant-based food product having a cream flavor, the fermented plant-based food product comprising an acetaldehyde content to diacetyl content ratio of less than 0.7, 0.6, 0.5, 0.25, 0.1, 0.075, 0.05, 0.025, or 0.01, or being 0, optionally wherein the fermented plant-based food product is a plant-based yogurt alternative, a plant-based cream alternative, a plant-based mature cream alternative, a plant-based butter alternative, a plant-based fat paste, a plant-based cheese alternative, a plant-based low-fat fresh cheese alternative, a plant-based milk beverage alternative, a plant-based processed cheese alternative, a plant-based cream dessert alternative, a plant-based cottage cheese alternative, or a plant-based kefir alternative.

13. A method of identifying bacteria capable of producing a fermented plant-based composition having a creamy flavor, the method comprising quantifying the acetaldehyde content and the diacetyl content in a fermented plant-based composition fermented in the presence of one or more bacteria, optionally wherein the fermented plant-based composition is stored, and the acetaldehyde content and diacetyl content are quantified during storage; and

The one or more bacteria are identified as being capable of producing a fermented plant-based composition having a creamy flavor if the acetaldehyde content and the diacetyl content are quantified such that the ratio of acetaldehyde content to diacetyl content is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.

14. A composition comprising one or more bacteria identified as being capable of producing a fermented plant-based composition having a creamy flavor according to the method of claim 13, optionally wherein the composition is a starter culture or a fermented plant-based food.

15. A bacterial strain, wherein the strain is:

(a) The lactobacillus rhamnosus strain deposited at the DSMZ under accession No. DSM33850 or a mutant thereof; or (b)

(B) The Streptococcus thermophilus strain deposited under DSMZ accession number DSM33651 or a mutant thereof.