US 20120288587A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0288587 A1 Cheng et al. (43) Pub. Date: Nov. 15, 2012 (54) METHOD FOR PREPARINGA Publication Classification PROTEINACEOUS VEGETABLE FLAVOR ENHANCER (51) Int. Cl. A23. 3/34 (2006.01) (75) Inventors: Taiben Cheng, Glendale Heights, IL (US); Dafne Diez de Medina, (52) U.S. Cl. .......................................................... 426/52 Naperville, IL (US) (57) ABSTRACT (73) Assignee: GRIFFITH LABORATORIES INTERNATIONAL, INC. Alsip, This invention includes a method of producing hydrolyzed IL (US) Vegetable protein from proteinaceous vegetable material using commercial enzymes and optimally also fresh baker's (21) Appl. No.: 13/106,517 yeast as source of proteases and peptidases. The invention further includes a method of Solubilizing and pasteurizing (22) Filed: May 12, 2011 proteinaceous vegetable material. Patent Application Publication US 2012/0288587 A1 2 ^ 2 % a ev a GD2x 2. US 2012/0288587 A1 Nov. 15, 2012 METHOD FOR PREPARINGA causing the release of intracellular glutaminases and pepti PROTEINACEOUS VEGETABLE FLAVOR dases into the medium. The released enzymes then help ENHANCER hydrolyze the proteinaceous material, increasing the rate of the overall hydrolysis process. BACKGROUND OF INVENTION 0007 Disruption of the cell walls of yeasts has been 0001 Vegetable materials rich in protein, such as oilseed achieved by treating the microorganisms with Volatile organic cakes, cereal glutenand Soy, are widely used as flavor enhanc Solvents (such as acetone or ethanol), or with quaternary ers in the preparation of dehydrated or liquid Soups, sauces ammonium salts or Surfactants (see, for example, U.S. Pat. and seasonings. Such use requires that the proteinaceous No. 2.536,171). However, this process requires the use of a material be hydrolyzed, whereby the plant proteins are large amount of yeast (in a ratio of 1.1:1 of yeast to proteina degraded to absorbable amino acids and short peptides. ceous material), which may adversely affect the flavor of the Chemical methods are generally used to perform the hydroly resulting hydrolyzate. Furthermore, the chemicals used in the sis. For example, the proteinaceous vegetable material can be disruption step may find their way into the final product, hydrolyzed with concentrated hydrochloric acid, and the potentially compromising its purity and quality. resulting hydrolyzate then neutralized with sodium hydrox 0008. In an alternative approach, fiber-hydrolyzing ide. Upon removal of insoluble fractions by filtration, the enzymes, such as hemicellulase, have been used to hydrolyze hydrolyzate is subjected to discoloration, concentration and/ the cell walls of the yeasts and cause release intracellular or drying, providing a material that can be used for flavoring yeast enzymes into the reaction mixture. Unfortunately, this or as a food additive. method affords only modest yields of glutamate in the final 0002 Although easily implemented in industrial opera hydrolyzate (0.2%-2.1% w/w MSG). See, for example, U.S. tions, this chemical hydrolysis process has particular limita Pat. No. 6,569,476. tions. Acidic hydrolysis causes partial or complete decompo 0009 Since enzymatic hydrolysis generally requires long sition of some essential amino acids. Furthermore, during the contact times, there is the potential for putrefaction of the acidic hydrolysis chlorinated by-products, such as chlorohy proteinaceous material by contaminating bacteria. One com drins, may beformed, raising possible health concerns. As an mon strategy to prevent putrefaction or development of bac additional issue, the use of sodium hydroxide to neutralize the terial contamination is to pre-treat the reaction mixture with acidic mixture leads to a high salt content in the final product, preservatives, such as sodium benzoate or Sodium chloride. which may not be suitable for use in foods prepared for However, these preservatives not only affect the flavor of the individuals seeking to minimize dietary salt intake. final product, they also drastically increase its sodium con 0003. In order to avoid the problems associated with tent. Furthermore, the use of preservatives in the hydrolysis chemical hydrolysis of proteinaceous vegetable material, process may be unacceptable to consumers interested in “all there has been an increased interest in using enzymatic meth natural products. ods to perform the hydrolysis. To liberate as many amino 0010 Alternatively, putrefaction may be avoided by auto acids as possible from the plant material, the enzymatic claving the proteinaceous material before hydrolysis. How hydrolysis process employs either a complex mixture of ever, this results in heat destruction of nutritious components endoproteases and exoproteases (International Patent Publi Such as vitamins B, C and E and may also introduce an cation No. WO94/25580), or a combination of endoproteases undesired flavor to the resulting material. and a broad-spectrum exoprotease (International Patent Pub 0011. There is thus a need for a method that allows for lication No. WO98/27827). efficient enzymatic hydrolysis of proteinaceous vegetable 0004. However, enzymatic hydrolysis is not without draw protein. Such a method should allow the preparation of a backs. For example, enzymatic reactions are generally not hydrolyzate with high glutamate and soluble peptide content, efficient, since they may lead to partial digestion of the pro as well as a favorable flavor profile. There is also a need to teinaceous material and low yields of free glutamate or MSG identify a method to process proteinaceous vegetable mate (monosodium glutamate), an amino acid that plays a key role rial to minimize putrefaction and/or bacterial contamination in the perceived flavor of the hydrolyzate. Also, as a result of during hydrolysis. The present invention addresses all of the partial digestion afforded by enzymatic reactions, larger these needs. peptides that can impart an unpleasant taste to the hydroly Zate may be formed. BRIEF DESCRIPTION OF THE DRAWINGS 0005. The low efficiency of enzymatic hydrolysis of pro 0012 For the purpose of illustrating the invention, there is teinaceous plant materials is primarily due to the high content depicted in the drawing following certain embodiments of the of fiber and complex carbohydrates in the plant material. invention. However, the invention is not limited to the precise Carbohydrates, such as cellulose or glucans, form a rigid and arrangements and instrumentalities of the embodiments stable network within the plant material, greatly reducing the depicted. exposure of the plant proteins to the proteases or peptidases. 0013 FIG. 1 is a flow diagram illustrating a method for Therefore, in order to improve the enzymatic hydrolysis of producing proteinaceous vegetable flavor enhancer products plant-derived material, it is necessary to break down the car bohydrate network using fiber-hydrolyzing enzymes to in accordance with the present invention. increase the exposure of the plant proteins to the proteases DEFINITIONS and peptidases of the hydrolysis process. 0006. In an effort to increase the efficiency of the enzy 0014. The definitions used in this application are for illus matic hydrolysis process, in the past microorganisms such as trative purposes and do not limit the scope used in the practice yeasts, bacteria and fungi have been added to the hydrolysis of the invention. mixture. In this modified enzymatic process, the cell walls of 0015. Unless defined otherwise, all technical and scien the microorganisms in the reaction mixture are disrupted, tific terms used herein generally have the same meaning as US 2012/0288587 A1 Nov. 15, 2012 commonly understood by one of ordinary skill in the art to 0023 The proteinaceous vegetable material useful in the which this invention belongs. Generally, the nomenclature method of the invention may be a plant protein material used and the laboratory procedures in chemistry, analytical including, but not limited to, soya, wheat germ, corn gluten, chemistry, and food chemistry are those well known and rice gluten, wheat gluten, potatoes, okra, alfalfa, oats, and the commonly employed in the art. like, or may be a fermented protein koji prepared from the 0016. The articles “a” and “an are used herein to refer to protein containing material and a carbohydrate. The carbo one or to more than one (i.e. to at least one) of the grammatical hydrate may be, for example, wheat flour, roasted wheat or object of the article. By way of example, “an element’ means wheat bran. Seeds (such as Sunflower, pumpkin, squash, one element or more than one element. poppy, sesame, flax, chia, celery, rapeseeds, cottonseeds, and 0017. As used herein, the term "vegetable” means a plant caraway), leguminous seeds (such as peas, peanuts, beans, based material originated from a living organism of the Plan Soybeans and lentils), nuts (such as walnuts, coconuts, pine tae kingdom. The term "vegetable' encompasses leaves, nuts, cashew nuts, hickory nuts, chestnuts, almonds, brazil seeds, roots, tubers, bulbs, flowers, fruits, stems, shoots, nuts, nuts and filberts), and grains (wheat, corn, oats, rye, rice and and any combination thereof. barley) are other proteinaceous vegetable materials that are 0018. As used herein, the term “proteolytic activity” refers useful within the methods of the invention. to enzymatic activity that leads to hydrolysis of peptide 0024. According to a method of the invention, the pro bonds. teinaceous