CN105285096B - A kind of leben and preparation method thereof rich in levulan levan - Google Patents
A kind of leben and preparation method thereof rich in levulan levan Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of leben rich in levulan levan, it includes the following steps: that (1) by lemon leukonid (Leuconostoc citreum) fermenting microbe fermented and cultured in skimmed milk sucrose culture medium, obtains acidified milk；(2) gained acidified milk is mixed with the blending liquid containing sweetening material and acid, homogeneous sterilizes to obtain the final product.The lemon leukonid that the present invention uses is one kind of lactic acid bacteria, and the bacterial strain of levan is produced with other, such as bacillus is compared, and metabolite has higher foodsafety.Preparation method of the invention greatlies simplify production stage, saves production cost, while reducing bring pollution risk in noncontinuity operation.The levan content that the preparation method prepares resulting leben is higher, and quality is stablized, and has broken the rare status using levulan levan as functional components beverage on existing market.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of leben and its system rich in levulan levan
Preparation Method.
Background technique
Polysaccharide (polysaccharide) is that a kind of be bonded by monosaccharide with glycosidic bond forms macromolecule polyalcohol, studies table
Bright, polysaccharide has the prebiotic function such as immunological regulation, antiviral, anticancer, hypoglycemic, blood pressure lowering.Polysaccharide in nature mainly divides
It is distributed in microorganism, higher plant, animal, lichens and algae, wherein microbe-derived polysaccharide espespecially its exocellular polysaccharide,
It is secreted into outside cell wall during growth metabolism for certain specified microorganisms (such as lactic acid bacteria, agrobacterium, rhizobium)
A kind of saccharide compound, wherein the polysaccharide depended on outside microorganism wall is referred to as capsular polysaccharide, and glue polysaccharide is then to seep
The form in growing environment exists thoroughly.
Levulan Levan is a kind of natural fructose polymer, is present in various plants and microbial fermentation product, though
It is so unwelcome microbial metabolic products in sugar-cane juice processing industry, but in anticancer, antitumor, antibacterial, antiviral, drop blood
The medical clinical fields such as sugar, reducing blood lipid, norcholesterol, the immune, weight-reducing of enhancing have high application value, are ground in recent years
The concern for the person of studying carefully.Report in relation to levan can be traced earliest in the 1930s, correlative study so far is excellent throughout yield
The multiple fields such as change, structure elucidation, the mechanism of action, functional characteristic, however, domestic beverage market is made a general survey of, although types of functionality
Beverage is grown thickly, such as red ox (supplement energy), pulsation (replenishing vitamins), but several as the beverage of main functional component using levan
Do not have.Moreover, the existing correlative study report containing polysaccharide drink, the polysaccharide origin being related to are mostly bacillus, produce gas
The metabolism of the harmful bacterias such as bacillus obtains, and application mode is also all artificially to make an addition in beverage, and the polysaccharide with certain application risk comes
Source and the application mode artificially added make this kind of product lack innovative and safety gradually.
Therefore, it is with levulan levan to produce using the higher microbial metabolism skimmed milk sucrose culture medium of safety
The leben of functional components, will be one of the trend of the following new type functional beverage development, which solves.
Summary of the invention
Therefore, the present invention lacks to solve existing market using levan as the leben of main functional component, and
Existing levan producing strains are mostly the problem of polysaccharide is mostly artificial addition in pathogenic bacteria and similar beverages, provide one kind and are rich in
The leben and preparation method thereof of levulan levan.
The inventors discovered that being CGMCC No.6431 lemon leukonid (Leuconostoc using deposit number
Citreum after) being metabolized skimmed milk sucrose culture medium, a large amount of levulan levan, acidified milk warp are contained in acidified milk obtained
Flavor after blending and tasting is rather good, can be received by most of consumer, so as to complete the present invention.
In order to solve the above technical problems, the technical solution that the present invention takes first is that: a kind of hair rich in levulan levan
The preparation method of kefir milk beverage, the preparation method the following steps are included:
(1) lemon leukonid (Leuconostoc citreum) fermenting microbe is sent out in skimmed milk sucrose culture medium
Ferment culture, obtains acidified milk；
(2) acidified milk obtained by step (1) is mixed with the blending liquid containing sweetening material and acid, homogeneous sterilizes to obtain the final product.
Wherein step (1) is by lemon leukonid (Leuconostoc citreum) fermenting microbe in degreasing lactosucrose
Fermented and cultured in culture medium, obtains acidified milk.
Wherein the lemon leukonid is the prior art, and preparation method is this field customary preparation methods, or
It is obtained by purchase.Lemon leukonid (Leuconostoc citreum) fermenting microbe is preferably deposit number and is
The lemon leukonid strain of CGMCC NO.6431, the bacterial strain are named as BD01707.The bacterial strain is on 08 13rd, 2012
It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation address: Chaoyang District, Beijing City
The institute 3 of North Star West Road 1, postcode: 100101.The preparation method of the skimmed milk sucrose culture medium is preferably comprised following step
It is rapid: 6~12% skimmed milk powder and 5~20% sucrose to be added in distilled water, the percentage is mass percent, is mixed
After completely dissolution, it sterilizes 5~20 minutes for 95~125 DEG C, cooling to obtain the final product.The temperature of the culture is preferably 25 DEG C~35
DEG C, it is more preferably 30 DEG C, the fermented and cultured is more preferably concussion fermented and cultured, the speed of oscillation is preferably 150~
250rpm is more preferably 200rpm, and incubation time is preferably 72h~110 hour, is more preferably 96 hours, inoculum concentration is preferable
Ground is 1.0%~3.0%, is more preferably 2.0%, and the percentage is percent by volume.
Wherein step (2) is to mix acidified milk obtained by step (1) with the blending liquid containing sweetening material and acid,
Matter sterilizes to obtain the final product.
Wherein the sweetening material is the sweetening material of this field routine, is more preferably white granulated sugar, in syrup and sweetener
It is one or more；Wherein, the sweetener is one of Aspartame, honey element and acesulfame potassium or a variety of；The sweet tea
The content of taste substance is that the sugariness of the leben is made to be converted into sweetness of cane sugar 5%~10%；The acid is
The acid of this field routine, preferably one of citric acid, malic acid, tartaric acid, lactic acid and acetic acid or a variety of；It is described
The content of acid be the 70 ° of T~80 ° T of titratable acidity for making the leben.Sweetening material and tart flavour of the present invention
The type and usage amount of agent all meet the regulation of Food Additives Used in China relevant laws and regulations.Wherein the acidified milk with blend
The volume ratio of liquid is preferably 1:0.69~1:2.02, is more preferably 1:1.33~1:2.02, it is therefore preferable to 1:2.02.Wherein
The pressure of the homogeneous is preferably 15~30MP, is more preferably 20~28MP, it is therefore preferable to 25MP；The temperature of the sterilizing compared with
It is goodly 95 DEG C~125 DEG C, is more preferably 98~110 DEG C, it is therefore preferable to 100 DEG C；The time of sterilizing is 5 minutes~20 minutes,
It is more preferably 10~18 minutes, it is therefore preferable to 15 minutes.
In order to solve the above technical problems, the technical solution that the present invention takes second is that: a kind of preparation as described in the present invention
The method leben obtained for being rich in levulan levan.
Leben as described above, wherein the content of levulan levan is preferably >=0.5%, the percentage
For mass percent.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that:
1, the bacterial strain that the present invention uses produces the lemon leukonid (Leuconostoc of levulan levan for one plant
Citreum), which is to be found to have the lemon leukonid for producing levan ability for the first time.
2, the lemon leukonid that the present invention uses is one kind of lactic acid bacteria, and bacterial strain (such as gemma bar of levan is produced with other
Bacterium etc.) it compares, metabolite has higher foodsafety.
3, levan contained in leben provided by the invention is that lemon leukonid metabolism sucrose generates,
Compared with the polysaccharide drink produced on the market with artificial rear addition manner, preparation process of the invention greatlies simplify production step
Suddenly, production cost is saved, while reducing bring pollution risk in noncontinuity operation.
4, leben provided by the invention, the fermentation medium that preparation institute's process uses are from a wealth of sources, at low cost
Honest and clean, natural safety improves the safety of food while reducing Material Cost.
5, the present invention is fermented natural fermented culture medium using the lemon leukonid for producing levulan levan, in conjunction with blending, adjust
The method of taste produces direct-edible functional beverage, breaks rare using levulan levan as functionality on existing market
The status of component beverage.
Biomaterial preservation information
The bacterial strain of lemon leukonid of the present invention was deposited in Chinese microorganism strain on 08 13rd, 2012
Preservation administration committee common micro-organisms center (CGMCC), preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postal
It compiles: 100101.The classification naming of the suggestion of the bacterial strain are as follows: lemon leukonid Leuconostoc citreum, the guarantor of the bacterial strain
Hiding number are as follows: CGMCC No.6431, the culture title of institute's preservation are as follows: BD01707.
Detailed description of the invention
Fig. 1 shows that the monosaccharide of lemon leukonid BD01707 polysaccharide forms chromatography measurement result.
Fig. 2 shows the infrared spectrum measurement result of lemon leukonid BD01707 polysaccharide.
Fig. 3 shows the nuclear magnetic resonance measuring result of lemon leukonid BD01707 polysaccharide.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
The selection of product specification.
The preparation of leben of the embodiment 1 rich in levulan levan
1, materials and methods
The preparation of skimmed milk sucrose culture medium: being 10% skimmed milk powder and 15% sucrose in water ratio and steaming by mass percent
Distilled water mixes, and after completely dissolution, sterilize 5min at 125 DEG C, is cooled to room temperature the sterile absorbent lactosucrose to get required concentration
Culture medium.
The preparation of seed (fermenting microbe): the lemon leukonid for being GCMCC No.6431 by deposit number
The freeze-dried powder of (Leuconostoc citreum) BD01707 is dissolved with a small amount of sterile distilled water, takes a ring to line with oese
On M17 agar medium (Merck Co. Germany) containing 2% (w/v) sucrose, 28 DEG C of aerobic cultures are taken out for 24 hours, are chosen with oese
Single colonie is taken to be put into M17 fluid nutrient medium (purchase from Merck Co. German) of the 20mL containing 2% (w/v) sucrose, 28 DEG C of 180rpm
Shaking table culture is taken out for 24 hours, culture 9, and 000rpm is centrifuged 10 minutes, discards supernatant, after thallus washs 2 times with sterile distilled water,
It is suspended with the sterile distilled water of former volume of culture, obtains the seed of fermentation.
The preparation of polysaccharide (levulan levan) in acidified milk: taking acidified milk to be centrifuged 10min with 15,000g, take supernatant,
3 times of volumes are added in the dehydrated alcohol of supernatant volume, stand overnight, 15,000g centrifugation 10min collect sediment and are dissolved in
After water, vacuum freeze drying to obtain the final product.
The preparation of polysaccharide (levulan levan) in leben: taking leben 15, and 000g is centrifuged 10min, takes
Clear liquid (the dead thallus of removal inactivation), is added the dehydrated alcohol of 3 times of volumes, stands overnight, and 15,000g centrifugation 10min, it is heavy to collect
Starch and after being dissolved in water, vacuum freeze drying to obtain the final product.
2, the preparation of the leben rich in levulan levan
(1) by resulting lemon leukonid BD01707 seed with (v/v) aseptic inoculation of inoculum concentration 2% in contain sucrose
The culture medium of 15% (v/v), skimmed milk 10% (w/w), 30 DEG C, 200rpm culture 96 hours, take acidified milk after fermentation, pass through
It measures, levan content is 1.6% (w/v) in the acidified milk.
(2) white granulated sugar, citric acid are dissolved completely in 50 DEG C of water to get blending liquid.By above-mentioned acidified milk with blend
Liquid carries out homogeneous after being sufficiently mixed uniformly with 1:2.02 (v/v) under 15MP pressure, and in 95 DEG C of sterilizing 20min, filling, obtains
The leben product A of embodiment.
Through detecting, 0.53% (w/ of levan content in the leben rich in levulan levan of the present embodiment production
V), sugariness is the sweetness of cane sugar containing 10%, 80 ° of T of acidity.
3, composition detection
(1) in lemon leukonid BD01707 acidified milk polysaccharide monosaccharide composition measuring
(a) standard sample configures:
Polysaccharide hydrolysis: it draws the polysaccharide sample solution that 100 μ L concentration are 4mg/mL~5mg/mL and is tried in the tool plug scale of 5mL
Guan Zhong is added the 4moL/LTFA of 100 μ L, fills N2Tube sealing hydrolyzes 2h in 110 DEG C of baking ovens；Lid is opened after cooling, adds 200 μ L methanol
After use N2Drying so repeats to add methanol and uses N2It blows 3 times, removes TFA, the dissolution of its residue water is settled to 5mL, with 0.45 μ
For sample introduction is analyzed after m micro-pore-film filtration.
(b) chromatographic condition
Chromatographic column: CarboPacPA203mmi.d. × 150mm；
Mobile phase: A, H2O；B,250mmol/L NaOH；C,1mol/L NaAc；
Ternary gradient elution: flow velocity: 0.5mL/min；Integrated pulsed amperometric detection device；
Au working electrode: Ag/AgCl reference electrode；
Sampling volume: 20 μ L；Column temperature: 30 DEG C.
(c) by the polysaccharide sample in (1) acidified milk the step of preceding method preparation, and with high-efficiency anion chromatography determination
The monosaccharide of polysaccharide forms.
Data result: the chromatography the result is shown in Figure 1 of lemon leukonid BD01707 polysaccharide sample glycosyl composition, this is more
Sugar has single absorption peak in 14.50min, and the retention time of the absorption peak is consistent with fructose mark product retention time.
Conclusion: lemon leukonid BD01707 polysaccharide is made of the single glycosyl of fructose, is a kind of levulan.
(2) in lemon leukonid BD01707 acidified milk polysaccharide structural determination
(a) FTIR spectrum (FTIR) measures
After polysaccharide sample and potassium bromide powder in the step of prepared by preceding method (1) acidified milk are sufficiently mixed, compacting
It is red to carry out Fourier using Fourier infrared spectrograph (Nicolet 6700, Thermo Fisher company, the U.S.) for slabbing
External spectrum (FTIR) measurement.
As a result as shown in Fig. 2, 3306cm-1The strong broad peak at place is intermolecular and intramolecular hydroxyl absorption peak, 2930cm-1With
2884cm-1For the absorption peak of c h bond stretching vibration in residue of fructose, 1634cm-1The absorption peak at place be combine water caused by,
1412-1217cm-1For the absorption peak of C-H angle vibration, 1120-1008cm-1The absorption peak at place is C-O-C and C-O-
The result of the stretching vibration of H.923cm-1And 810cm-1The absorption peak at place and the stretching vibration of furan nucleus are related.
Conclusion: the saccharide ring configuration of BD01707 polysaccharide is based on furan nucleus.
(b) nuclear magnetic resonance (NMR) measures
Polysaccharide sample in the step of prepared by preceding method (1) acidified milk is dissolved completely in weight by the concentration of 10mg/mL
In water, and application nuclear magnetic resonance chemical analyser (400 MHz of Avance III, Bruker company, Germany) carries out nuclear magnetic resonance
(NMR) it measures.As a result as shown in figure 3, in Fig. 3, A NMR-1H spectrum, shows that the chemical shift of seven protons of sample concentrates on
The region 3.4-4.2ppm, and in Fig. 3, B NMR-13C spectrum shows sample in 104.537 (C-2), 80.622 (C-5), 76.628
(C-3), there is resonance at 75.532 (C-4), 63.716 (C-6) and 60.234 (C-1) ppm.
Conclusion: FTIR with1H、13The data binding analysis of C spectrum shows that BD01707 polysaccharide is levulan levan.
The preparation of leben of the embodiment 2 rich in levulan levan
1, materials and methods
The preparation of skimmed milk sucrose culture medium: by mass percent be 12% skimmed milk powder and mass percent be 20%
Sucrose and distilled water mix, after completely dissolution, sterilize 20min at 95 DEG C, is cooled to room temperature to get the sterile of required concentration
Skimmed milk sucrose culture medium.
The preparation of polysaccharide (levulan levan) in the preparation of seed (fermenting microbe), acidified milk, polysaccharide in leben
The preparation of (levulan levan): with embodiment 1.
2, the preparation of the leben rich in levulan levan
(1) by resulting lemon leukonid BD01707 seed with the percent by volume of inoculum concentration be 0.5% aseptic inoculation
In the culture medium containing sucrose 20%, skimmed milk 12%, 35 DEG C, 100rpm culture 120 hours take acidified milk after fermentation, pass through
Measuring levan content in the acidified milk is 1.44%, and the percentage is mass percent.
(2) syrup, Aspartame, malic acid, tartaric acid, lactic acid are dissolved completely in 50 DEG C of water to get blending liquid.
Homogeneous is carried out under 30MP pressure after above-mentioned acidified milk and blending liquid are sufficiently mixed uniformly with 1:1.77 (v/v), and in 125 DEG C
Sterilize 5min, filling, obtains the leben product B of the present embodiment.
Through detecting, levan content 0.52% in the leben rich in levulan levan of the present embodiment production, sweet tea
Degree is the sweetness of cane sugar containing 5%, and 70 ° of T of acidity, the percentage is mass percent.
The preparation of leben of the embodiment 3 rich in levulan levan
1, materials and methods
The preparation of skimmed milk sucrose culture medium: by mass percent be 6% skimmed milk powder and mass percent be 5%
Sucrose and distilled water mix, and after completely dissolution, sterilize 15min at 100 DEG C, is cooled to room temperature to get the sterile of required concentration
Skimmed milk sucrose culture medium.
The preparation of polysaccharide (levulan levan) in the preparation of seed (fermenting microbe), acidified milk, polysaccharide in leben
The preparation of (levulan levan): with embodiment 1.
2, the preparation of the leben rich in levulan levan
(1) by resulting lemon leukonid BD01707 seed with (v/v) aseptic inoculation of inoculum concentration 4.0% in sucrose matter
Measure degree be 5%, the culture medium that skimmed milk mass percentage content is 6%, 20 DEG C, 300rpm cultivate 48 hours, hair
Acidified milk is taken after ferment, after measured, the mass percentage content of levan is 0.86% in the acidified milk.
(2) white granulated sugar, honey element, An Saimi, citric acid, acetic acid are dissolved completely in 50 DEG C of water to get blending liquid.
Homogeneous is carried out under 25MP pressure after above-mentioned acidified milk and blending liquid are sufficiently mixed uniformly with 1:0.69 (v/v), and in 120 DEG C
Sterilize 10min, filling, obtains the leben products C of the present embodiment.
Through detecting, the mass percent of levan contains in the leben rich in levulan levan of the present embodiment production
Amount is 0.51%, and sugariness is the sweetness of cane sugar containing 6.8%, 73 ° of T of acidity.
The preparation of leben of the embodiment 4 rich in levulan levan
1, materials and methods
The preparation of skimmed milk sucrose culture medium: by mass percent be 8% skimmed milk powder and mass percent be 10%
Sucrose according to a certain ratio with distilled water mix, after completely dissolution, sterilize 10min at 120 DEG C, is cooled to room temperature to get institute
Need the sterile absorbent lactosucrose culture medium of concentration.
The preparation of polysaccharide (levulan levan) in the preparation of seed (fermenting microbe), acidified milk, polysaccharide in leben
The preparation of (levulan levan): with embodiment 1.
2, the preparation of the leben rich in levulan levan
(1) by resulting lemon leukonid BD01707 seed with (v/v) aseptic inoculation of inoculum concentration 1.0% in contain sucrose
The culture medium that mass percentage content is 10%, skimmed milk mass percentage content is 8%, 25 DEG C, 150rpm culture 72 hours,
Acidified milk is taken after fermentation, and after measured, levan content is 1.26% (w/v) in the acidified milk.
(2) white granulated sugar, syrup, An Saimi, citric acid, lactic acid, acetic acid are dissolved completely in 50 DEG C of water to get blending
Liquid.Carry out homogeneous after above-mentioned acidified milk and blending liquid are sufficiently mixed uniformly with 1:1.33 (v/v) under 20MP pressure, and in
100 DEG C of sterilizing 15min, it is filling, obtain the leben product D of the present embodiment.
Through detecting, the mass percent of levan contains in the leben rich in levulan levan of the present embodiment production
Amount is 0.54%, and sugariness is the sweetness of cane sugar containing 8.5%, 77 ° of T of acidity.
The preparation of leben of the embodiment 5 rich in levulan levan
1, materials and methods
The preparation of skimmed milk sucrose culture medium: being 8% skimmed milk powder and mass percentage content by mass percentage content
It is mixed for 12% sucrose and distilled water, after completely dissolution, sterilize 12min at 115 DEG C, is cooled to room temperature to get required dense
The sterile absorbent lactosucrose culture medium of degree.
The preparation of polysaccharide (levulan levan) in the preparation of seed (fermenting microbe), acidified milk, polysaccharide in leben
The preparation of (levulan levan): with embodiment 1.
2, the preparation of the leben rich in levulan levan
(1) by resulting lemon leukonid BD01707 seed with (v/v) aseptic inoculation of inoculum concentration 3.0% in contain sucrose
The culture medium that mass percentage content is 12%, skimmed milk mass percentage content is 8%, 28 DEG C, 250rpm culture 88 hours,
Acidified milk is taken after fermentation, and after measured, the mass percentage content of levan is 1.48% in the acidified milk.
(2) by white granulated sugar, syrup, Aspartame, honey element, An Saimi, citric acid, malic acid, tartaric acid, lactic acid, vinegar
Acid is dissolved completely in 50 DEG C of water to get blending liquid.Above-mentioned acidified milk and blending liquid are sufficiently mixed with 1:1.69 (v/v)
Carry out homogeneous after even under 22MP pressure, and in 115 DEG C of sterilizing 12min, it is filling, obtain the leben product E of the present embodiment.
Through detecting, the mass percent of levan contains in the leben rich in levulan levan of the present embodiment production
Amount is 0.55%, and sugariness is the sweetness of cane sugar containing 7.6%, 75 ° of T of acidity.
The preparation of leben of the embodiment 6 rich in levulan levan
1, materials and methods
The preparation of skimmed milk sucrose culture medium: being 8% skimmed milk powder and mass percentage content by mass percentage content
It is mixed for 12% sucrose and distilled water, after completely dissolution, sterilize 5min at 125 DEG C, is cooled to room temperature to get required concentration
Sterile absorbent lactosucrose culture medium.
The preparation of polysaccharide (levulan levan) in the preparation of seed (fermenting microbe), acidified milk, polysaccharide in leben
The preparation of (levulan levan): with embodiment 1.
2, the preparation of the leben rich in levulan levan
(1) by resulting lemon leukonid BD01707 seed with (v/v) aseptic inoculation of inoculum concentration 3.0% in contain sucrose
The culture medium that mass percentage content is 12%, skimmed milk mass percentage content is 8%, 28 DEG C, 250rpm culture 88 hours,
Acidified milk is taken after fermentation, and after measured, the mass percentage content of levan is 1.35% in the acidified milk.
(2) by white granulated sugar, syrup, Aspartame, honey element, An Saimi, citric acid, malic acid, tartaric acid, lactic acid, vinegar
Acid is dissolved completely in 50 DEG C of water to get blending liquid.Above-mentioned acidified milk and blending liquid are sufficiently mixed with 1:1.41 (v/v)
Carry out homogeneous after even under 15MP pressure, and in 95 DEG C of sterilizing 20min, it is filling, obtain the leben product F of the present embodiment.
Through detecting, the mass percent of levan contains in the leben rich in levulan levan of the present embodiment production
Amount is 0.56%, and sugariness is the sweetness of cane sugar containing 9.1%, 71 ° of T of acidity.
Comparative example 1
By the inoculum concentration in embodiment 1, cultivation temperature, fermentation time, the speed and sucrose concentration for concussion of fermenting are one by one
It is adjusted, obtains the fermentation lotion of following set of distinct methods preparation, the content of levan is such as in fermentation lotion obtained by each group
Shown in table 1.
The content of levan in 1 distinct methods of table preparation gained fermentation lotion
It can be concluded that, by inoculum concentration in the preparation method of the skim-milk fermentation liquid, culture is warm from result shown in table 1
Degree, fermentation time, when the speed and sucrose concentration of concussion of fermenting are adjusted to except the present invention, fruit in gained fermentation lotion
The content of glycan levan can significantly reduce.
1 gained leben product taste of effect example and fancy grade are tested
Using leben product A, B, C, D and E prepared by above-described embodiment as experimental subjects, the taste of product is carried out
Test.50 people of test number.Trial test mode: it is tasted by the way of blank marking；Respectively to above-mentioned leben
The color of product A, B, C, D and E, sour-sweet ratio, flavor, mouthfeel, nutrition item are individually given a mark, and each single item full marks are 20 points, meter
It calculates average mark and its total score, statistical result is recorded in table 2.Meanwhile the opinion provided according to the whole fancy grade to product, system
The hobby number to each single-item is counted, statistical result is recorded in table 3.
2 leben product taste tests result data statistical form of table
3 product fancy grade test result data statistical form of table
It can be seen that in general from product taste tests and fancy grade statistical result, of the invention is rich in levulan
The leben of levan can be received in terms of product special flavour, mouthfeel, nutrition by most of consumer.
The stability experiment of levulan levan in 2 product of effect example
Resulting leben product A, B, C, D and E is prepared as experimental subjects, under normal temperature conditions using above-described embodiment
It places, the levulan levan content in different standing times in product is measured according to method described in embodiment 1, the results are shown in Table
4。
The variation of levulan levan content in 4 product of table
For leben of the invention during being stored at room temperature 90 days, fruit therein is poly- it can be seen from upper table result
Sugared levan does not degrade, and stable content is constant.
It should be understood that those skilled in the art can make the present invention various after having read above content of the invention
Change or modification, these equivalent forms also fall within the scope of the appended claims of the present application.
Claims (8)
1. a kind of preparation method of the leben rich in levulan levan, which is characterized in that itself the following steps are included:
(1) lemon leukonid (Leuconostoc citreum) fermenting microbe is fermented training in skimmed milk sucrose culture medium
It supports, obtains acidified milk；
(2) acidified milk obtained by step (1) is mixed with the blending liquid containing sweetening material and acid, homogeneous sterilizes to obtain the final product；
Wherein, step (1) the lemon leukonid fermenting microbe is the bright beading of lemon that deposit number is CGMCC NO.6431
Bacterial strain；
The preparation method of step (1) the skimmed milk sucrose culture medium is the following steps are included: skimmed milk powder and sucrose are mixed with water
Even dissolution, 95~125 DEG C sterilize 5~20 minutes, it is cooling to obtain the final product, in skimmed milk sucrose culture medium the content of skimmed milk powder be 6~
12%, the content of sucrose is 5~20%, and the percentage is mass percent.
2. preparation method as described in claim 1, which is characterized in that the inoculum concentration of step (1) described fermenting microbe is 0.5%
~4.0%, the percentage is percent by volume.
3. preparation method as described in claim 1, which is characterized in that step (1) fermented and cultured is concussion fermented and cultured,
The temperature of fermented and cultured is 20 DEG C~35 DEG C, and the concussion speed for shaking fermented and cultured is 100~300rpm, the time of fermented and cultured
It is 48~120 hours.
4. preparation method as described in claim 1, which is characterized in that step (2) sweetening material is white granulated sugar, syrup and sweet tea
One of taste agent is a variety of；Wherein, the sweetener is one of Aspartame, honey element and acesulfame potassium or a variety of；
The content of the sweetening material is that the sugariness of the leben is made to be converted into sweetness of cane sugar 5%~10%；The acid
Taste agent is one of citric acid, malic acid, tartaric acid, lactic acid and acetic acid or a variety of；The content of the acid is described to make
Leben 70 ° of T~80 ° T of titratable acidity.
5. preparation method as described in claim 1, which is characterized in that the volume ratio of step (2) acidified milk and blending liquid
Are as follows: 1:0.69~1:2.02.
6. preparation method as described in claim 1, which is characterized in that the pressure of step (2) described homogeneous is 15~30MPa；
The temperature of the sterilizing is 95 DEG C~125 DEG C；The time of sterilizing is 5~20 minutes.
7. a kind of preparation method as described in any one of claims 1 to 6 acidified milk obtained rich in levulan levan is drunk
Material.
8. leben as claimed in claim 7, which is characterized in that wherein content >=0.5% of levulan levan, institute
Stating percentage is mass percent.
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| CN110859256A (en) * | 2019-10-28 | 2020-03-06 | 国投中鲁果汁股份有限公司 | Levan-rich probiotic health-care sweet potato fermented milk beverage and preparation method thereof |
| CN113430143B (en) * | 2021-07-27 | 2022-12-30 | 光明乳业股份有限公司 | Leuconostoc citreum strain and application thereof |
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| JP2006166853A (en) * | 2004-12-17 | 2006-06-29 | Ajinomoto Co Inc | Methanation inhibitor for ruminant and feed composition |
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| CN103478839B (en) * | 2013-09-25 | 2015-06-24 | 光明乳业股份有限公司 | Glucan beverage and preparation method thereof |
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| CN105285096A (en) | 2016-02-03 |
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