CN102517236A - Lactobacillus reuteri and method for preparing Lactobacillus sugar with Lactobacillus reuteri biologically - Google Patents
Lactobacillus reuteri and method for preparing Lactobacillus sugar with Lactobacillus reuteri biologically Download PDFInfo
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- CN102517236A CN102517236A CN2011104498991A CN201110449899A CN102517236A CN 102517236 A CN102517236 A CN 102517236A CN 2011104498991 A CN2011104498991 A CN 2011104498991A CN 201110449899 A CN201110449899 A CN 201110449899A CN 102517236 A CN102517236 A CN 102517236A
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Abstract
Lactobacillus reuteri and a method for preparing Lactobacillus sugar with the strain of the Lactobacillus reuteri biologically belong to the technical field of biological engineering. The Lactobacillus reuteri SK24.003 provided by the invention is collected in China Center for Type Culture Collection, and the collection number is CCTCC NO: M2011397. The strain is obtained by screening a Chinese conventional fermented dairy product; during the fermentation of the strain, substrate cane sugar and cells of the Lactobacillus reuteri are in direct contact with each other; and under the action of the combination of a film and dextransucrase, the cane sugar is catalyzed and polymerized into the novel functional glucosan--the Lactobacillus sugar. As the strain provided by the invention is a generally recognized safe food-grade microbe, and the Lactobacillus sugar prepared biologically has properties such as favorable water-solubility, low heat quantity, sustained release of energy and the like, the invention can be applied to the fields of nutritional health foods, medicines, chemical engineering and the like.
Description
Technical field
One strain lactobacillus reuteri and with the biological sugared method of Roy that prepare of this bacterial strain, Roy's sugar is a kind of new function VISOSE, the invention belongs to technical field of bioengineering.
Background technology
VISOSE is meant the polysaccharide compound of being made up of 20 above glucose.According to the source, VISOSE can be divided into vegetable polysaccharides, animal polysaccharide and microbial polysaccharide; Form difference according to carbohydrate, VISOSE can be divided into homopolysaccharide and mixed polysaccharide again.The structure of VISOSE often because of configuration, the glycosylation mode of monose, have or not the difference of position etc. of hydroxyl in branch, the sugar, adjacent monosaccharide groups glucosides different, also make VISOSE become and have the active macromolecular substance of difference in functionality.The structure of function VISOSE is imitated and the active research focus that has become association area.
Milk-acid bacteria is one type of general designation that from the fermenting process of glucose or lactose, produces the bacterium of lactic acid, extensively is present in spontaneous fermentation milk-product, fermenting plant food and the human intestinal.In recent years, milk-acid bacteria is with its special physiologically active and trophic function and become the main source of probiotic bacterium, paid more and more attention.A large amount of research datas show that milk-acid bacteria can promote growth of animal, regulate the gi tract normal microflora, keep microecological balance, thereby improve gastrointestinal function; Improve food digestion rate and biological value; Reduce serum cholesterol, the control intracellular toxin; Suppress corrupt bacteria growing in the enteron aisle; Enhance immunity power etc.
The organic acid that milk-acid bacteria produces through fermentation, specific enzymes system, bacterium table have physiological function to the one-tenth material that grades, and can stimulate tissue development, to the generation effects such as nutritional status, physiological function, immunoreation and stress response of body.For example, some secondary metabolites such as exocellular polysaccharide, bacteriocin etc. not only have important technic function, also have multiple physiologically active, comprise conditioning enteron aisle, antitumor, enhancing immunity etc.Therefore, carry out the research that milk-acid bacteria prepares functional polysaccharide, can satisfy the process operation of traditional biological food and the demand that nutrition keeps, have crucial Research Significance and application market value as a kind of novel functional foodstuff auxiliary material simultaneously.
Summary of the invention
The object of the present invention is to provide lactobacillus reuteri CCTCC NO:M 2011397 and with the biological method for preparing the new function VISOSE of this bacterial strain, gained new function VISOSE-Roy's sugar can be applicable to fields such as nutritious and healthy food, medicine, chemical industry.
Technical scheme of the present invention: screening acquisition one strain is the bacterial strain of new function VISOSE-Roy's sugar with the sucrose bio-transformation from China's traditional zymotic milk preparation, classification called after lactobacillus reuteri (
Lactobacillus reuteri) SK 24.003, its deposit number is CCTCC NO:M 2011397.
With the biological methods for preparing Roy's sugar of lactobacillus reuteri CCTCC No. M 2011397, through preparatory incubation time, substratum, conversion incubation time, transform the selection of conditions such as inoculum size, sum up this bacterial strain and transform the biological sugared method of Roy for preparing of sucrose and be:
(1) bacterial strain of bio-transformation employing is CCTCC NO:M 2011397;
(2) substratum is:
Pre-culture medium is the MRS substratum of 5-10 g/L sucrose;
Transforming substratum is the MRS substratum of 80-100 g/L sucrose, pH6.8-7.0;
(3) preparatory culture condition: the 2%-4% inoculum size, 35 ~ 37 ℃ leave standstill cultivation 24 ~ 36h;
(4) transform culture condition: pre-incubated thalline is transferred to the 2%-4% inoculum size in the MRS substratum of the pH6.8-7.0 that contains 80-100g/L sucrose, and 35 ~ 37 ℃ leave standstill cultivation 24 ~ 48h.
Biological fermentation prepares the new function VISOSE--and the output of Roy's sugar reaches 20-35g/L; The glycosidic link mode of connection is α (1 → 4) and α (1 → 6) in its molecular structure, and branch's density is 8%-20%, and molecular mass is 10
6-10
8
Adopt IR to press the functional group and the glycosidic link mode of connection of KBr pressed disc method and nuclear magnetic resonance nmr technical measurement Roy sugar, wherein exist
1Chemical shift is the quantity that the peak value of 5.36ppm and 4.96ppm is represented connected anomeric carbon in Glucopyranose α-1,4 and α-1,6 glycosidic link respectively on the H NMR collection of illustrative plates, and branch's density is represented by the ratio of α-1,6 glycosidic link in the molecular chain fine structure.The mensuration of molecular weight adopts the method for high performance size exclusion chromatography (HPSEC) to measure.
Beneficial effect of the present invention: through a large amount of screening operations, from China various places traditional zymotic milk preparation source, separate the bacterial strain that obtaining the biological preparation of strain Roy sugar, through be accredited as lactobacillus reuteri (
Lactobacillus reuteri) SK 24.003, deposit number CCTCC NO:M 2011397.Substrate sucrose directly contacts with lactobacillus cell during this bacterium fermentation, combines at film under the effect of Sucrose:glucan alpha1, and sucrose is become new function VISOSE-Roy's sugar by catalyzed polymerization.This bacterium is suitable for liquid continuously ferments and cultivates and high-cell-density cultivation, and fermentation time is short, cost is low, output is high, is suitable for scale operation.The product of the present invention's preparation can be applicable to a plurality of fields such as food, medicine, chemical industry, has crucial commercial value and application prospect.
The biological material specimens preservation: a strain lactobacillus reuteri (
Lactobacillus reuteri) SK 24.003, depositary institution's title: Chinese typical culture collection center, be called for short CCTCC, address: Chinese Wuhan Wuhan University, deposit number is CCTCC NO:M 2011397, preservation date on November 18th, 2011.
Description of drawings
The molecular structure of Fig. 1 Roy sugar.
Fig. 2 Roy sugar
1H NMR figure.
Embodiment
For a better understanding of the present invention, below in conjunction with embodiment the present invention is done to further describe, protection of the present invention is not limited thereto.
Embodiment 1
Lactobacillus reuteri CCTCC NO:M 2011397 is received the MRS substratum of 10 g/L sucrose by 2% inoculum size; 35 ℃ leave standstill cultivation 24h; Be transferred to this pre-incubated thalline in the MRS substratum of the pH7.0 that contains 100g/L sucrose with 4% inoculum size then; 35 ℃ leave standstill cultivation 48h, and the content of Roy's sugar reaches 32.5g/L in the tunning.
Embodiment 2
Lactobacillus reuteri CCTCC NO:M 2011397 is received the MRS substratum of 5 g/L sucrose by 4% inoculum size; 37 ℃ leave standstill cultivation 36h; Be transferred to this pre-incubated thalline in the MRS substratum of the pH6.8 that contains 80g/L sucrose with 2% inoculum size then; 37 ℃ leave standstill cultivation 24h, and the content of Roy's sugar reaches 28.6g/L in the tunning.
Claims (5)
1. the bacterial strain of Roy's sugar is produced in a strain, classification called after lactobacillus reuteri (
Lactobacillus reuteri) SK 24.003, being preserved in Chinese typical culture collection center, deposit number is CCTCC NO:M 2011397.
2. one kind with the biological sugared methods of Roy for preparing of lactobacillus reuteri CCTCC NO:M 2011397, it is characterized in that:
(1) bacterial strain of bio-transformation employing is CCTCC NO:M 2011397;
(2) substratum is:
Pre-culture medium is the MRS substratum of 5-10 g/L sucrose;
Transforming substratum is the MRS substratum of 80-100 g/L sucrose, pH6.8-7.0;
(3) preparatory culture condition: the 2%-4% inoculum size, 35 ~ 37 ℃ leave standstill cultivation 24 ~ 36h;
(4) transform culture condition: pre-incubated thalline is transferred to the 2%-4% inoculum size in the MRS substratum of the pH6.8-7.0 that contains 80-100g/L sucrose, and 35 ~ 37 ℃ leave standstill cultivation 24 ~ 48h.
3. according to claim 2 a kind of with the lactobacillus reuteri CCTCC NO:M 2011397 biological methods that prepare Roy's sugar, it is characterized in that the output of Roy's sugar reaches 20-35g/L.
4. according to claim 2 a kind of with the lactobacillus reuteri CCTCC NO:M 2011397 biological methods that prepare Roy's sugar; The molecular chain structure characteristic that it is characterized in that Roy's sugar, glycosidic link mode of connection are α (1,4) and α (1; 6), and branch's density be 8%-20%.
5. according to claim 2 a kind of with the lactobacillus reuteri CCTCC NO:M 2011397 biological methods that prepare Roy's sugar, it is characterized in that the molecular mass of product Roy sugar is 10
6-10
8
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103329954A (en) * | 2013-07-16 | 2013-10-02 | 江南大学 | Method for utilizing Roy sugar to improve grain food quality |
| CN104988084A (en) * | 2015-02-28 | 2015-10-21 | 江南大学 | Lactobacillus gasseri producing inulin, and uses thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010128859A2 (en) * | 2009-05-08 | 2010-11-11 | Rijksuniversiteit Groningen | GLUCO-OLIGOSACCHARIDES COMPRISING (α1→4) AND (α1→6) GLYCOSIDIC BONDS, USE THEREOF, AND METHODS FOR PROVIDING THEM |
-
2011
- 2011-12-29 CN CN 201110449899 patent/CN102517236B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010128859A2 (en) * | 2009-05-08 | 2010-11-11 | Rijksuniversiteit Groningen | GLUCO-OLIGOSACCHARIDES COMPRISING (α1→4) AND (α1→6) GLYCOSIDIC BONDS, USE THEREOF, AND METHODS FOR PROVIDING THEM |
Non-Patent Citations (5)
| Title |
|---|
| 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 20020930 S. Kralj, et al. Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with alpha-(1->4) and alpha-(1->6) Glucosidic Bonds 4283-4291 1-5 第68卷, 第9期 * |
| 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 20111130 Slavko Kralj, et al. 4,6-alpha-Glucanotransferase, a Novel Enzyme That Structurally and Functionally Provides an Evolutionary Link between Glycoside Hydrolase Enzyme Families 13 and 70 8154-8163 1-5 第77卷, 第22期 * |
| S. KRALJ, ET AL.: "Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with α-(1→4) and α-(1→6) Glucosidic Bonds", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| SLAVKO KRALJ, ET AL.: "4,6-α-Glucanotransferase, a Novel Enzyme That Structurally and Functionally Provides an Evolutionary Link between Glycoside Hydrolase Enzyme Families 13 and 70", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| 张天琪等: "乳杆菌胞外多糖及其在酸乳中的应用", 《食品科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103329954A (en) * | 2013-07-16 | 2013-10-02 | 江南大学 | Method for utilizing Roy sugar to improve grain food quality |
| CN103329954B (en) * | 2013-07-16 | 2014-04-16 | 江南大学 | Method for utilizing Roy sugar to improve grain food quality |
| CN104988084A (en) * | 2015-02-28 | 2015-10-21 | 江南大学 | Lactobacillus gasseri producing inulin, and uses thereof |
| CN104988084B (en) * | 2015-02-28 | 2017-10-20 | 江南大学 | Lactobacillus gasseri of one plant of production synanthrin and application thereof |
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