Cacao cell agglomerates and methods of agglomerating the cacao cells

Abstract

A method of processing cellular biomass, e.g., cell culture cacao cells, by cellular agriculture is disclosed. A fluidized bed dryer is used to dry and agglomerate the cellular biomass. The process includes classifying the cell agglomerates to generate optimum sized cell agglomerates to increase the uniformity of the agglomerates. The cellular biomass can be fermented prior to agglomeration. The process also includes roasting the agglomerates. Compositions with the roasted agglomerates and cocoa products derived from the agglomerates are described.

Description

PT-2170-WO-PCTCACAO CELL AGGLOMERATES AND METHODS OF AGGLOMERATING THE CACAO CELLSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of European Application No. 25152663.8, filed January 17, 2025, which is incorporated by reference herein in its entirety.BACKGROUND

[0002] Cocoa is obtained from cacao beans that are harvested from cacao trees grown in tropical regions. The beans are processed to extract cocoa solids and cocoa butter, which are the primary components used in chocolate production. Cocoa beans, once processed, make the basis for chocolate candy and numerous desserts.

[0003] The global cacao production is facing serious challenges including land use, climate change, and sustainability while demand is rising. Cellular agriculture is a promising alternative to produce plant-based commodities such as coffee and cacao, which are conventionally produced by farming. Cellular agriculture is cell-culture based growth of plant cells in an external environment, generally a laboratory or manufacturing environment.

[0004] Cell-culture based chocolate is a new type of chocolate that involves growing cacao plant cells in a laboratory or a manufacturing facility instead of harvesting cacao pods from cacao farms. The process uses advanced cellular agriculture techniques to grow cacao cells. Cell-culture based cocoa has the potential to reduce many of the environmental impacts associated with traditional cacao farming, including deforestation, habitat loss, greenhouse gas emissions and lower carbon footprint. While promising, the true impact and cost of the various inputs needed to grow the cell cultures and the byproducts produced by such a system won’t be fully understood until these new methods are scaled up for mass production.

[0005] The flavor of cocoa is very complex, with various volatile and non-volatile compounds responsible for the resulting sensory properties. The chemical composition of cocoa is influenced by numerous variables, including cultivar, soil composition and growing climate conditions. Cocoa flavor and textures constitute the most important parameters for the consumer.

[0006] Production of the same complex flavors and textures in cell-culture based chocolate similar to those found in traditional chocolate derived from cacao beans remains a challenge.PT-2170-WO-PCTSUMMARY

[0007] The present disclosure also provides a method of processing a cellular biomass. The method comprises introducing a cellular biomass into a dryer, drying the cellular biomass in the dryer at selected drying parameters, wherein agglomerates comprising the cells from the cellular biomass are formed during the drying, classifying the size of the agglomerates by collecting agglomeration size data from agglomeration sensors in the dryer, and separating / sorting optimum cell agglomerates from sub-optimal cell biomass after a threshold level of agglomerates of optimum size are generated.

[0008] The dryer may be a fluidized bed dryer. The cellular biomass may be pre-dried prior to introduction into the dryer. The method may further comprise roasting the optimum cell agglomerates. The method may further comprise fermenting, alkalizing, pressing, refining, conching, tempering and / or molding. The method may further comprise reintroducing the sub-optimal cell biomass to fluidized bed dryer and / or discarding the sub-optimal cell biomass. The drying parameters may comprise temperature, air velocity, air moisture content and headspace gas composition. The temperature in the dryer may be between about 40°C and 160°C. The air velocity is between about 2m / s and about 25m / s. The air moisture content is between about 0% and about 60%. The headspace gas composition may be atmospheric gas composition. The optimum cell agglomerates may be at least about 1mm, preferably between about 1mm and 10mm. The optimum cell agglomerates comprise agglomerates that are substantially uniform in size. At least 70% of the agglomerates may have a diameter that is within 30% of the mean agglomerate size. The agglomeration sensors may comprise laser diffraction sensors and / or image analysis sensors. The roasting may comprise transfer of the optimum cell agglomerates to a fluidized bed roaster, drum roaster and / or hot air roaster. The roasting comprises heating the optimum cell agglomerates to a temperature of between about 105°C and about 130°C for a duration of between about 20 minutes and 60 minutes.

[0009] The disclosure also provides compositions containing optimum cell agglomerates, wherein the agglomerates comprise plant cells, wherein the plant cells are cell cultured cacao cells. The agglomerates comprise a diameter of at least about 1mm, preferably between about 1mm and 10mm. The optimum cell agglomerates may be roasted optimum cell agglomerates. The optimum cell agglomerates may be fermented optimum cell agglomerates. The composition may be a cocoa product, wherein the product is derived from the roasted optimum cell agglomerates. The cocoa product may be cocoa powder, chocolate bar, and / or chocolate cake.PT-2170-WO-PCT

[0010] The disclosure further provides a cellular biomass processing system comprising a dryer comprising an inlet for cellular biomass, agglomeration sensors, and at least one outlet for removal of optimum cell agglomerates, wherein the dryer comprises circuitry for receiving input comprising drying parameter data from a drying control parameter database, wherein agglomeration sensors comprise circuitry for determining level of agglomeration and wherein the dryer comprises component for separation of optimum cell agglomerates from sub-optimum sized agglomerates.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The drawings illustrate generally, by way of example, but not by way of limitation, various aspects discussed herein.

[0012] FIG. 1 shows a schematic diagram of a system for processing cell cultured plant cells.

[0013] FIG. 2 shows a schematic diagram of the steps in a method of processing cell cultured plant cells.

[0014] FIG. 3 shows a schematic diagram for the classification of the agglomerates to generate optimum uniform cell agglomerates.DETAILED DESCRIPTION

[0015] Reference will now be made in detail to certain aspects of the disclosed subj ect matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0016] This disclosure relates to a method of processing cell cultured plant cells, e.g., cell culture cacao cells, by cellular agriculture. The processing of cell cultured plant cells can occur after cultivation of the cells in nutrient media. The process may include separating the cell cultured plant cells from the nutrient media. The process can optionally include fermenting the cellular biomass. The fermentation step can occur after the cellular biomass is separated from the nutrient media used during cultivation.

[0017] The method can include generating optimum cell agglomerates from a cellular biomass that includes cell culture-based cacao cells. The cellular biomass may or may not have been fermented prior to agglomeration. The method can also include alkalization of the cellular biomass. The alkalization of the cellular biomass can occur prior to the agglomeration step orPT-2170-WO-PCTduring the agglomeration step. The disclosure also relates to a system, e.g., a fluidized bed dryer, for processing the cellular biomass and / or agglomerating the plant cells. The dryer can dry the cellular biomass and generate agglomerates during the drying process. The system can classify the agglomerates by size / shape and separate optimum sized cell agglomerates from sub-optimal cell biomass. The system may also include a roaster that may be incorporated within the dryer to roast the optimum sized cell agglomerates.

[0018] This disclosure also relates to a composition comprising cellular biomass comprising plant cells, e.g., cocoa cells. In one aspect, the composition comprises cocoa cell agglomerates, preferably optimum sized cell agglomerates. The composition may comprise roasted optimum sized plant cell agglomerates, preferably roasted cocoa cell agglomerates. The composition may comprise fermented and roasted optimum sized plant cell agglomerates, preferably fermented, and roasted cocoa cell agglomerates. The composition may comprise fermented, alkalized, and roasted optimum sized plant cell agglomerates, preferably fermented, alkalized, and roasted cocoa cell agglomerates. The composition may also comprise cocoa products derived from the fermented, alkalized and / or roasted plant cell agglomerates, e.g., cocoa powder comprising fermented, alkalized and / or roasted cocoa cell agglomerates.

[0019] The terms “cell biomass” and “cellular biomass” refers to cells derived from a cell culturing process outside of their natural environment, e.g., grown in a laboratory or in a manufacturing environment. These two terms will be used interchangeably. The cells are generally single cells and / or small clusters. Small clusters, as used herein, relate to association of cells during cultivation or harvesting and can be associated due to the presence of cellulose, hemicellulose, etc. on the cells. Small clusters are generally less than 1000 um. The cells in the cellular biomass may be fermented cells or unfermented cells. The cellular biomass may be fermented, for example, in the presence of a combination of yeast, bacteria and the like.

[0020] The term “agglomerates” as used herein refers to particles / clusters comprising cells from a cellular biomass that are assembled by adhesion of the cells / small clusters. The agglomerates are at least 1mm or larger. The cells in the agglomerates may be adhered or bridged together by proteins, sugars, carbohydrates, starch and the like. Agglomerates have improved hydration properties and flow characteristics and can be more easily wetted by aqueous solutions. Agglomerates can be processed in methods that are similar to methods for processing cocoa nibs.

[0021] The terms “size of the agglomerate” or “diameter” as used herein refers to the largest dimension of the agglomerate. In one aspect, in a spherical agglomerate the size of the agglomerate is the diameter of the sphere.PT-2170-WO-PCT

[0022] The term “optimum cell agglomerates” or “optimum size cell agglomerates” as used herein refers to agglomerates that are sized such that roasting the agglomerates results in desirable traits and does not lead to burning of the agglomerates. The optimum cell agglomerates can be, for example, at least 1mm or larger. The optimum cell agglomerates comprise uniformly sized agglomerates. Agglomerates of similar size can provide roasting uniformity and reducing the generation of burned or over roasted flavors.

[0023] The term “uniform size” or “uniformly sized” as used herein relates to agglomerates having a minimal or narrow size distribution range. A narrow size distribution range in an agglomerate sample / population provides more roasting uniformity. By uniform, it is meant that at least 60%, or at least 65%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates are within 40% of the mean agglomerate size; or that at least 60%, or at least 65%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates are within 30% of the mean agglomerate size; or that at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates are within 20% of the mean agglomerate size, or that at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates are within 10% of the mean agglomerate size. The difference in the diameter between the largest agglomerate and the smallest agglomerate is preferably minimized. The difference in the diameter between the largest agglomerate and the smallest agglomerate in a uniformly sized cell agglomerates is at most 10mm, or at most 8mm, or at most 5mm, or at most 3mm, or at most 2mm. The difference in the diameter between the largest agglomerate and the smallest agglomerate in the uniformly sized cell agglomerates can be between about 2mm and 10mm, or between 2mm and 8mm, or between 2 mm and 5mm.

[0024] The term “sub-optimal cell biomass” as used herein refers to cell biomass that has gone through at least one agglomeration cycle but does not contain the optimum cell agglomerates and is significantly smaller or larger in size than the median agglomerates. The sub-optimal cell biomass can be discarded and / or recycled back to the dryer to enter another agglomeration process if too small, or ground / broken if too large.

[0025] The term “cultivation” as used herein refers to generation or growth of plant cells, e.g., as a cell suspension, in a nutrient medium.

[0026] Cocoa products produced from cellular biomass can be significantly affected by the biomass particle size. Optimum cell agglomerates formed from the cellular biomass can lead to higher efficiency in subsequent processing steps and retain desirable flavors. Optimum sized cellPT-2170-WO-PCTagglomerates can mitigate the potential for unwanted burned / over roasted flavors and increase the ability to retain / develop desirable compounds and moisture levels. Smaller than optimum size (compared to traditional bean or nib cocoa roasting) particles are likely to be over roasted / burned. This can lead to undesirable flavors due to the lack of internal moisture content, a reduced ability to retain desirable volatile compounds and can also generate bitter, smoky, and burned flavors. Larger sized cellular biomass particles may have too much moisture and may not result in sufficient roasting to generate the desired profile.

[0027] The present description relates to a method of producing cell agglomerates, e.g., cocoa cell agglomerates, from cellular biomass. The cell agglomerates can be optimally sized agglomerates. Optimally sized agglomerates can include agglomerates of relatively uniform size. Optimally sized agglomerates can include at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates that are within 30% of the mean agglomerate size. Optimum size agglomerates may also comprise at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 20% of the mean agglomerate size. Optimum size agglomerates may also comprise at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 10% of the mean agglomerate size. Roasting of these optimally sized and cocoa cell agglomerates with a uniform size distribution can result in reducing and / or eliminating the undesirable sensory profiles, e.g., undesirable flavors and / or undesirable volatile compounds.

[0028] Advantageously, the invention described herein enables the agglomerate size to be controlled during a drying step such that subsequent processing steps such as roasting may be more efficient and retain desirable flavors. The cocoa product from the roasted cell agglomerates can have a reduced potential for unwanted burned / over roasted flavors due to a smaller particle size distribution range and due to an increased ability to retain / develop desirable compounds and moisture levels. The optimized process can enable efficient roasting due to the optimization of agglomerate size and flavor retention. The process may decrease roasting time and increase flavor retenti on / product! on .

[0029] Without being bound by any theory, it is thought that a fairly uniform particle size distribution in the agglomerates can ensure that there are not over or under sized agglomerates resulting in the aforementioned issues. In addition, agglomerates actively formed by an agglomeration process often exhibit a porous morphology as a result of their fractal structure, with the pores forming capillaries that can facilitate venting of steam or undesirable volatiles, and / or ingress of reagents or catalysts such as alkalizing agents.PT-2170-WO-PCTSystems for processing cellular biomass

[0030] In one aspect, the present description relates to a system for processing cellular biomass. The system comprises a dryer that can be configured to control the agglomeration of the cellular biomass, preferably, while drying to obtain optimum size and / or shapes of the agglomerates. The system may include a roaster to roast the optimum size agglomerates. The roaster may be separate from the dryer. Alternatively, the roaster may be integrated in the dryer / agglomerator.

[0031] In one aspect, the system may be manually operated to control the agglomeration by providing and / or adjusting the drying parameters. The level of agglomeration and removal of optimum cell agglomerates from the system may be provided manually by an operator, through the manual adjustment of operating conditions such as temperatures, flow rates and moisture content as desired by the operator.

[0032] The processing system used in the methods described herein may include, for example, a fluidized bed dryer / roaster, a hot air drum dryer and roaster, a forced convection continuous tumble roaster, a spray drying mechanism and the like. The present invention will be described in the context of a fluidized bed dryer, but it will be understood other types of processing systems with different types of dryers and / or roasters may also be used and are within the scope of this description.

[0033] Fig. 1 is a schematic diagram of an exemplary embodiment of a cellular biomass processing system wherein system (100) comprises fluidized bed dryer (120). Fluidized bed dryers are known in the art and are described, for example, in Jafari et al. (2023). Drying Technology in Food Processing - Unit Operations and Processing Equipment in the Food Industry - “Fluidized bed dryers: 4.1 Introduction, (pp. 67). Elsevier.

[0034] The system comprises an inlet or opening for receiving the cellular biomass into the dryer. The present disclosure will be described in the context of an automated system but the processes described herein may also be carried out in a manually operated system. An automated system and / or a manually operated system are within the scope of this description.

[0035] In one aspect, the cellular biomass enters dryer (120) through an inlet. The processing system may comprise a de-watering component and / or a pre-drying component (110). The cellular biomass added to dryer (120) may have been pre-dried in a pre-drying process using a variety of pre-drying devices prior to being added to dryer (120).

[0036] In one aspect, the cellular biomass can, optionally, be fermented prior to drying by entry into the inlet of dryer (120). The cellular biomass can be fermented by placing the cellularPT-2170-WO-PCTbiomass in a fermentation solution and providing conditions that enable fermentation of the cellular biomass and the appropriate starter cultures.

[0037] In one aspect, the processing system further comprises drying control (130) operably connected to and / or integrated within the dryer. Drying control (130) comprises a drying parameter database. Alternatively, the drying parameters can be provided to the dryer by manually setting and / or controlling the desired parameters.

[0038] The processing system comprises circuitry to receive input related to the desired drying parameters that can be applied during the drying of the cellular biomass in dryer (120). The drying control parameter database can be controlled or configured by a user to provide the desired drying parameter data to the dryer during the drying of the cellular biomass. The drying parameters selected can be dependent on the size of the cellular biomass, the type of cellular biomass, the level of hydration of cellular biomass, presence of solutes like sugars in the intercellular liquid, presence of fermentation solution and the like. The cellular biomass that has not been subject to pre-drying and is in a suspension can require different parameters than a cellular biomass that has been pre-dried to a wet form or a powder form. Drying control parameter database can include, for example, drying parameter data related to drying air temperature, drying air velocity, drying air moisture content / humidity, mixing rate or the mechanical shear forces applied when drying, time of drying and the like. The selection of the agglomerating conditions such as the type of binders, air temperature, flow rates and the like can be manipulated to generate the desired agglomerates with the desired porosity.

[0039] The processing system further comprises components to control and classify the agglomerates formed during the drying of cellular biomass. In one aspect, the processing system comprises agglomeration sensors (160) operably connected within the dryer to assess and classify the size of the agglomerates. Agglomeration sensors (160) can feed the agglomeration data collected by agglomeration sensors (160) into agglomeration size algorithm (150). In one aspect, agglomeration sensors (160) can include, for example, laser diffraction sensors and / or image analysis sensors that can measure the size of the particle agglomerates in fluidized bed dryer (120) and output the size data as agglomeration data.

[0040] The system comprises circuitry for performing agglomeration size algorithms (150). Agglomeration size algorithm (150) can receive and process agglomeration size data from agglomeration sensors (160). Furthermore, agglomeration size algorithm (150) can also be operably connected to an optimum cell agglomerated size database (140) in order to receive information regarding the desired optimum cell agglomerated size data.PT-2170-WO-PCT

[0041] Agglomeration size algorithm (150) can process / compare the input received regarding the agglomeration data from agglomeration sensors (170) and the desired optimum cell agglomerate size from the optimum cell agglomerate size database (140). The circuitry of the agglomeration size algorithm (150) can be operably connected to the system to provide removal control data to system (100) to remove agglomerates from dryer (120) when the agglomeration data aligns with the desired optimum cell agglomerate size data. By aligning, it is meant that the agglomerates in dryer (120) comprise a desired level of the optimum cell agglomerate size and / or uniformity based on the provided optimum cell agglomerate size and / or uniformity.

[0042] The processing system further comprises components for classifying the agglomerates to generate agglomerates that are more uniform in size. Classifying agglomerates can include separating or sorting the optimum cell agglomerates from the sub-optimal cell biomass. Classifying can include the use of, for example, sieves / screens, air classifiers, aspirators, sedimentation and the like. The classifying components can separate the optimum cell agglomerates from sub-optimum cell biomass.

[0043] The system further comprises at least two conduits to direct the optimum cell agglomerates into a first conduit and the sub-optimal cell biomass into a second conduit. The optimum cell agglomerates can be transferred in the first conduit to roaster (180). The optimum cell agglomerates may exit the dryer and be transferred to a roaster if the roaster is external to the system. Alternatively, if the roaster is integrated within the system, the optimum cell agglomerates may be transferred to the roaster. In one aspect, the optimum cell agglomerates may exit the dryer and be transferred to be alkalized in the system or to an external system. In one aspect, the drying and roasting can occur simultaneously. The optimum cell agglomerates can be roasted during the drying step.

[0044] The sub-optimal cell biomass can exit the dryer through the second conduit. The sub-optimal cell biomass may be discarded into receptacle (170) after leaving the dryer. Alternatively, the sub-optimal cell biomass may be recirculated back into the inlet of the dryer by operably connecting the outflow of the sub-optimal cell biomass with the inlet into dryer (120) as shown in Fig. 1. In the case of the cell biomass rejected due to being over the size limit, a size reduction step, such as a mill, can be used prior to recirculating into the agglomerator.

[0045] The processing system may further comprise roaster (180). Roaster (180) can be operably connected to the output of the optimum sized agglomerates. Roaster (180) may comprise circuitry to receive input regarding the roasting parameters to roast the agglomerates.PT-2170-WO-PCT

[0046] The processing system may optionally include components and / or circuitry to alkalize the agglomerates. The alkalization may occur before and / or during the agglomeration of the cellular biomass.Method of processing cellular biomass

[0047] In one aspect, the present description relates to a method of processing a cellular biomass to form agglomerates of a desired size and uniformity. The method may optionally include fermenting the cellular biomass prior to formation of the agglomerates. The method includes classifying the size of the cell agglomerates. Preferably, the cell agglomerate size is classified during drying of the cellular biomass. The method also includes separating optimum cell agglomerates from the sub-optimal cell biomass to generate a uniform population of cell agglomerates. The removal of sub-optimal cell biomass can allow for more optimal roasting of the cacao cell agglomerates. The method can include roasting the optimum cell agglomerates. The method can optionally include alkalization of the cellular biomass before or during the drying and / or roasting steps.

[0048] The method will be described with a fluidized bed dryer, but it will be understood that other drying systems may also be used. Fluidized bed dryers are described, for example, in Jafari et al. 2023. Fig. 2 shows a flowchart of an exemplary method (200) for processing the cellular biomass in a fluidized bed dryer.

[0049] In one aspect, the cellular biomass comprises cells that are derived from cellular agriculture, e.g., plant cell cultures. The cellular biomass can include a population of cells that have been grown under controlled conditions outside of their natural environment, e.g., in a laboratory or at a manufacturing facility. Generally, the cells are isolated from a living source, e.g., a plant, and maintained under carefully controlled conditions such as temperature, humidity, and media with the appropriate nutrients and growth factors and cofactors. Cellular agriculture of plant cells is known in the art and described for example, in Aisala, H. et al. J. Agric. Food Chem, 71( 7), 18478-18488. Plant cells derived from a variety of cellular agriculture methods may be used in the methods described herein.

[0050] In one aspect, the cellular biomass comprises cells from the cacao plant. The cells selected from the cacao plant are capable of reproducing and differentiating. Preferably, the cells are from roots, leaves, stems and the like. In one aspect, the cellular biomass comprises the cells from leaves. Cells from other parts of the plant may also be included. Cacao cells are cultivated, grown, replicated and / or then differentiated in a controlled environment to produce the cellular biomass. The present description will be described in the context of cacao cellular biomass, but itPT-2170-WO-PCTwill be understood that other cellular biomasses from other plants are also within the scope of this description.

[0051] In one aspect, the cellular biomass may be fermented. The fermenting step may be conducted after cultivation and prior to agglomeration. Fermentation is known in the art and can be conducted by any of the known methods in the art. The cellular biomass may be fermented with yeasts and bacteria, in temperature, time and oxygen availability conditions typical of those used in conventional cocoa bean fermentation. The fermentation may be conducted under aerobic conditions and / or or anaerobic conditions. In one aspect, the fermentation solution can include, for example, sugars, yeast, bacteria and the like. Fermentation can be conducted at a temperature of between about 23°C and 50°C, or between 25°C and 45°C, or between 25°C and 40°C, or between 25°C and 35°C, or between 25°C and 30°C, or between 30°C and 50°C, or between 30°C and 45°C, or between 30°C and 40°C. Fermentation can be conducted for at least 12 hours, or at least 1 day, or at least 2 days, or at least 3 days, or at least 4 days, or at least 5 days. Fermentation can be conducted for between 12 hours and 6 days, or between 1 days and 6 days, or between 12 hours and 5 days, or between 1 and 5 days, or between 12 hours and 4 days, or between 1 and 4 days, or between 12 hours and 3 days, or between 1 and 3 days, or between 12 hours and 2 days, or between 1 and 2 days, or between 36 hours and 6 days, or between 36 hours and 3 days, or between 36 hours and 3 days, or between 2 and 3 days.

[0052] The method described herein includes introducing a cellular biomass into a system (210), preferably a drying system, e.g., a fluidized bed dryer. The cellular biomass may be suspended in a liquid, e.g., a cell culture media, a buffered solution, a fermentation solution and the like. The cell suspension may be in a solution that includes nutrients, buffering salts, growth factors / hormones and the like. The cellular biomass may be wet, dry and / or in a powder form. The cellular biomass may include small particles and / or suboptimal cell biomass. The cellular biomass may be introduced into a dryer through an inlet or opening into the dryer of the processing system.

[0053] In one aspect, the method may include pre-drying the cellular biomass prior to introduction into the dryer. Pre-drying can include dewatering, filtering and / or at least partially drying the cellular biomass prior to introducing into the dryer. Pre-drying the cellular biomass can result in the reduction of the liquid in the input cellular biomass. Pre-drying the cellular biomass can increase the concentration of the cells in the input and decrease the amount of time needed for drying in the dryer. The pre-dried cellular biomass may be wet, dry and / or in a powder form. Predrying may be conducted at a temperature between 20 and 60°C, or between 20 and 50°C, orPT-2170-WO-PCTbetween 20 and 40°C, or between 20 and 30°C, or between 30 and 60°C, or between 30 and 50°C, or between 30 and 40°C, or between 40 and 60°C, or between 40 and 50°C, or between 50 and 60°C. Pre-drying may be conducted for a period of 1 and 90 min, or between 10 and 90 minutes, or between 30 and 90 minutes, or between 50 and 90 minutes, or between 70 and 90 minutes, or between 10 and 70 minutes, or between 30 and 70 minutes, or between 50 and 70 minutes, or between 10 and 50 minutes, or between 10 and 30 minutes. Pre-drying may be conducted using a variety of methods, e.g. filtration or centrifugation, and all are included in this description.

[0054] In one aspect, the method comprises drying (220) the cellular biomass in the fluidized bed dryer using the drying parameter data from the drying control parameter database. The drying control parameter database can be adjusted and / or programmed to adjust the drying parameters for the drying of the cellular biomass that has been introduced into the dryer. The drying parameters can determine rate of reduction in the moisture content of the cellular biomass. The rate of drying based on the drying parameters can determine the level of agglomeration and the resulting size of the cell agglomerates.

[0055] Drying parameters that can be controlled by the drying parameter database include, for example, drying time, air velocity, temperature, humidity, flow rate, mixing rate and the like. The selection of the combination of the drying parameters can determine the characteristics of the agglomerates such as the agglomeration size and the moisture content in the agglomerates.

[0056] The drying time of the cellular biomass can vary and all are within the scope of this description. The drying time can depend on, for example, the initial moisture content of the cellular biomass, the temperature used to dry the cellular biomass, the air flow in dryer and the like. In one aspect, the drying time can be at least 10 minutes, or at least 20 minutes, or at least 40 minutes, or at least 60 minutes, or at least 2 hours. The drying time can be between about 20min and about 120min, or between about 20 minutes and 90 minutes, or between 20 minutes and 60 minutes, or between 30 minutes and 120 minutes, or between 30 minutes and 90 minutes, or between 30min and about 60min.

[0057] The temperature at which the cellular biomass is dried can vary and all are within the scope of this description. The temperature can be between about 40°C and about 160°C, or between 40°C and 120°C, or between 40°C and 100°C, or between 40°C and 80°C, or between 40°C and 60°C, or between 60°C and 160°C, or between 60°C and 140°C, or between 60°C and 120°C, or between 60°C and about 100°C, or between 60°C and 80°C, or between 80°C and 160°C, or between 80°C and 140°C, or between 80°C and 120°C, or between 80°C and 100°C.PT-2170-WO-PCT

[0058] The average air velocity in the dryer for drying the cellular biomass can vary and all are within the scope of this description. The air velocity can be between about 2m / s and about 25m / s, or between about 2m / s and about 20m / s, or between about 2m / s and about 15m / s, or between about 2m / s and about lOm / s, or between about 3m / s and about 25m / s, or between about 3m / s and about 20m / s, or between about 3m / s and about 15m / s, or between about 3m / s and about lOm / s, or between about 5m / s and about 25m / s, or between about 5m / s and about 20m / s, or between about 5m / s and about 15m / s, or between about 5m / s and about lOm / s, or between about lOm / s and about 25m / s, or between about lOm / s and about 20m / s, or between about lOm / s and about 15m / s.

[0059] The air moisture content in the dryer for drying the cellular biomass can vary and all are within the scope of this description. The air moisture content in the dryer can be between about between 5% and 60%, or between 5% and 40%, or between 5% and about 30%, or between 5% and 20%, or between 10% and 60%, or between 10% and 40%, or between 10% and 30%, or between 10% and 20%, or between 15% and 60%, or between 15% and 40%, or between 15% and 30%, or between 20% and 60%, or between 20% and 40%, or between 30% and 60%, or between 30% and 40%.

[0060] The headspace gas composition in the dryer for drying the cellular biomass can vary and all are within the scope of this description. In one aspect, the agglomerates may be formed under atmospheric conditions. The headspace gas composition may also be a modified atmosphere. The amount of oxygen in the headspace gas composition can be between about 50 % and about 18% oxygen, or between 40% and 18%, or between 30% and 18%, or between 20% and 18%, or between 20% and about 19%.

[0061] In one aspect, the headspace gas composition may also include oxidizing gases. The oxidizing gases can enhance alkalization during the agglomeration of the cellular biomass. The oxidizing gases can include, for example, nitrous oxide, ozone and the like.

[0062] In one aspect, the method can include adding binding agents during the drying process to improve the agglomeration of the cellular biomass. The binding agents can include proteins, sugars, starches, carbohydrates, maltodextrins and the like. The binding agents may be added by spraying, mechanical dispersion, binding solution and the like. The amount of binding agents added can vary and all are within the scope of this description. In one aspect, the amount of binding agents can be at least 2wt%, or at least 5wt%, or at least 10wt% of the weight of the cellular biomass. The amount of binding agents can be between about 2wt% and 15wt% of the cellular biomass, or between 2wt% and 10wt%, or between 2wt% and 5wt%, or between about 5wt% andPT-2170-WO-PCT15wt%, or between 5wt% and 10wt%, or between 10wt% and 15wt% of the weight of the cellular biomass.

[0063] In one aspect, the method may include alkalizing the cellular biomass before or during agglomeration. The alkalizing step can generate alkalized agglomerates. In one aspect, drying the cellular biomass can result in alkalization of the biomass. Alkalizing can include the use of a combination of alkalizing agents such as potassium or sodium carbonate, potassium or sodium hydroxide, ammonia or calcium carbonate, oxidizing agents such as elevated oxygen partial pressure, ozone and other nitrous gasses, similarly to the alkalization process performed on conventional cocoa beans or nibs, as known in the art (methods are described in United States Patent Nos. 4,435,436, 4,784,866 and 5,009,917, and in European Patent No. 2068641, incorporated herein by reference).

[0064] The target pH value of the agglomerated cellular biomass can vary and all are within the scope of this description. In one aspect, the pH can be between about 7 and about 10, or between 7 and 9, or between 7 and 8, or between 8 and 10, or between 8 and 9, or between 9 and 10.

[0065] Drying the cellular biomass in the fluidized bed dryer can result in the formation of cell agglomerates during the drying process. The combination of the drying parameters can determine the characteristics of the agglomerates such as the agglomeration size and the moisture content in the agglomerates. The agglomerates can be at least 1mm, or at least 2 mm, or at least 3 mm, or at least 5 mm, or at least 8mm, or at least 10mm. The agglomerates can be between about 1 mm and about 10 mm, or between 2 mm and about 10mm, or between 3 mm and about 10mm, or between 5 mm and about 10mm, or between 8mm and about 10mm. The agglomerates can be between about 1 mm and about 8 mm, or between 2 mm and about 8 mm, or between 2 to 7mm, or between 3 mm and about 8 mm, or between 3 to 7mm, or between 5 mm and about 8 mm. The agglomerates can be between about 1 mm and about 5 mm, or between 2 mm and about 5 mm, or between 3 mm and about 5 mm, or between about 1 mm and about 3 mm, or between 2 mm and about 3 mm.

[0066] In one aspect, the method further comprises classifying / controlling the size of the agglomerates based on the agglomeration data collected by the agglomeration sensors. The agglomeration sensors can determine the level of agglomeration in the dryer and provide the agglomeration size data as input to an agglomeration size algorithm. The agglomeration size algorithm can receive the agglomeration data from the agglomeration sensors and compare it with the optimum cell agglomerate size data from the optimum cell agglomerate size database (230).PT-2170-WO-PCTIf the agglomeration data is within the threshold of the optimum cell agglomerate size data, the agglomeration size algorithm outputs removal control data which controls the removal of the sub-optimal cellular biomass and optimum cell agglomerates from the fluidized bed dryer (240). If the agglomeration data is outside the threshold of the optimum cell agglomerate size data, the agglomeration size algorithm continues to monitor the agglomeration data.

[0067] In one aspect, the removal control data enable the dryer to separate the optimum cell agglomerates from the sub-optimal cell biomass (250 and 260). The optimal cell agglomerates that are inside the thresholds of the optimum cell agglomerate size data can be removed for transfer to a roaster (260). The optimum cell agglomerates can be transferred to a roaster in order to be further processed by roasting. The sub-optimal cell biomass may be discarded or recirculated to enter the fluidized bed dryer again at the inlet of the dryer to go through another round of processing or reduced in size to form the optimum cell agglomerates, as appropriate (250).

[0068] In one aspect, the optimum cell agglomerates comprise at least about 80wt%, or at least about 85wt%, or at least 90wt%, or at least about 95wt% of the cellular biomass that was provided to the inlet of the dryer. The optimum cell agglomerates comprise between about 80wt% and 99wt%, or between 80wt% and about 95wt%, or between 80wt% and about 90wt%, or between 80wt% and about 85wt%, or between 85wt% and about 99wt%, or between 85wt% and 95wt%, or between 85wt% and 90wt%, or between 90wt% and 99wt%, or between 90wt% and 95wt%, or between 95wt% and 99wt% of the cellular biomass that was provided to the inlet of the dryer.

[0069] The agglomerates can be classified to separate the suboptimal cell biomass from the optimum cell agglomerates. A variety of classification methods may be used to achieve a desired output size distribution of the agglomerates. The methods may include, for example, sieving and / or screening the agglomerates to separate and collect the optimum size agglomerates from the sub-optimum size agglomerates. In one aspect, the method may include using an air classifier to separate and collect the optimum size agglomerates from the sub-optimum size agglomerates as shown in Fig. 3. Air classification is also described in Walas (1990) “Chemical Process Equipment-Selection and Design-Air Classifiers, Ch. 12, incorporated herein by reference. The methods may include using sedimentation to separate and collect the optimum size agglomerates from the sub-optimum size agglomerates. The classification method may include a combination of sieving / screening, air classifiers and / or sedimentation to separate and collect the optimum size agglomerates from the sub-optimum size agglomerates.PT-2170-WO-PCT

[0070] In one aspect, Fig. 3 shows a schematic of a method for separating the optimum size agglomerates from the sub-optimum size agglomerates. Fig. 3 is a schematic diagram of an exemplary process of separating the optimum cell agglomerates from the sub-optimal cell biomass. Fig 3, for example, shows an aspirator 300 (a.k.a. density separator). In Fig. 3, by blowing air through the material, the smaller agglomerates (dust / fines) get blown away with the air, and thus separated from the bulk material, leaving behind only the larger / denser agglomerates. By controlling the air velocity, the size of the agglomerates that are removed can be controlled. In one aspect, the method can separate the agglomerates by density and / or weight by controlling the air flow. In Fig. 3, classifier 300 includes all of the agglomerates at step 310. In step 320, agglomerated particles smaller than the threshold are removed. The undesirable particles sizes are removed in the fluidized bed reactor exhaust. The agglomerates are classified in a batch process. The optimum cell agglomerates retained in step 310 can be roasted. Alternatively, the agglomerates may be classified in a continuous process.

[0071] In one aspect, the step of classifying the optimum cell agglomerates can minimize and / or decrease the size variation, e.g. diameter variation, of the agglomerates present in the optimum cell agglomerates. Decreasing and / or minimizing the variation in the diameters of the agglomerates present can increase the uniformity of the agglomerates in the optimum cell agglomerates. In one aspect, the difference in the diameter between the largest agglomerate and the smallest agglomerate in the optimum cell agglomerates is minimized. The difference in the diameter between the largest agglomerate and the smallest agglomerate in the optimum cell agglomerates is at most 10mm, or at most 8mm, or at most 5mm, or at most 3mm, or at most 2mm. The difference in the diameter between the largest agglomerate and the smallest agglomerate in the optimum cell agglomerates can be between about 2mm and 10mm, or between 2mm and 8mm, or between 2 mm and 5mm.

[0072] In one aspect, the percentage of agglomerates in the composition having a diameter between about 1 mm and about 10 mm can be at least 50%, or at least 60%, preferably at least 70%, or at least 80%, or at least 90%, or at least 95%. The percentage of agglomerates between about 2 mm and about 5 mm can be at least 50%, or at least 60%, preferably at least 70%, or at least 80%, or at least 90%, or at least 95%. The percentage of agglomerates between about 2 mm and about 4 mm can be at least 50%, or at least 60%, preferably at least 70%, or at least 80%, or at least 90%, or at least 95%.

[0073] Optimum size agglomerates that are uniform in size can have at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates withinPT-2170-WO-PCT30% of the mean agglomerate size; or optimum size agglomerates that are uniform in size can have at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 20% of the mean agglomerate size; or optimum size agglomerates that are uniform in size can have at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 10% of the mean agglomerate size.

[0074] In one aspect, sub-optimal cell biomass that are smaller than the optimum cell agglomerates may be extracted via a fluidized bed exhaust gas in the fluidized bed dryer. These may be fed back into the inlet of the fluidized bed dryer, re-hydrated and agglomerated again. Sub-optimal cell biomass that are larger than the optimum cell agglomerates may be extracted via the base of the fluidized bed. These may be fed back into the inlet of the fluidized bed dryer. The larger agglomerates may be broken down into sizes which may be re-hydrated and agglomerated again.

[0075] In one aspect, ultrasound may additionally be used such that the biomass agglomerates in sonic nodes. Ultrasound can lead to single cells that resonate and come together to form agglomerates. Ultrasound can be used to further control the particle size during the drying process. The ultrasound used may be in the range of 20 to 2400 kHz, or between 50 to 200 kHz.

[0076] The method further comprises transferring the optimum cell agglomerates to a roaster to conduct a roasting step. The roasting protocol can be optimized to size of the agglomerates. Optimum cell agglomerates that comprise larger size agglomerates can have a higher intensity of roasting, e.g. higher temperature and / or longer roasting times, than optimum cell agglomerates with smaller size agglomerates. Preferably, the roasting can achieve a high degree of Maillard reaction with minimal burning. The roasting of the agglomerates can remove moisture and develop flavors. The flavors can be developed through Maillard reactions (reducing sugars react with amino acids), caramelization (sugars polymerize) and pyrolysis (molecules breakdown) reactions. These are the reactions that can give cocoa cells the characteristic roasted and chocolatey notes. Driving the Maillard reaction during roasting can increase the roasted / cocoa notes and minimize burning (off notes).

[0077] The roasting can be done in a continuous system, batch system and the like. The roasting can comprise a roasting profile that includes, for example, a combination of time, temperature. The roasting profile can include increases and decreases in temperature and the duration of the roasting at each of these temperatures. Roasting can also include the use of hot air velocity, pressure and the like. During roasting, the temperature can be the same during the entire duration of the roasting. Alternatively, the temperature can be increased or decreased for varyingPT-2170-WO-PCTlengths of time during the roasting. In one aspect, the roasting profile can be tuned, for example, based on the size of the agglomerates and / or the moisture in the agglomerates to generate desired flavors.

[0078] The roasting of the optimum cell aggregates may be conducted at temperatures between about 105 and about 130°C, or between 110 and about 130°C, or between 115 and about 130°C, or between 120 and about 130°C, or between 125 and about 130°C, or between 105 and about 125°C, or between 110 and about 125°C, or between 115 and about 125°C, or between 120 and about 125°C, or between 105 and about 120°C, or between 110 and about 120°C, or between 115 and about 120°C, or between 105 and about 115°C, or between 110 and about 115°C, or between 105 and about 110°C, or between 105 and about 110°C.

[0079] The total roasting time of the optimum cell aggregates may be conducted for a duration of between about 20 and about 60 min, or between 20 and 55 minutes, or between 20 and 50 minutes, or between 20 and 45 minutes, or between 20 and 40 minutes, or between 20 and 35 minutes, or between 20 and 30 minutes, or between 20 and 25 minutes, or between 25 and about 60 min, or between 25 and 55 minutes, or between 25 and 50 minutes, or between 25 and 45 minutes, or between 25 and 40 minutes, or between 25 and 35 minutes, or between 25 and 30 minutes, or between 30 and about 60 min, or between 30 and 55 minutes, or between 30 and 50 minutes, or between 30 and 45 minutes, or between 30 and 40 minutes, or between 30 and 35 minutes, or between 35 and about 60 min, or between 35 and 55 minutes, or between 35 and 50 minutes, or between 35 and 45 minutes, or between 35 and 40 minutes, or between 40 and about 60 min, or between 40 and 55 minutes, or between 40 and 50 minutes, or between 40 and 45 minutes, or between 45 and about 60 min, or between 45 and 55 minutes, or between 45 and 50 minutes, or between 50 and about 60 min, or between 50 and 55 minutes, or between 55 and 60 minutes.

[0080] The method further comprises roasting the optimum cell aggregates to acquire and retain a desirable sensory profile. Desirable sensory profile can include desirable texture, flavor profile, volatile compounds and the like. The efficiency of roasting may be balanced with the need for a minimum agglomerate size, to enable the retention of volatile flavours and regulate the heat transfer rate through the cell-mass particles. This may help to prevent over-roasting / buming and therefore minimizes undesirable flavors. If the agglomerates are too small, they are likely to burn, giving off flavors. In addition, if the agglomerates are too small, or the roasting conditions are too long, the aroma may be lost, even under conditions of low temperatures.PT-2170-WO-PCT

[0081] In one aspect, the agglomerates are further processed as used for processing with cocoa nibs. The method may comprise grinding the roasted optimum cell agglomerates to produce agglomerate powder, e.g., agglomerate cocoa powder. The agglomerates and / or the products derived from the agglomerates may be pressed, refined, conched, tempered and molded. All of these processes are known in the art.

[0082] In one aspect, the cell lines used for cultivating the cellular biomass may include a significant amount of fat. Thus, the resulting agglomerates may include a significant amount of fat. In this case, standard nib grinding technology such as knife mills and ball mills can be used to create a cocoa liquor from the agglomerates, which can later be pressed to expel part of the cocoa butter and further ground in hammer mills into powder (as in conventional production). If the cell line used to create the cellular biomass is poor in fat, then agglomerates may be processed by directly be grinding using a hammer or gap mill and go straight to powder. The agglomerate processing may not include milling into liquor first nor pressing the butter out.

[0083] The method may comprise incorporating the agglomerate cocoa powder into food products. The cocoa powder, cocoa liquor, cocoa butter generated from the agglomerates may be used in food products similar to the cocoa products derived from cocoa nibs.

[0084] Compositions

[0085] In one aspect, the present description relates to a composition comprising roasted plant cell agglomerates, preferably roasted cacao cell agglomerates. The compositions described herein comprise the optimum cell agglomerates described herein. Preferably, the compositions comprise roasted optimum cell agglomerates.

[0086] In one aspect, the agglomerates can be at least 1mm, or at least 2 mm, or at least 3 mm, or at least 5 mm, or at least 8mm, or at least 10mm. The agglomerates can be between about 1 mm and about 10 mm, or between 2 mm and about 10mm, or between 3 mm and about 10mm, or between 5 mm and about 10mm, or between 8mm and about 10mm. The agglomerates can be between about 1 mm and about 8 mm, or between 2 mm and about 8 mm, or between 2 to 7mm, or between 3 mm and about 8 mm, or between 3 to 7mm, or between 5 mm and about 8 mm. The agglomerates can be between about 1 mm and about 5 mm, or between 2 mm and about 5 mm, or between 3 mm and about 5 mm, or between about 1 mm and about 3 mm, or between 2 mm and about 3 mm.

[0087] In one aspect, the desired agglomerate size / shape is controlled to alter the rate of heat transfer through the agglomerate during roasting, such that desired flavours are retained and / orPT-2170-WO-PCTcreated during roasting. Without being bound by any theory, it is thought that the smaller the agglomerate, the easier it is for volatiles to diffuse out, and thus loose aromas. Some of the aromas (like acetic acid, which is commonly produced during fermentation) can be exhausted to eliminate a vinegar smell, while retaining the cocoa smell. Diffusion rate can depend on porosity (which we can control with the agglomeration conditions) and pore depth (i.e. agglomerate diameter). The factors affecting diffusion rates are well known by those in the art.

[0088] In one aspect, the composition comprises cell agglomerates that are substantially uniform in size. The difference in the diameter between the largest agglomerate and the smallest agglomerate in the optimum cell agglomerates is at most 10mm, or at most 8mm, or at most 5mm, or at most 3mm, or at most 2mm. The difference in the diameter between the largest agglomerate and the smallest agglomerate in the optimum cell agglomerates can be between about 2mm and 10mm, or between 2mm and 8mm, or between 2 mm and 5mm.

[0089] In one aspect, the percentage of agglomerates in the composition having a diameter between about 1 mm and about 10 mm can be at least 50%, or at least 60%, preferably at least 70%, or at least 80%, or at least 90%, or at least 95%. The percentage of agglomerates between about 2 mm and about 5 mm can be at least 50%, or at least 60%, preferably at least 70%, or at least 80%, or at least 90%, or at least 95%. The percentage of agglomerates between about 2 mm and about 4 mm can be at least 50%, or at least 60%, preferably at least 70%, or at least 80%, or at least 90%, or at least 95%.

[0090] Optimum size agglomerates that are uniform in size can have at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 30% of the mean agglomerate size; or optimum size agglomerates that are uniform in size can have at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 20% of the mean agglomerate size; or optimum size agglomerates that are uniform in size can have at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the agglomerates within 10% of the mean agglomerate size.

[0091] The cell agglomerates can be a variety of shapes and all are within the scope of this description. The shapes can be regular, smooth, irregular, bumpy, uneven and the like. In one aspect, the shapes can be spherical, oblong and may be bumpy. The agglomerates may not be needle like and / or pointy. In one aspect, the longest dimension should not be greater than 10 times the shortest dimension, or greater than 8 times the shortest dimension, or greater than 6 times the shortest dimension, or greater than 4 times the shortest dimension.PT-2170-WO-PCT

[0092] In one aspect, the composition comprises cell agglomerates that are roasted, fermented and / or alkalized. The cell agglomerates and / or the cocoa powder derived from the cell agglomerates can have a variety of colors similar to traditional cocoa powders, e.g. brown, dark, reddish and the like. Measured by the wet method on the Hunter color coordinate scale or CIE 1976 (CIELAB) color system, the cocoa powders derived from this composition can range from an L value between 12 and 20 for slightly alkalized powders; between 6 and 12 for highly alkalized powders, and below 6 for “black” powders.

[0093] In one aspect, the present description relates to a composition comprising cocoa products derived from the agglomerates. The cocoa products can include cocoa powder, cocoa liquor, cocoa butter and the like derived from the roasted optimum cell agglomerates described herein.

[0094] The cocoa products in the compositions described herein can have a sensory profile comparable to the roasted cocoa products derived from the cacao beans. The cocoa powder can, for example, have fruity, roasted and chocolaty notes. It can have bitterness levels similar to the traditional cocoa powders from cacao beans. It can lack the off notes that can sometimes be present traditional cocoa powders from cacao beans.

[0095] Advantageously, the roasted cocoa cell agglomerates of the present invention can be used like any other cocoa materials in the manufacture of food and beverage compositions. These may include, by way of illustration only, milk, dark, and white chocolate and compound compositions (for use, amongst others, in confectionary, as bars, in truffles and pralines, or as inclusions, coatings, or fillings), drinking chocolate, flavored milks (dairy and non-dairy), flavored syrups, bakery products (such as cakes, cookies and pies), diet bars and meal substitutes, sports and infant nutrition, ice-cream products, dairy products, puddings, mousses, sauces, and breakfast cereals.

[0096] In one aspect, the fully processed agglomerates can be converted to a confectionery bar in a manner similar to processing cocoa nibs. The agglomerates may have a reduced fat content. The agglomerates may be further processed by the addition of fat, e.g., shea butter. In one aspect, the processing of the cocoa product may include, for example, processing with 30% w / w processed agglomerates, 39.3% shea butter, 0.7% lecithin and 30% sucrose, in a benchtop melangeur at 50°C for 2h, followed by tempering and molding into tablets.

[0097] In one aspect, the fully processed agglomerates may be ground in a gap mill to make cocoa powder. The cocoa powder can be added to milk to make chocolate milk.PT-2170-WO-PCT

[0098] Representative features of the present invention are set out in the following clauses, which stand alone or may be combined, in any combination, with one or more features disclosed in the text of the specification.

[0099] The present invention is as set out in the following clauses:

[0100] Clause 1 : A method of processing a cellular biomass comprising:a. introducing a cellular biomass into a dryer;b. drying the cellular biomass in the dryer at selected drying parameters, wherein agglomerates comprising the cells from the cellular biomass are formed during the drying;c. classifying the size of the agglomerates by collecting agglomeration size data from agglomeration sensors in the dryer; andd. separating / sorting optimum cell agglomerates from sub-optimal cell biomass after a threshold level of agglomerates of optimum size are generated.

[0101] Clause 2: The method of clause 1, wherein the dryer is a fluidized bed dryer.

[0102] Clause 3: The method of any one of the preceding clauses, wherein the cellular biomass is pre-dried prior to introduction into the dryer.

[0103] Clause 4: The method of any one of the preceding clauses, wherein the method further comprises roasting the optimum cell agglomerates.

[0104] Clause 5: The method of any one of the preceding clauses, wherein the method further comprises fermenting, alkalizing, pressing, refining, conching, tempering and / or molding.

[0105] Clause 6: The method of any one of the preceding clauses, wherein the method further comprises reintroducing the sub-optimal cell biomass to fluidized bed dryer and / or discarding the sub-optimal cell biomass.

[0106] Clause 7: The method of any one of the preceding clauses, wherein the drying parameters comprise temperature, air velocity, air moisture content and headspace gas composition.

[0107] Clause 8: The method of clause 7, wherein the temperature in the dryer is between about 40°C and 160°C.

[0108] Clause 9: The method of any one of the preceding clauses, wherein the air velocity is between about 2m / s and about 25m / s.

[0109] Clause 10: The method of any one of the preceding clauses, wherein the air moisture content is between about 5% and about 60%.PT-2170-WO-PCT

[0110] Clause 11: The method of any one of the preceding clauses, wherein the headspace gas composition is atmospheric gas composition.

[0111] Clause 12: The method of any one of the preceding clauses, wherein the optimum cell agglomerates are at least about 1mm, preferably between about 1mm and 10mm.

[0112] Clause 13: The method of any one of the preceding clauses, wherein the optimum cell agglomerates comprise agglomerates that are substantially uniform in size.

[0113] Clause 14: The method of clause 13, wherein at least 70% of the agglomerates have a diameter that is within 30% of the mean agglomerate size.

[0114] Clause 15: The method of any one of the preceding clauses, wherein agglomeration sensors comprise laser diffraction sensors and / or image analysis sensors.

[0115] Clause 16: The method of any one of the preceding clauses, wherein the roasting comprises transfer of the optimum cell agglomerates to a fluidized bed roaster, drum roaster and / or hot air roaster.

[0116] Clause 17: The method of any one of the preceding clauses, wherein the roasting comprises heating the optimum cell agglomerates to a temperature of between about 105°C and about 130°C for a duration of between about 20 minutes and 60 minutes.

[0117] Clause 18: A composition comprising optimum cell agglomerates, wherein the agglomerates comprise plant cells, wherein the plant cells are cell cultured cacao cells.

[0118] Clause 19: The composition of clause 18, wherein the agglomerates comprise a diameter of at least about 1mm, preferably between about 1mm and 10mm.

[0119] Clause 20: The composition of clauses 18-19, wherein the optimum cell agglomerates are roasted optimum cell agglomerates.

[0120] Clause 21 : The composition of clauses 18-20, wherein the optimum cell agglomerates are fermented optimum cell agglomerates.

[0121] Clause 22: The composition of clauses 18-21, wherein the composition is a cocoa product and wherein the product is derived from the roasted optimum cell agglomerates.

[0122] Clause 23: The composition of clauses 18-22, wherein the cocoa product is cocoa powder.

[0123] Clause 24: A cellular biomass processing system comprising a dryer comprising an inlet for cellular biomass, agglomeration sensors, and at least one outlet for removal of optimum cell agglomerates, wherein the dryer comprises circuitry for receiving input comprising drying parameter data from a drying control parameter database, wherein agglomeration sensorsPT-2170-WO-PCTcomprise circuitry for determining level of agglomeration and wherein the dryer comprises component for separation of optimum cell agglomerates from sub-optimum sized agglomerates.

[0124] Clause 25: The system of clause 24, wherein the dryer is a fluidized bed dryer.

[0125] Clause 26: The system of clauses 24-25, further comprising a roaster.

[0126] Clause 27: The system of clauses 24-26, wherein the agglomerations sensors comprise laser diffraction sensors and / or image analysis sensors.

[0127] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.EXAMPLESProphetic Example

[0128] For biomass production, cacao cells are cultivated in a wave bioreactor using 50L wave bags with final working volumes of 25 L. Cultivation is adjusted as follows: temperature 24°C, aeration 300 mL / min, in darkness. Plant cell biomass is harvested by filtering with Miracloth and subsequently washed with sterile water.

[0129] The collected plant cells are consequently inoculated with a starter culture consisting of yeasts (Saccharomyces cerevisae), lactic acid bacteria (Lactobacillus plantarum) and acetic acid bacteria (Acetobacter aceti), at a 5% w / w inoculation rate. A fermentable sugar blend of sucrose: glucose: fructose at a 2:1:1 ratio is added to sterilized water at 22g / L concentration. The sugar solution is added at the biomass at a ratio of 1 part inoculated biomass to 4 parts sugar solution in a conical fermenter. The fermentation is allowed to develop for 60 hours, first 30 hours anaerobically and then 30 hours aerobically, at a controlled temperature of 30°C, under continuous aeration with 0.3 L / min / Kg of biomass microfiltered atmospheric air, and gentle, constant agitation in absence of light. The fermented biomass was consequently dewatered by re-filtering with Miracloth, and rinsed with sterile water, re-filtered with Miracloth and partially dried in an oven at 60°C until a moisture content of approximately 42% was achieved.

[0130] The resulting fermented biomass had a moisture content of approximately 42% w / w and was then introduced in a Conti FB 20 / 4 (Neuhaus) fluidized bed via a weigh feeder. To aid with both the agglomeration and alkalization a 45% w / w solution of potassium hydroxide was sprayed onto the fluidized bed using top nozzles.PT-2170-WO-PCT

[0131] The process parameters are summarized below:Table 1

[0132] The resulting alkalized aggregates have a moisture content of approximately 15% w / w. The material is then passed through vibrating sieves to only select agglomerates with a size of 2-4mm, and the residual over and under material was reworked as appropriate.

[0133] The agglomerates were consequently alkalized in the conventional manner, by infusing them with 270g of a 7% aqueous solution of potassium hydroxide per Kg of biomass and holding for 1.5h at 80°C. Finally, the alkalized agglomerates were roasted in a batch tray oven at 120°C for 120 min.

[0134] Cocoa product 1: The fully processed agglomerates are converted to a confectionery bar in a manner similar to processing cocoa nibs, taking care to account for the reduced fat contentPT-2170-WO-PCTof the agglomerates. In short, the processed cocoa product included processing 30% w / w processed agglomerates, 39.3% shea butter, 0.7% lecithin and 30% sucrose, in a benchtop melangeur at 50°C for 2h, followed by tempering and molding into tablets.Cocoa product 2: The fully processed agglomerates are also ground in a gap mill to make cocoa powder. The cocoa powder can be added to milk at 2% w / w along with 8% w / w sucrose to make chocolate milk.

[0135] In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0136] Values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range were explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0137] Unless expressly stated, ppm (parts per million), percentage, and ratios are on a by weight basis. Percentage on a by weight basis is also referred to as wt% or % (wt) below.

Claims

PT-2170-WO-PCTCLAIMSWhat is claimed is:

1. A method of processing a cellular biomass comprising:a. introducing a cellular biomass into a dryer, preferably a fluidized bed dryer; b. drying the cellular biomass in the dryer at selected drying parameters comprising temperature, air velocity, air moisture content and headspace gas composition to form agglomerates;c. classifying the size of the agglomerates by collecting agglomeration size data from agglomeration sensors in the dryer, wherein the agglomeration sensors comprise laser diffraction sensors and / or image analysis sensors; andd. separating optimum size cell agglomerates from sub-optimal cell biomass after a threshold level of agglomerates of optimum size are generated when a threshold is reached at which at least 70% of the agglomerates have a diameter within 30% of a mean agglomerate size, wherein the optimum size cell agglomerates have a diameter between about 1 mm and about 10 mm.

2. The method of any one of the preceding claims, wherein the cellular biomass is pre-dried prior to introduction into the dryer.

3. The method ofclaim 1 or claim 2, wherein the method further comprises roasting the optimum size cell agglomerates.

4. The method of any one of the preceding claims, wherein the method further comprises fermenting, alkalizing, pressing, refining, conching, tempering and / or molding.

5. The method of any one of the preceding claims, wherein the method further comprises reintroducing the sub-optimal cell biomass to the fluidized bed dryer and / or discarding the sub-optimal cell biomass.

6. The method of any one of the preceding claims, wherein the temperature is between about 40°C and 160°C, the air velocity is between about 2 m / s and about 25 m / s, the air moisture content is between about 5% and about 60%, and the headspace gas composition is atmospheric.PT-2170-WO-PCT7. The method of any one of the preceding claims, wherein the headspace gas composition is atmospheric gas composition.

8. The method of any one of the preceding claims, wherein the optimum size cell agglomerates have a diameter between about 1 mm and about 10 mm.

9. The method of any one of the preceding claims, wherein the optimum size cell agglomerates comprise agglomerates that are substantially uniform in size, wherein at least 70% of the agglomerates have a diameter that is within 30% of the mean agglomerate size.

10. The method of any one of the preceding claims, wherein agglomeration sensors comprise laser diffraction sensors and image analysis sensors.

11. The method of any one of the preceding claims, wherein the roasting comprises heating the optimum size cell agglomerates to a temperature of between about 105°C and about 130°C for a duration of between about 20 minutes and 60 minutes.

12. A composition comprising optimum size cell agglomerates, wherein the agglomerates comprise plant cells, wherein the plant cells are cell cultured cacao cells, wherein the agglomerates have a diameter between about 1 mm and about 10 mm and at least 70% of the agglomerates have a diameter within 30% of a mean agglomerate size, and wherein the agglomerates are optionally roasted and / or fermented.

13. The composition of claim 12, wherein the agglomerates are roasted at a temperature of between about 105°C and about 130°C for a duration of between about 20 minutes and 60 minutes.

14. The composition of claim 12 or 13, wherein the composition is a cocoa product and wherein the product is derived from the roasted optimum cell agglomerates, preferably the cocoa product is cocoa powder.

15. A cellular biomass processing system comprising a dryer, preferably a fluidized bed dryer, comprising an inlet for cellular biomass, agglomeration sensors, and at least one outlet for removalPT-2170-WO-PCTof optimum cell agglomerates, wherein the dryer comprises circuitry for receiving input comprising drying parameter data from a drying control parameter database, wherein agglomeration sensors comprise circuitry for determining level of agglomeration, the agglomeration sensors comprise laser diffraction sensors and / or image analysis sensors configured to determine agglomerate size distribution in the dryer and wherein the dryer comprises a component for separation of optimum cell agglomerates from sub-optimum sized agglomerates when a threshold is reached at which at least 70% of the agglomerates have a diameter within 30% of a mean agglomerate size.

16. The system of claim 15, wherein the agglomeration sensors comprise both laser diffraction sensors and image analysis sensors.

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