Tobacco-derived cellulose sugar

Abstract

This document provides a method for forming cellulose sugars from the stems or roots of a *Nicotiana* species, the method comprising i) receiving tobacco material, said tobacco material comprising at least one of harvested stem and root material of a *Nicotiana* species; ii) deligating said tobacco material to form a tobacco-derived pulp; and iii) hydrolyzing said tobacco-derived pulp to form a hydrolyzed tobacco product comprising residual solids and a liquid, said liquid comprising at least one tobacco-derived cellulose sugar. This document also provides cellulose sugars derived from hydrolyzed tobacco material, said tobacco material comprising at least one of harvested tobacco stem and root material of a *Nicotiana* species. A tobacco product is also provided, said tobacco product incorporating said cellulose sugar derived from hydrolyzed tobacco material or a product derived from said cellulose sugar.

Description

Technical Field

[0001] This invention relates to products made from or derived from tobacco, or otherwise incorporated with tobacco or tobacco components. Of particular interest are components or parts obtained from or derived from the stems or roots of species of the genus *Nicotiana*. Background Technology

[0002] Cigarettes, cigars, and tobacco pouches are popular smoking products that use tobacco in different forms. These smoking products are used by heating or burning tobacco to produce a substance (such as smoke) that can be inhaled by the smoker. Popular smoking products (such as cigarettes) have a generally cylindrical rod-like structure and include a charge, tube, or column of flammable smoking material, such as shredded tobacco (e.g., in the form of desiccated tobacco) wrapped in wrapping paper, forming what is called a "tobacco rod." Typically, cigarettes have a cylindrical filter element aligned end-to-end with the tobacco rod. The filter element typically contains a tow of plasticized cellulose acetate surrounded by a paper material called a "wrapper." Some cigarettes incorporate a filter element with multiple sections, one of which may contain activated carbon particles. The filter element is typically attached to one end of the tobacco rod using a wrapping material called a "tip paper." It has also become desirable to perforate the tip paper and wrapper to provide dilution of the inhaled mainstream smoke with ambient air. Cigarettes are used by the smoker by lighting one end of the cigarette and burning the tobacco rod. Next, the smoker receives the mainstream smoke into his / her mouth by inhaling from the opposite end of the cigarette (e.g., the end of the filter).

[0003] Tobacco used in cigarette manufacturing is typically used in blends. For example, some popular tobacco blends, often referred to as “American blends,” contain a mixture of smoked tobacco, burley tobacco, and oriental tobacco, and in many cases, some processed tobacco, such as reconstituted tobacco and processed tobacco stems. The exact amount of each type of tobacco in the blend used to manufacture a particular brand of cigarette varies between brands. However, for many tobacco blends, smoked tobacco constitutes a relatively large proportion of the blend, while oriental tobacco constitutes a relatively small proportion. See, for example, *The Tobacco Encyclopedia*, edited by Voges, pp. 44-45 (1984), *The Design of Cigarettes*, 3rd ed., p. 43 (1990), and *Tobacco Production, Chemistry and Technology*, edited by Davis et al., p. 346 (1999).

[0004] Tobacco can also be enjoyed in so-called "smokeless" forms. Particularly popular smokeless tobacco products are consumed by inserting some form of processed tobacco or tobacco-containing blends into the user's mouth. See, for example, the types, ingredients, and processing methods of smokeless tobacco blends described in the following documents: U.S. Patent No. 1,376,586 to Schwartz; U.S. Patent No. 3,696,917 to Levi; U.S. Patent No. 4,513,756 to Pittman et al.; U.S. Patent No. 4,528,993 to Sensabaugh, Jr. et al.; U.S. Patent No. 4,624,269 to Story et al.; U.S. Patent No. 4,991,599 to Tibbetts; U.S. Patent No. 4,987,907 to Townsend; U.S. Patent No. 5,092,352 to Sprinkle, III et al.; U.S. Patent No. 5,387,416 to White et al.; and U.S. Patent No. 6,668 to Williams. 839; Williams' U.S. Patent No. 6,834,654; Atchley et al.'s U.S. Patent No. 6,953,040; Atchley et al.'s U.S. Patent No. 7,032,601; and Atchley et al.'s U.S. Patent No. 7,694,686; Williams' U.S. Patent Publication No. 2004 / 0020503; Quinter et al.'s U.S. Patent Publication No. 2005 / 0115580; Strickland et al.'s U.S. Patent Publication No. 2005 / 0244521; Strickland et al.'s U.S. Patent Publication No. 2006 / 0191548; Holton, Jr. et al.'s U.S. Patent Publication No. 2007 / 0062549; Holton, Jr.U.S. Patent Publication No. 2007 / 0186941, et al.; U.S. Patent Publication No. 2007 / 0186942, Strickland et al.; U.S. Patent Publication No. 2008 / 0029110, Dube et al.; U.S. Patent Publication No. 2008 / 0029116, Robinson et al.; U.S. Patent Publication No. 2008 / 0029117, Mua et al.; U.S. Patent Publication No. 2008 / 0173317, Robinson et al.; U.S. Patent Publication No. 2008 / ... US Patent Publication No. 0196730; US Patent Publication No. 2008 / 0209586, Neilsen et al.; US Patent Publication No. 2008 / 0305216, Neilsen et al.; US Patent Publication No. 2009 / 0025738, Neilsen et al.; US Patent Publication No. 2009 / 0025739, Neilsen et al.; US Patent Publication No. 2009 / 0065013, Neilsen et al.; US Patent Publication No. 2009 / 02938 ... U.S. Patent Publication No. 2010 / 0018540; U.S. Patent Publication No. 2010 / 0018541 by Gerardi et al.; U.S. Patent Publication No. 2010 / 0291245 by Gao et al.; U.S. Patent Publication No. 2011 / 0139164 by Mua et al.; U.S. Patent Publication No. 2011 / 0174323 by Coleman, III et al.; U.S. Patent Publication No. 2011 / 0247640 by Beeson et al.; U.S. Patent Publication No. 2011 / 02593 by Coleman, III et al. 53; U.S. Patent Publication No. 2012 / 0037175, Cantrell et al.; U.S. Patent Publication No. 2012 / 0055494, Hunt et al.; U.S. Patent Publication No. 2012 / 0103353, Sebastian et al.; U.S. Patent Publication No. 2012 / 0125354, Byrd et al.; U.S. Patent Publication No. 2012 / 0138073, Cantrell et al.; and U.S. Patent Publication No. 2012 / 0138074, Cantrell et al.; PCT, Arnarp et al. WO 04 / 095959; PCT WO 05 / 063060 by Atchley et al.; PCT WO 05 / 004480 by Engstrom; PCT WO 05 / 016036 by Bjorkholm; PCT WO 05 / 041699 by Quinter et al. and PCT WO 10 / 132444 by Atchley; each of these references is incorporated herein by reference.

[0005] One type of smokeless tobacco product is called "snuff." In Europe, particularly in Sweden, representative types of wet snuff products (often referred to as "snus") have been manufactured by or through companies such as Swedish Match AB, Fiedler & Lundgren AB, Gustavus AB, Skandinavisk Tobakskompagni A / S, and Rocker Production AB. In the United States, snus products are sold under the trademarks of RJ Reynolds Tobacco Company: Camel Snus Frost, Camel Snus Original, and Camel Snus Spice. See also, for example, Bryzgalov et al., 1N1800 Life Cycle Assessment, Comparative Life Cycle Assessment of General Loose and Portion Snus (2005). Additionally, certain quality standards associated with snus manufacturing have been compiled into the so-called GothiaTek standards.Representative smokeless tobacco products are also sold under the following brand names: Oliver Twist from the Oliver Twist A / S family; Copenhagen Wet Tobacco, Copenhagen Pouch Tobacco, Skoal Bandits, Skoal Pouches, Skoal Dry, Rooster, Red Seal Long Tobacco, Husky, and Revel Menthol Tobacco from Smokeless Tobacco Co.; Marlboro Snus and Taboka from Philip Morris; and Levi Garrett, Peachy, and Taylor's from American Snuff Company, LLC. Pride, Kodiak, Hawken Wintergreen, Grizzly, Dental, Kentucky King, and Mammoth Cave; Camel Snus, Camel Orbs, Camel Sticks, and Camel Strips from RJ Reynolds Tobacco Company. Other exemplary smokeless tobacco products for sale include products with the following names: Kayak moist snuff and Chattanooga Chew chewing tobacco from Swisser International, Inc.; and Redman chewing tobacco from Pinkerton Tobacco Co. LP.

[0006] The expectation is to provide additional uses for parts of the tobacco plant that are typically considered waste. Specifically, it will be advantageous to develop products derived from tobacco biomass, specifically products derived from the stems or roots of species in the genus *Nicotiana*. Summary of the Invention

[0007] This invention provides a method for forming cellulose sugars from the stems or roots of a *Nicotiana* species, comprising i) receiving tobacco material comprising at least one of harvested stem and root material of a *Nicotiana* species; ii) deligating the tobacco material to form a tobacco-derived pulp; and iii) hydrolyzing the tobacco-derived pulp to form a hydrolyzed tobacco product comprising residual solids and a liquid, the liquid comprising at least one tobacco-derived cellulose sugar. In various embodiments, the tobacco material may comprise at least about 90% dry weight of harvested stem and root material of a *Nicotiana* species.

[0008] In various embodiments, the step of deligating the tobacco material may include milling at least one of the harvested stem and root materials of a Nicotiana species to form tobacco-derived pulp. The step of deligating the tobacco material may further include, for example, rinsing and dehydrating the tobacco-derived pulp. In some embodiments, the step of deligating the tobacco material may further include adjusting the pH of the tobacco-derived pulp to a value in the range of about 4.5 to about 5.5. The method of the present invention may further include drying the tobacco-derived pulp to a moisture content of at least 10% or less.

[0009] In various embodiments, the step of hydrolyzing tobacco-derived pulp may include enzymatic saccharification of the tobacco-derived pulp in the presence of at least one enzyme. In some embodiments, the method may include a two-step saccharification process comprising adding at least one enzyme to the enzymatically hydrolyzed tobacco-derived pulp to reduce the amount of xylose in the hydrolyzed tobacco product.

[0010] In various embodiments, the method may further include evaporating a liquid containing at least one tobacco-derived cellulose sugar to form a condensed syrup. The condensed syrup may contain, for example, at least about 80% by weight of glucose. In some embodiments, the condensed syrup may contain, for example, about 15% by weight of xylose or less.

[0011] In various embodiments, the method may further include using a nickel catalyst to convert glucose derived from the original tobacco material into sorbitol via hydrogenation. In some embodiments, the method may further include converting glucose derived from the original tobacco material into levulinic acid via hydrolysis.

[0012] In various embodiments, the method may further include adding yeast to the hydrolyzed tobacco product and allowing the hydrolyzed tobacco product and yeast to ferment and form a fermented pulp containing glycerol. The average yield of glycerol produced may, for example, be at least about 10% by weight of the total weight of the delignified tobacco-derived pulp.

[0013] In some embodiments, the method may further include separating residual solids and liquid; adding yeast to the liquid; and allowing the liquid and yeast to ferment and form a fermented product containing ethanol. In some embodiments, the method may further include separating residual solids and liquid; adding a high-protein culture medium to the liquid; and allowing the liquid and the high-protein culture medium to ferment and form a fermented product containing lactic acid.

[0014] In various embodiments, the method may further include incorporating tobacco-derived cellulose sugars into the tobacco product. The tobacco product may be, for example, a smoking article.

[0015] The present invention also provides cellulose sugars derived from hydrolyzed tobacco material comprising at least one of harvested tobacco stalk and root material from a tobacco species. Prior to hydrolysis, the tobacco material may comprise at least about 90% by dry weight of harvested tobacco stalk and root material from a tobacco species.

[0016] In several embodiments, a tobacco product may be provided, which incorporates cellulose sugars derived from hydrolyzed tobacco material or downstream products derived from cellulose sugars. Downstream products of the cellulose sugars may be selected from the group consisting of: high-fructose tobacco syrup, glycerol, levulinic acid, lactic acid, and combinations thereof.

[0017] This invention includes (but is not limited to) the following embodiments:

[0018] Example 1: A method for forming cellulose sugars from the stems or roots of a species of the genus *Nicotiana*, comprising:

[0019] i) Receiving tobacco material, said tobacco material comprising at least one of stem material and root material from harvested tobacco species;

[0020] ii) Deligninating tobacco material to form tobacco-derived pulp; and

[0021] iii) Hydrolyzing tobacco-derived pulp to form a hydrolyzed tobacco product comprising residual solids and liquid, said liquid comprising at least one tobacco-derived cellulose sugar.

[0022] Example 2: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the tobacco material comprises at least about 90% by dry weight of harvested stem and root material of a tobacco species.

[0023] Example 3: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the step of deligating the tobacco material comprises grinding at least one of the harvested stem and root materials of a tobacco species to form tobacco-derived pulp.

[0024] Example 4: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the step of deligating the tobacco material further comprises washing and dehydrating the tobacco-derived pulp.

[0025] Example 5: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the step of deligating the tobacco material further comprises adjusting the pH of the tobacco-derived pulp to a value in the range of about 4.5 to about 5.5.

[0026] Example 6: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising drying the tobacco-derived pulp to a moisture content of at least 10% or less.

[0027] Example 7: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the step of hydrolyzing the tobacco-derived pulp includes enzymatic saccharification of the tobacco-derived pulp in the presence of at least one enzyme.

[0028] Example 8: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising adding at least one enzyme to the hydrolysis step (iii) to reduce the amount of xylose in the hydrolyzed tobacco product.

[0029] Example 9: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising evaporating a liquid containing at least one tobacco-derived cellulose sugar to form a viscous syrup.

[0030] Example 10: A method for forming cellulose sugar from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the condensed syrup contains at least about 80% by weight of glucose.

[0031] Example 11: A method for forming cellulose sugars from the stems or roots of tobacco species according to any of the foregoing or subsequent examples, further comprising using a nickel catalyst to convert glucose into sorbitol by hydrogenation.

[0032] Example 12: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising converting glucose into levulinic acid by hydrolysis.

[0033] Example 13: A method for forming cellulose sugar from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the condensed syrup contains about 15% by weight xylose or less.

[0034] Example 14: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising adding yeast to the hydrolyzed tobacco product, and allowing the hydrolyzed tobacco product and yeast to ferment and form a fermented pulp containing glycerol.

[0035] Example 15: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, wherein the average yield of glycerol produced is at least about 10% by weight of the tobacco-derived pulp.

[0036] Example 16: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising separating residual solids and liquids; adding yeast to the liquid; and allowing the liquid and yeast to ferment and form a fermented product containing ethanol.

[0037] Example 17: A method for forming cellulose sugars from the stems or roots of a tobacco species according to any of the foregoing or subsequent examples, further comprising separating residual solids and liquids; adding a high-protein culture medium to the liquid; and allowing the liquid and the high-protein culture medium to ferment and form a fermented product containing lactic acid.

[0038] Example 18: A method for forming cellulose sugars from the stems or roots of a species of the genus *Nicotiana* according to any of the foregoing or subsequent examples, further comprising incorporating tobacco-derived cellulose sugars into a tobacco product.

[0039] Example 19: A method for forming cellulose sugars from the stems or roots of a species of the genus *Nicotiana* according to any of the foregoing or subsequent examples, wherein the tobacco product is a smoking product.

[0040] Example 20: A cellulose sugar derived from hydrolyzed tobacco material, said tobacco material comprising at least one of harvested tobacco stem material and tobacco root material from a tobacco species.

[0041] Example 21: Cellulose sugar according to any of the foregoing or subsequent examples, wherein the tobacco material comprises at least about 90% by dry weight of at least one of the stem and root materials of a harvested tobacco species.

[0042] Example 22: A tobacco product incorporating a cellulose sugar or a downstream product of a cellulose sugar according to any of the foregoing or subsequent examples, wherein the downstream product of the cellulose sugar is selected from the group consisting of: high fructose tobacco syrup, glycerol, levulinic acid, lactic acid, and combinations thereof.

[0043] These and other features, aspects, and advantages of this disclosure will become apparent from the following detailed description together with the accompanying drawings, which are briefly described below. The invention includes any combination of two, three, four, or more of the embodiments pointed out above, as well as any combination of two, three, four, or more features or elements set forth in this disclosure, regardless of whether such features or elements are explicitly combined in the description of a particular embodiment herein. This disclosure is intended to be read in its entirety such that any separable feature or element of the invention disclosed in any of its various aspects and embodiments should be considered as intended to be composable unless the context clearly indicates otherwise. Attached Figure Description

[0044] To provide an understanding of embodiments of the invention, reference is made to the accompanying drawings, which are not necessarily drawn to scale, and wherein reference numerals refer to components of exemplary embodiments of the invention. The drawings are merely illustrative and should not be construed as limiting the invention.

[0045] Figure 1 It is a flowchart describing a method for forming cellulose sugars and optional downstream products from tobacco biomass materials;

[0046] Figure 2 It is an exploded perspective view of a smoking product in the form of a cigarette, showing the ignitable smoking material, packaging material components, and filter elements of the cigarette;

[0047] Figure 3 This is a top view taken across the width of an embodiment of a smokeless tobacco product, showing an outer bag filled with tobacco material; and

[0048] Figure 4 This is a cross-sectional view of an electronic smoking article according to an exemplary embodiment of the present disclosure, the electronic smoking article comprising a cylindrical tube and a control body and including a reservoir housing. Detailed Implementation

[0049] The invention will now be described more fully below. However, the invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and these embodiments will fully convey the scope of the invention to those skilled in the art. Unless the context clearly requires otherwise, the singular forms “a / an” and “the” as used in this specification and claims include a plurality of indicators. References to “dry weight %” or “dry basis” refer to weight based on dried components (i.e., all components except water).

[0050] This invention provides a method for forming cellulose sugars from the stems or roots of tobacco species, comprising i) receiving tobacco material comprising at least one of harvested stem and root material of tobacco species; ii) deligating the tobacco material to form tobacco-derived pulp; and iii) hydrolyzing the tobacco-derived pulp to form a hydrolyzed tobacco product comprising residual solids and liquid, the liquid comprising at least one tobacco-derived cellulose sugar. In several embodiments, the method may further comprise converting the tobacco-derived cellulose sugar into downstream products, as discussed in more detail below. This invention also provides cellulose sugars derived from hydrolyzed tobacco material comprising at least one of harvested tobacco stem and root material of tobacco species.

[0051] As in Figure 1 As shown and discussed in more detail below, tobacco biomass (loaded at operation 100) may undergo a pulping process at operation 102, and in some embodiments, a delignification process at operation 104. The delignified products may include, for example, lignin, xylose, and cellulose pulp. Lignin and xylose may be separated from the pulp, and lignin may be recovered, for example, at operation 106. Optionally, for example, at operation 108, xylose may undergo a conversion and recovery process to produce xylose syrup, glycerol, lactic acid, acetic acid, and combinations thereof. Additionally, for example, at operation 110, cellulose pulp may undergo a hydrolysis and sugar recovery process to produce various products, including xylose, glucose, and glucose syrup. At operation 112, glucose may further undergo a conversion process to produce high-fructose tobacco syrup. At operation 114, glucose may also undergo various conversion processes to produce bioproducts, including glycerol, lactic acid, sorbitol, levulinic acid, ethanol, and combinations thereof, as discussed in more detail below.

[0052] Tobacco materials

[0053] As in Figure 1As shown in operation 100, for example, the preparation of tobacco material according to the invention may include harvesting plants from species of the genus *Nicotiana*, and in some embodiments, separating certain components from the plant, such as stems and / or roots, and physically processing these components. Although the whole tobacco plant or any of its components (e.g., leaves, flowers, pedicels, roots, stems, etc.) may be used in the invention, it is advantageous to use the stems and / or roots of the tobacco plant. The remainder of the specification focuses on the use of the stems and / or roots of the plant, but the invention is not limited to such embodiments.

[0054] Tobacco stems and / or roots can be separated into individual parts (e.g., roots separated from the stem, and / or root portions separated from each other, such as large, medium, and small root portions), or stems and roots can be combined. “Stem” means the stem remaining after the leaves (including pedicels and leaf blades) have been removed. “Root” and various specific root portions applicable to the invention can be defined and classified as described, for example, in the following references: Mauseth, *Botany: An Introduction to Plant Biology*, 4th edition, Jones and Bartlett Publishers (2009); and Glimn-Lacy et al., *Botany Illustrated*, 2nd edition, Springer (2006), which are incorporated herein by reference. Harvested stems and / or roots are typically washed, ground, and dried to produce material that can be described as particulate (i.e., chopped, crushed, ground, granulated, or powdered). As used herein, stems and / or roots may also refer to stems and / or roots that have undergone an extraction process to remove water-soluble material. The cellulose material remaining after the stem and / or root material has undergone the extraction process (i.e., pulp) is also suitable for use in this invention.

[0055] The root and stem of tobacco plants have a higher weight percentage of cellulose content than tobacco stalks. Therefore, the sugar yield potential of tobacco plant roots and stems is higher than that of tobacco stalks. Furthermore, tobacco stalks represent a valuable starting material for the preparation of tobacco reconstituted sheets and expanded stem materials used in tobacco products. Using tobacco stalks as a source of cellulose sugars would reduce the supply of tobacco stalks available for other tobacco manufacturing processes. Tobacco stalks and roots represent tobacco material not otherwise used in tobacco manufacturing, and therefore represent excellent raw materials for the preparation of tobacco-derived cellulose sugars. Additional tobacco raw materials not otherwise used in tobacco manufacturing are so-called tobacco dust (i.e., very small particulate tobacco material collected during cigarette manufacturing) and so-called destemmed dust (i.e., tobacco-derived material collected during the destemming of tobacco leaves). Tobacco dust and destemmed dust can also be used to produce cellulose sugars.

[0056] Although tobacco material may comprise material from any part of a tobacco species, the majority of the material typically comprises material obtained from the stems and / or roots of the plant. For example, in some embodiments, the tobacco material comprises at least 90% by dry weight, at least 92% by dry weight, at least 95% by dry weight, or at least 97% by dry weight of harvested stem and root material from a tobacco species.

[0057] The manner in which stems and / or roots are provided in this form can vary. For example, material obtained from the stems of a tobacco plant can be processed separately from material obtained from the roots of a tobacco plant. Alternatively, material from different parts of the stems and / or roots can be processed separately (e.g., material derived from different parts of the roots can remain separate throughout the processing). In some embodiments, material from different parts of a tobacco plant can be combined and processed together to form a single homogeneous tobacco material. In some embodiments, material from different parts of a tobacco plant is processed separately and then combined at some stage of processing to obtain a single tobacco product.

[0058] Preferably, the physical processing step includes crushing, grinding, and / or pulverizing the stems and / or roots from tobacco plants into particulate form using equipment and techniques for grinding, milling, etc. In some preferred embodiments, the stems and / or roots are dried prior to the physical processing step, and thus the stems and / or roots are relatively dry in form during grinding or milling. For example, the stems and / or roots can be ground or milled when the moisture content is less than about 15% by weight or less than about 5% by weight. In such embodiments, hammer mills, cutter heads, pneumatically controlled mills, etc., can be used.

[0059] Tobacco material provided after being crushed, ground, and / or pulverized from tobacco stems and / or roots can be of any size. Tobacco material may be such that the average width and / or length of its portions or blocks is between about 1 / 16 inch and about 2 inches, between about 1 / 4 inch and about 1 inch, or between about 1 / 4 inch and about 1 / 2 inch. In some embodiments, the average width and / or length of the tobacco material is greater than or equal to about 1 / 8 inch, greater than or equal to about 1 / 4 inch, greater than or equal to about 1 / 2 inch, greater than or equal to about 1 inch, or greater than or equal to about 2 inches.

[0060] In some embodiments, tobacco material may be treated with water to extract water-soluble components of the tobacco material therefrom. In some preferred embodiments, particulate or powdered tobacco material may be combined with water to form a moistened aqueous material (e.g., in the form of a suspension or slurry), and the resulting material is typically heated to achieve the extraction of various compounds. The water used to form the moistened material may be pure water (e.g., tap water or deionized water) or a mixture of water and a suitable co-solvent (such as certain alcohols). In some embodiments, the amount of water added to form the moistened material may be at least about 50% by weight, or at least about 60% by weight, or at least about 70% by weight, based on the total weight of the moistened material. In some cases, the amount of water may be described as at least about 80% by weight, or at least about 90% by weight.

[0061] The resulting extract may contain a certain level of solid (insoluble) material entrained in the liquid. Therefore, "extract" is intended to refer to material obtained after the stems and / or roots have come into contact with water, and may contain soluble components and dispersed solid components dissolved therein. Following the extraction process, the extracted liquid component is typically filtered to remove at least some of the solids. In other words, some or all of the tobacco material portion that is insoluble in an aqueous solvent is removed. The filtration process may involve passing the liquid through one or more filter screens to remove particulate matter of a selected size. The screens may be, for example, stationary, vibrating, rotating, or any combination thereof. The filter may be, for example, a filter press or pressure filter. In some embodiments, the filtration method used may involve microfiltration, ultrafiltration, and / or nanofiltration. Filter aids may be used to provide effective filtration, and the filter aids may contain any material commonly used for this purpose. For example, some commonly used filter aids include cellulose fibers, perlite, bentonite, diatomaceous earth, and other silica-containing materials. Alternative methods may also be used to remove the solid components, such as centrifugation or sedimentation / deposition of the components and siphoning of the liquid. See, for example, the processes and products described in: U.S. Patent Application Publication No. 2012 / 0152265 and U.S. Patent Application Publication No. 2012 / 0192880, Dube et al., which are incorporated herein by reference in their entirety. The extracted solid components may be used as starting tobacco materials in the various embodiments of the invention described herein.

[0062] The exact composition of the tobacco material produced according to the present invention can vary. The composition can depend in part on whether the tobacco material is prepared from tobacco stems, roots, or a combination thereof. Tobacco material prepared solely from material obtained from tobacco stems can exhibit different characteristics than tobacco material prepared solely from material obtained from tobacco roots. Similarly, tobacco material prepared from material obtained from certain portions of one of these components can exhibit different characteristics than material obtained from other portions of that component (e.g., tobacco material prepared from medium-sized root material can differ from tobacco material prepared from large-root material). For example, in some embodiments, the volatile compound content of tobacco material derived from tobacco stems is higher than that of tobacco material derived from tobacco roots.

[0063] The selection of tobacco species used in the process of this invention may vary; and specifically, one or more types of tobacco may be different. The types of tobacco used as the source of tobacco stems and / or roots for derived tobacco materials may be different. The tobaccos that can be used include smoked tobacco or Virginia tobacco (e.g., K326), Burley tobacco, sun-cured tobacco (e.g., Indian Kurnool and Oriental tobacco, including Katerini, Prelip, Komotini, Xanthi and Yambol tobacco), Maryland tobacco, dark tobacco, dark-fired tobacco, dark air-cured tobacco (e.g., Passanda, Cubano, Jatin and Bezuki tobacco), light-cured tobacco (e.g., North Wisconsin and Galpao tobacco), Indian air-cured tobacco, Red Russian and Rustica tobacco, as well as a variety of other rare or special tobaccos. Descriptions of various types of tobacco, their growth habits, and harvesting habits are presented in *Tobacco Production, Chemistry and Technology*, Davis et al. (eds.) (1999), which are incorporated herein by reference. Representative types of plants within the genus *Nicotiana* are described in: Goodspeed, *The Genus Nicotiana*, *Chonica Botanica* (1954); U.S. Patent No. 4,660,577, Sensabaugh, Jr. et al.; U.S. Patent No. 5,387,416, White et al., and U.S. Patent No. 7,025,066, Lawson et al.; U.S. Patent Application Publication No. 2006 / 0037623, Lawrence, Jr., and U.S. Patent Application Publication No. 2008 / 0245377, Marshall et al.; each of which is incorporated herein by reference.

[0064] The specific tobacco species used in this invention may vary. Of particular interest are *N. alata*, *N. arentsii*, *N. excelsior*, *N. forgetiana*, *N. glauca*, *N. glutinosa*, *N. gossei*, *N. kawakamii*, *N. knightiana*, *N. langsdorffi*, *N. otophora*, *N. setchelli*, *N. sylvestris*, *N. tomentosa*, *N. tomentosiformis*, *N. undulata*, and *N. sanderae*. Also of interest are N.africana, N.amplexicaulis, N.benavidesii, N.bonariensis, N.debneyi, N.longiflora, N.maritina, N.megalosiphon, N.occidentalis, N.paniculata , N.plumbaginifolia, N.raimondii, N.rosulata, N.rustica, N.simulans, N.stocktonii, N.suaveolens, N.tabacum, N.umbratica, N.velutina and N.wigandioides. Other plants derived from the genus *Nicotiana* include *N. acaulis*, *N. acuminata*, *N. attenuata*, *N. benthamiana*, *N. cavicola*, *N. clevelandii*, *N. cordifolia*, *N. corymbosa*, *N. fragrans*, *N. goodspeedii*, *N. linearis*, *N. miersii*, *N. nudicaulis*, *N. obtusifolia*, *N. occidentalis* subsp. *Hersperis*, *N. pauciflora*, *N. petunioides*, *N. quadrivalvis*, *N. repanda*, *N. rotundifolia*, *N. solanifolia*, and *N. spegazzinii*. *Nicotiana* species can be derived using genetic modification or hybridization breeding techniques (e.g., tobacco plants can be genetically engineered or hybridized to increase or decrease the production of certain components or otherwise alter certain characteristics or properties).See, for example, the types of plant genetic modifications described in the following: U.S. Patent No. 5,539,093 to Fitzmaurice et al.; U.S. Patent No. 5,668,295 to Wahab et al.; U.S. Patent No. 5,705,624 to Fitzmaurice et al.; U.S. Patent No. 5,844,119 to Weigl; U.S. Patent No. 6,730,832 to Dominguez et al.; U.S. Patent No. 7,173,170 to Liu et al.; U.S. Patent No. 7,208,659 to Colliver et al.; and U.S. Patent No. 7,230,160 to Benning et al.; U.S. Patent Application Publication No. 2006 / 0236434 to Conkling et al.; and PCT WO 2008 / 103935 to Nielsen et al.

[0065] One or more plant components from tobacco species can be used in their immature form. That is, the plant can be harvested before it reaches a stage that is normally considered ripe or mature. Thus, for example, the tobacco plant can be harvested when it is about to bud, is just beginning to form leaves, or is just beginning to flower.

[0066] Plant components from the genus *Nicotiana* can be used in their mature form. That is, plants can be harvested when they reach a point traditionally considered ripe, overripe, or mature. Thus, for example, Oriental tobacco plants can be harvested using tobacco harvesting techniques conventionally used by farmers, Burley tobacco plants can be harvested, or Virginia tobacco leaves can be harvested or picked based on stem position.

[0067] After harvest, tobacco species or parts thereof can be used in unprocessed form (e.g., tobacco can be used without any curing process). For example, unprocessed tobacco can be frozen, freeze-dried, irradiated, yellowed, dried, cooked (e.g., roasted, fried, or boiled), or otherwise stored or treated for later use. Such tobacco can also be subjected to aging conditions.

[0068] Transforming tobacco materials into cellulose materials

[0069] In various embodiments of the invention, tobacco material can be converted into cellulosic material, for example, through delignification of tobacco material. Delignification of tobacco material can involve multiple operations. As an initial step, tobacco biomass can undergo a pulping process. Pulp can be produced from the virgin material mechanically or chemically, for example, as in… Figure 1 As shown at operation 102.

[0070] For example, mechanical pulping technology can be used to produce tobacco stalk and / or root pulp. In the mechanical pulping process, virgin tobacco material can be cut, for example, and then fed between refiners, where the fragments are shaped into fibers between rotating metal discs. See, for example, mechanical pulping apparatus and processes disclosed in: U.S. Patent No. 4,421,595 to Huusari and U.S. Patent Nos. 7,237,733, WO 2010 / 023363, CA 1074606, and CN 201268810 to Vikman, all of which are incorporated herein by reference in their entirety. In some embodiments, the virgin tobacco stalks and / or roots may be pretreated with water for several hours. The water-to-stalk weight ratio may be from about 10:1 to about 5:1 (e.g., about 7:1). Pretreatment of the tobacco stalks softens them and removes water-soluble extracts. The pretreated mixture can then be drained to a consistency of about 20%. As used herein, the term "consistency" is defined as the percentage of solids in a mixture. This pretreated stalk can then be refined under atmospheric pressure by passing it multiple times through a stalk-cutting machine. See, for example, the machines discussed below: U.S. Patent No. 3,661,192 to Nicholson et al.; U.S. Patent No. 3,861,602 to Smith et al.; U.S. Patent No. 4,135,563 to Maucher; and U.S. Patent No. 5,005,620 to Morey, each of which is incorporated herein by reference. In various embodiments, the machine can be calibrated such that the target size of the stalk fragments passed through each time is reduced. This stalk-cut pulp can then be refined to various levels, for example, in a PFI mill. See, for example, the methods and apparatus discussed below: U.S. Patent No. 6,773,552 to Albert et al.; and U.S. Application Publication No. 2010 / 0036113 to Mambrim Filho et al., each of which is incorporated herein by reference.

[0071] As in Figure 1As shown at operation 104, for example, tobacco-derived pulp can undergo a deligninization process. Mechanical pulping does not separate lignin from cellulose fibers, therefore the yield is typically relatively high (i.e., above 95%). Several processes can be used to deligninate mechanically pulped tobacco material. For example, the pulp can be washed with water and dewatered at least once. The pulp can be dewatered by wet grading, centrifugation, filtration, or similar liquid separation processes. Centrifuges or other similar equipment can facilitate the separation of pulp and syrup (i.e., solid and liquid). See, for example, the equipment disclosed in: U.S. Patent No. 521,104 to Davis, U.S. Patent No. 3,168,474 to Stallman et al., U.S. Patent No. 5,713,826 to West, and U.S. Patent No. 7,060,017 to Collier, each of which is incorporated herein by reference in its entirety. For example, a basket centrifuge can be used to assist in pulp dewatering and lignin syrup recovery operations. Additionally, the pulp can be rinsed once or multiple times, and the pH can be adjusted to a range of about 4.5 to about 5.5. In a preferred embodiment, the pH can be adjusted to about 4.8. The pulp can be dewatered after each rinse.

[0072] In some embodiments, chemical pulping processes can be used to pulp and deligneinize tobacco biomass. Chemical pulping processes separate lignin from cellulose fibers by dissolving lignin in the cooking liquor, allowing the lignin binding the cellulose fibers together to be washed away from the cellulose fibers without significantly degrading them. Three main chemical pulping processes are known in the art. Soda pulping involves cooking fragments of raw material in a sodium hydroxide cooking liquor. The kraft paper process, evolved from soda pulping, involves cooking fragments of raw material in a solution of sodium hydroxide and sodium sulfide. The acid sulfite process involves using sulfite and bisulfate ions during cooking. The kraft paper process is the most commonly used method for chemical wood pulping; however, the soda process can also be used to produce some hardwood pulp. Any chemical pulping process, including (but not limited to) the three examples listed above, can be used to produce tobacco pulp from virgin tobacco material.

[0073] The cooking liquor may contain a strong base. As used herein, a strong base refers to a basic compound (or combination of such compounds) capable of deprotonating a very weak acid in an acid-base reaction. For example, strong bases suitable for use in this invention include (but are not limited to) one or more of the following: sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, ammonium hydroxide, ammonium bicarbonate, and ammonium carbonate. In some embodiments, the weight of the strong base may be greater than about 5%, greater than about 25%, or greater than about 40% of the weight of the tobacco added. In some embodiments, the weight of the strong base may be less than about 60% or less than about 50% of the weight of the tobacco added. In still other embodiments, the weight of the strong base may be from about 5% to about 50% or from about 30% to about 40% of the weight of the tobacco added. In other embodiments, various other chemicals and their weight ratios may also be used in the chemically pulped tobacco.

[0074] Besides combining tobacco with a strong alkali, tobacco added in chemical pulping can also include heated tobacco and a strong alkali. Heating the tobacco and strong alkali can increase the efficiency of chemical pulping. In this respect, increasing the cooking temperature or time will result in an increased reaction rate (lignin removal rate).

[0075] In various embodiments, organic solvent processes can be used to deliquinate tobacco biomass. Organic solvent (solvent-based) pulping is a chemical pulping method in which biomass is deliquinated via an organic solvent or a solvent-water system. See, for example, an organic solvent process described in: Ziaie-Shirkolaee et al., Study on Cellulose Degradation during Organosolv Delignification of Wheat Straw and Evaluation of Pulp Properties, Iranian Polymer Journal 16(2), 2007, pp. 83-96, which is incorporated herein by reference. Organic solvents use the acid decomposition of the pulp in the presence of a solvent (e.g., ethanol or butanol) to dissolve and remove lignin from the pulp. See, for example, U.S. Patent Application Publication No. 2013 / 0172628, which is incorporated herein by reference.

[0076] In some embodiments, methods for producing tobacco-derived cellulosic materials may include one or more additional operations. See, for example, U.S. Patent Application Publication No. 2013 / 0276801 by Byrd Jr. et al., which is incorporated herein by reference in its entirety. For example, the tobacco input may undergo additional processing steps prior to pulping, and / or the deligation method may include additional treatment steps (e.g., drying the tobacco input, depigmenting the tobacco input, milling the tobacco input, etc.). In some embodiments, these additional steps may be performed to artificially remove pith (which contains lignin) from the tobacco input and / or tobacco pulp, and thus, for example, reduce the amount of chemicals necessary to deliquinate the tobacco input during a chemical pulping process. Water may be mixed with the tobacco pulp to form a pulp, and the pulp may be filtered, for example, to remove some non-cellulosic materials, such as pith, parenchyma, and tissue, from the tobacco pulp. Additional treatment steps (e.g., milling the tobacco input) may be performed to increase the surface area of ​​the tobacco input, thereby increasing the efficiency of the pulping and / or bleaching operations. Steam- or water-based pre-hydrolysis of tobacco stalks prior to pulping can, for example, reduce the amount of chemicals required in bleaching operations. Anthraquinones can be used, for example, in chemical pulping methods in an attempt to provide higher yields by protecting carbohydrates from destruction by strong alkalis during delignification. Other processing steps known in the pulping and delignification fields can be used to form cellulosic materials from the input virgin tobacco.

[0077] In some embodiments, the aqueous mixture of lignin and xylose with pulp can be separated via separation processes known in the art, and lignin can be recovered at operation 106. For example, water can be boiled down to produce a more concentrated lignin syrup. In some embodiments, an acid or base can be added to the lignin syrup to form lignin salts. In some embodiments, a Group I or Group II metal (e.g., magnesium, calcium, potassium, etc.) can be added to the lignin syrup to form a solid (e.g., calcium hydroxide) via precipitation.

[0078] In some embodiments, such as at operation 108, xylose separated from the pulp may undergo optional conversion and recycling processes to produce xylose syrup, glycerol, lactic acid, acetic acid, and combinations thereof. The conversion and recycling processes may be similar to those described below regarding glucose conversion and recycling processes; however, the yield of xylose-derived bioproducts may be relatively low. In fact, in various embodiments, the conversion and recycling of xylose may primarily result in the formation of acetic acid.

[0079] Formation of tobacco-derived cellulose sugars

[0080] After the tobacco biomass is deligated, the cellulose material undergoes at least one saccharification process, such as, for example, in Figure 1The operation is shown at point 110. Any form of hydrolysis known in the art can be used to break down carbohydrates in tobacco-derived cellulosic materials into constituent sugar molecules. In some embodiments, a salt of a weak acid or a weak base (or both) can be dissolved in water during hydrolysis. For example, acid-base catalytic hydrolysis can also be used. In various embodiments, the cellulosic material can undergo enzymatic hydrolysis to form glucose. See, for example, a discussion of the enzymatic hydrolysis of cellulose presented in the following: Zhang, Yi-Heng Percival et al., "Toward an Aggregated Understanding of Enzymatic Hydrolysis of Cellulose: Noncomplexed Cellulase Systems," Wiley InterScience. Biotechnology and Bioengineering, Vol. 88, No. 7, December 30, 2004, pp. 797-824. Generally, cellulase systems can hydrolyze cellulosic materials. Insoluble cellulose materials can undergo three processes simultaneously: (i) chemical and physical changes in the residual solid-phase cellulose; (ii) primary hydrolysis of the solid-phase cellulose to form intermediates comprising cellobiose, soluble cellodextrin, and glucose; and (iii) secondary hydrolysis involving the hydrolysis of the soluble intermediates into lower molecular weight intermediates and ultimately into a liquid-phase product comprising glucose. Therefore, hydrolyzing tobacco-derived cellulose materials can produce a hydrolyzed tobacco product (i.e., a liquid paste) comprising residual solids and a liquid, the liquid containing at least one tobacco-derived cellulose sugar. In various embodiments, the at least one tobacco-derived cellulose sugar may include glucose, xylose, and combinations thereof.

[0081] In various embodiments, the enzyme used for saccharification may include CTec 2 (manufactured by Novozymes A / S). CTec 2 is an effective cellulase / hemicellulase for sugar production from biomass. In some embodiments, the enzyme used for saccharification may comprise an enzyme manufactured by DSM Food Specialties BV (Netherlands), such as an enzyme for food processing. Any cellulase / hemicellulase known in the art can be used in the enzymatic saccharification process described herein. Preferred enzyme concentrations and time periods for enzymatic hydrolysis are typically recommended by the manufacturer. In various embodiments, the enzyme concentration may range from about 1% by weight to about 10% by weight, or from about 2% by weight to about 5% by weight. In a preferred embodiment, the enzyme concentration may be about 3% by weight of the total material undergoing enzymatic hydrolysis. In various embodiments, significant hydrolysis (i.e., about 50% to about 75% conversion of the starting cellulose material) can be achieved within about 48 hours.

[0082] In various embodiments, residual solids can be removed from a liquid paste by centrifugation, filtration, or other liquid / solid separation methods. Centrifuges or other similar devices can facilitate solid-liquid separation. See, for example, devices disclosed in: U.S. Patent No. 521,104 to Davis, U.S. Patent No. 3,168,474 to Stallman et al., U.S. Patent No. 5,713,826 to West, and U.S. Patent No. 7,060,017 to Collier, each of which is incorporated herein by reference in its entirety.

[0083] In some embodiments, the separated liquids can be thickened to form a syrup. An evaporator can be used, for example, to thicken the liquid products. In some embodiments, a mechanical vapor recompression (MVR) evaporator can be adapted to assist in thickening the syrup. See, for example, the evaporators and processes disclosed in: U.S. Patent No. 4,303,468 to Laguilharre et al., U.S. Patent No. 3,396,086 to Starmer and U.S. Patent No. 4,530,737 to Ostman; and U.S. Patent Application Publication No. 2014 / 0262730 to Zimmer, each of which is incorporated herein by reference. A concentrated syrup derived from hydrolyzed tobacco products may contain at least about 80% by weight of glucose, at least about 85% by weight of glucose, or at least about 90% by weight of glucose. In some embodiments, the concentrated syrup may contain about 20% by weight or less xylose, about 15% by weight or less xylose, about 10% by weight or less xylose, or about 5% by weight or less xylose.

[0084] In various embodiments, enzymes may be added to the saccharification process to reduce the amount of xylose in the liquid product. Enzymes may, for example, remove hemicellulose prior to final saccharification. In various embodiments, enzymes may be selected from the group consisting of: (Manufactured by American BioSystems, Inc.) HC&HC2500 (manufactured by Novozymes) Xylanase (produced by Dyadic Inc.) and combinations thereof. It is an enzyme with high specificity for hemicellulose and only a small amount of cellulase activity. HC&HC 2500 is an enzyme that is highly specific to hemicellulose and also has very little cellulase activity. Xylanase is an enzyme that has been shown to have both high cellulase and high xylanase activity. The preferred enzyme concentration is usually recommended by the manufacturer.

[0085] Downstream products

[0086] As discussed above, the present invention provides cellulose sugars derived from hydrolyzed tobacco material comprising at least one of harvested tobacco stem and root material from tobacco species. The tobacco-derived cellulose sugar material produced according to the present invention is suitable for the production of a variety of products, including (but not limited to) humectants, acids, flavorings, ethanol, and other glucose-derived products. See, for example, in... Figure 1 The products listed in the process flow diagram are shown below. The various products produced from tobacco-derived glucose are discussed in more detail below.

[0087] High fructose tobacco syrup

[0088] As described above, tobacco-derived cellulose materials can undergo enzymatic hydrolysis to form, for example, glucose. Then, commercially available immobilized glucose isomerases (e.g., those produced by Novozymes) can be used. Type IT) produces high-fructose tobacco syrup. For example, in... Figure 1 As shown in operation 112, this isomerization, for example, can convert glucose, which is not very sweet, into fructose, the sweetest natural sugar. Syrups obtained from this process can compete with sucrose (cane sugar) in many food applications. For example, because high-fructose syrup is cheaper than sucrose, almost all soft drink manufacturers use high-fructose syrup. Tobacco products may also incorporate high-fructose syrup, for example, for flavoring.

[0089] Glucose isomerase (D-glucose ketone isomerase) isomerizes glucose to fructose. Glucose isomerization to fructose is part of the glycolytic cycle that converts glucose to pyruvate. This is accomplished by isomerizing aldehyde (hemiacetal) glucose to ketone (as hemiacetal) fructose, yielding another phosphate ester. Isomerization utilizes the easy breaking of the CH bond involving the carbon adjacent to the carbonyl carbon. In the next step, the bond between carbon 3 and carbon 4 of the fructose is broken.

[0090] For example, Novo Industries has developed [products] from Bacillus coagulans. Glucose isomerase, for commercial use. When a procedure for enzyme immobilization is developed that allows for the repeated use of the same batch of enzyme, a commercial process for producing fructose from glucose becomes feasible. In this immobilized enzyme process, microorganisms perform the direct isomerization of glucose. This glucose isomerase is primarily a xylose isomerase, therefore xylose or xylose-containing compounds must be added for enzyme induction.

[0091] glycerin

[0092] like Figure 1 As shown in operation 114, for example, glycerol can be produced from tobacco-derived glucose. Figure 1 As shown in operation 108, glycerol can also be produced, for example, from xylose recovered from tobacco pulp after lignin removal. Glycerol is suitable for a wide variety of industries and products. For example, in food and beverages, glycerol can be used as a humectant, solvent, sweetener, and combinations thereof. Glycerol can also be used to help preserve food, as a filler, and / or as a thickener. Glycerol is also suitable for medical, pharmaceutical, and personal care products (e.g., for providing lubrication and / or as a humectant). Various tobacco products may also incorporate glycerol in the form of a humectant, sweetener, solvent, and combinations thereof.

[0093] As described above, tobacco-derived cellulose materials can undergo enzymatic hydrolysis to form glucose. The enzymes used for saccharification may include, for example... CTec 3 (manufactured by Novozymes). CTec 3 is an efficient cellulase / hemicellulase for saccharification from biomass. In various embodiments, yeast may be added to the saccharification process to produce hydrolyzed tobacco products containing glycerol. For example, rapid-start baker's yeast may be used. In various embodiments, approximately 10 g of yeast may be used per liter of tobacco pulp slurry, wherein the tobacco pulp slurry is approximately 5% to 10% solids (e.g., approximately 9% solids). Nutrients and hops (i.e., food for the yeast) may also be used to supplement the process. The hydrolyzed tobacco products and yeast may be allowed to ferment, thereby forming a fermented pulp slurry containing glycerol. In various embodiments, the average yield of glycerol produced is at least approximately 10% by weight of the starting cellulose material. A weak base anion may be used on the solution to remove any lactic acid impurities.

[0094] In various embodiments, pH adjusters or buffers may be added to the saccharification process to maintain the pH within an optimal range, depending on the cellulose material and yeast used. For example, the pH may be maintained in the range of about 5 to about 10 during the saccharification process. Exemplary reagents include metal hydroxides, metal carbonates, metal bicarbonates, and mixtures thereof. Specific exemplary materials include sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, and sodium bicarbonate. The amount of pH adjuster or buffer used in enzymatic hydrolysis may vary, but is typically up to about 5% dry weight, and some embodiments may be characterized by a pH adjuster / buffer content of at least about 0.5% dry weight (e.g., from about 1% dry weight to about 5% dry weight).

[0095] In some embodiments, low doses of bisulfite (e.g., about 2000 ppm or less) may be added periodically throughout the reaction to prevent acetaldehyde from volatilizing and to increase glycerol production by the yeast. The bisulfite may be, for example, in the form of Na₂S₂O₅. During fermentation, a pH controller and alkali may be used to maintain the pH between 6.7 and 7.3 to stabilize the bisulfite concentration within a pH range tolerable by the yeast.

[0096] Sorbitol

[0097] As described above, tobacco-derived cellulose materials can undergo enzymatic hydrolysis to form glucose. In some embodiments, the glucose can be fused to form a syrup. The fused glucose syrup can be converted to sorbitol by hydrogenation, for example, using a nickel catalyst. Sorbitol is suitable as a sugar substitute. Sorbitol can also be used as a non-irritating laxative. Sorbitol is also suitable for medical applications and healthcare, food, and cosmetic uses, such as as a humectant and / or thickener. Sorbitol is also suitable as a humectant in tobacco products (e.g., smoking articles and / or smokeless tobacco products). In some aspects, sorbitol can be used to form transparent gels because it has a sufficiently high refractive index for transparent formulations.

[0098] A high-yield and efficient method for the production of sorbitol from glucose is known. Since 1942, this method has been used... Nickel is used to produce sorbitol via batch hydrogenation. See Zhang, J, *Ind. Eng. Chem. Res.*, 2013, 52, 11800, which is incorporated herein by reference. Improvements to the batch hydrogenation of glucose primarily involve changes to nickel sponge-type catalysts, such as the addition of molybdenum, chromium, and iron. Commercially available catalysts specifically designed for the production of sorbitol from glucose include (but are not limited to) BASF Actimet. TM C, WRGrace 3111, and Johnson Matthey "sponge metal catalyst 1". These catalysts typically contain about 2% by weight of molybdenum. Nickel catalysts are popular in industrial settings due to their low cost. However, they are not entirely stable in reaction environments.

[0099] During hydrogenation, nickel is applied to the product, and its removal increases costs, particularly in high-volume production or when USP-grade products are desired. Consequently, there has been a trend towards using ruthenium catalysts on various supports for the production of sorbitol. See Zhang, J, *Industrial and Engineering Chemical Research*, 2013, 52, 11802-11804, which is incorporated herein by reference. However, due to improvements in the leaching behavior of some nickel catalysts and their lower cost, nickel catalysts will likely remain the preferred option for the hydrogenation of glucose to sorbitol for some time until they are replaced.

[0100] Continuous processes are also commonly used for sorbitol production. A typical continuous process for producing sorbitol from glucose is the subject of U.S. Patent No. 4,322,569 to Chou, which is incorporated herein by reference. The precursor is fed at a controlled rate over a fixed bed of catalyst, where hydrogen is supplied at high pressure to hydrogenate glucose to sorbitol in high yield.

[0101] As revealed in Zhang, J., *Industrial and Engineering Chemical Research*, 2013, 52, 11805 (incorporated herein by reference), it has also been found that glucose can be converted to sorbitol without the use of gaseous hydrogen. Using Pd / C as a catalyst and sodium formate as a reducing agent, a yield of up to 71.7% and a sorbitol production selectivity of 94.1% were achieved. Formic acid is a byproduct of the production of levulinic acid from tobacco raw materials, and sodium formate is simply the sodium salt of said byproduct.

[0102] levulinic acid

[0103] As mentioned above, tobacco-derived cellulose materials can undergo enzymatic hydrolysis to form glucose. See, for example... Figure 1 Operation 114. In some embodiments, glucose can be fused to form a syrup. The fused glucose syrup can be converted into levulinic acid by hydrolysis. Levulinic acid can act as a precursor for pharmaceuticals, plasticizers, and various other additives. Potential biofuels can be prepared from levulinic acid. Various tobacco products can incorporate levulinic acid. For example, e-cigarette products can incorporate levulinic acid into a floating precursor composition.

[0104] The methods used to produce levulinic acid from biomass can be roughly grouped into four categories: (1) concentrated acid / low temperature hydrolysis; (2) dilute acid / high temperature hydrolysis; (3) continuous process; and (4) two-step continuous process.

[0105] The McKenzie organic synthesis procedure provides an example of concentrated acid / low-temperature hydrolysis, described in Organic Syntheses, Coll. Vol 1, p. 335 (1941), which is incorporated herein by reference. This type of hydrolysis has several disadvantages, such as the generation of large amounts of rotten matter, difficult handling, low yield, and being environmentally unfriendly due to its highly acidic waste stream.

[0106] Second-generation biomass hydrolysis to produce levulinic acid is a dilute acid / high-temperature hydrolysis reaction. Due to its high-temperature conditions, these reactions are carried out in pressure vessels. A table of 46 acid-catalyzed batch production methods for levulinic acid can be seen, for example, in: Girsuta, B, "Levulinic Acid from Lignocellulosic Biomass," paper, University of Groningen, 2007, which is incorporated herein by reference. Thirty-four of these methods utilize high temperatures (above 100°C) and low acid concentrations.

[0107] Continuous processes have been the preferred choice for the industrial production of levulinic acid for some time. Earlier continuous processes involved high-temperature hydrolysis in a continuous flow through a tubular furnace. More recent production processes utilize reactive extruders to raise the temperature of the acidic slurry and initiate the hydrolysis reaction in a more controlled manner. See, for example, U.S. Patent No. 5,859,263 to Ghorpade et al., which is incorporated herein by reference.

[0108] A more in-depth analysis of the chemical reactions constituting the process of biomass hydrolysis and levulinic acid production reveals that it is advantageous to separate the carbohydrate hydrolysis step from levulinic acid production. Biomass exposed to relatively high temperatures (e.g., about 210°C) for a short period (e.g., about 12 seconds) hydrolyzes to hexoses and pentoses in high yields. Continuous removal of pentose reaction products and adjustment of reaction conditions (e.g., about 190°C for about 20 minutes) can provide optimal levulinic acid production from hexoses.

[0109] Work at the University of Wisconsin offers another promising future direction for the industrial-scale production of levulinic acid. See, for example, U.S. Patent No. 8,389,749 to Dumesic, which is incorporated herein by reference. It is recognized that it is advantageous not only to separate the hydrolysis of biomass into sugars and levulinic acid, but also to continuously separate the intermediate hydroxymethylfurfural from an aqueous hydrolysis environment. This is achieved through a two-phase reactor design in which hydroxymethylfurfural is continuously extracted into an organic solvent (2-sec-butylphenol) and conveyed to another two-phase reaction vessel, in which the reaction conditions for hydroxymethylfurfural to levulinic acid have been optimized, and the resulting product is continuously removed. Alternative solvents, such as γ-valerolactone, are also being investigated as more environmentally acceptable reaction media for this process. See, for example, Alonso, D., *Energy Environ. Sci.*, 2013, 6, 76-80, which is incorporated herein by reference.

[0110] lactic acid

[0111] As described above, tobacco-derived cellulose materials can undergo enzymatic hydrolysis to form a hydrolyzed tobacco product (i.e., a liquid paste) comprising residual solids and liquid, said liquid containing at least one tobacco-derived cellulose sugar. As in Figure 1 As shown in operation 114, for example, the process may further include separating residual solids and liquid, adding a high-protein medium to the liquid, and allowing the liquid and high-protein medium to ferment and form a fermented product containing lactic acid. Figure 1 As shown in operation 108, lactic acid can also be produced by the conversion and recycling of xylose, for example.

[0112] Lactic acid fermentation is carried out by lactic acid bacteria that convert simple carbohydrates such as glucose, sucrose, or galactose into lactic acid. Lactic acid bacteria (LAB) (also known as the order Lactobacilli) are a branch of Gram-positive, non-spore-forming cocci, cocci, or bacilli with a DNA base composition of less than 53 mol% G+C (guanine + cytosine). They are typically non-respiratory and lack catalytic enzymes. They primarily ferment glucose into lactic acid, or into lactic acid, CO2, and ethanol. In pure lactic acid fermentation, one molecule of glucose is ultimately converted into two molecules of lactic acid. In heterogeneous lactic acid fermentation, carbon dioxide and ethanol are produced in addition to lactic acid in a process called the phosphatidylcholine pathway. Although many genera of bacteria produce lactic acid as a primary or secondary fermentation end product, the term "lactic acid bacteria" is conventionally reserved for genera within the order Lactobacillus, which, in addition to *Carnobacterium*, *Enterococcus*, *Oenococcus*, *Tetragenococcus*, *Vagococcus*, and *Weisella*, also include *Lactobacillus*, *Leuconostoc*, *Pediococcus*, *Lactococcus*, and *Streptococcus*. Two common applications of lactic acid fermentation are, for example, in the production of yogurt and kimchi.

[0113] Lactic acid can be used to act on monomers that produce polylactic acid (PLA), which, as a biodegradable polymer, has many applications. For example, tobacco-derived PLA can be incorporated into materials suitable for forming outer bags containing tobacco compositions in smokeless tobacco products. Tobacco-derived PLA can be incorporated into filters, for example. Tobacco products may also include a composition of lactic acid and a floating precursor in the tobacco composition as a preservative, curing agent, and / or flavoring agent. Lactic acid can also be used in pharmaceutical technology to generate water-soluble lactates from otherwise insoluble active ingredients. Lactic acid is suitable for use in topical preparations and cosmetics to adjust acidity and their disinfecting and keratolytic properties. Lactic acid is suitable as a food preservative, curing agent, and flavoring agent. Lactic acid is also suitable for use in detergents (as a soap scum remover) and antibacterial agents. Lactic acid is used in combination with ammonium bicarbonate. In mosquito attractants, lactic acid is generally considered beneficial as a relatively environmentally safe and natural ingredient in many different products.

[0114] ethanol

[0115] As described above, tobacco-derived cellulose materials can undergo enzymatic hydrolysis to form a hydrolyzed tobacco product (i.e., a liquid paste) comprising residual solids and liquid, said liquid containing at least one tobacco-derived cellulose sugar. As in Figure 1 As shown in operation 114, for example, the process may further include separating residual solids and liquid, adding yeast to the liquid, and allowing the liquid and yeast to ferment and form a fermented product containing ethanol.

[0116] Ethanol (also known as ethyl alcohol) is the primary type of alcohol found in alcoholic beverages. Ethanol is also used as a solvent, disinfectant, fuel, and the active fluid in modern thermometers. Ethanol used as an industrial feedstock or solvent is typically produced by the acid-catalyzed hydration of ethylene. The vast majority of ethanol used in alcoholic beverages and as fuel is produced through fermentation.

[0117] Regarding ethanol fermentation, certain yeast species (such as *Saccharomyces cerevisiae*) produce ethanol and carbon dioxide when they metabolize sugars (such as glucose) under low-oxygen conditions. This process can be carried out, for example, at about 35°C to 40°C. The toxicity of ethanol to yeast limits the concentration of ethanol obtainable through brewing. Therefore, supplemental or distillation processes can be used to obtain higher ethanol concentrations.

[0118] As discussed above, cellulose-containing materials (such as the roots and / or stems of tobacco plants) typically contain other polysaccharides, including hemicellulose. Hydrolysis of hemicellulose yields pentose sugars, such as xylose. Saccharomyces cerevisiae, the yeast most commonly used in ethanol production, cannot metabolize xylose. Therefore, in some embodiments, the concentration of xylose in the hydrolysate can be reduced by enzymatic treatment prior to undergoing ethanol fermentation.

[0119] Uses of tobacco-derived cellulose sugar materials in tobacco products

[0120] As described above, the cellulose sugar material produced according to the present invention is suitable for producing a variety of products derived from tobacco glucose, including (but not limited to) humectants, fillers, flavoring agents, and various acids. The cellulose sugar material and other downstream products produced by the process according to the present invention are suitable as components incorporated into, for example, tobacco products. Tobacco products incorporating the material of the present invention may vary and may include any product configured or adapted to deliver tobacco or some of its components to a user. Exemplary tobacco products include smoking articles (e.g., cigarettes), smokeless tobacco products, and flotation generating devices containing tobacco material or other plant materials that do not burn during use.

[0121] In various embodiments, products derived from cellulose sugar materials (specifically, tobacco glucose) may be incorporated into smoking articles in the form of flavorings, fillers, and / or humectants in flammable puff filling materials. In some embodiments, lactic acid derived from the cellulose sugar materials of the present invention may be used to produce polylactic acid, which may be suitable for use in filtration products, including, for example, filters in smoking articles.

[0122] Reference Figure 2 This illustration shows a smoking article 10 in the form of a cigarette and having certain representative components of a smoking article, said components which may contain products derived from cellulose sugar materials of the present invention. The cigarette 10 comprises a generally cylindrical rod 12 surrounding a flammable puffing filler material (e.g., about 0.3 to about 1.0 g of flammable puffing filler material, such as tobacco material) contained in a wrapping material 16. The rod 12 is conventionally referred to as a “tobacco strip”. The ends of the tobacco strip 12 are open to expose the flammable puffing filler material. The cigarette 10 is shown having an optional strip 22 (e.g., a printed coating comprising a film-forming agent such as starch, ethyl cellulose, or sodium alginate) applied to the wrapping material 16, and said strip surrounding the cigarette rod in a direction transverse to the longitudinal axis of the cigarette. The strip 22 may be printed on the inner surface of the wrapping material (i.e., facing the flammable puffing filler material), or less preferably, on the outer surface of the wrapping material.

[0123] At one end of the tobacco strip 12 is a ignition end 18, and a filter element 26 is positioned at the mouth end 20. The filter element 26 is positioned adjacent to one end of the tobacco strip 12 such that the filter element and the tobacco strip are axially aligned end-to-end, preferably adjacent to each other. The filter element 26 may have a generally cylindrical shape, and its diameter may be substantially equal to the diameter of the tobacco strip. The end of the filter element 26 allows air and smoke to pass through it. A plug 28 covers the filter element, and a fitting material (not shown) covers a portion of the outer covering material 16 of the plug and the rod 12, thereby securing the rod to the filter element 26.

[0124] The filter element of the present invention typically comprises multiple longitudinally extending segments. Each segment may have different characteristics and may include various materials capable of filtering or absorbing particulate matter and / or gaseous compounds. Typically, the filter element of the present invention comprises 2 to 6 segments, often 2 to 4 segments. In a preferred embodiment, the filter element includes a mouth end segment, a tobacco end segment, and a compartment therebetween. This filter arrangement is sometimes referred to as a “compartment filter” or a “plug / space / plug” filter. As described in more detail below, the compartment may be divided into two or more compartments.

[0125] In various embodiments, the filter element may comprise an adsorbent in the form of activated carbon material, wherein activated carbon capable of removing at least one gaseous component of the mainstream smoke is incorporated into the filter element. In some embodiments, the filter element 26 may include vents 30 extending through a tipping paper (not shown) and a wrapping plug 28, and thus providing air dilution of the mainstream smoke. The vents 30 may be via a single line of perforations configured to extend circumferentially around the filter element 26, or may include perforations in several lines. As will be understood, the exact number and size of the vents 30 will vary depending on the desired level of air dilution.

[0126] In various embodiments, products derived from cellulose sugar materials (specifically, tobacco glucose) may be incorporated into smokeless tobacco products as flavoring agents, fillers, and / or humectants in smokeless tobacco blends. In some embodiments, lactic acid derived from the cellulose sugar materials of the present invention may be used to produce polylactic acid, which may be suitable for use in nonwoven products, including, for example, nonwoven fabrics suitable for pouch products.

[0127] The smokeless tobacco product of the present invention may take different forms. In one specific embodiment, the product is in the form of a snuff-type product containing particulate tobacco material and flavoring agents, fillers, and / or humectants derived from tobacco glucose. The methods and approaches for formulating snuff-type tobacco blends will be readily apparent to those skilled in the art of snuff tobacco product manufacturing. For example, such as... Figure 3 As shown, an exemplary bagged product 300 may include an external water-permeable container 320 in the form of a bag, the container 320 containing a particulate mixture 315 suitable for oral use. The orientation, size, and type of external water-permeable bags shown herein, as well as the type and properties of compositions suitable for oral use, should not be construed as limiting them.

[0128] In various embodiments, moisture-permeable bags or pouches can be used as containers for the use of the internal composition. Such bags or pouches (e.g.) Figure 3The composition / construction of the container bag 320 in the illustrated embodiments can be varied as indicated herein. For example, suitable pouches, bags, or containers (which may be modified according to the invention) for the manufacture of smokeless tobacco products are available under the following trademarks: CatchDry, Ettan, General, Granit, Goteborgs Rape, Grosnus White, Metropol Kaktus, Mocca Anis, Mocca Mint, Mocca Wintergreen, Kicks, Probe, Prince, Skruf, and TreAnkrare. Products of bag types similar in shape and form to the various embodiments of the bagged products described herein are commercially available as ZONNIC (distributed by Niconovum AB). Additionally, in Example 1 of PCT WO 2007 / 104573 by Axelsson et al., which is incorporated herein by reference, a bag-type product that is substantially similar in shape and form to various embodiments of a bagged product is described as a snuff pack composition EJ, which is produced using excipient ingredients and processing conditions suitable for manufacturing bagged products as described herein.

[0129] The amount of material contained in each bag may vary. In a smaller embodiment, the dry weight of the material in each bag is at least about 50 mg to about 150 mg. In a larger embodiment, the dry weight of the material in each bag preferably does not exceed about 300 mg to about 500 mg. The material in each bag may contain flavoring agents, fillers, and / or humectants derived from glucose produced from the virgin tobacco material.

[0130] In some embodiments, each pouch / container may have a flavoring component disposed therein, as described in more detail in U.S. Patent No. 7,861,728 to Holton, Jr. et al., which is incorporated herein by reference. The flavoring component may contain a flavoring derived from tobacco glucose as discussed above. Other components may be contained in each pouch if desired. For example, a flavor strip, block, or sheet of at least one flavor water-dispersible or water-soluble material (e.g., a breathable edible film type material) may be disposed in each pouch along with at least one capsule or, without at least one capsule. Such strips or sheets may be folded or crumpled to facilitate their inclusion in the pouch. See, for example, the types of materials and techniques described in: U.S. Patent No. 6,887,307 to Scott et al. and U.S. Patent No. 6,923,981 to Leung et al.; and the European Food Safety Authority Journal (2004) 85,1-32; which are incorporated herein by reference.

[0131] In various embodiments, the outer water-permeable pouch may contain PLA derived from lactic acid derived from tobacco glucose. Descriptions of various components and their constituent parts of snuff-type products are also set forth in U.S. Patent Application Publication No. 2004 / 0118422 by Lundin et al., which is incorporated herein by reference. Furthermore, see, for example, U.S. Patent No. 4,607,479 by Linden; U.S. Patent No. 4,631,899 by Nielsen; U.S. Patent No. 5,346,734 by Wydick et al.; and U.S. Patent No. 6,162,516 by Derr, and U.S. Patent Publication No. 2005 / 0061339 by Hansson et al.; each of these documents is incorporated herein by reference. Additionally, see the type of pouch described in U.S. Patent No. 5,167,244 by Kjerstad, which is incorporated herein by reference. Small snuff-type products are manufactured using equipment such as SB 51-1 / T, SBL 50, and SB 53-2 / T, which are available from Merz Verpackungmaschinen GmBH. Small snuff pouches can be supplied as individual pouches, or multiple pouches (e.g., 2, 4, 5, 10, 12, 15, 20, 25, or 30 pouches) can be connected or linked together (e.g., end-to-end), allowing individual pouches or separate portions to be easily removed from the pouch's single-piece bundle or matrix for use.

[0132] This invention is not limited to smokeless tobacco products of the snuff type. For example, mixtures of tobacco materials and flavorings, fillers and / or humectants derived from tobacco glucose can also be incorporated into various forms of smokeless tobacco, such as loose wet snuff, loose dry snuff, chewing tobacco, granulated tobacco blocks, extruded tobacco strips or blocks, finely pulverized or milled agglomerates of powdered blocks and components, flake blocks (e.g., which can be formed in a fluidized bed by agglomerating tobacco blend components), molded tobacco blocks (e.g., formed in the approximate shape of a coin, cylinder, bean, cube, etc.), blocks containing tobacco gum, products incorporating mixtures of edible materials with tobacco blocks and / or tobacco extracts, and products incorporating tobacco (e.g., in the form of tobacco extracts) carried on a solid inedible substrate, etc. For example, smokeless tobacco products may take the form of: compressed tobacco pellets, multilayer extruded blocks, extruded or formed rods or bars, compositions (having one type of tobacco blend surrounded by different types of tobacco blends), rolls of adhesive films, readily water-soluble or water-dispersible films or strips (see, for example, U.S. Patent Application Publication No. 2006 / 0198873, Chan et al.), or capsule-like materials having an outer shell (e.g., a flexible or rigid outer shell that may be transparent, colorless, translucent, or highly colored) and an inner region containing tobacco or tobacco flavoring (e.g., a Newtonian or thixotropic fluid incorporating some form of tobacco).

[0133] In some embodiments, the smokeless tobacco product of the present invention may be in the form of a lozenge, tablet, microtab, or other tablet-type product. See, for example, the types of lozenge formulations and techniques for formulating or manufacturing lozenges described in the following: U.S. Patent No. 4,967,773 to Shaw; U.S. Patent No. 5,110,605 to Acharya; U.S. Patent No. 5,733,574 to Dam; U.S. Patent No. 6,280,761 to Santus; U.S. Patent No. 6,676,959 to Andersson et al.; U.S. Patent No. 6,248,760 to Wilhelmsen; and U.S. Patent No. 7,374,779; Wil U.S. Patent Publication No. 2001 / 0016593 to Helmsen; U.S. Patent Publication No. 2004 / 0101543 to Liu et al.; U.S. Patent Publication No. 2006 / 0120974 to Mcneight; U.S. Patent Publication No. 2008 / 0020050 to Chau et al.; U.S. Patent Publication No. 2009 / 0081291 to Gin et al.; and U.S. Patent Publication No. 2010 / 0004294 to Axelsson et al. are incorporated herein by reference.

[0134] Depending on the type of smokeless tobacco product being processed, in addition to tobacco materials and flavorings, fillers, and / or humectants derived from tobacco glucose, the tobacco product may also include one or more additional components. For example, tobacco materials and flavorings, fillers, and / or humectants derived from tobacco glucose may be processed, blended, formulated, combined, and / or mixed with other materials or ingredients such as other tobacco materials or flavorings, fillers, binders, pH adjusters, buffers, salts, sweeteners, colorants, disintegrants, humectants, and preservatives (any of which may be encapsulated components). See, for example, those representative components, combinations of components, relative amounts of those components and ingredients relative to tobacco, and methods and means of using those components, as set forth in the following: U.S. Patent Publication No. 2011 / 0315154 of Mua et al., U.S. Patent Publication No. 2007 / 0062549 of Holton, Jr., and U.S. Patent No. 7,861,728 of Holton, Jr., each of which is incorporated herein by reference.

[0135] In various embodiments, products derived from cellulose sugar materials (specifically, tobacco glucose) can be incorporated into smokeless tobacco products in the form of flavorings, fillers, and / or humectants used in e-cigarette products. Numerous smoking products, flavor generators, and pharmaceutical inhalers have been proposed that utilize electrical energy to vaporize or heat volatile materials or attempt to provide, to a large extent, the sensation of a cigarette, cigar, or tobacco pouch without burning tobacco. See, for example, various alternative smoking articles, floating delivery devices, and heat sources described in the background art below: U.S. Patent No. 7,726,320 to Robinson et al., U.S. Patent Publication No. 2013 / 0255702 to Griffith Jr. et al., U.S. Patent Publication No. 2014 / 0000638 to Sebastian et al., U.S. Patent Publication No. 2014 / 0060554 to Collett et al., U.S. Patent Publication No. 2014 / 0096781 to Sears et al., U.S. Patent Publication No. 2014 / 0096782 to Ampolini et al., and U.S. Patent Application No. 14 / 011,992 to Davis et al., filed August 28, 2013, all of which are incorporated herein by reference in their entirety.

[0136] Figure 4An exemplary embodiment of an electronic smoking article 200 is shown. As shown therein, a control body 202 may be formed of a control body housing 201, which may include a control component 206, a flow sensor 208, a battery 210, and an LED 212. A cylinder 204 may be formed of a cylinder housing 203 surrounding a reservoir housing 244, which is in fluid communication with a liquid delivery element 236 adapted to deliver a float precursor composition stored in the reservoir housing to a heater 234 by capillary action or otherwise. An opening 228 may be present in the cylinder housing 203 to allow the formed float to drain from the cylinder 204. Such components represent components that may be present in a cylinder and are not intended to limit the scope of cylinder components covered by this disclosure. The cylinder 204 may be adapted to engage the control body 202 by a press-fit engagement between a control body protrusion 224 and a cylinder receiver 240. This type of connection facilitates a stable connection between the control body 202 and the cylinder 204, as well as an electrical connection between the battery 210 and control component 206 in the control body and the heater 234 in the cylinder. The cylinder 204 may also include one or more electronic components 250, which may include ICs, memory components, sensors, etc. The electronic components 250 may be adapted to communicate with the control component 206. Various components of the electronic smoking device according to this disclosure are selectable from those described in the art and commercially available components.

[0137] In various embodiments, the floating precursor composition may comprise tobacco-derived cellulose sugars or downstream products of tobacco-derived sugars, said downstream products being selected from the group consisting of glucose, glucose syrup, high-fructose tobacco syrup, glycerol, lactic acid, levulinic acid, sorbitol, and combinations thereof. Exemplary formulations for use with floating precursor materials that may be used according to this disclosure are described in: U.S. Patent No. 7,217,320 to Robinson et al.; U.S. Patent Publication No. 2013 / 0008457 to Zheng et al.; U.S. Patent Publication No. 2013 / 0213417 to Chong et al.; U.S. Patent Publication No. 2014 / 0060554 to Collett et al.; and U.S. Patent Publication No. 2014 / 0000638 to Sebastian et al., the disclosures of which are incorporated herein by reference in their entirety. Other floating precursors that may be incorporated into the cellulose sugars and their downstream products described herein include those already incorporated into the following: RJ Reynolds Vapor Company. Products, Lorillard Technologies' BLU TMProducts from Mistic Ecigs, MISTIC MENTHOL, and VYPE from CNCreative Ltd. So-called "smoke juice" e-cigarettes from Johnson Creek Enterprises LLC are also anticipated.

[0138] experiment

[0139] The aspects of the invention are illustrated more fully by the following examples, which are set forth to illustrate certain aspects of the invention and should not be construed as limiting the invention.

[0140] Example 1

[0141] Glucose is produced from virgin tobacco material. Three sets of delignification and saccharification are performed. Each set consists of milled (mechanically pulped) tobacco stalk biomass, which undergoes two delignification processes in a large delignification inlet tank with an approximate working volume of about 60 gallons. 14 kg (dry weight) of tobacco stalks are used in each delignification process. After milling the tobacco stalk biomass, 10 kg of delignified pulp remains after each delignification.

[0142] The delignified pulp is then rinsed, dehydrated, and rinsed again. The pH is adjusted to 4.8, and the pulp is then dehydrated again to obtain pulp ready for enzymatic saccharification. Two additions of saccharification are performed, starting at a 5% solids and 5% enzyme loading level on a dry weight basis. After this 24-hour run, the second addition of pulp is added to the paste, and an additional 24-hour run is performed.

[0143] The residual solids were removed from the paste, and the liquid was transferred to an evaporator for thickening. The resulting concentrated glucose syrup had a sugar composition of 84.4% by weight glucose and 15.6% by weight xylose.

[0144] Example 2

[0145] A glucose syrup with a reduced xylose concentration is produced from raw tobacco material. The process described in Example 1 above is followed to produce the glucose syrup. However, in order to remove xylose from the syrup, xylanase is used to remove hemicellulose before final saccharification. Three different types of xylanase are used ( HC&HC 2500 Xylanase and its combinations).

[0146] HC&HC 2500 is the only enzyme that releases xylose without glucose. However, xylose release is lower than expected. Adding another xylanase showed good hydrolysis of hemicellulose, but also exhibited the same cellulose hydrolysis. However, in this step, the combination of xylanases enabled the removal of half of the still usable hemicellulose, which reduced xylose in the glucose syrup by about 50%, but did not eliminate xylose.

[0147] Example 3

[0148] Use commercially available immobilized glucose isomerase (also known as Sweetzyme type) High-fructose tobacco syrup was produced. Experiments were conducted at 55°C and an initial tobacco glucose concentration of 13%. The enzyme was hydrated before being added (45 g) to 4 L of glucose-containing Tris-buffered medium (pH 7.5). The fructose yield was approximately 0.5 g g⁻¹. The final glucose and fructose concentrations were 275 g / L and 225 g / L, respectively. This reaction showed a good fit between the experimental data and model predictions (Gaily, MH et al., Production of fructose from highly concentrated date extracts using Saccharomyces cerevisiae., Biotechnology Letters, 2013).

[0149] Example 4

[0150] Glycerin is produced from virgin tobacco material. Tobacco stalk biomass is ground (mechanically pulped) and subjected to delignification in a large delignification inlet tank with a working volume of approximately 60 gallons. The delignified pulp is then washed, dewatered, and washed again. The pH is adjusted to 4.8, and the pulp is then dewatered again to obtain pulp ready for enzymatic saccharification. A strong loading (3% by weight) is used. CTec 3 (manufactured by Novozymes) saccharifies delignified tobacco pulp to provide tobacco glucose for glycerol production.

[0151] Tobacco pulp (9% solids) was hydrolyzed for 24 hours before adding 10 g / L of rapid-starting baker's yeast. Nutrients and hops were also added to the medium. Low doses of bisulfite (2000 ppm) in the form of Na₂S₂O₅ were added periodically throughout the reaction to prevent acetaldehyde volatilization and increase glycerol production by the yeast. The pH was maintained between 6.7 and 7.3 using a pH controller and AmOH₂ to stabilize the bisulfite concentration within a pH range tolerable to the yeast. Over 100 g of glycerol was produced at an average yield of 13% by weight of pulp. A weak base anion was used on the solution to remove any lactic acid impurities.

[0152] Example 5

[0153] Sorbitol is produced from tobacco-derived glucose. The process described in Example 1 above is followed to produce glucose syrup. The conditions outlined below are used to design the reduction of tobacco-derived glucose to sorbitol: Gallezot, P. *Journal of Catalysis* 146(1994) 93 and Hoffer, BW., "Tuning Raney-type and Supported Ni Catalysts for Commercial Hydrogenation Reactions," Delft University of Technology, Delft, Netherlands, 2003. Key parameters such as catalyst type, precursor concentration, H2 pressure, reaction temperature, and purge cycle are adjusted for use in the reaction.

[0154] Specifically, we reduced the conversion of tobacco glucose to sorbitol as follows. Tobacco-derived glucose (Burley 34.9, approximately 100 g, 0.55 M) was present in H₂O, yielding a solution volume of 250 mL. The pH was adjusted to 6.5 by adding 0.1 M acetic acid and Mo alloy Raney nickel (WRGrace 3202, 3 g). The reaction mixture was placed in a 600 mL stainless steel Parr stirred autoclave equipped with a thermocouple-controlled heating mantle and a purging system adapted by a Parr oscillating hydrogenation device. The reaction vessel was exposed to three vacuum evacuation / H₂ filling cycles to remove oxygen from the system. The purging system was disconnected, and hydrogen was added to a pressure of 800 psig (5.5 mPa). Stirring was started (1000 rpm), and the reactants were rapidly heated to 120 °C. The reaction required the hydrogen pressure level to be returned from 580 psi (4 mPa) to 800 psi three times over a period of 2.5 h, during which H₂ absorption had ceased. The reaction vessel was rapidly cooled to 25°C in a water bath, and the H2 pressure was carefully released. The vessel was purged under vacuum to reduce the amount of H2 dissolved in the solution. The autoclave was opened, with the magnetic Raney nickel held at the bottom of the lower half of the autoclave containing a strong magnet, and the reaction solution was decanted into an Erlenmeyer flask. The catalyst was washed with distilled H2O, and the washings were combined with the product. The catalyst was immersed in water. The aqueous solution was vacuum filtered through a diatomaceous earth pad over a sintered funnel to remove any non-magnetic nickel particles.

[0155] The reaction was repeated twice, and due to the nature of the catalyst washing process, the final solutions had slightly different volumes and therefore different sorbitol concentrations. HPLC analysis showed that the sorbitol concentration in round 1 was 150 g / L, and in round 2 it was 173 g / L. The resulting solutions were clear and white.

[0156] Example 6

[0157] Levulopyric acid is produced from tobacco biomass. The process described in Example 1 above is followed to produce glucose from virgin tobacco material. The hydrolysis portion of levulopyric acid synthesis is suitable for the conditions outlined below: Du, X., Angewandte Chemie, 2011, 123, 7961-7965 and Runge, T., Research in Industrial and Engineering Chemistry, 2012, 51, 3265-3270, which are incorporated herein by reference. Most modern literature uses levulopyric acid as a starting material for other reactions; therefore, few separation procedures exist in modern literature. Edwards' U.S. Patent No. 4,612,391, incorporated herein by reference, describes a process in which acidic and basic solutions are extracted with organic solvents to purify the product. This process is adapted by replacing the environmentally unfriendly chlorinating solvent described therein with ethyl acetate.

[0158] The levulinic acid sample was prepared as follows. Glucose (44.6 g, 0.248 mol) was dissolved in 0.5 M H₂SO₄ (230 mL) and loaded into a 500 mL stainless steel stirred autoclave lined with Teflon. The autoclave was rapidly heated to 170 °C (approximately 30 min) and stirred vigorously for 45 min after reaching the target temperature. The outer jacket was removed, and the autoclave was rapidly cooled in a water bath. When the apparatus reached 40 °C, it was placed in a ring rack, and the top was removed. The reaction mixture was poured into a 500 mL Erlenmeyer flask and transported to the laboratory. HPLC analysis at this point yielded a levulinic acid concentration of 15 g / L to 16 g / L. The pH of the acidic solution was adjusted to 1 by adding CaO (5 g) to remove H₂SO₄ and insoluble precipitates. The resulting CaSO₄ was filtered off, the filter cake was washed with DI H₂O, and the aqueous solution containing levulinic acid and formic acid was extracted with ethyl acetate (3 × 100 mL). The organic matter was washed with brine, and the ethyl acetate was removed by rotary evaporation. Additional water was added to the residue to ensure that all formic acid was removed as an azeotrope. The residue was dissolved in 3N NaOH (150 mL) and washed with ethyl acetate to remove non-carboxylic acidic organic compounds via Na salt formation. The alkaline aqueous solution was acidified to pH 1 with concentrated HCl, and the acidic solution was extracted with ethyl acetate (3 × 100 mL) to remove water-soluble salts and recover the product. The organic solution was dried by washing with a saturated brine solution, and the organic solvent was removed by rotary evaporation. The products of repeated reactions were combined, and the crude levulinic acid was treated with activated carbon to remove color.

[0159] Example 7

[0160] Lactic acid is produced from tobacco biomass. The process described in Example 1 above is followed to produce glucose from the virgin tobacco material.

[0161] A successful lactic acid yield is defined as an observed conversion of tobacco-derived glucose to lactic acid greater than 90%. Partially purified tobacco glucose (82 g / L) was fermented in a 12 L reactor with a high-protein medium until the sugar was completely removed. The pH was maintained at 6.5 ± 0.2 using Ca(OH)2 and a pH controller. The temperature was maintained at 38 °C. The medium was inoculated with a 10 g / L bacterial composition including live cultures of *Streptococcus thermophilus*, *Lactobacillus bulgaricus*, *Lactobacillus acidophilus*, *Bifidus*, *Lactobacillus casei*, and *Lactobacillus rhamnosus*. Fermentation was completed in less than 48 hours.

[0162] After completion, the pH of the fermentation broth is increased to 10.15 using Ca(OH)2, and the reactor is then heated to 50°C to kill any present organic matter and coagulate the protein material, thus improving filtration. Calcium lactate is then decomposed with sulfuric acid, and calcium sulfate (precipitate) is removed. The lactic acid solution is concentrated to 15%, bleached with activated carbon, and then concentrated to 33%. This process can be improved by using a more refined tobacco glucose (e.g., purified with activated carbon before use) and fermenting for 5 to 10 days in a very low-protein medium.

[0163] Example 8

[0164] Ethanol is produced from tobacco biomass. As described in Example 1 above, tobacco-derived cellulose material undergoes enzymatic hydrolysis to form a hydrolyzed tobacco product (i.e., a liquid paste) comprising residual solids and liquid, the liquid containing glucose. The residual solids and liquid are separated, yeast is added to the liquid, and the liquid and yeast are allowed to ferment and form a fermented product containing ethanol.

[0165] Regarding the fermentation of tobacco-derived glucose, a 20-liter reactor vessel was filled with 14 L of tobacco stem-derived glucose / xylose at 22 Brix (produced according to Example 1). Nutrients and buffers were added, followed by the addition of yeast (a proprietary yeast flocculant strain developed by BPI). The syrup was allowed to ferment for 48 hours, after which HPLC analysis indicated approximately 100 g / L of ethanol and almost no residual glucose. The analysis also showed the production of approximately 13 g / L of glycerol as a natural 'byproduct' of ethanol fermentation.

[0166] Regarding ethanol distillation, a distillation column was designed and constructed for the recovery and purification of ethanol from tobacco stems. The column was constructed from 2" stainless steel tubing and then packed with stainless steel sieve-type packing. The stripping section (below the feed) was 50 cm long, and the rectifying section (above the feed) was 100 cm long. The feed and bottoms outflow were controlled by a variable-speed peristaltic pump with two heads: one head pumping the feed to the middle of the column, and the other head pumping the bottoms out of the reboiler. The reflux rate was determined based on the feed concentration.

[0167] Regarding ethanol recovery, 14 L of tobacco stem glucose / xylose at 25 Brix was fermented to 95 g / L ethanol, and then boiled in a condenser to remove approximately 50% of the volume. This produced 6 L of residual fermentation broth containing a concentrated ethanol solution and a concentrated residual xylose / glycerol solution. 7 L of condensate was recovered, with the first 3.5 L containing 20% ​​ethanol and the second 3.5 L containing 5% ethanol. HPLC analysis of the 14 L indicated the production of approximately 12 g / L of glycerol as a recoverable byproduct.

[0168] Next, a xylose-metabolizing yeast strain from Bio Process Innovations (West Lafayette, IL) was added to the concentrated fermentation broth, and air was added to remove xylose (which is converted into yeast cell clumps) and essentially leave only glycerol (along with the yeast and unconsumed nutrients). Some inhibitors in the concentrated fermentation broth prevent the xylose-metabolizing yeast from growing and consuming xylose. It is possible that the high-temperature 'boiling' process for 4 hours caused the formation of inhibitors, or simply that the concentration of the fermentation broth made some of the inhibitors remaining from the enzymes (or fermentation byproducts) high enough to more or less stop the fermentation. Ultimately, 7L was discarded without recovering the glycerol.

[0169] By combining 2L of 5% ethanol (second bottle) with 3.5L of 20% ethanol, the 3.5L of 20% ethanol was diluted to approximately 14% ethanol. The column was heated, with 1L of water and 1L of 5% ethanol (second bottle) in the reboiler. Once the column was hot, the condensation rate was measured at approximately 2 to 3 ml / min, which was significantly lower than the design rate of 30 ml / min. Therefore, the column was insulated, and the condensation rate was increased to 5.5 ml / min. The reflux rate was set at 2.5 ml / min, and the column was run for approximately 12 hours, during which a net yield of 1.3L of 165% ethanol was produced. Adding another 'spearhead' type heater to the reboiler should increase the boiling rate of the condensate by 150% to approximately 15 ml / min.

[0170] Those skilled in the art will appreciate many modifications and other embodiments of the invention, which have the benefits of the teachings presented in the foregoing description. Therefore, it should be understood that the invention is not limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terminology is used herein, it is used in a general and descriptive sense only and not for limiting purposes.

Claims

1. A method for forming a floatable precursor composition configured for use in electronic smoking articles, comprising: i) Receiving tobacco material, said tobacco material comprising at least one of stem material and root material from harvested tobacco species; ii) Deligninate the tobacco material to form tobacco-derived cellulose pulp and lignin; iii) Separating the tobacco-derived cellulose pulp from the lignin; iv) Hydrolyze the tobacco-derived cellulose pulp to form a hydrolyzed tobacco product comprising residual solids and liquid, wherein the liquid comprises at least one tobacco-derived cellulose sugar; v) Hydrolyze the at least one tobacco-derived cellulose sugar to form levulinic acid; and vi) Incorporate the levulinic acid into the floating precursor composition.

2. The method according to claim 1, wherein the tobacco-derived cellulose sugar is glucose.

3. The method of claim 2, further comprising, prior to the step of hydrolyzing the at least one tobacco-derived cellulose sugar to form levulinic acid, thickening the glucose to form a syrup.

4. The method according to claim 2, wherein the step of hydrolyzing the at least one tobacco-derived cellulose sugar to form levulinic acid comprises at least one of the following: (1) hydrolysis at low temperature with concentrated acid; and (2) hydrolysis at high temperature with dilute acid.

5. The method of claim 1, wherein the step of deligating the tobacco material further comprises adjusting the pH of the tobacco-derived cellulose pulp to a value in the range of 4.5 to 5.

5.

6. The method of claim 1, wherein the step of hydrolyzing the tobacco-derived cellulose pulp comprises a primary enzymatic saccharification of the tobacco-derived cellulose pulp in the presence of at least one enzyme.

7. A method for forming a floatable precursor composition configured for use in electronic smoking articles, comprising: i) Receiving tobacco material, said tobacco material comprising at least one of stem material and root material from harvested tobacco species; ii) Deligninate the tobacco material to form tobacco-derived cellulose pulp and lignin; iii) Separating the tobacco-derived cellulose pulp from the lignin; iv) Hydrolyze the tobacco-derived cellulose pulp to form a hydrolyzed tobacco product comprising residual solids and liquid, wherein the liquid comprises at least one tobacco-derived cellulose sugar; v) Separate the residual solids and the liquid containing at least one tobacco-derived cellulose sugar; vi) Add a high-protein culture medium to the liquid containing at least one tobacco-derived cellulose sugar; vii) Allowing the liquid containing at least one tobacco-derived cellulose sugar and the high-protein culture medium to ferment and form a fermented product containing lactic acid; and viii) The lactic acid is incorporated into the floating precursor composition.

8. The method according to claim 7, wherein the at least one tobacco-derived cellulose sugar is glucose.

9. The method according to claim 7, wherein the at least one tobacco-derived cellulose sugar is xylose.

10. The method of claim 7, wherein the step of allowing fermentation of the liquid containing at least one tobacco-derived cellulose sugar and the high-protein culture medium comprises a pure lactic acid fermentation process.

11. The method of claim 7, wherein the step of allowing fermentation of the liquid containing at least one tobacco-derived cellulose sugar and the high-protein culture medium comprises a heterologous lactic acid fermentation process.

12. The method of claim 7, wherein the step of deligating the tobacco material further comprises adjusting the pH of the tobacco-derived cellulose pulp to a value in the range of 4.5 to 5.5.

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