Researchers from the Zhengzhou Tobacco Research Institute of the China National Tobacco Corporation used a distillation extraction device, a gas chromatograph and a gas chromatography/mass spectrometer to analyze the main neutral aroma components of flue-cured tobacco stems and leaves. The study showed that Yunnan The neutral aroma components with high content in tobacco stems mainly include benzyl alcohol, phenylacetaldehyde, β-phenylethyl alcohol, etc.
The research literature of Shandong Agricultural University reviewed several aroma enhancement pathways and their research progress in the artificial fermentation process of tobacco leaves at home and abroad, including microorganisms, enzymes, sugars and organic acids, enzyme Rad reaction products and ammoniation and other methods. Research shows that in recent years, microorganisms and enzymes have obvious aroma-enhancing effects due to their low input amounts. Microbial fermentation to produce natural flavors has become a research and development hotspot for food additives in many countries around the world. Tobacco can produce natural rose aroma components such as benzyl alcohol and phenylethanol during the fermentation process of Bacillus, Bacillus subtilis, Pseudomonas and other microorganisms. It is an ideal method to use foreign aid to ferment tobacco leaves and increase the aroma of tobacco leaves.
Foreign scholars have done a lot of research on artificial fermentation of tobacco leaves. They found that among the microorganisms on the leaves of flue-cured tobacco, bacteria dominate, and actinomycetes and molds are less. Among bacteria, Bacillus is the dominant flora, and among molds, Aspergillus is the dominant flora. The number of microorganisms on the leaf surface of fine varieties of flue-cured tobacco is larger and there are more types. It can be seen that microorganisms are one of the reasons that cannot be ignored in promoting the fermentation of tobacco leaves and improving the aroma of tobacco leaves. This is very attractive to scholars at home and abroad who are engaged in tobacco leaf aroma research.
As for the research on the process of producing natural-grade phenylethanol spices by microbial fermentation, in recent years, foreign countries have successfully developed phenylalanine and fluorophenylalanine as raw materials, using the microbial beer yeast Saccaromyces cerevisiae or ( and) a process for preparing natural grade phenylethyl alcohol by transforming it into Kluyveromyces sinensis fermentation process. Although this method is a feasible method for producing natural-grade phenylethanol spices, its raw material cost is too high and is not suitable for industrial production. Therefore, people are also looking for a cheap natural material that can replace phenylalanine and fluorophenylalanine as a starting material for microbial fermentation to minimize production costs and meet the needs of industrial production of food flavor additives.
2. Introduction to major domestic and foreign patents
After years of innovative research and development, Huabao Aromatherapy Technology Development (Shanghai) Co., Ltd. has successfully completed the production of tobacco (including tobacco stems and tobacco powder). ) as the raw material for the production of spice phenylethanol by microbial fermentation, and obtained the invention patent authorization on July 28, 2004 (ZL Patent No. 02137575.5, Authorization Announcement No. CN1159447C). The process includes using tobacco, an agricultural by-product, as a precursor for biotransformation, adding an appropriate culture medium, and selecting an appropriate bacterial strain (such as one of Candida utilis, Saccharomyces cerevisiae, and Kluyveromyces sinensis). Fermentation culture is carried out under certain fermentation process conditions to degrade the lignin, pectin and polyphenolic compounds in tobacco and convert them into phenylethanol, which is then purified by extraction or ion exchange resin adsorption and separation. The obtained phenylethyl alcohol has a pure natural flavor and can be used as a natural flavor in food. The entire production process is simple and has broad industrial development prospects, finding a green and environmentally friendly application path for agricultural by-product tobacco.
Patent US5919991, authorized by SUBBIAH VEN Company in the United States on July 6, 1999, relates to a fermentation method for producing phenylethanol, which uses L-phenylalanine as raw material through the microorganisms Saccaromyces cerevisiae and Gram Kluyveromyces sinensis fermentation culture and ion exchange resin adsorption separation method are used to produce natural grade phenylethanol spices.
The European patent EP1158042 published by German Bluemke Wilfried (GKSS-Forschungszentrum Geesthacht Gmbh) on November 28, 2001 uses biotechnology such as microbial strains or enzyme catalysts to prepare phenylethanol spices, which uses L-phenylalanine. Acid is used as raw material, and natural-grade phenethyl alcohol flavoring is produced through the microbial fermentation process of Kluyveromyces sinensis.
Japanese patent JP9224650 published by Japan KAGOME Company in September 1997 involves a method using rice wine and fluorophenylalanine as raw materials through the microbial fermentation process of Saccharomyces cerevisiae, the brewing (beer) yeast in alcoholic beverages. Preparation of natural grade β-phenylethyl alcohol fragrance.
The patent JP2000041655 published by Japan Kirin Brewery in early 2000 also involves the modification of rice wine and the use of a microbial fermentation process that combines the combined action of brewing (beer) yeast Saccharomyces and decarboxylase to produce high-purity phenylethanol spices.
The 1998 patent EP0822250 published by PENOD RICARD of France provides a method for extracting high-purity phenylethyl alcohol from its distillation residue in a plant producing phenylethyl alcohol by fermentation.
List of major domestic and foreign patents related to the production of natural phenethyl alcohol flavorings by microbial fermentation (see Appendix 1)
Overview of the above-mentioned main domestic and foreign process technologies related to the production of natural phenethyl alcohol flavorings by microbial fermentation , it is not difficult to see that at present, foreign countries generally use phenylalanine and fluorophenylalanine as raw materials to produce phenylethanol through microbial fermentation and transformation. Although this method is indeed a feasible method for producing natural phenylethanol spices, due to the expensive raw materials such as phenylalanine and fluorophenylalanine used in this method, the production cost is high, and it is not suitable for large-scale industrial production. . Therefore, through innovative research and development, domestic enterprises use cheap and natural tobacco waste resource materials as starting materials for microbial fermentation. This not only opens up an industrial path for the deep processing of agricultural and sideline products tobacco, but also finds a safe, reliable and effective method for the spice industry. Useful resources that are conducive to environmentally friendly and ecologically sustainable development, thereby reducing production costs to the greatest extent, effectively improving the ecological environment, and better meeting the industrial production needs of food flavor additives.