A method of modifying the content of certain chemical compounds in tobacco materials is provided, the method including treatment of a tobacco plant or portion thereof with at least one enzyme. For example, the method may modify the content of tobacco smoke toxicant precursors in tobacco materials, which can result in a modification in toxicant production when the tobacco material is exposed to elevated temperatures. The type of tobacco plant or portion thereof treated according to the invention can be, for example, a tobacco seed, a tobacco seedling, an immature live plant, a mature live plant, a harvested plant, or a plant derivative. Smoking articles and other tobacco products including such enzyme-treated tobacco materials are also provided.

Bacteriophage against Geobacillus are provided, and methods of making and using the bacteriophage also are provided.

Methods of modifying the tobacco-specific nitrosamine content of a tobacco material are described herein. One exemplary method comprises contacting a tobacco material with a composition comprising salt, sugar, enzyme, lactic acid bacteria, yeast, or a combination thereof to reduce the total bacterial content; curing the tobacco material; and fermenting the tobacco material in the presence of one or more microorganisms. The method can provide a fermented tobacco material having a tobacco-specific nitrosamine content that is reduced relative to a fermented tobacco material that has not been subjected to the disclosed method steps. In certain embodiments, the tobacco-specific nitrosamine content of the fermented tobacco material is no more than that of the cured tobacco material. Tobacco-containing products including such treated tobacco materials are also provided.

Methods and compositions are provided for expressing an endolysin in a bacterium or in a tobacco plant or plant part. Expression of the endolysin can reduce the total bacterial load on the plant and in tobacco products produced from the plant. Accordingly, the level of tobacco specific nitrosamines (TSNA) in a cured tobacco product can be decreased when the tobacco leaves used in the product were harvested from a plant expressing endolysin or when bacteria modified to express endolysin is used to treat the tobacco. Thus, bacteria and tobacco plants and plant parts are provided that express an endolysin. Further provided are methods for decreasing the microbial load on a tobacco plant and in a tobacco product by applying a solution of bacteria modified to express an endolysin or by modifying a tobacco plant to express endolysins.

Smokeless tobacco products including such modified tobacco plant parts are also provided.

Bacteriophage against Geobacillus are provided, and methods of making and using the bacteriophage also are provided.

Methods of modifying the tobacco-specific nitrosamine content of a tobacco material are described herein. One exemplary method comprises contacting a tobacco material with a composition comprising salt, sugar, enzyme, lactic acid bacteria, yeast, or a combination thereof to reduce the total bacterial content; curing the tobacco material; and fermenting the tobacco material in the presence of one or more microorganisms. The method can provide a fermented tobacco material having a tobacco-specific nitrosamine content that is reduced relative to a fermented tobacco material that has not been subjected to the disclosed method steps. In certain embodiments, the tobacco-specific nitrosamine content of the fermented tobacco material is no more than that of the cured tobacco material. Tobacco-containing products including such treated tobacco materials are also provided.

A tobacco composition for use in a smoking article or a smokeless tobacco composition is provided, the composition including a component derived from a seed of the Nicotiana species. The component can be a seed of the Nicotiana species or a portion thereof in particulate form or in the form of seed isolate derived from a seed of the Nicotiana species. In certain embodiments, the seed isolate is in the form of a chemically transformed seed isolate, the chemical transformation being selected from hydrogenation, acid/base reaction, hydrolysis, thermal treatment, and enzymatic treatment. The invention also provides smoking articles and smokeless tobacco compositions that include the seed components described herein, and methods for preparing a component derived from a seed of the Nicotiana species for addition to a tobacco composition.

Methods of modifying the tobacco-specific nitrosamine content of a tobacco material are described herein. One exemplary method comprises contacting a tobacco material with a composition comprising salt, sugar, enzyme, lactic acid bacteria, yeast, or a combination thereof to reduce the total bacterial content; curing the tobacco material; and fermenting the tobacco material in the presence of one or more microorganisms. The method can provide a fermented tobacco material having a tobacco-specific nitrosamine content that is reduced relative to a fermented tobacco material that has not been subjected to the disclosed method steps. In certain embodiments, the tobacco-specific nitrosamine content of the fermented tobacco material is no more than that of the cured tobacco material. Tobacco-containing products including such treated tobacco materials are also provided.

A method of modifying the content of certain bacteria in uncured tobacco material is provided, the method including contacting an uncured tobacco material with a treatment solution, wherein the treatment solution is selected from the group consisting of: (i) a solution comprising salt, sugar, or a combination thereof; (ii) a solution comprising one or more enzymes; and (iii) a solution comprising one or more probiotics, wherein said contacting provides a treated tobacco material having a reduced total bacterial content following harvest. Also provided is a method of reducing TSNA content, the method including growing a tobacco plant in soil treated with a chloride source; harvesting the tobacco plant; and curing the harvested tobacco plant to give a cured tobacco material, providing a treated, cured tobacco material having a TSNA content that is reduced as compared to a cured tobacco material that has been grown in soil not treated with a chloride source. Smoking articles and smokeless tobacco products including such treated tobacco materials are also provided.

Methods for obtaining natural colorants from plant material (e.g., tobacco material) and for using such natural colorants to dye various substrates are provided. Natural colorants are obtained using particular enzymes and particular conditions (e.g., time, temperature, and pH profiles). Such colorants can be used to dye substrates, for example, using conventional dyeing techniques or using unique in situ methods.