Patent Application
20200008445
An Application Data Sheet is filed concurrently with this specification as part of the present application. Each application that the present application claims benefit of or priority to as identified in the concurrently filed Application Data Sheet is incorporated by reference herein in its entirety and for all purposes.
The present invention relates to increasing the amount of protein in acidic liquid foods such as acidic beverages and salad dressings.
As the human population continues to increase, there is a growing need for additional food sources, particularly food sources that are inexpensive to produce and nutritious. Moreover, the current reliance on meat as the staple of many diets, at least in the most developed countries, contributes significantly to the release of greenhouse gases. There is a need for new foodstuffs that are less harmful to the environment to produce.
Requiring only water and sunlight to grow, algae have long been looked to as a potential source of food. While certain types of algae, primarily seaweed, do indeed provide important foodstuffs for human consumption, the promise of algae as a foodstuff has not been fully realized. Algal powders made with algae grown photosynthetically in outdoor ponds or photobioreactors are commercially available but have a deep green color (from the chlorophyll) and a strong, unpleasant taste. When formulated into food products or as nutritional supplements, these algal powders impart a visually unappealing green color to the food product or nutritional supplement and have unpleasant fish, seaweed or other flavors.
There are several species of algae that are used in foodstuffs today, most being macroalgae such as kelp, purple laver (Porphyra, used in nori), dulse (Palmaria palmate) and sea lettuce (Ulva lactuca). Microalgae, such as Spirulina (Arthrospira platensis) are grown commercially in open ponds (photosynthetically) for use as a nutritional supplement or incorporated in small amounts in smoothies or juice drinks (usually less than 0.5% w/w). Other microalgae, including some species of Chlorella are popular in Asian countries as a nutritional supplement. Poor flavor is a major factor that has impeded the widespread adoption of microalgae in food. WO2010/045368, WO2010/120923, PCT/US13/65369, and PCT/US14/013405 disclose methods of making and using microalgal biomass as a food. These references disclose the growth of microalgae, especially Chlorella protothecoides, in a dark environment, to produce a non-green microalgal biomass.
Low pH food products such as beverages (including fruit and tomato juices), dressings, condiments, tomato soups, tomato sauces, etc. are difficult to supplement with added protein (e.g. soy protein isolate or whey protein) because the protein tends to aggregate and precipitate when heated in pasteurization or hot-fill steps. The problem is worse for low pH products than for neutral pH products because the low pH destabilizes proteins. Thus, alternate and improved methods for supplementing foods, especially low-pH foods, with protein are needed.
Protein has been shown to induce satiety. Consuming a diet high in protein can facilitate reaching fitness and nutritional goals. Nutritionists and bodybuilders often recommend diets that are high in protein to help build muscle and/or lose fat. Supplementing low pH foods like fruit juices, dressings, and sauces with protein can increase the nutritional density or can transform nutritionally deficient foods into satiety inducing, nontraditional sources of protein. Supplementing low pH foods with microalgal flour and/or microalgal protein allows for this transformation without substantially changing the flavor, mouthfeel, texture or viscosity of the food product. This marks a valuable improvement for consumers seeking protein-rich food products that are flavorful and pleasant to eat or drink. Supplementing low pH food products with microalgal proteinor microalgal flour provides consumers with a variety of foods that retain their flavor and texture and reap the benefits of the additional protein.
In accordance with an embodiment of the present invention, a method for making a high-protein food product includes making a protein-supplemented acidic liquid by combining an acidic liquid with high-protein microalgal flour, or combining a non-acidic liquid with high-protein microalgal flour and an acid, to arrive at a microalgal protein-supplemented liquid having a pH of 4.6 or less, wherein the protein-supplemented liquid comprises at least 0.1%, 1%, 2%, 5%, 10% or 15% microalgal protein from the microalgal flour; subjecting the protein-supplemented liquid to a heating step so as to inactivate microbes. After the heating step, the product does not have a gritty texture resulting from protein precipitation from the heating step and/or the viscosity of the liquid increases by no more than 0.5, 1, 2, 5, 7 or 10% after the heating step. The high-protein microalgal flour comprises at least 55% microalgal cell protein by dry weight.
Optionally, the protein-supplemented liquid is a fruit juice, salad dressing or sauce.
The high-protein microalgal flour can be comprised predominantly of intact microalgal cell bodies of heterotrophically cultivated microalgae.
Optionally, at least one heating step comprises sterilizing the protein-supplemented liquid by HTST and/or hot-filling a container.
The heating step can include sterilizing the protein-supplemented liquid by LTLT; and/or hot-filling a container.
The HTST step can include heating to between 60-140° C. for between 5 to 60 seconds or heating to between 70-100° C. for between 5 to 60 seconds.
Optionally, the food product includes at least 5, 10, 20, 30, 40, or 50% protein.
The heating step can have temperature and time parameters such that if a 10% high-protein microalgal flour solution at pH 6 where heated in the heating step, the viscosity of the solution would increase by no more than 5%, whereas the viscosity of a 10% whey or soy protein solution at pH 6 would increase by at least 10%.
The heating step can have temperature and time parameters such that at least a 1, 2, 3, 4, or 5 log reduction in bacterial load is achieved.
The microalgal flour can comprise, consist of or consist essentially of dried Chlorella. The Chlorella can be non-green, yellow or yellow-white in color. The Chlorella can also be a color-mutant. The Chlorella can be of the species Chlorella protothecoides.
Alternately, the microalgal flour comprises dried Prototheca, optionally Prototheca moriformis.
In a related embodiment, the liquid lacks gritty or sandy texture due to protein aggregation.
Optionally, the microalgal flour has cells with, on average, less than 20%, 15%, or 14% lipid. The microalgal flour can also have cells with, on average, 5-20, 7-14, 8-13, or 10-13% lipid.
In a related embodiment, a low-pH food product is produced according to one of the methods mentioned above. In some cases, the food product comprises 1%, 2%, 3%, 4% or 5% microalgal flour, or a range of from 1-5%, 1-4%, 1-3%, 1-2%, 2-5%, 2-4%, 2-3%, 3-5%, 3-4%, or 4-5% microalgal flour. In some cases, the low-pH food product induces satiety upon consumption, and in some cases can aid in losing weight and/or building muscle when consumed as part of a high-protein diet.
In some cases, the low pH food product is a fruit beverage comprising microalgal flour at a concentration of 3-5% by weight.
The features of the invention will be more readily understood by reference to the following detailed description, taken with reference to the accompanying drawings.
As used herein, in connection with a range, the term “between” shall be inclusive of the endpoints.
The terms “microalgal powder” and “microalgal flour” are used interchangeably and mean a particulate dried cultured microalgae cell product with particles suitably sized for effective dispersion in a liquid.
In connection with microalgal cells of a microalgal flour, “intact” shall mean that the cells have not been treated with a disruption technique such as bead-milling designed to expose and release intracellular components. As a result, the cell walls of the microalgal cells are essentially continuous so as to contain the intracellular proteins.
The present invention is based on the discovery that acidic foods can be supplemented with microalgal protein in a way that allows for the use of microbial control techniques (e.g., pasteurization, HTST and hot-filling) without substantial precipitation or aggregation of the protein and without large changes in viscosity of the food. Not only is the microalgal protein supplementation ecologically and nutritionally advantageous, it also bypasses rheological and associated sensory problems resulting from heat-treating acidic liquid foods that have been supplemented with whey, soy protein, and many other unmodified protein concentrates.
As a result of the microalgal protein supplementation, the food product will comprise at least 0.1, 0.5 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, or 30% microalgal protein from the microalgal flour. For example, the food product can comprise 1-5, 5-10, 10-15, or 15-20% microalgal protein.
Because the microalgal cells are intact, the intracellular proteins will be protected from aggregating with the intracellular proteins of other cells, even after harsh pathogen inactivation steps. The cell walls will render the cells as a whole stably dispersible in the liquid. The cells can be intact such that at least 70, 75, 80, 85, 90, or 95% of the intracellular proteins remain within the cell at various stages of the process including in the completed food product.
The microalgal biomass can be combined with other ingredients to form a low pH food product (step 110). The other ingredients can be inherently acidic as in the case of a fruit juice, coffee or tomato product, or can be adjusted to have a low pH by including an acid as an ingredient. For example, vinegar, or solid citric acid can be added. The acidic food product can be, for example, a fruit juice, vegetable juice, hot and sour soup, a pickle (vegetable or otherwise), salad dressing, coffee beverage, or sauce (including tomato sauce). The final pH after addition of all ingredients can be between 3.0 and 6.0; e.g., between 3.3 and 5.5, or between 4 and 5. In a specific embodiment, the pH is 4.6 or less. This meets the definition of an acid food or an acidified food under Title 21 part 114 of the US Code of Federal Regulations (CFR).
The low pH food is then subjected to a microbial inactivation step (step 120). Although pasteurization (including HTST) is most commonly used, other techniques can be used including high pressure treatment, pulsed electric field, ultrasound, and ultraviolet light. For those microbial inactivation techniques that cause protein precipitation (e.g., in whey and soy protein-supplemented foods), the use of the high-protein microalgal biomass can prevent unwanted changes in viscosity and organoleptic properties. The inactivation step can result in 1, 2, 3, 4, or 5 log, or greater reduction in microbial counts. The inactivation step may meet US Food and Drug Administration guidelines and regulations for processing juice including those related to CFR Title 21 , including part 120 and related HACCP (hazard analysis and critical control points) including regulations and guidelines for 5 log pathogen reduction.
For example, where HTST is used, the liquid may move through a heating zone while subjected to temperatures between 71.5 and 74° C. for about 15 to 30 seconds. For example, the US standard for flash pasteurization of milk can be used; i.e., 71.7° C. for 15 seconds (shown to give 5 log reduction in harmful bacteria). Inactivation can be determined by microbial testing or using an appropriate model such as alkaline phosphatase inactivation.
In an embodiment, the liquid is treated at between 60-140° C. for 5-60 seconds, 70-100° C. for 5-60 seconds, 70-90° C. for 6 to 60 seconds, 70-80° C. for 5 to 60 seconds, 70-90° C. for 10 to 40 seconds, or 70-80° C. for 10 to 40 seconds.
Alternately, the liquid is pasteurized at a low temperature for a long time (LTLT). For example, the liquid can be treated under batch pasteurization conditions of 63° C. FOR 30-60 minutes. The conditions can be altered provided that sufficient bacterial inactivation is achieved. In various embodiments, the liquid is heated at a temperature of between 55-69° C. for 10 to 120 minutes, or between 60-68° C. for 15 to 100 minutes, from 61-67° C. for 20 to 80 minutes, or from 62-66° C. for 25 to 60 minutes. Time and temperature conditions can be chosen or optimized to give a 5 log reduction in pathogens.
The food product can then be placed in an appropriate container (step 130). In an embodiment, HTST and hot-filling is used on an acid food or acidified food. Because the food is at a low-pH, expensive aseptic filling is not necessary. For avoidance of doubt, aseptic filling is technically compatible with the embodiments of the invention and can be practiced; however, it is more expensive.
The steps described above can be used in numerous combinations resulting in a low-pH liquid food product that is supplemented with 0.5% or more microalgal protein yet does not become sandy, gritty, chalky, or increase substantially in viscosity after a microbial inactivation step. In specific embodiments, the viscosity increases after the pathogen inactivation step by no more than 0.5, 1, 2, 5, 7 or 10%. The heating step can have temperature and time parameters such that at least a 1, 2, 3, 4, or 5 log reduction in bacterial load is achieved.
In an embodiment, the pathogen inactivation step is a heating step having temperature and time parameters such that if a 10% high-protein microalgal flour solution at pH 6.0 were heated in the heating step, the viscosity of the solution would increase by no more than 5%, whereas the viscosity of a 10% whey or soy protein solution at pH 6 would increase by at least 10%.
In an embodiment, the pathogen inactivation step is a heating step having temperature and time parameters such that if a 10% high-protein microalgal flour solution at pH 4.0 were heated in the heating step, the viscosity of the solution would increase by no more than 5%, whereas the viscosity of a 10% whey or soy protein solution at pH 6 would increase by at least 10%.
In an embodiment, the pathogen inactivation step is a heating step having temperature and time parameters such that if a 10% high-protein microalgal flour solution at pH 3.3 were heated in the heating step, the viscosity of the solution would increase by no more than 5%, whereas the viscosity of a 10% whey or soy protein solution at pH 6 would increase by at least 10%.
The use of intact cells protects proteins without the need for adding hydrocolloid stabilizers such as pectin, carrageenan, plant gums or the like.
In a specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 50% protein by dry cell weight, less than 200 ppm of chlorophyll and less than 5% DHA. The flour is mixed with a low-pH liquid such as fruit juice, sauce or salad dressing so that the acidic liquid comprises at least 1% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate at least 3 logs of pathogens and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in the dark so as to have at least 55% protein by dry cell weight, less than 100 ppm of chlorophyll and less than 3% DHA. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 2% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate at least pathogens by at least 4 logs and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 60% protein by dry cell weight, less than 50 ppm of chlorophyll and less than 2% DHA. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 4% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate pathogens by at least 5 logs and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 60% protein by dry cell weight, less than 50 ppm of chlorophyll and less than 2% DHA. The microalgal flour comprises intact cells such that at least 85% of the intracellular protein in the cells remains in the cells. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 4% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate pathogens by at least 5 logs and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 60% protein by dry cell weight, less than 20 ppm of chlorophyll and less than 1% DHA. The microalgal flour comprises intact cells such that at least 90% of the intracellular protein in the cells remains in the cells. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 5% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate pathogens by at least 5 logs and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 60% protein by dry cell weight, less than 20 ppm of chlorophyll and less than 1% DHA. The microalgal flour comprises intact cells such that at least 90% of the intracellular protein in the cells remains in the cells. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 7% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate pathogens by at least 5 logs and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 60% protein by dry cell weight, less than 20 ppm of chlorophyll and less than 1% DHA. The microalgal flour comprises intact cells such that at least 90% of the intracellular protein in the cells remains in the cells. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 10% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is heat treated under conditions sufficient to inactivate pathogens by at least 5 logs and hot filled. The viscosity of the liquid increases by less than 10% after the heat treatment.
In another specific embodiment, a microalgal flour of Chlorella protothecoides is produced heterotrophically in a dark environment so as to have at least 60% protein by dry cell weight, less than 20 ppm of chlorophyll and less than 1% DHA. The microalgal flour comprises intact cells such that at least 90% of the intracellular protein in the cells remains in the cells. The flour is mixed with a low-pH liquid such as fruit juice or salad dressing so that the acidic liquid comprises at least 10% microalgal protein and has a pH of 4.6 or less. The microalgal protein-supplemented acidic liquid is HTST treated under conditions sufficient to inactivate pathogens by at least 5 logs and hot filled. The viscosity of the liquid increases by less than 10% after the HTST treatment.
Crude protein concentration for the microalgal flour was determined by the Dumas method applying an adjustment factor of 6.25.
We tested the viscosity of acidic solutions of 10% high-protein Chlorella protothecoides flour comprising about 65% crude protein by dry cell weight (“whole algal protein” or “WAP”) under conditions that simulate typical food processing conditions. The solutions were heated for 0 min, 10 min and 20 min at 70° C. We used 10% whey and soy protein as comparators. The experiment was performed at pH 3.3, 4.0 and 6.0
The instrument used in the experiment was the Brookfield LV DV-II+ Pro with the small sample adapter or the UL adapter. The different adapters allowed measurement of viscous (but pourable) to low viscosity fluids. We used the UL adapter for the low viscosity protein solutions, and this required about 25-30mL of fluid. The small sample adapter is for the more viscous protein solutions, and this required 10-20mL of fluid. The instrument was also measuring all the protein solutions at a consistent 22° C. with help from a thermosel connected to a controlled water bath set to 22° C.
The procedure was to measure the viscosity of the solutions at different shear rates, and then compare the proteins where the fluids have constant viscosity. In the experiment, the different protein solutions show constant viscosity between shear rate 61-66 s−1.
As can be seen in
Likewise, as can be seen from Table 1, at pH 4.0, soy protein was especially unstable at pH 3.3. and 6.0, whey protein was especially unstable pH 4.0 and 6.0, but the WAP was stable under all three conditions, with a viscosity that did not exceed 4.7.
Components of a protein beverage were experimentally combined in various proportions to achieve a surprising and unexpected result. Microalgal flour was introduced to numerous fruit juices and concentrates in various combinations to identify compatible and appealing low pH fruit beverages. In one experiment, juices or concentrates of passion fruit, blueberry and raspberry were combined with microalgal flour; in another experiment, juices or concentrates of raspberry, cranberry, and lemon were combined with microalgal flour. In further experiments, other flavors were tested. In yet another experiment, juices or concentrates of passion fruit and mango were combined with microalgal flour; in another experiment, juices or concentrates of mango and pineapple were combined with microalgal flour.
In additional experiments, protein levels as well as flavors were adjusted. In at least three formulations, microalgal flour comprised 3%, 4%, or 5%, by weight, of the respective batches, resulting in beverages that could be characterized as strong and savory, sweet and sour, or balanced.
An exemplary low pH beverage with tropical flavors is provided below. In this beverage, juices, concentrates, and flavors of several fruits were combined with microalgal flour to create an appealing juice drink with 5 grams of protein per 240 gram serving and a final pH of 4.2.
Low pH fruit beverages made with pea protein or rice protein instead of algal flour were unacceptable because the non-algal protein precipitated out and the texture of the beverage was chalky and gritty. Additionally, the viscosity of the low pH beverages made with pea protein or rice protein increased significantly, making the beverages unacceptable.
The low pH tropical fruit flavored beverage was made by the following procedure. Microalgal flour (3% by dry weight of the finished formula) was mixed with cane sugar (2% by dry weight of the finished formula). To this dry mixture, water (16% dry weight of the finished formula), orange juice (38.74% dry weight of the finished formula), and apple juice (35.38% dry weight of the finished formula) were added and combined using a high speed mixer, and allowed to sit for 30 minutes. After 30 minutes, tropical concentrate (4.6% dry weight of the finished formula), mango flavor (0.08% dry weight of the finished formula), and tropical flavor (0.1% dry weight of the finished formula) were added to the protein-water-juice formulation. The pH of the formulation was measured and adjusted using citric acid (0.1% dry weight of the finished formula) until the combination reached the desired level of acidity, pH 4.2). The formulation was heated to approximately 88° C. for 45 seconds, poured into containers, and cooled to approximately 4° C. As discussed above, beverages made with non-algal proteins were unacceptable because the precipitated pea protein and rice protein made the beverage chalky and gritty. Viscosity of the beverage was minimally impacted by the addition of algal protein and was very similar to typical juices that were not fortified with protein. The texture of the formulation was smooth and very similar to typical juices that were not fortified with protein.
Although the above discussion discloses various exemplary embodiments of the invention, it should be apparent that those skilled in the art can make various modifications that will achieve some of the advantages of the invention without departing from the true scope of the invention.
| Number | Date | Country | |
|---|---|---|---|
| 62018417 | Jun 2014 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 14752770 | Jun 2015 | US |
| Child | 16447809 | US |
