CN102807940A - Health-maintaining un-distilled liquor made of longan, phyllanthus emblica, olive, hawthorn and passion fruit and technology thereof - Google Patents
Health-maintaining un-distilled liquor made of longan, phyllanthus emblica, olive, hawthorn and passion fruit and technology thereof Download PDFInfo
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- CN102807940A CN102807940A CN2012103150514A CN201210315051A CN102807940A CN 102807940 A CN102807940 A CN 102807940A CN 2012103150514 A CN2012103150514 A CN 2012103150514A CN 201210315051 A CN201210315051 A CN 201210315051A CN 102807940 A CN102807940 A CN 102807940A
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Abstract
The invention provides a method for preparing health-maintaining liquor from longan, phyllanthus emblica, olive, hawthorn and passion fruit. The method comprises the steps of: adding a yeast with flocculation optimization characteristics and a fermentation assistant into longan juice for fermentation; and then adding phyllanthus emblica juice, olive juice and/or hawthorn juice to obtain the health-maintaining wine.
Description
Technical field
The invention belongs to fermentation technique and brewing technical field, particularly, the present invention relates to non-liquor of health and technology thereof with longan and Niuganguo, olive, hawthorn, passion fruit.
Background technology
The application is the part continuation application of one Chinese patent application 201210027412 and 201210027411.
With the fruit is the fermenting raw materials wine brewing, the fragrance that can bring strong various individuals such as fruital to have a good opinion.For example, it is the method for raw material production liquor that the inventor's Chinese patent 200910114150 discloses with longan and sugar cane juice, and this wine can have the delicate fragrance of longan and sugarcane.And for example, it is the method for raw material production cognac that the inventor's Chinese patent 200910114151 discloses with longan and sugar cane juice, and this wine also can have the delicate fragrance of longan and sugarcane.
Yet the raw material that these wine use is expensive than grain, causes cost higher.Although some yeast saccharomyces cerevisiae itself can bring some unique local flavors to wine, this type yeast often but loses some required in the wine brewing industry good operational characteristiies, like flocculence.Set out with these yeast and to carry out mutagenesis screening, although can improve operational characteristic, flavor characteristics tends in mutagenesis, lose.
For this reason, the inventor is through long-term and arduous research, developed the new technology of using grain and longan as the fermenting raw materials liquor.The peculiar flavour that the wine of this prepared not only can keep yeast saccharomyces cerevisiae to bring, thus reduce even need not the use of comparatively expensive raw material, and have good operational characteristic, and make things convenient for brewing process, further practiced thrift cost.More unexpected is; Employed zymic flocculence changes along with the variation of sugar degree in the fermentating wine in this technology, and flocculating rate is low when sugar degree is high, is of value to abundant fermentation; And flocculating rate is high when sugar degree is low, is convenient to the separation of wine liquid after the fermentation ends.
In addition, mostly prior art is to ferment as raw material after the direct making beating of various pulp or squeezing the juice; Yet the inventor further discovers, some pulp is like Niuganguo, olive, hawthorn, passion fruit pulp; Directly slurrying is as raw material, and step by fermentation is not easy its local flavor is brought in the wine that last fermentation processes; Often need a large amount of pulp to pull an oar, cost is higher.The inventor directly with grain and longan fermented system after drinking, directly adds the fruit juice of these pulp, and the wine article that are prepared into not only have longan and local flavor that yeast has, and more can keep the local flavor of above-mentioned pulp.
Summary of the invention
The technology of preparation longan that the technical problem that the present invention will solve is to provide new and the health wine of Niuganguo, olive and/or hawthorn.In addition, the present invention also provides the health wine with this explained hereafter.
Particularly, in first aspect, the invention provides the method for the health wine of fermentative prepn longan and Niuganguo, olive, hawthorn and/or passion fruit, it comprises:
(1) get rice and pulverize after, soak and boil, add αDian Fenmei then and liquefy, obtain the liquefaction material;
(2) get longan and pulverize after, add water and cellulase and under acidic conditions, handle, must longan juice;
(3) the liquefaction material of step (1) acquisition and the longan juice of step (2) acquisition are mixed, be adjusted to acidity, add saccharifying enzyme and carry out saccharification, obtain saccharification liquid;
(4) the saccharification liquid that obtains to step (2) adds yeast and fermentation assistant with flocculation optimization characteristics and ferments, and obtains fermented liquid;
(5) supernatant of the wine liquid of filtration step (4) acquisition gets longan local flavor wine; With
(6) the longan local flavor wine that obtains to step (5) adds Niuganguo fruit juice, olive fruit juice, haw juice and/or passion fruit fruit juice, obtains health wine.
In this article, the health wine of longan and Niuganguo, olive, hawthorn and/or passion fruit be raw material with longan, simultaneously one or more are raw material in Niuganguo, olive, hawthorn and the passion fruit to be selected from, the health wine for preparing.
In this article, the flocculation optimization characteristics refer to yeast in that to contain under the sugared environment flocculating rate low, and flocculating rate is high under the sugar-free environment.This specific character is especially useful for the wine brewing process.Because at the fermentation initial stage, sugar degree is high in the fermented liquid, do not have flocculation character or the low yeast of flocculating rate this moment, will make it in fermented liquid, keep suspended state as far as possible, thereby make that fermentation is more abundant, improves fermentation efficiency; When fermentation ends, sugar degree is low in the fermented liquid, and have flocculation character or the high yeast of flocculating rate this moment, will be deposited to the fermentor tank bottom easily, is convenient to fermentation termination back and collects the wine body.
Preferably in the method for first aspect present invention, the weight ratio of rice and longan is 1～10: 0.5～5, is preferably 3～8: 0.8: 1.2, and more preferably 5: 1.
In the present invention, longan can directly be squeezed the juice, but uses cellulose treatment behind the preferred thin up, and more preferably the add-on of cellulase is 1～10U/ gram longan raw material.Preferably in the method for first aspect present invention, in step (2), tart pH is 4.5～6.5, is preferably 5.0～6.0, more preferably 5.5.Tart is regulated and can be carried out with lactic acid.
In the present invention, liquefaction and saccharification can be carried out according to this area conventional grain liquefaction and saccharifying.Wherein used reagent has been very sophisticated commodity all like glycase and saccharifying enzyme.Preferably in the method for first aspect present invention, the condition of liquefaction is: the αDian Fenmei consumption is 5～15U/ gram rice raw material, in 50～60 ℃ of stirrings 15～60 minutes, is warming up to 85～97 ℃ then and continues to stir 20～60 minutes.Also preferred in the method for first aspect present invention, the condition of saccharification is: add saccharifying enzyme 100～300U/ in 50～65 ℃ and restrain rice raw material, be incubated 45～120 minutes.
Preferably in the method for first aspect present invention, in step (3), being adjusted to acidity is that lactic acid is regulated pH to 4.0～4.8, preferably regulates pH to 4.1～4.5, more preferably regulates pH to 4.2.
Preferably in the method for first aspect present invention; Yeast with flocculation optimization characteristics is the yeast that comprises flocculation protein variant gene; The nucleotide sequence of said flocculation protein variant gene is shown in SEQ ID No:1, and the aminoacid sequence of its encoded protein variant is shown in SEQ ID No:2.Wild-type is flocculated proteic aminoacid sequence shown in GenBank accession number CAA55024.1, its can give do not have the flocculation character yeast with the flocculation character.Although the flocculation character that wild-type flocculation albumen is given to yeast is stronger; But this character contain under the sugared environment also very strong; Caused in the fermenting process yeast also to be deposited in the fermentor tank bottom easily like this, caused fermentation efficiency low, therefore do not obtained in the practice promoting.The inventor in our fermentation has found accidentally that through studying for a long period of time a strain has the yeast of flocculation optimization characteristics, through commissioned research, finds that it is the sudden change that at least three place's amino acid sites have taken place with respect to wild-type flocculation albumen flocculation albumen.The protein variant gene that should flocculate imports other existing yeast, as not having the yeast of flocculation character, can give this yeast flocculation optimization characteristics, and not change the original fermented flavour of this yeast.Therefore, make the existing yeast have fermented flavour but not have flocculation optimization characteristics can be able to utilize again.
Preferably in the method for first aspect present invention, fermentation assistant is Fermaid K.This is the fermentation assistant very useful to yeast fermentation, commercialization at present.
In the method for first aspect present invention, fermenting process can carry out according to the fermentation process of routine.Usually the judge index of fermentation ends is to ferment to residual sugar content＜4 gram/L.Preferably in the method for first aspect present invention, fermentation is carried out in two steps, and wherein the leavening temperature of the first step is higher than the leavening temperature in second step.More preferably wherein the leavening temperature of the first step is that 23～35 ℃ and second leavening temperatures that go on foot are 18～22 ℃.In addition more preferably wherein the fermentation time of the first step be 6～48 hours and second the step fermentation time be 9～12 days.
In the method for first aspect present invention, the supernatant of the wine liquid that filtration step (4) obtains can be directly as the wine article, and it has the longan local flavor; Also can after the supernatant of the wine liquid that filtration step (4) obtains, distill.Except blending with Niuganguo fruit juice, olive fruit juice, haw juice and/or passion fruit fruit juice, the method for first aspect present invention can also comprise the step of filtration and/or ageing.
In the method for first aspect present invention; Niuganguo fruit juice, olive fruit juice, haw juice and/or passion fruit fruit juice can use conventional juice extracting technology preparation; Therefore but, preferably will pulverize pulp and add the water mixing and produce fruit juice because water cut is lower in these pulp.
In second aspect, the invention provides the health wine of the method preparation of first aspect present invention.This wine can keep original zymic fermented flavour; And have Niuganguo, olive, hawthorn and/or passion fruit etc. and be not easy to ferment and introduce the local flavor in the wine; And be convenient to the later stage purifying of wine liquid; Because the yeast major sedimentary in the fermentor tank bottom, only need be got the supernatant of wine liquid, can obtain clarifying wine body through simple filtering.
The present invention has advantage and effect: when sugar degree is high in fermentation primary fermentation liquid, yeast will make it in fermented liquid, keep suspended state as far as possible, thereby make that fermentation is more abundant, improve fermentation efficiency; When sugar degree is low in the fermentation ends fermented liquid, yeast will be deposited to the fermentor tank bottom easily, be convenient to collect the wine body after fermentation stops; Can give yeast with the flocculation optimization characteristics, thereby can utilize the good and flocculation character not good yeast of original fermentation character, make full use of the yeast resource of present fermentation plant and brewery; Wine of the present invention has sugarcane delicate fragrance and can inherit the good zymic characteristic of original fermentation character, and fragrance is attractive, can reach the raw material similar effect expensive with adding cost; Wine of the present invention has used grain as fermentation raw material, has reduced production cost; The wine article further have Niuganguo, olive, hawthorn and/or passion fruit etc. and are not easy to ferment and introduce the local flavor in the wine.
For the ease of understanding, below will the present invention be described in detail through concrete embodiment.What need particularly point out is that specific examples only is in order to explain, not constitute limitation of the scope of the invention.Obviously those of ordinary skill in the art can explain according to this paper, within the scope of the invention the present invention is made various corrections and change, and these corrections and change are also included in the scope of the present invention.In addition, the present invention has quoted open source literature, and these documents also are in order more clearly to describe the present invention, and their full text content is all included the present invention in and carried out reference, just look like they full text in specification sheets of the present invention repeated description the same excessively.
Embodiment
Following this paper will describe invention through concrete embodiment.As not specializing part, listed method is implemented in the reference that can be quoted according to brewing technology that those skilled in the art were familiar with and recombinant gene handbook and this paper.In addition, employed material and facility all can be bought from market among the embodiment.
The zymic of embodiment 1 of fine qualityization of flocculence obtains
Gene order (SEQ ID No:1 according to our design; The aminoacid sequence of its encoded protein variant is SEQ ID No:2), entrust the synthetic flocculation protein variant gene that we find of Guangdong Jida Genetic Pharmaceutical Engineering Research Center Co., Ltd. and be built in the yeast saccharomyces cerevisiae through commercial sources.Clone's process is carried out with reference to the operational guidance of " molecular cloning experiment guide " and used commercialization reagent, and concise and to the point process is following:
Pass through automatic dna synthesizer; The nucleic acid fragment of synthetic flocculation protein variant gene; With T4 polynucleotide kinase (available from TaKaRa company) 5 ' end of these nucleic acid fragments is carried out phosphorylation; Wait then behind these nucleic acid fragments of mixed in molar ratio in 75 ℃ of sex change 5 minutes, annealing is cooled to 12 ℃ then, adds T4 dna ligase (available from TaKaRa company) and connects 12 hours.Then; Get the above-mentioned connection product of 1 μ L and in 50 μ L reaction volumes, carry out pcr amplification; Wherein forward primer (has been introduced EcoR I restriction enzyme site) shown in the SEQ IDNo:3 of sequence table, reverse primer (introduced Xba I restriction enzyme site) shown in the SEQ ID No:4 of sequence table; Reaction conditions is: with 94 ℃ of sex change 2 minutes, extend with 94 ℃ of sex change 30 seconds, 63 ℃ of annealing 60 seconds and 72 ℃ then and carried out 30 circulations in 90 seconds, extended 12 minutes with 72 ℃ at last and be cooled to 4 ℃.
The above-mentioned PCR product of agarose gel electrophoresis; Reclaim the fragment of about 4.5kb size, with EcoR I and this fragment of Xba I double digestion, and be connected with the T4 dna ligase through the pYESS2/CT of these two endonuclease digestions plasmid (can available from Invitrogen company); Electrotransformation changes the pYESS-fln plasmid over to yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) QN732 strain and (is prone to overcharge bio tech ltd available from Guangxi; It is the non-yeast of wadding a quilt with cotton property with fixed attention, but in the wine that fermentation produces light chocolate flavor is arranged) in, be coated on the selective medium and cultivate; After getting the yeast that grows and extracting total DNA; With above-mentioned forward primer and reverse primer pcr amplification, agarose gel electrophoresis find the to have an appointment fragment of 4.5kb size shows that will the gene shown in the SEQ ID No:1 of sequence table having imported yeast saccharomyces cerevisiae has suffered.Simultaneously, there is the pYESS2/CT plasmid electricity of wild-type flocculation protein gene to be transformed into this non-yeast of wadding a quilt with cotton property with fixed attention with the clone pYESS2/CT empty plasmid, respectively as negative control and positive control.
We detect liquid (50mmol/L Trisodium Citrate, 5mmol/L EDTA, 20mmol/L CaCl with the 20mL flocculation
2, pH 4.5; And add the material of following final concentration respectively: do not add (sugar-free), 1M glucose) place 50mL to shake bottle, add above-mentioned each yeast cell 1 respectively and restrain (weight in wet base), with 30 ℃, 100r/min shaking culture 2 hours.Then; Get the 5mL cell suspending liquid to the 10mL test tube; At the liquid level 200 μ L that take a sample, vertically left standstill 5 minutes at once, then at the liquid level 200 μ L that take a sample; Measure the OD600 value before and after vertically leaving standstill respectively, calculate flocculating rate (flocculating rate=(the OD600 value after 1-vertically leaves standstill/vertically leave standstill preceding OD600 value) * 100%).The result is as shown in table 1, and yeast saccharomyces cerevisiae QN732 does not have flocculation character basically; After having cloned wild-type flocculation protein gene, no matter still containing under the sugared environment at sugar-free, yeast all will have flocculation character; The yeast that flocculation protein variant gene of the present invention changes over to does not have flocculation character containing under the sugared environment, will keep the suspended state fermentation, and under the sugar-free environment, will have flocculation character, is deposited to the fermentor tank bottom easily, is convenient to fermentation and stops the back collection.
Each yeast of table 1 is containing sugar or is not containing the flocculating rate under the sugared condition
| Flocculation detect liquid the zymic gene | Empty carrier | Flocculation protein variant gene | Wild-type flocculation protein gene |
| Sugar-free | 7％ | 66％ | 73％ |
[0037]
| Contain glucose | 6% | 12% | 58% |
The preparation of embodiment 2 longan wines
500 gram rice are ground; Add 700 gram water logging bubbles 1 hour, boil then, add 2 kg of water and αDian Fenmei (can match grey hair ferment Products Co., Ltd) 10U/ and restrain rice raw material available from Nanning; Stirred 30 minutes in 55 ℃; Be warming up to 95 ℃ then and continue to stir 30 minutes, make rice liquefaction, get the rice liquefier.Getting fresh longan pulp 100 grams pulverizes; Liquid that after pulverizing, obtains and solid residue add 400 gram water and cellulase (can match grey hair ferment Products Co., Ltd available from Nanning) 2U/ gram longan pulp raw material; Add lactic acid and regulate pH to 5.0,, get longan juice in 50 ℃ of stirrings 60 minutes.After rice liquefier and longan juice mixed, add lactic acid and regulate pH to 4.2, add saccharifying enzyme (can match grey hair ferment Products Co., Ltd available from Nanning) 150U/ in 58 ℃ and restrain rice raw material, be incubated 90 minutes, removal is pickled with grains or in wine and obtains saccharification liquid behind the slag.
With above-mentioned saccharification liquid pack into the sterilization fermentor tank; The clone who adds embodiment 1 has yeast 2 gram (dry weight) and the fermentation assistant Fermaid K of flocculation protein variant gene (can available from the outstanding rabbit industry and trade in Shanghai ltd) 5 to restrain; In 28 ℃ of fermentations 24 hours; Temperature is controlled at 18～22 ℃ then, sealing and fermenting 9～12 days.When residual sugar＜4 gram/L, fermentation ends owing at the bottom of this moment, yeast was deposited on jar basically, therefore supernatant part simple filtration can be obtained clarifying wine body, obtains longan wine.
Blending of embodiment 3 Niuganguo fruit juice
Get fresh Niuganguo pulp 100 grams and pulverize, liquid that after pulverizing, obtains and solid residue add 100 gram water, the after-filtration that stirs, and filtrating is Niuganguo fruit juice.The longan wine 1L that gets embodiment 2 preparation mixes with 50mL Niuganguo fruit juice, ageing 3 months or more than.Have longan delicate fragrance and Niuganguo fragrance in longan that obtains and the Niuganguo health wine.
Blending of embodiment 4 olive fruit juice
Get new fresh olive pulp 100 grams and pulverize, liquid that after pulverizing, obtains and solid residue add 100 gram water, the after-filtration that stirs, and filtrating is olive fruit juice.The longan wine 1L that gets embodiment 2 preparation mixes with 30mL olive fruit juice, ageing 3 months or more than.Have strong longan delicate fragrance in longan that obtains and the olive health wine, and light olive fragrance and chocolate flavor are arranged.Blending of embodiment 5 haw juices
Get fresh haw pulp 100 grams and pulverize, liquid that after pulverizing, obtains and solid residue add 100 gram water, the after-filtration that stirs, and filtrating is haw juice.The longan wine 1L that gets embodiment 2 preparation mixes with the 25mL haw juice, ageing 3 months or more than.Have strong longan delicate fragrance in longan that obtains and the hawthorn health wine, and light hawthorn fragrance and chocolate flavor are arranged.
Blending of embodiment 6 passion fruit fruit juice
Get fresh passion fruit pulp 100 grams and pulverize, liquid that after pulverizing, obtains and solid residue add 50 gram water, the after-filtration that stirs, and filtrating is passion fruit fruit juice.The longan wine 1L that gets embodiment 2 preparation mixes with 10mL passion fruit fruit juice, ageing 3 months or more than.Have longan delicate fragrance and passion fruit fragrance in longan that obtains and the passion fruit health wine, the local flavor that yeast brings has necessarily to be covered.
Claims (10)
1. the method for the health wine of fermentative prepn longan and Niuganguo, olive, hawthorn and/or passion fruit, it comprises:
(1) get rice and pulverize after, soak and boil, add αDian Fenmei then and liquefy, obtain the liquefaction material;
(2) get longan and pulverize after, add water and cellulase and under acidic conditions, handle, must longan juice;
(3) the liquefaction material of step (1) acquisition and the longan juice of step (2) acquisition are mixed, be adjusted to acidity, add saccharifying enzyme and carry out saccharification, obtain saccharification liquid;
(4) the saccharification liquid that obtains to step (2) adds yeast and fermentation assistant with flocculation optimization characteristics and ferments, and obtains fermented liquid;
(5) supernatant of the wine liquid of filtration step (4) acquisition gets longan local flavor wine; With
(6) the longan local flavor wine that obtains to step (5) adds Niuganguo fruit juice, olive fruit juice, haw juice and/or passion fruit fruit juice, obtains health wine.
2. the described method of claim 1, wherein the weight ratio of rice and longan is 1～10: 0.5～5, is preferably 3～8: 0.8: 1.2, more preferably 5: 1.
3. claim 1 or 2 described methods, wherein tart pH is 4.5～6.5 in the step (2), is preferably 5.0～6.0, more preferably 5.5.
4. the described method of claim 1, wherein tart pH is 4.0～4.8 in the step (3), is preferably 4.1～4.5, more preferably 4.2.
5. the described method of claim 1, the yeast that wherein has the flocculation optimization characteristics is the yeast that comprises flocculation protein variant gene, the nucleotide sequence of said flocculation protein variant gene is shown in SEQ ID No:1.
6. the described method of claim 1, wherein fermentation assistant is Fermaid K.
7. the described method of claim 1 is wherein fermented and is carried out in two steps, and wherein the leavening temperature of the first step is higher than the leavening temperature in second step.
8. the described method of claim 7, wherein the leavening temperature of the first step is that the leavening temperatures in 23～35 ℃ and second step are 18～22 ℃.
9. the described method of claim 7, wherein the fermentation time of the first step is that the fermentation time in 6～48 hours and second step is 9～12 days.
10. the health wine of the arbitrary described method of claim 1～9 preparation.
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Cited By (6)
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| CN102978063A (en) * | 2012-11-29 | 2013-03-20 | 李恩东 | Healthcare wine prepared from longans and passion fruits or dragon fruits and process thereof |
| CN103013754A (en) * | 2012-12-17 | 2013-04-03 | 青岛森淼实业有限公司 | Method for brewing health-care wine |
| CN104531463A (en) * | 2014-11-30 | 2015-04-22 | 柳州联海科技有限公司 | Technology for biologically brewing passion fruit and mango wine |
| CN104531462A (en) * | 2014-11-30 | 2015-04-22 | 柳州联海科技有限公司 | Technology for biologically brewing magno, dragon fruit and passion fruit mixed wine |
| CN105039103A (en) * | 2015-08-07 | 2015-11-11 | 融安县飞煌原生草药材专业合作社 | Passion fruit juice wine |
| CN107384680A (en) * | 2017-07-21 | 2017-11-24 | 广西壮族自治区农业科学院农产品加工研究所 | A kind of passion fruit longan witches' broom and its brewing method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102978063A (en) * | 2012-11-29 | 2013-03-20 | 李恩东 | Healthcare wine prepared from longans and passion fruits or dragon fruits and process thereof |
| CN103013754A (en) * | 2012-12-17 | 2013-04-03 | 青岛森淼实业有限公司 | Method for brewing health-care wine |
| CN104531463A (en) * | 2014-11-30 | 2015-04-22 | 柳州联海科技有限公司 | Technology for biologically brewing passion fruit and mango wine |
| CN104531462A (en) * | 2014-11-30 | 2015-04-22 | 柳州联海科技有限公司 | Technology for biologically brewing magno, dragon fruit and passion fruit mixed wine |
| CN105039103A (en) * | 2015-08-07 | 2015-11-11 | 融安县飞煌原生草药材专业合作社 | Passion fruit juice wine |
| CN107384680A (en) * | 2017-07-21 | 2017-11-24 | 广西壮族自治区农业科学院农产品加工研究所 | A kind of passion fruit longan witches' broom and its brewing method |
| CN107384680B (en) * | 2017-07-21 | 2020-10-09 | 广西壮族自治区农业科学院农产品加工研究所 | Passion fruit and longan fruit wine and brewing method thereof |
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Application publication date: 20121205 |