Summary of the invention
The object of the present invention is to provide a kind of with natural matter, as the precursor of agricultural byproducts tobacco as bio-transformation, by the technology of fermenting and producing perfume phenylethanol, not only opened up an industrial approach, and found a kind of reliable resource for perfume industry for the deep processing of agricultural byproducts tobacco.
Design of the present invention is such:
The contriver is that raw material is produced spices through microbial fermentation according to oneself practice and working experience with the agricultural byproducts tobacco, finds the evolutionary path of a green for tobacco.Tobacco is mainly used in and produces cigarette at present, and Smoking is harmful to your health, but formed hobby one fashion of people can not be broken away from for a long time, needs long transition period, particularly before tobacco is not found new application approach as yet, tobacco grower's economic interests will be influenced directly.In a period of transition, tobacco industry has considerable inferior material-offal, offal, can also produce spices by microbial fermentation as raw material.In a word, it is that raw material is produced phenylethyl alcohol with the agricultural byproducts tobacco that the present invention intends by preferred fermented bacterium and technological condition for fermentation, adsorb through extraction or ion exchange resin then and extract and concentrate the production perfume phenylethanol, thereby found the approach of a green for the development of tobacco industry.
The present invention also is achieved in that
The contriver is by a large amount of tests, find that following microorganism has good ferment effect to tobacco, xylogen in the tobacco, pectin and polyphenolic compound can be degraded and change into spices such as phenylethyl alcohol, said microorganism can be a kind of in Candida utilis (Candida units Lodder et Kueger), yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), the Chinese kluyveromyces (Kluyveromyce sinensis), but is good with yeast saccharomyces cerevisiae.
Concrete bacterial classification source is as follows:
Candida utilis (Candida units Lodder et Kueger)
1. northeast Science Institute Dalian branch is found, China Committee for Culture Collection of Microorganisms's preservation,
Preserving number is CGMCC, No2.120;
2. Institute of Micro-biology of the Chinese Academy of Sciences finds, China Committee for Culture Collection of Microorganisms's preservation, and preserving number is
CGMCC,No2.1180:
3. Kyoto Univ Japan finds, China Committee for Culture Collection of Microorganisms's preservation, and preserving number is
CGMCC,No2.1495。
Yeast saccharomyces cerevisiae (Saccharomyces cerevisiae)
Chinese Academy of Sciences Shanghai plant give birth to find China Committee for Culture Collection of Microorganisms's preservation, preserving number
Be CGMCC, No2.502;
2. Institute of Micro-biology of the Chinese Academy of Sciences finds, China Committee for Culture Collection of Microorganisms's preservation, and preserving number is
CGMCC,No2.742;
3. Institute of Micro-biology of the Chinese Academy of Sciences finds, China Committee for Culture Collection of Microorganisms's preservation, and preserving number is
CGMCC,No2.1419。
China's kluyveromyces (Kluyveromyce sinensis)
1. Institute of Micro-biology of the Chinese Academy of Sciences finds, China Committee for Culture Collection of Microorganisms's preservation, and preserving number is
CGMCC,No2.1535。
The concrete zymotechnique and the extracting method of product are described below:
1, zymotechnique
Seed culture: utilize in the above-mentioned bacterial classification any, adopt conventional yeast liquid aerobic fermentation method to cultivate, be carbon source (said glucide can be one or more in glucose, sucrose, lactose, fructose, maltose, the starch) promptly with the carbohydrate goods and materials, with peptone, casein or yeast extract paste is nitrogenous source, and add an amount of inorganic salt and trace element, be deployed into solution as seed culture medium with deionized water, obtain the liquid yeast bacterial classification through cultivation, stand-by.Because the conventional method of the culturing process of barms system is so narration is conformed to the principle of simplicity.
The fermentating culturing process of tobacco:
The composition of fermention medium: the tobacco with pulverizing is raw material (comprising inferior material offal and offal that present tobacco industry stays), because tobacco itself is a good nitrogenous source, also contain 5~20% starch and reducing sugar simultaneously, so tobacco also is a good carbon source, add an amount of glucide and an amount of inorganic salt and trace element and deionized water again, place fermentor tank, form substratum slurries, regulate the PH=7.0 of slurries earlier with oxalic acid or oxalate, then with slurries at 15psig, under 116~121 ℃, sterilized 15~20 minutes, be cooled to≤more above-mentioned liquid yeast bacterial classification is linked in the sterilized fermention medium after 30 ℃, inoculum size is 5%~10% (V/V), under 30 ℃ of aerobic conditions, stir fermentation 24~72 hours, wherein the feeding amount of sterile air is every liter of substratum slurries 1.0~1.5L/min, take a sample after the fermentation ends or in the fermenting process, with the content of phenylethyl alcohol in gas-chromatography-GC-MS (being called for short the GC-MS instrument) detection fermented liquid, evaluate the technique effect of fermenting process.
Wherein the tobacco of said pulverizing is that tobacco powder is broken to 20~40 sieve apertures, add-on 50~60g/L; Said an amount of glucide is one or more in glucose, sucrose, lactose, fructose, maltose, the starch, and add-on is 20~30g/L; Said inorganic salt comprise FeSO
4, MgSO
4, KCl, CaCl
2, KH
2PO
4All kinds of compound, add-on is respectively 0.1~0.5g/L; All the other are deionized water; Said trace element is then brought into the adding of inorganic salt and tobacco, needn't add in addition.
The purpose that adds the carbohydrate carbon source in the fermention medium is in order to promote yeast amplification rapidly after inoculation, to reach 2.0 * 10 at least with yeast in every milliliter of substratum slurries of dilution-plate method mensuration
8Individual, yeast just utilizes carbon source and nitrogenous source in the tobacco to grow afterwards, promptly by saccharomycetic fermentation culture, and the degraded xylogen in the tobacco, pectin and polyphenolic compound, and change into material such as phenylethyl alcohol, thereby obtain to be rich in the fermented liquid of phenylethyl alcohol.
The zymotechnique of tobacco is the core of fermentative Production phenylethyl alcohol, and after a large amount of experiments, the present invention preferentially recommends following zymotechnique:
Substratum is formed: pulverize tobacco 50g/L; Glucose 20g/L; Inorganic salt: FeSO
40.5g/L, MgSO
40.5g/L, KCl0.1g/L, CaCl
20.1g/L, KH
2PO
40.125g/L; All the other are deionized water, place fermentor tank, under agitation constitute the substratum slurries of homogeneous.Regulate substratum slurries PH=7.0 with oxalic acid, then 15psig, 121 ℃ down sterilization to obtain the sterile culture based slurries in 15 minutes stand-by.
Above-mentioned culture medium after sterilization slurries are cooled to≤30 ℃ after, insert the liquid yeast bacterial classification, inoculum size was 10% (V/V), 30 ℃ of ferment under aerobic conditions 48 hours, wherein sterile air feeding amount is every liter of substratum slurries 1.5L/min, and mixing speed is 80~150r/min.Used yeast can be any in Candida utilis, yeast saccharomyces cerevisiae, the Chinese kluyveromyces, but is good with yeast saccharomyces cerevisiae.Sampling can reach more than the 50ml/L with the content that the GC-MS instrument detects phenylethyl alcohol in the fermented liquid after the fermentation ends.
2, fermented liquid gives processing
The fermented liquid that above-mentioned gained is contained phenylethyl alcohol is regulated PH=5~6.5 with oxalic acid earlier, adds a spot of flocculating aids then, puts a jar solids removed by filtration impurity (comprising mycelium and other insolubles) behind the stirring and evenly mixing, obtains fermented supernatant fluid and filter cake.Wherein said flocculating aids is a kind of in powdered diatomite, argentine, lime carbonate or the paper pulp, and its add-on is 0.5~2% (weight percent).The purpose of regulating PH=5~6.5 of fermented liquid with oxalic acid is to prevent the self-dissolving of yeast filament under alkalescence or neutrallty condition and the precipitation of impelling protein matter, to keep the quality of fermented liquid.The purpose that adds flocculating aids is to improve the filtering feature of fermented liquid, and it can be filtered by general vacuum filter easily, avoids using high speed centrifugation to filter, and helps industrialization promotion and uses.
3, the processing of gained filter cake
Because often be entrained with 10% left and right sides tunning in the filtering fermentation liquor gained filter cake, its treatment process has two.One reclaims the phenylethyl alcohol of carrying secretly in the filter cake with the solvent leaching, gained reclaims liquid and does the extraction agent use for next step, said solvent is a kind of in normal hexane, hexanaphthene, chloroform, tetracol phenixin, benzene,toluene,xylene, the sherwood oil, they also are benzene extraction alcoholic acid extraction agents from fermented supernatant fluid, its consumption is suitable with the fermented supernatant fluid amount, be solvent/fermented supernatant fluid=1 (volume ratio), because the relative filter cake of consumption of solvent is a large amount of, so the rate of recovery of the phenylethyl alcohol in the filter cake can be up to more than 98%; Its dual-purpose 95% industrial spirit or methyl alcohol are made washing composition, and with the phenylethyl alcohol in the washing and recycling filter cake, washing composition consumption 0.5~1.0L/kg filter cake, gained washings and fermented supernatant fluid merge the back to be handled for going on foot down, and phenylethyl alcohol washing and recycling rate can reach about 85%.
4, from fermented supernatant fluid, extract phenylethyl alcohol
The method of extracting phenylethyl alcohol from fermented supernatant fluid has two.One, extract phenylethyl alcohol with liquid-liquid extraction method, processing condition during extraction are: be organic phase (used solvent during the leaching of above-mentioned filter cake with the extraction agent, the phenylethyl alcohol that wherein contains recovery), fermented supernatant fluid is a water, organic phase/water=1 (volume ratio), 5~40 ℃ of extraction temperature, the PH=5 of water~6.5, the concentration of salting-out agent is 0.5~2% (weight percent), the one-level percentage extraction reaches about 95%, with the load organic phases (extraction liquid) of extracting and separating gained through underpressure distillation, the phenylethyl alcohol product.Wherein said that extraction agent is a normal hexane, hexanaphthene, chloroform, tetracol phenixin, benzene, toluene, dimethylbenzene, a kind of in the sherwood oil, said salting-out agent are sodium-chlor, Repone K, a kind of in the ammonium sulfate, but be good with ammonium sulfate, not only can consumption when using ammonium sulfate less, and raffinate can also be used by as fertilizer sources, and extraction liquid is in underpressure distillation acquisition phenylethyl alcohol product, recyclable solvent is for recycling.They are two years old, the spent ion exchange resin absorption method is extracted phenylethyl alcohol, its processing condition comprise, at first fermented supernatant fluid (comprising the washings of integrating with) reduction vaporization is concentrated into 1/20~1/30 (volume ratio), obtain the concentrated solution that ferments, the concentrated solution that will ferment then injects the ion exchange resin column (carrying out selective adsorption separates) of activated processing (being that ordinary method is handled so narration is omitted), use earlier the deionized water wash-out, elutriant connects nucleic acid-protein detector (280nm) and fraction collector, be eluted to till the no peak, use 70% alcoholic solution wash-out again, for extremely, the flow velocity of elutriant is 5~10ml/min until no peak.Collect elutriant gas chromatograph-mass spectrometer (the being called for short the GC-MS instrument down together) check and analysis of deionized water wash-out gained, wherein do not contain phenylethyl alcohol, and the strong peak of unique separation appears with the elutriant of 70% alcoholic solution wash-out gained, detecting its main component through the GC-MS instrument is phenylethyl alcohol.As seen spent ion exchange resin absorption and purification phenylethyl alcohol also is a good feasible method.After will concentrating with the elutriant of 70% alcoholic solution wash-out at last, obtain the phenylethyl alcohol product through underpressure distillation.It is good that said ion exchange resin is recommended with trade mark JK-108 through preferred the present invention, and said 70% alcoholic solution is meant 70% ethanol or 70% methanol solution.
Embodiment
Further illustrate content of the present invention below in conjunction with embodiment
Embodiment 1
Adopt Candida utilis to carry out the liquid aerobic fermentation of tobacco, concrete zymotechnique is as follows:
Seed culture:
Seed culture medium consist of peptone 2% (weight percent, down with), yeast extract paste 1%, glucose 2%, inorganic salt: FeSO
4, KH
2PO
4, KCl, MgSO
4, CaCl
2Be 0.1%, place 200 milliliters culturing bottle, add to 100 milliliters with distilled water, PH=6.0 with oxalic acid or oxalate regulator solution, sterilization back (being cooled to room temperature) adds the Candida utilis bacterial strain, carry out seed culture according to a conventional method, obtain liquid Candida utilis bacterial classification (being called for short the liquid yeast bacterial classification), stand-by.
The shake flask fermentation of tobacco:
The culture medium prescription of shake flask fermentation is as follows: get and be crushed to 20-40 sieve aperture tobacco (or offal, offal) 50g/L, glucose (or in other glucide any) 20g/L, inorganic salt: FeSO
40.5g/L, KH
2PO
40.125g/L, KCl0.1g/L, MgSO
40.5g/L, CaCl
20.1g/L, place the fermentation flask of 2L, add to 1L with deionized water, transfer PH=7.0 with oxalic acid or oxalate, constitute substratum slurries, then with this substratum in 15psig, 116~121 ℃ sterilization about 15 minutes down, be cooled to≤30 ℃, insert the liquid yeast bacterial classification, inoculum size is 5~10% (V/V), leavening temperature is 30 ℃, rotating speed is 220r/min, and fermentation time is 24-72 hour, after the fermentation ends, sampling uses the GC-MS instrument to measure the content of phenylethyl alcohol in the fermented liquid, and the result is as shown in table 1.Wherein said oxalate is sodium oxalate or potassium oxalate, and said leavening temperature is the optimum growth temperature of yeast for 30 ℃, is proved by practice.
By table 1 as seen, preferred fermentation condition is:
Inoculum size is good with 10% (V/V), leavening temperature is 30 ℃, fermentation time was advisable with 48 hours (increases the content that fermentation time can increase target product on a small quantity, but the mycelial self-dissolving of part yeast makes the fermented liquid quality variation, be unfavorable for later separation, weigh advantages and disadvantages or be advisable with 48 hours), the content of phenylethyl alcohol can reach 44ml/L in the fermented liquid at this moment.
Embodiment 2
Except adopting yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), outside bacterial classification.All the other processing condition are all with embodiment 1.After the fermentation ends, sampling uses the GC-MS instrument to measure the content of phenylethyl alcohol in the fermented liquid, and the result is as shown in table 2.
Same as seen by table 2: preferred fermentation condition is:
Inoculum size is good with 10% (V/V), and leavening temperature is 30 ℃, and fermentation time was advisable with 48 hours, and the content of phenylethyl alcohol can reach 50ml/L in the fermented liquid at this moment.
Embodiment 3
Except adopting Chinese kluyveromyces to do the bacterial classification, all the other processing condition are all with embodiment 1.
After the fermentation ends, sampling uses the GC-MS instrument to measure the content of phenylethyl alcohol in the fermented liquid, and the result is as shown in table 3.
By table 3 as seen, Shi Yi fermentation condition is:
10% (V/V) of inoculum size is good, and leavening temperature is 30 ℃, and fermentation time was advisable with 48 hours, and the content of phenylethyl alcohol can reach 46ml/L in the fermented liquid at this moment.
By embodiment 1~3 as seen: under same condition, when adopting yeast saccharomyces cerevisiae to make bacterial classification, can obtain better technique effect.For this reason just is that bacterial classification carries out scale-up further to illustrate content of the present invention with the yeast saccharomyces cerevisiae.
Embodiment 4
Scale-up: in the fermentor tank of a 5L, with the yeast saccharomyces cerevisiae is bacterial classification, the cultural method of its seed, the proportioning of fermentation culture, and the sterilising conditions of fermentation culture based slurries etc. is all with embodiment 1, putting tank volume is 3L, fermentation condition is: inoculation is 10% (V/V), and 30 ℃ of leavening temperatures, the flow of sterile air are 3~4.5L/min, mixing speed is 150r/min, fermentation time is 48 hours, and after the fermentation ends, sampling is 50ml/L with the content that the GC-MS instrument detects phenylethyl alcohol in the fermented liquid, regulate PH=5~6.5 of fermented liquid with oxalic acid, add flocculating aids 0.5~2.0% (weight percent) simultaneously, i.e. 15~60g, said flocculating aids is a powdered diatomite, argentine, a kind of in lime carbonate or the paper pulp, behind the stirring and evenly mixing, put a jar filtration, can get fermented supernatant fluid and filter cake two portions, wherein fermented supernatant fluid is 3L, the unit content of phenylethyl alcohol is 45ml/L, and total amount is 135ml.The content of carrying phenylethyl alcohol in the filter cake secretly is 15ml.When extracting phenylethyl alcohol with extraction process, filter cake 3L organic solvent soaking extraction reclaims the phenylethyl alcohol of wherein being carried secretly, and the rate of recovery is more than 98%, obtains 3L leach liquor a (organic solvent that contains phenylethyl alcohol) after filtering once more, and is stand-by.Said organic solvent is normal hexane or hexanaphthene.
Fermented supernatant fluid extracts phenylethyl alcohol with extraction process, and its extraction process condition is:
(be that organic solvent leach liquor recited above a) is called for short organic phase O, fermented supernatant fluid is called for short water A to extraction agent, (down together).Organic phase O/ water A=1 (volume ratio), 25~30 ℃ of extraction temperature, the PH=6.5 of aqueous phase, the concentration of salting-out agent is 2% (weight percent), said salting-out agent are sodium-chlor or Repone K, the one-level percentage extraction is 95%, the unit content of phenylethyl alcohol is 47.7ml/L in the extraction phase (being the organic phase of load), and total amount is 143.1ml, behind vacuum fractionation, can get phenylethyl alcohol product 135ml, total yield can reach 90% (so that the phenylethyl alcohol total content is benchmark in the fermented liquid).Organic phase recycles after reclaiming.
Embodiment 5
Carry out according to the 4 identical conditions of enforcement, obtain fermented supernatant fluid and filter cake.Wherein the content of phenylethyl alcohol is similarly 45ml/L in the fermentation clear liquid, and total amount is 135ml; The phenylethyl alcohol amount of carrying secretly is 16ml in the filter cake.Filter cake reclaims phenylethyl alcohol with 3L tetracol phenixin (or chloroform) leaching, and the rate of recovery is 95%, obtains 3L leach liquor b after filtration, and wherein the phenylethyl alcohol total content is 15.2ml, gained leach liquor b for below do the extraction agent use.
From the fermented supernatant fluid phenylethyl alcohol of purifying, the processing condition during its extraction are with extraction process:
Extraction agent (organic phase) is leach liquor b recited above, organic phase/water=1 (volume ratio), 40 ℃ of extraction temperature, the PH=5.0 of aqueous phase, the concentration of salting-out agent ammonium sulfate is 0.5% (weight percent), extract through one-level, percentage extraction is 94%, the unit content of phenylethyl alcohol is 47.3ml/L in the organic phase, total amount is 142ml, can get phenylethyl alcohol 132ml behind vacuum fractionation, total yield can reach 88% (is benchmark with phenylethyl alcohol total content in the fermented liquid), and the organic phase after the recovery is for recycling.
Embodiment 6
Undertaken by embodiment 4 identical conditions, obtain fermented supernatant fluid and filter cake.Wherein the fermented supernatant fluid volume is 3L, the unit content of phenylethyl alcohol is 45.2ml/L, total amount is 135.6ml, the content of phenylethyl alcohol is 13.4ml in the filter cake, filter cake 3L dimethylbenzene (or benzene, toluene) leaching recovery phenylethyl alcohol, the rate of recovery can reach 96%, after filtration 3L leach liquor c, wherein the total content of phenylethyl alcohol is 12.9ml, gained leach liquor c for below do extraction agent use.
The extraction process phenylethyl alcohol of from fermented supernatant fluid, purifying, the processing condition during its extraction are:
Extraction agent (organic phase) is leach liquor c recited above, compare: organic phase/water=1 (volume ratio), 15 ℃ of extraction temperature, the PH=6.0 of aqueous phase, the concentration of salting-out agent ammonium sulfate is 1% (weight percent), and through the one-level extraction, percentage extraction is 96%, the unit content of phenylethyl alcohol is 47.7ml/L in the organic phase, and total content is 143ml.Behind vacuum fractionation, can get phenylethyl alcohol 134ml, total yield can reach 89.3% (is benchmark with phenylethyl alcohol total content in the fermented liquid), and recovered solvent is for recycling.
Embodiment 7
Ferment by embodiment 4 identical technologies, obtain fermented supernatant fluid and filtrate, wherein the fermented supernatant fluid volume is 3L, and the unit content of phenylethyl alcohol is 44.8ml/L, and total content is 134.4ml; Phenylethyl alcohol content is 15.6ml in the filter cake.
Filter cake reclaims the phenylethyl alcohol of wherein carrying secretly with sherwood oil leaching, and the rate of recovery is 98%, after filtration 3L leach liquor d, wherein the total content of phenylethyl alcohol is 15.3ml, gained leach liquor d for below do the extraction agent use.
With the extraction process phenylethyl alcohol of purifying from fermented supernatant fluid, the processing condition during its extraction are:
Extraction agent (organic phase) is above-mentioned said leach liquor d, compare: organic phase/water=1 (volume ratio), 5 ℃ of extraction temperature, the PH=5.5 of aqueous phase, the concentration of salting-out agent ammonium sulfate is 1.5% (weight percent), through the one-level extraction, percentage extraction is 95%, and the phenylethyl alcohol unit content is 47.7ml/L in the organic phase, total content is 143ml, behind vacuum fractionation, can get phenylethyl alcohol product 135ml, total yield reaches 90% (total content with phenylethyl alcohol in the fermented liquid is a benchmark).The sherwood oil that reclaims is for recycling.
Embodiment 8
Ferment by embodiment 4 identical conditions, obtain fermented supernatant fluid and filter cake, wherein the volume of fermented supernatant fluid is 3L, and the total content of phenylethyl alcohol is 135ml, and the phenylethyl alcohol total content is 15ml in the filter cake.The phenylethyl alcohol that filter cake is wherein carried secretly with 95% industrial spirit or methyl alcohol 0.3L washing and recycling, the rate of recovery can reach more than 85%, promptly contains the above phenylethyl alcohol of 12.8ml in the washings.Gained 0.3L washings and fermented supernatant fluid are merged back reduction vaporization concentrated (stating as follows).
Adopt the ion exchange resin absorption method to handle fermented liquid supernatant liquid:
Earlier JK.108 ion exchange resin is packed in the chromatography column of a Φ 3cm * 50cm, it is stand-by that method is routinely handled the activation back; Then fermented supernatant fluid (containing the washings of integrating with) underpressure distillation is concentrated into 1/30, be about to 3.3L fermented supernatant fluid and washings and concentrate the concentrated solution 110ml that obtains fermenting, inject aforesaid chromatography column, realize adsorption separation process: use earlier the deionized water wash-out, elutriant connects nucleic acid-protein detector (280nm) and fraction collector, when being eluted to no peak, use 70% ethanol (or 70% methyl alcohol) eluant solution again, be eluted to till the no peak.With deionized water wash-out gained elutriant, detect with the GC-MS instrument, wherein do not contain phenylethyl alcohol; Elutriant with 70% ethanol (or 70% methyl alcohol) wash-out gained has a strong peak of unique separation, detect with the GC-MS instrument, its composition is a phenylethyl alcohol, with 70% ethanol (or the 70% methyl alcohol) elutriant collected after underpressure distillation, can get phenylethyl alcohol product 130ml, total recovery is 88% (is benchmark with phenylethyl alcohol total content in the fermented liquid).
In a word, by embodiment 4~8 as seen: no matter be extraction process, or the ion exchange resin absorption method, all be the good method of separation and purification phenylethyl alcohol from fermented liquid, can satisfy need of industrial production.
Fermentation result when table 1 bacterial classification is Candida utilis
Sequence number | Inoculum size (V/V) | Leavening temperature | Fermentation time | Phenylethyl alcohol content (ml/L) |
1 | 5% | 30℃ | 24 | 18 |
2 | 48 | 25 |
3 | 72 | 28 |
4 | 8% | 30℃ | 24 | 30 |
5 | 48 | 36 |
6 | 72 | 39 |
7 | 10% | 30℃ | 24 | 38 |
8 | 48 | 44 |
9 | 72 | 46
* |
*Increase fermentation time, can improve the content of target product, but the mycelial self-dissolving of a small amount of yeast takes place in lengthy fermentation inevitably under the condition of PH=7, influence the quality of fermented liquid, strengthen the difficulty of subsequent disposal, therefore behind the comprehensive evaluation ferment effect, recommend fermentation time to be advisable with 48 hours.
Fermentation result when table 2 bacterial classification is yeast saccharomyces cerevisiae
Sequence number | Inoculum size (V/V) | Leavening temperature | Fermentation time | Phenylethyl alcohol content (ml/L) |
1 | 5% | 30℃ | 24 | 20 |
2 | 48 | 30 |
3 | 72 | 33 |
4 | 8% | 30℃ | 24 | 32 |
5 | 48 | 40 |
6 | 72 | 42 |
7 | 10% | 30℃ | 24 | 40 |
8 | 48 | 50 |
9 | 72 | 51 |
Fermentation result when table 3 bacterial classification is Chinese kluyveromyces
Sequence number | Inoculum size (V/V) | Leavening temperature | Fermentation time | Phenylethyl alcohol content (ml/L) |
1 | 5% | 30℃ | 24 | 19 |
2 | 48 | 26 |
3 | 72 | 30 |
4 | 8% | 30℃ | 24 | 31 |
5 | 48 | 38 |
6 | 72 | 43 |
7 | 10% | 30℃ | 24 | 39 |
8 | 48 | 46 |
9 | 72 | 48 |