CN110305912A - Purposes of the fragrance of method and extraction that fragrance is extracted from jasmine Marasmius bacterial strain in tobacco - Google Patents

Purposes of the fragrance of method and extraction that fragrance is extracted from jasmine Marasmius bacterial strain in tobacco Download PDF

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Publication number
CN110305912A
CN110305912A CN201910386587.7A CN201910386587A CN110305912A CN 110305912 A CN110305912 A CN 110305912A CN 201910386587 A CN201910386587 A CN 201910386587A CN 110305912 A CN110305912 A CN 110305912A
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fragrance
jasmine
marasmius
bacterial strain
culture
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CN201910386587.7A
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CN110305912B (en
Inventor
王猛
赵瑞琳
吴涛
朱杰
曾旭
张伟
任晋伟
刘安琪
贺茂强
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China Tobacco Yunnan Industrial Co Ltd
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China Tobacco Yunnan Industrial Co Ltd
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    • A—HUMAN NECESSITIES
    • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
    • A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
    • A24B3/00—Preparing tobacco in the factory
    • A24B3/12—Steaming, curing, or flavouring tobacco
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B9/00—Essential oils; Perfumes
    • C11B9/02—Recovery or refining of essential oils from raw materials
    • C11B9/022—Refining
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Mycology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Fats And Perfumes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Seasonings (AREA)

Abstract

The invention belongs to field of biotechnology, more particularly to the purposes of the method for extraction fragrance and the fragrance of extraction in tobacco from jasmine Marasmius bacterial strain.This method comprises the following steps: (1) carrying out constant temperature incubation from the culture vessel for taking fungus block access that fluid nutrient medium is housed in jasmine Marasmius bacterial strain flat board；(2) liquid component is obtained by filtration in the culture solution that step (1) obtains, then liquid component is extracted and is enriched with by adsorbent material；(3) it is eluted with adsorbent material of the solvent to step (2) to get the eluent containing the fragrance is arrived；(4) it removes the eluent solvent of step (3) to obtain the fragrance.In addition, the fragrance for also disclosing above method extraction is used to improve the rich purposes with fragrance musical tone of cigarette perfume of tobacco.The present invention is separately cultured from jasmine cuticle destroying angel for the first time, obtains jasmine Marasmius bacterial strain, and extract fragrance from jasmine Marasmius bacterial strain, extracting method is simple, it is easy to accomplish.

Description

The method of fragrance and the fragrance of extraction are extracted from jasmine Marasmius bacterial strain in tobacco Purposes
Technical field
The invention belongs to field of biotechnology, more particularly to the method and extraction of fragrance are extracted from jasmine Marasmius bacterial strain Purposes of the fragrance in tobacco.
Background technique
Fragrance be can pleasant mental and body, pressure of releiving, improve mood, each side of life is had penetrated into the application of fragrance Face, for example, food, animal husbandry, daily cosmetics industry, pharmaceuticals industry etc..The exploitation of fragrance matter, which is one, as a result, is worth The direction furtherd investigate.
Nowadays the source of fragrance matter is mainly there are three aspect, on the one hand the volatile component in fragrant plant, from It is extracted in plant；On the other hand from have produce Xiang Gong can the fungies such as saccharomycete, mould in directly acquire flavor component or It is obtained indirectly in the method for bioconversion；Third is realized by chemistry route, and artificial synthesized method is carried out using chemical reagent Directly obtain the fragrance matter of high-purity.And it is very limited to produce fragrant new plant source, and the plant cultivation period is long, essence yield It is low.And using natural aromatic ingredient as object of reference, the path technical requirements with higher of artificial synthesized new fragrance, technique is multiple Miscellaneous, risk is high.So using the biofermentation technique of aroma-producing microbe, fragrance matter is obtained by the technologies such as isolating and purifying Method is more carried out concern the more because it has the characteristics such as efficient, green, environmental protection.
Summary of the invention
The present invention provides from newfound jasmine Marasmius (Marasmius jasminodorus) bacterial strain and extracts fragrance Method, the bacterial strain are latest find, which is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, Bacterial strain deposit number CGMCC17072；Its source Saving specimen is in Institute of Microorganism, Academia Sinica's fungus specimen museum, collection number For HMAS279932.The fragrance extracted from above-mentioned jasmine Marasmius is for being also the scope of protection of the invention in tobacco.
The present invention provides a kind of method identifying fragrance bacterial strain and its volatile products and extracting with analysis.
Method provided by the invention includes the following steps: to separate bacterium from the fructification of fragrance bacterium using single spore separation method Kind, expand culture mycelia using maltose yeast powder fluid nutrient medium and is obtained by mycelia through liquid fermentation, Solid Phase Extraction and elution To the main eluent in aroma compounds, removes solvent and obtain the fragrance.
Technical scheme is as follows:
First aspect present invention discloses the method that fragrance is extracted from jasmine Marasmius bacterial strain, includes the following steps:
(1) constant temperature is carried out from the culture vessel for taking fungus block access that fluid nutrient medium is housed in jasmine Marasmius bacterial strain flat board Culture；
(2) liquid component is obtained by filtration in the culture solution that step (1) obtains, then by liquid component by adsorbent material into Row extracts and enrichment；
(3) it is eluted with adsorbent material of the solvent to step (2) to get the eluent containing the fragrance is arrived；
(4) it removes the eluent solvent of step (3) to obtain the fragrance.
Preferably, fluid nutrient medium described in step (1) is maltose yeast powder fluid nutrient medium, the malt sugar yeast Powder liquid culture medium includes following component: 20g maltose, 3g yeast powder, 3g KH2PO4、0.01g VB1With 1000ml water.
Preferably, step (2) adsorbent material is macroreticular resin HP20.
Preferably, step (3) solvent is dehydrated alcohol.
Preferably, the condition of step (1) described culture is 25-30 DEG C, is cultivated 7-12 days.
Preferably, culture vessel is is placed in shaking table by step (1) culture, the revolving speed 150- of the shaking table 200r/min。
Second aspect of the present invention disclose the fragrance that the method is extracted be used to improve tobacco cigarette perfume is rich and fragrance The purposes of musical tone
Beneficial effects of the present invention:
1, the fructification of jasmine Marasmius is smaller, and cap part is relatively thin, and common method for tissue separation is difficult to obtain phase Sterile tissue block is cultivated.So the present invention obtains bacterium by the sprouting of single spore using the method for single spore separation Strain, and the process of spore germination has certain randomness, is not easy to obtain bacterial strain.
2, pass through the morphology to wild mushroom and the molecular biological based on the general bar code gene ITS sequence of fungi Analysis identifies that the bacterium is jasmine Marasmius (Marasmius jasminodorus), which has under fructification and hypha form There is strong floral scent, fragrance is unique.
2, the present invention is separately cultured from jasmine cuticle destroying angel for the first time, obtains jasmine Marasmius bacterial strain, and from jasmine Fragrance is extracted in jasmine Marasmius bacterial strain, which is colourless oil liquid.Extracting method is simple, it is easy to accomplish.
3, the culture medium that the present invention selects is maltose yeast powder fluid nutrient medium, can promote jasmine using this culture medium Marasmius growth, keeps the fragrance of jasmine Marasmius more rich, detects more flavor components.Culture medium raw material is cheap and easy to get, It is easy to operate.
4, the present invention extracts fragrance from jasmine Marasmius bacterial strain, and bacterial strain uses therefor is easy to save, activation and fermentation, source are rich Richness, it is low in cost.
Specific embodiment
The following examples embody the process that the present invention describes, but the invention is not limited to these examples.
Embodiment 1: the extraction of fragrance
By the jasmine cuticle destroying angel of field acquisition, cap is taken to be partially disposed in sterile culture in alcolhol burner flame vicinity In ware, cap is upward downward for lamella.Cap is taken out after standing overnight, and acquires basidiospore in culture dish.Use a small amount of sterile water The basidiospore on culture dish is rinsed, spore suspension is obtained.By preparing different spore suspension concentration gradients, obtaining concentration is suitable for Be diluted after spore suspension, and be applied in solid plate, 25 DEG C are protected from light culture, and timing is rechecked, after picking spore germination Mycelia into other culture dishes, after purification i.e. obtain related strain.
Use maltose yeast powder fluid nutrient medium (composition are as follows: 20g maltose, 3g yeast powder, 3g KH2PO4、0.01g VB1With 1000ml water) expand culture mycelia.Fluid nutrient medium is prepared with 250ml conical flask, connecing liquid measure is about 125ml.Ultra-clean In platform, (10 pieces are connect in each culture bottle from the fungus block for cutting about 0.5cm in raw plates and covering with mycelia with sterilized scalpel Fungus block), it accesses in new culture solution, culture bottle will be put into shaking table, and adjusting temperature is 25 DEG C, and revolving speed 150r/min is cultivated 10 days.
It is filtered obtained culture solution to obtain liquid, carries out Solid Phase Extraction by adsorbent material of macroreticular resin HP20, Preliminary extract and enrichment is carried out to flavor component；Macroreticular resin HP20 is eluted with dehydrated alcohol, obtain containing it is savory at The ethanol eluate divided removes dehydrated alcohol and obtains fragrance material.
Embodiment 2: obtained fragrance applications are in tobacco
Embodiment 1 is obtained fragrance to compound with gel-type vehicle, in terms of additive amount carries fragrant gel by 100g fragrance/KG, To be not added with the load perfume (or spice) gel of the fragrance as control sample.Then fragrant gel is carried with the additive amount of 1.5mg/mm in cigarette filter rod Applied when molding.
Analyses Methods for Sensory Evaluation Results shows that, relative to control sample is not added with, adding the fragrance cigarette has apparent faint scent fragrant Rhythm, flue-cured tobacco cigarette perfume, the fragrance of a flower, faint scent in aromatic style and the target improvements such as fragrant and sweet are obvious, and cigarette perfume is rich and fragrance musical tone is bright It is aobvious to be promoted.In terms of qualitative characteristics: oral stimulation and dry sensation, flue gas sophistication and flue gas concentration are changed compared to control cigarette Kind, aroma quality is promoted, and musical tone harmony is balanced, and aftertaste is comfortable.
Sense organ comparison index quantification evaluation is carried out using YC/T 497-2014 " cigarette Chinese-style cigarette sensory evaluation method ", To be not added with the load perfume (or spice) gel cigarette of jasmine Marasmius fragrance as control sample, the results are shown in Table 1.
1 blank control sample of table and test cigarette style sensory evaluation data

Claims (7)

1. extracting the method for fragrance from jasmine Marasmius bacterial strain, which comprises the steps of:
(1) from taken in jasmine Marasmius bacterial strain flat board fungus block access equipped with fluid nutrient medium culture vessel in carry out constant temperature training It supports；
(2) liquid component is obtained by filtration in the culture solution that step (1) obtains, then mentions liquid component by adsorbent material It takes and is enriched with；
(3) it is eluted with adsorbent material of the solvent to step (2) to get the eluent containing the fragrance is arrived；
(4) it removes the eluent solvent of step (3) to obtain the fragrance.
2. the method according to claim 1, wherein fluid nutrient medium described in step (1) is malt sugar yeast Powder liquid culture medium, the maltose yeast powder fluid nutrient medium includes following component: 20g maltose, 3g yeast powder, 3g KH2PO4、0.01gVB1With 1000ml water.
3. the method according to claim 1, wherein step (2) adsorbent material is macroreticular resin HP20.
4. the method according to claim 1, wherein step (3) solvent is dehydrated alcohol.
5. being cultivated the method according to claim 1, wherein the condition of step (1) described culture is 25-30 DEG C 7-12 days.
6. the method according to claim 1, wherein step (1) culture is shaken for culture vessel to be placed on In bed, the revolving speed 150-200r/min of the shaking table.
7. the rich use with fragrance musical tone of cigarette perfume that the fragrance extracted according to the method for claim 1 is used to improve tobacco On the way.
CN201910386587.7A 2019-05-09 2019-05-09 Method for extracting spice from jasmine Marasmius pauciflorus strain and application of extracted spice in tobacco Active CN110305912B (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2020358A1 (en) * 1990-07-04 1992-01-05 Thomas Andrew Reisner Process for the preparation and manufacture of a vegetable-based substitute for anchovy paste
TW201000637A (en) * 2008-06-02 2010-01-01 Omya Development Ag Nucleic acids and methods for detecting turfgrass pathogenic fungi
EP3082729A1 (en) * 2013-12-20 2016-10-26 Unilever N.V. Sunless tanning
CN107624513A (en) * 2017-10-25 2018-01-26 山东惠民齐发果蔬有限责任公司 It is a kind of rich in the edible and medical fungi cultural hypha method of polysaccharide and application
CN109722393A (en) * 2019-01-04 2019-05-07 云南中烟工业有限责任公司 A kind of tobacco yellow soil source bacterium and its separation method and application

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2020358A1 (en) * 1990-07-04 1992-01-05 Thomas Andrew Reisner Process for the preparation and manufacture of a vegetable-based substitute for anchovy paste
TW201000637A (en) * 2008-06-02 2010-01-01 Omya Development Ag Nucleic acids and methods for detecting turfgrass pathogenic fungi
EP3082729A1 (en) * 2013-12-20 2016-10-26 Unilever N.V. Sunless tanning
CN107624513A (en) * 2017-10-25 2018-01-26 山东惠民齐发果蔬有限责任公司 It is a kind of rich in the edible and medical fungi cultural hypha method of polysaccharide and application
CN109722393A (en) * 2019-01-04 2019-05-07 云南中烟工业有限责任公司 A kind of tobacco yellow soil source bacterium and its separation method and application

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ACHARYA, KRISHNENDU等: "A new species of Marasmius sect. Sicci from India", 《MYCOTAXON》 *
LITTIBED KHOLA等: "Marasmius (Basidiomycota, Agaricales) in Dry Deciduous Dipterocarp Forest at Dong-Yai Community Forest, Thailand", 《NU. INTERNATIONAL JOURNAL OF SCIENCE》 *
刘安琪等: "一个产香大型真菌鉴定及其挥发性成分分析", 《微生物学报》 *
刘选明等: "一株香茅草内生细菌Pantoea ananatis CcSh-1的分离、鉴定及产香条件优化研究", 《湖南大学学报(自然科学版)》 *

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