CN106263010B - A kind of method that bioconversion improves reconstituted tobacco organoleptic quality - Google Patents
A kind of method that bioconversion improves reconstituted tobacco organoleptic quality Download PDFInfo
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- CN106263010B CN106263010B CN201610781986.XA CN201610781986A CN106263010B CN 106263010 B CN106263010 B CN 106263010B CN 201610781986 A CN201610781986 A CN 201610781986A CN 106263010 B CN106263010 B CN 106263010B
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- 101001065065 Aspergillus awamori Feruloyl esterase A Proteins 0.000 claims abstract description 15
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- 235000001785 ferulic acid Nutrition 0.000 description 13
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 description 13
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 description 9
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- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 description 7
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 5
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- 150000001413 amino acids Chemical class 0.000 description 1
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- 238000009395 breeding Methods 0.000 description 1
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- 239000012982 microporous membrane Substances 0.000 description 1
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- 150000002989 phenols Chemical class 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
- A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
- A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
- A24B15/18—Treatment of tobacco products or tobacco substitutes
- A24B15/20—Biochemical treatment
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Manufacture Of Tobacco Products (AREA)
Abstract
The present invention, which provides a kind of bioconversion, improves the method for reconstituted tobacco organoleptic quality, includes the following steps：(1) bacterial strain activates：Food methanol bacillus (VJ4 1) is inoculated on slant medium, inoculates seed culture medium, transfers and seed culture fluid is made；(2) compound bio conversion pretreatment：In the extraction process of reconstituted tobacco, seed culture fluid described in step 1) is inoculated into reconstituted tobacco aqueous solution, and add the complex enzymes such as Penicillium citrinum feruloyl esterase, zytase, glucose oxidase, catalase, phenol oxidase, it is uniformly mixed, fermentation obtains reconstituted tobacco bioconversion broth；(3) reconstituted tobacco is produced through reconstituted tobacco normal process steps, in the process, more than 75 DEG C, enzyme of the invention and bacterium inactivation roll to obtain reconstituted tobacco temperature.Biotechnology produced by the invention promotes the reconstituted tobacco of flavouring essence quality, reduces irritation, reduces blue foreign smell, slightly fragrance grace, milk fragrance, improves reconstituted tobacco organoleptic quality.
Description
Technical field
The present invention relates to tobacco industry field, the method for specifically a kind of bioconversion improvement reconstituted tobacco organoleptic quality.
Background technology
Reconstituted tobacco mainly adds in adhesive and other additives etc. by offal, fragment, offal or discarded tobacco leaf and forms.Cigarette
End, fragment, offal are the by-products of tobacco industry, in the past to make Disposal more.China is tobacco big country, there is few hundred thousand tonnes of every year
Offal, fragment, offal resource are discarded, and both cause environmental pollution, while are also the waste to natural resources.Research shows that cigarette
The ingredient type and tobacco leaf ingredient contained in end, fragment, offal is basically identical, but there are miscellaneous gas weight, irritation is big and mouthfeel
Discomfort etc. inhales taste defect.
The cell wall substance of reconstituted tobacco mainly has cellulose, hemicellulose, pectin, lignin etc., if cell wall substance
With content of starch height, tobacco leaf can be caused coarse, flue gas stimulation is big, and discord is suitable；Excessive cellulose in cell wall substance, can assign
It gives flue gas spine and burning heat sensation and generates cough of choking；And the thermal cracking of lignin has a negative impact to cigarette quality, content is got over
Height, quality of tobacco is poorer, has a strong impulse smell, blue foreign smell weight, inhales that acrid flavour is peppery, puckery mouth.Lignin is also to make simultaneously
The main reason for paper industrial discharge black liquor COD and coloration are formed, is one of microorganism generally acknowledged at present aromatic compound difficult to degrade.
Biotransformation method is the chemical reaction carried out under the living things catalysis such as cell or enzyme, is both referred to (micro- using microorganism
Biology growth cell, microorganism resting cell), enzyme (pure enzyme or thick enzyme from microorganism or animals and plants) or plant it is thin
Born of the same parents pass through the method for substance needed for simple bioconversion or biosynthesis preparation.Compared with chemical catalyst, biocatalyst
Specificity is stronger, catalytic activity higher, condition are more easy to control.Microorganism can generate huge high active enzyme in breeding
System, such as polysaccharide hydrolysis enzyme, protease, esterification enzyme, redox enzymes and lyases, in enzymatic catalysis, chemistry
In effect and microbial body under the synergistic effect of complicated metabolism, plant material can be made to decompose, degrade, aoxidize, restore, gather
Close, coupling, conversion the effects that, form complicated low molecular compound, including various perfume compounds, as alcohols, aldehydes,
Ketone, acids, lipid, phenols, pyran, pyridines and terpenes etc., these aroma substances are also undoubtedly tobacco or for cigarette
The aroma component of the additive of grass.
Feruloyl esterase can cut off the crosslinking between one polysaccharide of polysaccharide, one lignin of polysaccharide in cell wall, be conducive to cell wall
The degradation of polysaccharide and the release of lignin in substance, can be by ferulic acid using the raw material of feruloyl esterase process vegetal
The separate out from the structure of plant cell wall, so as to destroy the skeleton structure of cell wall so that lignin, hemicellulose with
And the connection between cellulose is partly broken, structure becomes more loose than before processing.Ferulic acid is present in cured tobacco leaf.Fiber
Element, starch, protein can generate glucose, oligosaccharide, small peptide, amino acid etc. by biological enzyme directionally hydrolyzing, can effectively reduce volume
The irritation of cigarette mitigates miscellaneous gas and improves and inhales taste.Water-soluble sugar content height makes the jealous alcohol of tobacco leaf and fragrance is graceful, but carbohydrate
It is high to close object content, releasing content of coke tar is big.
Using reconstituted tobacco aqueous solution as substrate, using Penicillium citrinum feruloyl esterase, zytase, glucose oxidase, mistake
Hydrogen oxide enzyme, phenol oxidase synergistic effect, and using food methanol bacillus methylotrophicus (VJ4-
1) it converts, decomposes the macromoleculars pungent such as lignin, cellulose, protein in reconstituted tobacco, discharge ferulic acid, and convert
Ferulic acid is the fragrance precursors substance such as vanillic aldehyde, make production reconstituted tobacco reduce irritation, reduce blue foreign smell, fragrance it is graceful,
Slightly milk fragrance improves reconstituted tobacco organoleptic quality, has not been reported.
Invention content
In view of the deficiencies of the prior art, the present invention provides a kind of biotechnology promoted reconstituted tobacco flavouring essence quality method,
Using food methanol bacillus methylotrophicus (VJ4-1) and Penicillium citrinum feruloyl esterase, xylan
The complex enzymes such as enzyme, glucose oxidase, catalase, phenol oxidase act synergistically, using reconstituted tobacco aqueous solution as substrate into
Row bioconversion, then produce reconstituted tobacco through conventional industrial processes.Wherein it is thin to discharge reconstituted tobacco for Penicillium citrinum feruloyl esterase
Ferulic acid in cell wall makes reconstituted tobacco cell wall open structure, is conducive to the degradation of polysaccharide and lignin in cell wall substance
Release, recycle combined oxidation enzyme oxidation lignin degrading, food methanol bacillus (VJ4-1) ferulic acid can be converted
Vanillic aldehyde is generated, enzyme system can decompose, convert the macromolecular pungent in reconstituted tobacco or the fragrance precursor of difficult volatilization
Substance is small molecule aroma substance or reduced sugar etc.；And the polysaccharases such as feruloyl esterase and zytase have synergistic effect,
These enzymes and the compound bio of microorganism conversion, make production reconstituted tobacco reduce irritation, reduce blue foreign smell, flue gas and it is suitable,
Fragrance grace, slightly milk fragrance, improve reconstituted tobacco organoleptic quality.
The present invention, which provides a kind of bioconversion, improves the method for reconstituted tobacco organoleptic quality, includes the following steps：
(1) bacterial strain activates：Food methanol bacillus methylotrophicus (VJ4-1) is inoculated into horse
On bell potato dextrose agar slant culture medium, under the conditions of 25~38 DEG C, pH5~8, inclined-plane is obtained after 24~96h of quiescent culture
Culture inoculates seed culture medium, under the conditions of 25~38 DEG C, 100~150rpm/min of rotating speed shaken cultivation 12~
Seed culture fluid is made after continuously transferring 1~3 time in 36h；
Described food methanol bacillus methylotrophicus (VJ4-1) preserving number is CCTCC NO：
2012004, it is preserved in Wuhan University's China typical culture collection center on January 16th, 2012；
The slant medium：Potato immersion liquid 200mL/L, glucose 20g/L, agar 13g/L.The seed training
Support base：Maltose 50g/L, peptone 10g/L, sodium chloride 5g/L.
(2) compound bio conversion pretreatment：In the extraction process of reconstituted tobacco, the seed culture fluid described in step 1) is pressed
Mass ratio 2~18% is inoculated into reconstituted tobacco aqueous solution, and adds 0.15~0.4% tangerine blueness for accounting for reconstituted tobacco mass ratio
The complex enzymes such as mould feruloyl esterase, zytase, glucose oxidase, catalase, phenol oxidase, are uniformly mixed,
In pH to 4.5~6.0, under the conditions of 30~45 DEG C, ferment 1~12 hour, obtain reconstituted tobacco bioconversion broth；
(3) reconstituted tobacco routine production process：The reconstituted tobacco bioconversion broth of the step 1), in 55~65 DEG C, pH
40~60min of extraction reaction, dish spiral shell centrifuge melt cinder under natural conditions, contain being concentrated in vacuo at 75~85 DEG C of extract liquor admittedly
Amount 40%, obtains concentrate, and in the process, temperature is more than 75 DEG C, enzyme of the invention and bacterium inactivation；And residue is then by using dry
Dry machine implements 100~120 DEG C of drying process, inactivator and bacterium, and the tobacco substrate of 5~15% water content of mass ratio is made；It will be dense
Contracting liquid sprays to tobacco-based on piece and rolls into the reconstituted tobacco for promoting flavouring essence quality.
Further, Penicillium citrinum feruloyl esterase, zytase, glucose oxidase, hydrogen peroxide described in step (2)
The complex enzymes such as enzyme, phenol oxidase, be food-grade, enzyme activity 500u/g~10000u/g, account for respectively reconstituted tobacco mass ratio 0~
0.1%.
Beneficial effects of the present invention：
The present invention carries out bioconversion using functional microorganism and complex enzyme to reconstituted tobacco, passes through Penicillium citrinum ferulic acid ester
Ester bond in enzyme hydrolysis offal, the Ferulic acid methylester of offal cell wall, oligosaccharide ferulic acid ester and polysaccharide ferulic acid ester, release Ah
Wei's acid, is conducive to the degradation of polysaccharide and the release of lignin in cell wall substance, destroys the skeleton structure of cell wall, make wooden
Connection between element, hemicellulose and cellulose is partly broken, and structure becomes more loose than before processing, is cooperateed with zytase
Effect, degradation of cell wall substance；Glucose oxidase, catalase, phenol oxidase synergistic effect oxidative degradation lignin；Work(
Energy microbial food methanol bacillus methylotrophicus (VJ4-1) can convert ferulic acid generation vanillic aldehyde,
Its complicated enzyme system can decompose, convert macromolecular pungent in reconstituted tobacco or the difficult fragrance precursor substance that volatilizees is
Small molecule aroma substance or reduced sugar etc.；The reconstituted tobacco of production, drops in all these enzymes and the conversion of the compound bio of microorganism
Low irritant, reduce blue foreign smell, flue gas and it is suitable, fragrance is graceful, slightly milk fragrance, improve reconstituted tobacco organoleptic quality.
Present invention process mild condition is easy to operate, realizes enzyme and microorganism conversion and reconstituted tobacco flavouring essence quality is carried
It rises, it is environmentally protective, there is the prospect of industrial applications.
Specific embodiment
The technical solution in the present invention is clearly and completely described below by specific embodiment.It should be appreciated that
The described specific embodiment in this part can only be used to explain the present invention, be not intended to limit the present invention.
Embodiment 1
A kind of method that bioconversion improves the reconstituted tobacco of organoleptic quality, includes the following steps：
(1) bacterial strain activates：Food methanol bacillus methylotrophicus (VJ4-1) is inoculated into horse
On bell potato dextrose agar slant culture medium, under the conditions of 28 DEG C, pH5, slant culture is obtained after quiescent culture 28h, then connect
Kind is to seed culture medium, the shaken cultivation 12h under the conditions of 28 DEG C, rotating speed 115rpm/min, and seed, which is made, after continuously transferring 2 times trains
Nutrient solution；
The slant medium：Potato immersion liquid 200mL/L, glucose 20g/L, agar 13g/L.The seed training
Support base：Maltose 50g/L, peptone 10g/L, sodium chloride 5g/L.
(2) compound bio conversion pretreatment：In the extraction process of reconstituted tobacco, the seed culture fluid described in step 1) is pressed
Mass ratio 5% is inoculated into reconstituted tobacco aqueous solution, and adds the 0.18% Penicillium citrinum ferulic acid for accounting for reconstituted tobacco mass ratio
The complex enzymes such as esterase, zytase, glucose oxidase, catalase, phenol oxidase, are uniformly mixed, in pH5,30
Under the conditions of DEG C, ferment 4 hours, obtain reconstituted tobacco bioconversion broth；The Penicillium citrinum feruloyl esterase, zytase, grape
The complex enzymes such as carbohydrate oxidase, catalase, phenol oxidase, are food-grade, and enzyme activity 1000u/g respectively accounts for reconstituted tobacco mass ratio
0.05%, 0.05%, 0.03%, 0.03%, 0.02%；
(3) reconstituted tobacco routine production process：Reconstituted tobacco bioconversion broth obtained by the step 2), in 55 DEG C, pH
Extraction reaction 55min, dish spiral shell centrifuge melt cinder under natural conditions, will be concentrated in vacuo to solid content 40% at 78 DEG C of extract liquor,
Concentrate is obtained, in the process, temperature is more than 78 DEG C, enzyme of the invention and bacterium inactivation；And residue is then by using drying machine reality
102 DEG C of drying process, inactivator and bacterium are applied, the tobacco substrate of 12% water content of mass ratio is made；Concentrate is sprayed into tobacco
The reconstituted tobacco for promoting flavouring essence quality is rolled on substrate.
Embodiment 2
A kind of method that bioconversion improves the reconstituted tobacco of organoleptic quality, includes the following steps：
(1) bacterial strain activates：Food methanol bacillus methylotrophicus (VJ4-1) is inoculated into horse
On bell potato dextrose agar slant culture medium, under the conditions of 30 DEG C, pH5.5, slant culture is obtained after quiescent culture 36h, then
Seed culture medium is inoculated into, seed is made after continuously transferring 3 times in the shaken cultivation 18h under the conditions of 30 DEG C, rotating speed 120rpm/min
Culture solution；
The slant medium：Potato immersion liquid 200mL/L, glucose 20g/L, agar 13g/L.The seed training
Support base：Maltose 50g/L, peptone 10g/L, sodium chloride 5g/L.
(2) compound bio conversion pretreatment：In the extraction process of reconstituted tobacco, the seed culture fluid described in step 1) is pressed
Mass ratio 9% is inoculated into reconstituted tobacco aqueous solution, and adds the 0.3% Penicillium citrinum ferulic acid ester for accounting for reconstituted tobacco mass ratio
The complex enzymes such as enzyme, zytase, glucose oxidase, catalase, phenol oxidase, are uniformly mixed, in pH to 5.5,
Under the conditions of 32 DEG C, ferment 6 hours, obtain reconstituted tobacco bioconversion broth；The Penicillium citrinum feruloyl esterase, zytase,
The complex enzymes such as glucose oxidase, catalase, phenol oxidase, are food-grade, and 800u/g accounts for reconstituted tobacco mass ratio respectively
0.1%, 0.1%, 0.05%, 0.03%, 0.02%；
(3) reconstituted tobacco routine production process：Reconstituted tobacco bioconversion broth obtained by the step 2), in 58 DEG C, pH
Extraction reaction 55min, dish spiral shell centrifuge melt cinder under natural conditions, will be concentrated in vacuo to solid content 40% at 80 DEG C of extract liquor,
Concentrate is obtained, in the process, temperature reaches 80 DEG C, and enzyme of the invention and bacterium inactivate；And residue is then by using drying machine reality
105 DEG C of drying process, inactivator and bacterium are applied, the tobacco substrate of 13% water content of mass ratio is made；Concentrate is sprayed into tobacco
The reconstituted tobacco for promoting flavouring essence quality is rolled on substrate.
Embodiment 3
A kind of method that bioconversion improves the reconstituted tobacco of organoleptic quality, includes the following steps：
(1) bacterial strain activates：Food methanol bacillus methylotrophicus (VJ4-1) is inoculated into horse
On bell potato dextrose agar slant culture medium, under the conditions of 32 DEG C, pH6, slant culture is obtained after quiescent culture 72h, then connect
Kind is to seed culture medium, the shaken cultivation 30h under the conditions of 32 DEG C, rotating speed 125rpm/min, and seed, which is made, after continuously transferring 2 times trains
Nutrient solution；
The slant medium：Potato immersion liquid 200mL/L, glucose 20g/L, agar 13g/L.The seed training
Support base：Maltose 50g/L, peptone 10g/L, sodium chloride 5g/L.
(2) compound bio conversion pretreatment：In the extraction process of reconstituted tobacco, the seed culture fluid described in step 1) is pressed
Mass ratio 12% is inoculated into reconstituted tobacco aqueous solution, and adds the 0.25% Penicillium citrinum ferulic acid for accounting for reconstituted tobacco mass ratio
The complex enzymes such as esterase, zytase, glucose oxidase, catalase, phenol oxidase, are uniformly mixed, pH6.0,
Under the conditions of 32 DEG C, ferment 8 hours, obtain reconstituted tobacco bioconversion broth；The Penicillium citrinum feruloyl esterase, zytase, Portugal
The complex enzymes such as grape carbohydrate oxidase, catalase, phenol oxidase, are food-grade, and enzyme activity 1500u/g accounts for reconstituted tobacco matter respectively
Measure 0.08%, 0.08%, 0.04%, 0.03%, the 0.02% of ratio；
(3) reconstituted tobacco routine production process：Reconstituted tobacco bioconversion broth obtained by the step 2), in 62 DEG C, pH
Extraction reaction 45min, dish spiral shell centrifuge melt cinder under natural conditions, will be concentrated in vacuo to solid content 40% at 82 DEG C of extract liquor,
Concentrate is obtained, in the process, temperature reaches 82 DEG C, and enzyme of the invention and bacterium inactivate；And residue is then by using drying machine reality
110 DEG C of drying process, inactivator and bacterium are applied, the tobacco substrate of 11% water content of mass ratio is made；Concentrate is sprayed into tobacco
The reconstituted tobacco for promoting flavouring essence quality is rolled on substrate.
Test example 1：Sensory evaluating smoking's Piglet s colibacillosis
Bioconversion prepared by embodiment 1,2,3 improves the reconstituted tobacco of organoleptic quality and untreated control sample
Cigarette is rolled into, (22 DEG C+1, relative humidity 60%+2%) balance 48h is put into climatic chamber, expert is asked to smoke panel test group
It smokes panel test.
It smokes panel test repeatedly through the group of smokeing panel test, table 1 shows：Bioconversion prepared by the present invention improves the reconstruction cigarette of organoleptic quality
Under the biotransformation of leaf, complex enzyme and function stem, degradation is decomposed, the egg in conversion reconstituted tobacco extract liquor and substrate
The macromolecular substances such as white matter, lignin, pectin, starch, cellulose are reduced sugar, vanillic aldehyde and its fragrance precursor substance, make life
The reconstituted tobacco of production reduce irritation, reduce blue foreign smell, flue gas and it is suitable, fragrance is graceful, slightly milk fragrance, improve reconstituted tobacco sense
Official's quality.
1. biotechnology of table promotes sensory evaluating smoking's Contrast on effect of the reconstituted tobacco of flavouring essence quality
Test example 2：The detection of reconstituted tobacco content of lignin
Content of lignin in reconstituted tobacco is detected using the classical way Klason methods for measuring content of lignin, it is former
Reason is to remove cellulose with 72% sulfuric acid, the lignin of indissoluble is filtered, washing, it is dry weigh, calculates and etc. it is available its
Content, the content of lignin that bioconversion of the invention improves in the reconstituted tobacco of organoleptic quality have reduction, as a result such as table 2.
The content of lignin of the reconstituted tobacco of 2. integrated approach General Promotion quality of table
| Experimental group | Content of lignin | Lignin reduced rate |
| Blank control | 3.32% | / |
| 1 group of embodiment | 2.82% | 15.06% |
| 2 groups of embodiment | 2.80% | 15.66% |
| 3 groups of embodiment | 2.75% | 17.16% |
Test example 3：Reconstituted tobacco bioconversion broth vanillic aldehyde HPLC is detected
Reconstituted tobacco bioconversion broth with high performance liquid chromatography detection after 0.22 μm of filtering with microporous membrane, uses Agilent
Eclipse XDB-C8 chromatographic columns (5 μm, 4.6*150mm), mobile phase are glacial acetic acid=30/70 of methanol/0.5%, flow velocity 1mL/
Min, Detection wavelength 280nm, sample size 10 μ L, detection time 40min.The standard sample that 1g/L is prepared with 50% methanol solution is molten
Liquid is diluted to a series of various concentrations, and each 10 μ L of sample introduction map to standard sample concentration with peak area, draw standard curve.
The concentration of vanillic aldehyde in 3. reconstituted tobacco bioconversion broth of table
| Experimental group | Vanillic aldehyde concentration (mg/L) |
| Untreated control | / |
| 1 group of embodiment | 121 |
| 2 groups of embodiment | 262 |
| 3 groups of embodiment | 209 |
The above description is merely a specific embodiment, but protection scope of the present invention is not limited thereto, any
Belong to those skilled in the art in the technical scope disclosed by the present invention, the change or replacement that can be readily occurred in all should
It is included within the scope of the present invention.Therefore, protection scope of the present invention should be subject to the protection scope in claims.
Claims (1)
1. a kind of method that bioconversion improves reconstituted tobacco organoleptic quality, it is characterised in that include the following steps：
(1) bacterial strain activates：Food methanol bacillus methylotrophicus (VJ4-1) is inoculated into potato
On dextrose agar slant culture medium, under the conditions of 25~38 DEG C, pH5~8, inclined-plane culture is obtained after 24~96h of quiescent culture
Object inoculates seed culture medium, 12~36h of shaken cultivation under the conditions of 25~38 DEG C, 100~150rpm/min of rotating speed, even
Continue and seed culture fluid is made after transferring 1~3 time；
Described food methanol bacillus methylotrophicus (VJ4-1) preserving number is CCTCC NO：
2012004, it is preserved in Wuhan University's China typical culture collection center on January 16th, 2012；
The slant medium is：Potato immersion liquid 200mL/L, glucose 20g/L, agar 13g/L；The seed culture
Base：Maltose 50g/L, peptone 10g/L, sodium chloride 5g/L；
(2) compound bio conversion pretreatment：In the extraction process of reconstituted tobacco raw material, the seed culture fluid described in step 1) is pressed
Mass ratio 2~18% is inoculated into reconstituted tobacco aqueous solution of raw material, and add account for reconstituted tobacco material quality than 0.15~
0.4% Penicillium citrinum feruloyl esterase, zytase, glucose oxidase, catalase, phenol oxidase complex enzyme, stirring
It is uniformly mixed, in pH to 4.5~6.0, under the conditions of 30~45 DEG C, ferments 1~12 hour, obtain reconstituted tobacco raw material biology and turn
Change liquid；The Penicillium citrinum feruloyl esterase, zytase, glucose oxidase, catalase, phenol oxidase are food
Grade, enzyme activity 500u/g~10000u/g, account for respectively reconstituted tobacco material quality than 0.08%, 0.08%, 0.04%,
0.03%th, 0.02%；
(3) reconstituted tobacco routine production process：The reconstituted tobacco raw materials through biotransformation liquid of the step 2), in 55~65 DEG C, pH
40~60min of extraction reaction, dish spiral shell centrifuge melt cinder under natural conditions, contain being concentrated in vacuo at 75~85 DEG C of extract liquor admittedly
Amount 40%, obtains concentrate, in the process, temperature is more than 75 DEG C, the Penicillium citrinum feruloyl esterase, zytase, grape
Carbohydrate oxidase, catalase, phenol oxidase and the food methanol bacillus methylotrophicus
(VJ4-1) it inactivates；And residue then implements 100~120 DEG C of drying process, inactivator and bacterium by using drying machine, and mass ratio 5 is made
The tobacco substrate of~15% water content；By concentrate spray to tobacco-based on piece roll into promoted flavouring essence quality reconstituted tobacco.
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