The present disclosure describes methods of obtaining and/or deriving proteins from plants of the Nicotiana species and methods for incorporation of such proteins into various products. For example, a method for obtaining a protein-enriched material from a plant of the Nicotiana species or portion thereof is provided, comprising: extracting one or more proteins from the plant material into a solvent to form a liquid protein-containing extract; separating a solid extracted plant material from the liquid protein-containing extract; clarifying the liquid protein-containing extract to form a clarified protein-containing extract and a solids fraction; and treating the clarified protein-containing extract so as to provide a protein-enriched material comprising at least about 60% protein by weight.

There is described herein a mutant plant or part thereof having at least partially reduced expression or activity of at least two heavy metal ATPases (HMAs), said two HMAs comprising, consisting or consisting essentially of: (i) polypeptides having at least 65% sequence identity to SEQ ID NO:1 and SEQ ID NO:2; (ii) polynucleotides encoding the polypeptides set forth in (i); or (iii) polynucleotides having at least 65% sequence identity to SEQ ID NO:3 and SEQ ID NO:4 encoding HMAs; wherein the expression or activity of one of the HMAs set forth in (i) or (ii) or (iii) is partially reduced or lost and the expression or activity of one of the HMAs set forth in (i) or (ii) or (iii) is lost as compared to a control plant; and wherein the mutant plant or part thereof exhibits at least a 27% reduction, as compared to the control plant, in the accumulation of cadmium in leaf when the mutant plant is grown in the field in the presence of naturally or non-naturally occurring cadmium.

A method of preparing a whitened tobacco material is provided, the method including the steps of (i) extracting a tobacco material with an aqueous solution to give a tobacco pulp and a tobacco extract; (ii) treating the tobacco pulp with at least one of a caustic reagent and an oxidizing agent for a time and at a temperature sufficient to lighten the color of the tobacco pulp to give a whitened tobacco pulp; (iii) clarifying the tobacco extract to remove higher molecular weight components; and (iv) combining the whitened tobacco pulp with a clarified tobacco extract to form a whitened tobacco material. The whitened tobacco material can be isolated and incorporated into a smokeless tobacco product. The invention also provides a smokeless tobacco product incorporating a whitened tobacco material. The smokeless tobacco product may be a snus-type formulation contained within a sealed pouch.

A method of preparing a whitened tobacco material is provided, the method including the steps of (i) extracting a tobacco material with an aqueous solution to give a tobacco pulp and a tobacco extract; (ii) treating the tobacco pulp with at least one of a caustic reagent and an oxidizing agent for a time and at a temperature sufficient to lighten the color of the tobacco pulp to give a whitened tobacco pulp; (iii) clarifying the tobacco extract to remove higher molecular weight components; and (iv) combining the whitened tobacco pulp with a clarified tobacco extract to form a whitened tobacco material. The whitened tobacco material can be isolated and incorporated into a smokeless tobacco product. The invention also provides a smokeless tobacco product incorporating a whitened tobacco material. The smokeless tobacco product may be a snus-type formulation contained within a sealed pouch.

The invention provides a tobacco product including a flavorful tobacco composition in the form of an extract of a fire-cured tobacco material. Exemplary tobacco products include smoking articles, smokeless tobacco compositions, and aerosol-generating devices that do not burn tobacco. The invention also provides a process for preparing a smokeless tobacco composition, the method including: mixing a fire-cured tobacco material having a first benzo[a]pyrene concentration with water to produce an aqueous slurry; maintaining the slurry for a time and at a temperature sufficient to form a fire-cured tobacco extract, the aqueous fire-cured tobacco extract exhibiting a second benzo[a]pyrene concentration lower than the first benzo[a]pyrene concentration; separating the aqueous fire-cured tobacco extract from a residual pulp material, and mixing the aqueous fire-cured tobacco extract with a tobacco or non-tobacco plant material to form a smokeless tobacco composition.

The invention provides a tobacco product including a flavorful tobacco composition in the form of an extract of a fire-cured tobacco material. Exemplary tobacco products include smoking articles, smokeless tobacco compositions, and aerosol-generating devices that do not burn tobacco. The invention also provides a process for preparing a smokeless tobacco composition, the method including: mixing a fire-cured tobacco material having a first benzo[a]pyrene concentration with water to produce an aqueous slurry; maintaining the slurry for a time and at a temperature sufficient to form a fire-cured tobacco extract, the aqueous fire-cured tobacco extract exhibiting a second benzo[a]pyrene concentration lower than the first benzo[a]pyrene concentration; separating the aqueous fire-cured tobacco extract from a residual pulp material, and mixing the aqueous fire-cured tobacco extract with a tobacco or non-tobacco plant material to form a smokeless tobacco composition.

Immobilized diluents in a smoking article are provided, wherein diluents can be immobilized through absorption and/or adsorption of the diluents into immobilizing materials, such as sorbents like silica gels. By immobilizing diluents, the diluents can be available for vaporization, while still being protected from migration and/or loss of the diluents in a smoking article.

An article for use with an apparatus for heating aerosol generating agent to volatilize at least one component of the aerosol generating agent, the article including a support layer having a first surface, wherein at least a portion of the first surface is rough and an aerosol generating agent on the portion of the first surface that is rough.

The present disclosure provides approaches for reducing tobacco-specific nitrosamines (TSNAs) in tobacco. Some of these approaches include genetically engineering tobacco plants to increase one or more antioxidants, increase oxygen radicle absorbance capacity (ORAC), or reduce nitrite. Also provided are methods and compositions for producing modified tobacco plants and tobacco products therefrom comprising reduced TSNAs.

The present disclosure provides approaches for reducing tobacco-specific nitrosamines (TSNAs) in tobacco. Some of these approaches include genetically engineering tobacco plants to increase one or more antioxidants, increase oxygen radicle absorbance capacity (ORAC), increase phenylalanine, or reduce nitrite. Also provided are methods and compositions for producing modified tobacco plants and tobacco products therefrom comprising reduced TSNAs.