CN106256910A - Shadow yeast fermentation is utilized to remove the method for arabinose in Testa Tritici oligosaccharide - Google Patents
Shadow yeast fermentation is utilized to remove the method for arabinose in Testa Tritici oligosaccharide Download PDFInfo
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- CN106256910A CN106256910A CN201510343018.6A CN201510343018A CN106256910A CN 106256910 A CN106256910 A CN 106256910A CN 201510343018 A CN201510343018 A CN 201510343018A CN 106256910 A CN106256910 A CN 106256910A
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Abstract
The present invention relates to a kind of utilize shadow yeast fermentation to remove the method for arabinose in Testa Tritici oligosaccharide, belong to field of fermentation engineering.It is an object of the invention to improve the bacillus bifidus cultivation effect of Testa Tritici oligosaccharide.Inventive feature be with shadow yeast (Sporobolomyces singularis) as fermented bacterium, with Testa Tritici oligosaccharide as fermentation substrate.Shadow yeast thalline is inoculated in Testa Tritici oligosaccharide sugar liquid in proportion, fermentation temperature 25 DEG C, fermentation time 8h under the conditions of 160r/min, and arabinose clearance is up to 33%.In Vitro Fermentation experimental data shows, the gas production before Testa Tritici oligosaccharide is relatively removed after removing arabinose reduces, pH value reduces, lactic acid increases.After these declaratives remove arabinose, Testa Tritici oligosaccharide is notable to the probiotics cultivation effect such as bacillus bifidus and lactic acid bacteria.
Description
Technical field
The present invention relates to one utilize shadow yeast (Sporobolomyces singularis) method of arabinose in Testa Tritici oligosaccharide of removing of fermenting.Belong to field of fermentation engineering.
Background technology
Result of study shows: araboxylan is structural polysaccharide main in Testa Tritici, it is to be formed by connecting by xylan backbone and side chain, wherein β-D-xylopyranose connects and composes xylan backbone through β-(1-4)-glycosidic bond, and side chain is to be made up of α-L-arabinofuranose.α-L-arabinofuranose can independent C-2 and the C-3 position in substituted beta-D-xylose residues, it is possible to disubstituted in C-2 and C-3 position.α-L-arabinofuranose side chain is to pass through 1-2 with the α-L-arabinofuranose monosaccharide molecule of 2 or more than 2,1-3,1-5 is bonded, simultaneously possibly together with a certain amount of ferulic, by form and the covalently bound (Zheng Xueling of AX of esterification, Li Limin, Yao Huiyuan. the Structural Analysis of Purified Fraction of Water Soluble Pentosan [J].Food and biotechnology journal, 2005,24 (2): 6-9).
Containing the pentosan of about 20% in Testa Tritici, wherein most is araboxylan, is the good source preparing oligosaccharide.Comparing with cotton seed hulls etc. with other oligosaccharide raw materials for production such as corn cobs, production pretreatment is simple, waste water little, low cost and other advantages to utilize Testa Tritici production oligosaccharide to have, thus has the biggest market prospect.
Testa Tritici oligosaccharide is after xylanase hydrolysis, and owing to containing relatively araban in oligosaccharide, this can affect the functional of Testa Tritici oligosaccharide.According to our previous studies result, the probiotics cultivation effect of Testa Tritici oligosaccharide less substantially (Zhou Yingchun. Testa Tritici oligosaccharide production technology optimization and Study on functional properties thereof, master thesis, He'nan University of Technology 2014).Thus, it is an object of the invention to partly or entirely remove the arabinose in Testa Tritici oligosaccharide, functional to improve it.
But there is presently no to see and remove the report of arabinose in Testa Tritici oligosaccharide.Comparing with chemical method, microbe fermentation method has that high specificity, reaction condition be gentle, low cost and the advantage such as low in the pollution of the environment, thus has feasibility.Through early-stage Study and data check, the present invention uses shadow yeast to be that strain fermentation removes arabinose in Testa Tritici oligosaccharide.Up to now, the most do not inquire and utilize yeast to remove the report of arabinose in oligosaccharide.
Summary of the invention
It is an object of the invention to provide and a kind of remove the method for arabinose in Testa Tritici oligosaccharide.
Present disclosure is a kind of method utilizing shadow yeast fermentation to remove arabinose, it is characterised in that: using shadow yeast as strain, the part arabinose in Testa Tritici oligosaccharide is removed in fermentation, thus it is functional to improve its probiotics cultivation effect etc..
The method of removal arabinose of the present invention, its step is as follows: (1) preparation glucose 6%, peptone 1%, the Shake flask medium of yeast extract 0.5%, boils to being completely dissolved；(2) by the shake-flask culture for preparing based on 121 DEG C of sterilizing 15 min, it is cooled to room temperature；(3) shadow yeast accesses in Shake flask medium 25 DEG C, and 120 r/min cultivate 3 ~ 4 days and centrifugal obtain thalline；(4) the centrifugal thalline obtained is accessed in Testa Tritici oligosaccharide sugar liquid according to 1 ~ 3% mass ratio, at 25 DEG C, shaking speed 160 r/min bottom fermentation 8 h；(5) the rear centrifugal segregation that fermented precipitates, and supernatant is the Testa Tritici oligosaccharide sugar liquid removing part arabinose.
Testa Tritici oligosaccharide of the present invention produces obtain through high-temperature acid hydrolysis, xylanase enzymolysis, activated carbon decolorizing, the ion exchange operation such as deionization and concentration, and sugar liquid solid content is 3-10%.The assay method of solid content uses hand-held saccharometer, and its measuring principle is solid content and refractive power has dependency.
The Testa Tritici oligosaccharide sugar liquid of removal part arabinose of the present invention can obtain Testa Tritici oligosaccharide dry powder through concentration and spray drying.
It is to pass through Syrups by HPLC that the present invention removes the effect of arabinose, concrete grammar is as follows: be first the hydrolysis of Testa Tritici oligosaccharide, with mass ratio 1:1, the sugar liquid of solid content about 2% is added 1 mol/L sulfuric acid solution, boiling water bath 1.5 h respectively, neutralize with BaCO3, standby.Sugar liquid after hydrolysis carries out arabinose assay, first, after hydrolyzed solution dilutes by volume ratio 50/50 acetonitrile/water, draw hydrolyzed solution with syringe, be placed in sample bottle through 0.22 μm membrane filtration, in efficient liquid phase, analyze component, calculate the clearance of arabinose.Liquid phase chromatogram condition is as follows: detector (RID) temperature: 35 DEG C；Column temperature: 35 DEG C；Sample size: 10 μ L；Flow velocity: 0.5 mL/min；Chromatographic column model: XBRIDGE Amide (4.6 × 250 mm, 3.5 μm).By analysis, after the fermentation method of the present invention removes Arab, the arabinose clearance in Testa Tritici oligosaccharide can reach 25-33%(mass percent).
The functional authorization of the Testa Tritici oligosaccharide after removal part arabinose of the present invention uses extracorporeal simulating experiment, i.e. use Rycroft etc. external intestinal microbial population enrichment procedure (Rycroft C.E., Jones M.R.,
Gibson G.R., et al. A comparative in vitro evaluation of the fermentation
properties of prebiotic oligosaccharides[J]. Journal of Applied Microbiology,
2001,91 (05): 878-887).
Concrete operations are as follows: In Vitro Fermentation culture medium loads 10 milliliters of vial mesohigh sterilizings of band rubber cap, and Testa Tritici oligosaccharide sample is degerming through 0.22 μm micropore water system membrane filtration, then add in sterilising medium in 5g/L ratio.Taking fresh excreta 5.0 g of healthy volunteer, add 45 mL normal saline, mixing, equivalent is seeded in In Vitro Fermentation culture medium, 37 DEG C of quiescent culture.Simulation fermentation is when 2h the most in vitro, measures fermentation liquid pH value and gas production.Fermentation liquid pH value uses pH meter directly to measure.The mensuration of gas production is inserted perpendicularly into asepsis injector on the rubber cap of vial, observes the scale of syringe on fermentation liquid every 2h and keeps a record.Short-chain fat acidity test uses HPLC to analyze the lactic acid in sample, acetic acid, propanoic acid, the content of butanoic acid.
Contrast is removed the sugar liquid before and after arabinose and is simulated the change of the pH value in sweat and gas production in vitro, and utilizes the short-chain fat acid content after the in-vitro simulated fermentation of high effective liquid chromatography for measuring 24 hours.Result is shown in accompanying drawing 1-3.
It can be seen that the present invention's utilizes shadow yeast to remove before the Testa Tritici oligosaccharide after part arabinose relatively removes from accompanying drawing 1-3, gas production substantially reduces, and pH value substantially reduces, and in short-chain fatty acid, lactic acid content substantially increases.According to the Changing Pattern of intestinal microbial population microorganism, it is that lactic acid bacteria and bifidobacteria and ratio increase during simulation experiment in vitro that the above changes most probable reason.These data show, after inventive method later processes, the Testa Tritici oligosaccharide after namely part removes arabinose has more preferable probiotics cultivation effect.
Accompanying drawing explanation
Fig. 1 be in in-vitro simulated sweat gas production with fermentation time variation diagram；
Fig. 2 is that in-vitro simulated fermentation liquid pH value is with fermentation time variation diagram；
Fig. 3 is 24 hours fermentation liquid Short-Chain Fatty Acids changes of contents figures of in-vitro simulated fermentation.
Claims (5)
1. utilize microbe fermentation method to remove the method for arabinose in Testa Tritici oligosaccharide, it is characterised in that: with shadow yeast (Sporobolomyces singularis) as strain, the part arabinose in Testa Tritici oligosaccharide is removed in fermentation, thus it is functional to improve its probiotics cultivation effect etc..
2. a kind of described in claim 1 utilizes microbe fermentation method to remove the method for arabinose in Testa Tritici oligosaccharide, and its step is as follows:
Preparation glucose 6%, peptone 1%, the Shake flask medium of yeast extract 0.5%, boils to being completely dissolved；
By the shake-flask culture for preparing based on 121 DEG C of sterilizing 15 min, it is cooled to room temperature；
Shadow yeast test tube strains is accessed in Shake flask medium, cultivate 3 ~ 4 days under the conditions of cultivation temperature 25 DEG C, shaking speed 120 r/min, be centrifuged and obtain thalline；
The centrifugal thalline obtained is accessed in Testa Tritici oligosaccharide sugar liquid according to 1 ~ 3% mass ratio, at 25 DEG C, shaking speed 160 r/min bottom fermentation 8 h；
The rear centrifugal segregation that fermented precipitates, and supernatant is the Testa Tritici oligosaccharide sugar liquid removing part arabinose.
Testa Tritici oligosaccharide described in 1 produces obtain through high-temperature acid hydrolysis, xylanase enzymolysis, activated carbon decolorizing, the ion exchange operation such as deionization and concentration the most as requested.
In Testa Tritici oligosaccharide sugar liquid sugar liquid described in 2, solid content is 3-10% the most as requested.
The Testa Tritici oligosaccharide sugar liquid of the removal part arabinose described in 2 can obtain Testa Tritici oligosaccharide dry powder through concentration and spray drying the most as requested.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1557956A (en) * | 2004-01-15 | 2004-12-29 | 华南理工大学 | Method for dispelling monosaccharide component from oligosaccharide |
| WO2014207087A1 (en) * | 2013-06-26 | 2014-12-31 | Abengoa Bioenergia Nuevas Tecnologias S.A. | Production of advanced fuels and of chemicals by yeasts on the basis of second generation feedstocks |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1557956A (en) * | 2004-01-15 | 2004-12-29 | 华南理工大学 | Method for dispelling monosaccharide component from oligosaccharide |
| WO2014207087A1 (en) * | 2013-06-26 | 2014-12-31 | Abengoa Bioenergia Nuevas Tecnologias S.A. | Production of advanced fuels and of chemicals by yeasts on the basis of second generation feedstocks |
Non-Patent Citations (5)
| Title |
|---|
| VIRGIL GREENE LILLY ET AL.: "The utilization of D- and L-arabinose by fungi", 《AMERICAN JOURNAL OF BOTANY》 * |
| 包怡红: "《木聚糖酶工程菌的构建》", 31 December 2005, 东北林业大学出版社 * |
| 周迎春: "小麦麸皮低聚糖生产工艺优化及其功能特性研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 张艳艳: "卷须链霉菌D-10木聚糖酶酶解玉米芯汽爆液制备低聚木糖的研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》 * |
| 王丽琼: "《粮油加工技术》", 31 January 2008, 中国农业出版社 * |
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