WO2020228410A1 - 一株高加索酸奶乳杆菌msr101及其应用 - Google Patents
一株高加索酸奶乳杆菌msr101及其应用 Download PDFInfo
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- WO2020228410A1 WO2020228410A1 PCT/CN2020/079861 CN2020079861W WO2020228410A1 WO 2020228410 A1 WO2020228410 A1 WO 2020228410A1 CN 2020079861 W CN2020079861 W CN 2020079861W WO 2020228410 A1 WO2020228410 A1 WO 2020228410A1
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- WIPO (PCT)
- Prior art keywords
- msr101
- lactobacillus
- lactobacillus kefiri
- strain
- present
- Prior art date
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Classifications
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B23—MACHINE TOOLS; METAL-WORKING NOT OTHERWISE PROVIDED FOR
- B23K—SOLDERING OR UNSOLDERING; WELDING; CLADDING OR PLATING BY SOLDERING OR WELDING; CUTTING BY APPLYING HEAT LOCALLY, e.g. FLAME CUTTING; WORKING BY LASER BEAM
- B23K26/00—Working by laser beam, e.g. welding, cutting or boring
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- B—PERFORMING OPERATIONS; TRANSPORTING
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- B23K—SOLDERING OR UNSOLDERING; WELDING; CLADDING OR PLATING BY SOLDERING OR WELDING; CUTTING BY APPLYING HEAT LOCALLY, e.g. FLAME CUTTING; WORKING BY LASER BEAM
- B23K31/00—Processes relevant to this subclass, specially adapted for particular articles or purposes, but not covered by only one of the preceding main groups
- B23K31/003—Processes relevant to this subclass, specially adapted for particular articles or purposes, but not covered by only one of the preceding main groups relating to controlling of welding distortion
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- C—CHEMISTRY; METALLURGY
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/24—Processes using, or culture media containing, waste sulfite liquor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B23—MACHINE TOOLS; METAL-WORKING NOT OTHERWISE PROVIDED FOR
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- C—CHEMISTRY; METALLURGY
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Definitions
- the invention belongs to the technical field of Lactobacillus, and specifically relates to a strain of Lactobacillus kefiri MSR101 and its application.
- Probiotic microorganisms are "living microorganisms" that can bring various health benefits to the host, including immune system regulation, gastrointestinal microbiota rearrangement and inhibition of harmful microorganism growth (Maleki Kakelar et al., 2019). Lactic acid bacteria are the most important probiotics and are compatible with the human digestive system because of their natural resistance to low pH and high bile salt conditions (Shehata et al., 2016, Riaz Rajoka et al., 2018).
- probiotics have become a research hotspot in the field of lactic acid bacteria, including Lactobacillus and Bifidobacterium. (Bao et al., 2010). Recently, there has been a new understanding of the consumption of Lactobacillus strains isolated from traditional Chinese dairy products with various health-promoting effects. In order to provide the host with the necessary conditions for health, the isolated Lactobacillus strain must have the ability to pass through the physical and chemical barriers of the gastrointestinal tract (Ramos et al., 2018). In addition, as a probiotic, the Lactobacillus strain must have the ability to survive in sufficient numbers during the production and storage of the intended product.
- the purpose of the present invention is to provide a strain of Lactobacillus kefiri MSR101 and its application.
- the lactobacillus of the present invention has acid resistance, bile salt resistance, phenol resistance, antibiotic resistance, antioxidant activity, cell surface hydrophobicity, adhesion to intestinal epithelial cells and cholesterol-lowering functions.
- the present invention provides a strain of Lactobacillus kefiri MSR101, the deposit number is CGMCCNo.17506.
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in the anti-hyperbile salt.
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in phenol tolerance.
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in antioxidant.
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in reducing cholesterol concentration.
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in resistance to penicillin, ampicillin, streptomycin and tetracycline.
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in the treatment of hypercholesterolemia.
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in the treatment of high bile salt diseases.
- the present invention provides a strain of Lactobacillus kefiri MSR101.
- the lactobacillus of the present invention has acid resistance, bile salt resistance, phenol resistance, antibiotic resistance, antioxidant activity, cell surface hydrophobicity, adhesion to intestinal epithelial cells and cholesterol-lowering functions.
- the test results show that the Lactobacillus kefiri MSR101 strain is in a stable phase after 48 hours of culture and the EPS output is up to 753mg/L.
- the Lactobacillus kefiri MSR101 strain shows high hydrophobicity (70%) to xylene.
- the highest adhesion rate of HT-29 to intestinal epithelial cells is 34.4%, the clearance rate of superoxide anion free radicals is 65.5%, the clearance rate of DPPH free radicals is 43.3%, and the removal rate of cholesterol is 26.2%.
- Figure 1 is a diagram of the phylogenetic tree analysis results provided by the present invention.
- Figure 2 is a result diagram of antibiotic resistance provided by the present invention.
- Figure 3 is a graph showing the production of extracellular polysaccharides provided by the present invention.
- Lactobacillus caucasus yoghurt Lactobacillus kefiri MSR101 This strain is preserved in the General Microbiology Center of China Microbial Culture Collection Management Committee, the address is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, the preservation number is CGMCC No.17506, the preservation time is 2019 April 1.
- the present invention provides a strain of Lactobacillus kefiri MSR101, the deposit number is CGMCC No. 17506.
- Lactobacillus kefiri MSR101 is an anaerobic, gram-positive bacteria, rod-shaped bacteria.
- Lactobacillus kefiri MSR101 of the present invention is Gram-positive, catalase-negative, rod-shaped and mesophilic. Its 16s rRNA gene sequence is shown in SEQ ID NO.1:
- the sequence shows that it is 99% similar to the sequence of known species available in the NCBI database, and the sequence is deposited in GeneBank under the accession number MK491609.
- Antibiotic sensitivity, tolerance under intestinal conditions low pH, bile salt tolerance and 0.2% to 0.4% phenol resistance), hydrophobicity, antioxidant capacity, lower cholesterol and adhesion to HT-29, etc. It has beneficial effects.
- the Lactobacillus kefiri MSR101 is resistant to penicillin, ampicillin, streptomycin and tetracycline. In addition, it also exhibited adhesion (34.3%) and antioxidant activity (67%) to HT-29 cells. Shows a high survival rate (>80%) under gastrointestinal conditions.
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in the anti-hyperbile salt.
- the strain of the invention grows well under high bile salt conditions (0.3%, 0.5% and 1% bile (w/v)).
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in phenol tolerance.
- the strain of the present invention has good survival ability at a phenol concentration of 0.2% to 0.4%.
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in antioxidant.
- the strain of the present invention has antioxidant activity (67%).
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in reducing cholesterol concentration.
- the cholesterol-lowering ability of the strain of the present invention is 26.2 ⁇ 0.6%.
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in resistance to penicillin, ampicillin, streptomycin and tetracycline.
- the present invention also provides the application of Lactobacillus kefiri MSR101 of the above technical scheme in the treatment of hypercholesterolemia.
- the present invention also provides the application of Lactobacillus kefiri MSR101 described in the above technical scheme in the treatment of high bile salt diseases.
- the experimental samples come from traditional Chinese dairy products from the Cambodia Autonomous Region of Qinghai province, China. There are 20 samples in total, including yak milk (4), horse milk wine (5), cheese (4) and kefir grains (7). The samples were collected in sterile tubes and stored in a mini freezer at -15°C. The laboratory immediately conducted the test after receiving the sample.
- the samples were serially diluted in phosphate buffered saline (PBS, pH 7.2), and a small aliquot (50 ⁇ l) of each batch of dilution was spread on the decanter supplemented with 0.05% L-cysteine (MRSc) Man-Rogosa-Sharpe (Merck, China) agar plate.
- the plate was incubated anaerobically at 37°C for 72h. Select colonies with different morphologies and re-streak several generations on the newly prepared MRS c agar plate to separate and purify a single colony.
- the results of the phylogenetic tree analysis are shown in Figure 1.
- the phylogenetic tree is constructed on the basis of the 16S rRNA sequence. Genome probability is determined by the number of 1000 repeats and expressed as a percentage value.
- the filled circle indicates that the strain is from NCBI, and the open circle indicates that the isolated Lactobacillus strain was used for developmental tree construction.
- the survival ability of lactobacilli at pH 2.0, pH 2.5, pH 3.0 and pH 6.5 was evaluated according to the method described in (Lee et al., 2011). At the same time, using MRS c broth medium containing 0.3%, 0.5% and 1% bile (w/v), the bile salt tolerance of lactobacilli that survived for 3 hours under acidic conditions was measured according to the method (Sabir et al., 2010b). Acceptability. MRS c broth medium without bile salts was used as a control. According to formula (1), the resistance of lactobacilli to acidic conditions and bile salt conditions was evaluated by counting on MRS c agar plates.
- N 1 is the total number of live bacterial cells in the MRS c broth after treatment
- N 0 is the total number of live bacterial cells in the MRS c broth before treatment.
- phenol tolerance was performed by the method previously reported (Shehata et al., 2016) with a slight modification.
- the 1% lactic acid bacteria isolate was cultured for 24 hours and then inoculated into MRsc fermentation broth supplemented with 0.2% phenol and 0.4% phenol. MRSc broth without phenol was used as a control. After 24 hours of incubation at 37°C, the absorbance (A) OD630nm was measured according to the formula to evaluate phenol resistance.
- Phenol tolerance (%) (A1/A0) ⁇ 100
- a 1 is the absorbance of the culture after treatment
- a 0 is the absorbance of the culture before treatment.
- Lactobacillus isolates were distributed in stripes on the surface of Columbia blood medium (Sigma, China) with 5% sheep blood, and the plates were cultured at 37°C for 72 hours. After incubation, the plate was checked for hemolytic activity.
- Lactobacillus isolates The antibiotic resistance of Lactobacillus isolates was evaluated by the previously reported (Riaz Rajoka et al., 2017a) agar overlap diffusion method. All lactobacillus isolates were tested for resistance to 10 antibiotics, including penicillin, erythromycin, ampicillin, streptomycin, tetracycline, vancomycin, gentamicin, kanamycin, chloramphenicol and gram Linmycin. The method is to cover the freshly prepared MRS c agar plate with 50 ⁇ l of active Lactobacillus culture, and incubate the plate at 4°C for 1h, place the antibiotic plate on the plate, and incubate the plate at 37°C for 24h, then measure The diameter of the suppression zone. The results of antibiotic resistance are shown in Figure 2 and Table 1. The Lactobacillus kefiri MSR101 is resistant to penicillin, ampicillin, streptomycin and tetracycline.
- Lactobacillus cultures were grown in flasks of freshly prepared 100ml MRS c broth supplemented with 3% (w/v) glucose and incubated at 37°C for 72h.
- the bacterial cells were removed by centrifugation (7000 ⁇ g, 10 min), and two volumes of pre-cooled ethanol were added to one volume of supernatant for precipitation of extracellular polysaccharides (EPS).
- EPS extracellular polysaccharides
- the EPS precipitate was recovered by centrifugation (10000 ⁇ g, 35min) at 4°C, dialyzed (6000Da to 8000Da) for 48h, and then lyophilized.
- glucose as a standard solution, the total amount of sugar was measured by the phenol sulfuric acid method (Nikolic et al., 2012).
- the EPS-producing L.kefiri MSR101 strain was originally isolated from the kefir grains in the Cambodia Autonomous Region of Qinghai province.
- the MSR101 strain showed a sticky appearance, indicating that it is an EPS-producing strain.
- the L. kefiri MSR101 strain was in the stable phase after 48 hours of culture and the EPS amount reached the maximum value of 753 mg/L, and then entered the death phase.
- the results showed that the MSR101 strain grew rapidly within 32 hours of culture, and the pH value of the medium dropped rapidly. After 48h, the growth rate of the strain decreased, and the pH value of the medium dropped slightly. The pH value was about 3.1 ⁇ 0.5 at 48h.
- Lactobacillus kefiri MSR101 strain shows high hydrophobicity (70%) for xylene; xylene is a standard hydrocarbon for measuring the hydrophobicity of microbial cells.
- ⁇ A 0 and ⁇ A are the auto-oxidation rates of pyrogallol before and after adding the sample and deionized water, respectively.
- the scavenging rate of superoxide anion radical is 65.5%.
- the DPPH free radical scavenging ability of Lactobacillus was measured by a previously reported method (Son et al., 2018). That is, three (3) ml of Lactobacillus cell suspension and 3 ml of 0.5 mM DPPH free radical ethanol solution are mixed, and incubated for 30 minutes in the dark at room temperature. After incubation, the reaction mixture was centrifuged (10000 ⁇ g, 20 min at 4°C), and the absorbance of the supernatant at 517 nm was measured.
- the DPPH removal capacity is calculated by the following formula:
- OD Sample and OD Control are the absorbance of sample and distilled water, respectively, mixed with DPPH solution.
- the scavenging rate of Lactobacillus kefiriMSR101 on DPPH free radicals is 43.3%.
- each lactobacillus isolate (1% v/v) was inoculated into 10 ml of freshly prepared MRS c broth medium containing 0.2% sodium thioacetate (Sigma, China), 0.3% With bovine bile (Sigma, China) and 100 ⁇ l/ml water-soluble cholesterol (Sigma, China), each test tube was incubated at 37°C for 24h. The cells were removed by centrifugation (10,000 ⁇ g, 35 min at 4°C).
- the phthalaldehyde method of measuring cholesterol was used to determine the amount of cholesterol in used broth and non-inoculated sterile broth.
- the cholesterol removal rate of strain Lactobacillus kefiri MSR101 was 26.2%.
- DMEM Dulbecco's Modified Eagle Medium
- 2mM L-glutamine 10% heat-inactivated fetal calf serum
- 100 ⁇ g streptomycin/ml 100 ⁇ g streptomycin/ml
- 1% non-essential amino acids 100Ul
- HT-29 cells were seeded into a 24-well culture plate at a concentration of 2.5 ⁇ 10 5 cells/well, cultured for 3 days, and the medium was changed every day. The cells were incubated at 37°C in a 5% CO 2 atmosphere.
- the overnight culture of the lactobacillus isolate was centrifuged, washed twice with PBS (0.1M, pH7.2), and resuspended in the same buffer to an appropriate concentration of diluent (absorbance OD6300.2, about 2 ⁇ 10 8 CFU/ml). Then add the bacterial cells to each cell well, incubate at 37°C for 4h, wash the cells with PBS (0.1M, pH7.2) after incubation, and lyse with 0.1% Triton X-100 solution. The cell lysate was serially diluted and spread on the MRS c agar plate. Incubate at 37°C for 3 days. Calculate the bacterial adhesion rate:
- Adhesion rate% (attached bacteria/total added bacteria) ⁇ 100
- strain Lactobacillus kefiri MSR101 had the highest adhesion rate in HT-29 cells of 34.4%.
- Lactobacillus MSR101 was isolated from Kefir product samples collected in the Cambodia Autonomous Region of Qinghai Republic, China. It is gram positive, catalase negative, rod-shaped and mesophilic. Based 16s rRNA gene sequence (GCTTGGCGTCGTGCTATACATGCAAGTCGAACGCGTTTCCGTTATTGATTTTAGAGTGTTGCATTTGAATGATTTAACACGAAACGAGTGGCGAACTGGTGAGTAACACGTGGGTACCTGCCCTTGAAGTAGGGGATAACACTTGGAAACAGGTGCTAATACCGTATAACAACCAAAACCACATGG TTTTGGTTTAAAAGATGGCTTCGGCTATCACTTTAGGATGGACCCGGGCGTATTAGCTTGTTGGTAAGGTAATGGCCTACCAAGGCAATGATACGTAGCCGACCTAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGT
- Antibiotic sensitivity tolerance under intestinal conditions (low pH, bile salt tolerance and 0.2% to 0.4% phenol resistance), hydrophobicity, antioxidant capacity, lower cholesterol and adhesion to HT-29, etc.
- the strain of the present invention is resistant to penicillin, ampicillin, streptomycin and tetracycline. In addition, it also exhibited adhesion (34.3%) and antioxidant activity (67%) to HT-29 cells. They showed a high survival rate (>80%) under gastrointestinal conditions, indicating their potential in probiotic applications (as shown in Table 2).
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Abstract
提供了一株高加索酸奶乳杆菌(Lactobacillus kefiri)MSR101,其保藏编号为CGMCC No.17506,所述菌株具有耐酸、耐胆汁盐、耐酚、耐抗生素、抗氧化活性,细胞表面疏水性,对肠上皮细胞的粘附能力和降低胆固醇的功能。
Description
本申请要求于2019年05月10日提交中国专利局、申请号为CN201910388000.6、发明名称为“一株高加索酸奶乳杆菌MSR101及其应用”的中国专利申请的优先权,其全部内容通过引用结合在本申请中。
本发明属于乳酸杆菌技术领域,具体涉及一株高加索酸奶乳杆菌Lactobacillus kefiri MSR101及其应用。
益生菌微生物是“活微生物”,可以为宿主带来各种健康益处,包括免疫系统的调节,胃肠道微生物群的重排和抑制有害微生物的生长(Maleki Kakelar等,2019)。乳酸菌是最重要的益生菌,并且与人体消化系统具有相容性,因为它们具有对低pH值和高胆汁盐条件的天然抗性(Shehata等,2016,Riaz Rajoka等,2018)。
近10年来,益生菌已成为乳酸菌领域的研究热点,其中包括乳酸杆菌属和双歧杆菌属。(Bao等,2010)。最近,对从具有各种健康促进作用的中国传统乳制品中分离的乳杆菌菌株的消耗量有了新的认识。为了提供给宿主健康所需的条件,分离的乳杆菌菌株必须具备通过胃肠道的物理和化学屏障的能力(Ramos等,2018)。此外,作为益生菌,在预期产品的生产和储存期间,乳杆菌菌株必须具有存活足够数量的能力。
发明内容
有鉴于背景技术中存在的问题,本发明的目的在于提供一株高加索酸奶乳杆菌Lactobacillus kefiri MSR101及其应用。本发明所述乳杆菌具有耐酸性,耐胆汁盐性,耐酚性,抗生素耐性,抗氧化活性,细胞表面疏水性,对肠上皮细胞的粘附能力和降低胆固醇的功能。
本发明提供了本发明提供了一株高加索酸奶乳杆菌Lactobacillus kefiri MSR101,保藏编号为CGMCCNo.17506。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在抗高胆汁盐中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在苯酚耐受中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在抗氧化中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在降低胆固醇浓度中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在抗青霉素、氨苄青霉素、链霉素和四环素中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在治疗高胆固醇血症中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在治疗高胆汁盐疾病中的应用。
有益效果:本发明提供了一株高加索酸奶乳杆菌Lactobacillus kefiri MSR101。本发明所述乳杆菌具有耐酸性,耐胆汁盐性,耐酚性,抗生素耐性,抗氧化活性,细胞表面疏水性,对肠上皮细胞的粘附能力和降低胆固醇的功能。试验结果表明,高加索酸奶乳杆菌Lactobacillus kefiri MSR101菌株培养48h后处于稳定期且EPS产量最高达753mg/L,高加索酸奶乳杆菌Lactobacillus kefiri MSR101菌株对二甲苯显示出较高的疏水性(70%),对肠上皮细胞HT-29的最高粘附率为34.4%,对超氧化物阴离子自由基的清除率为65.5%,对DPPH自由基清除率为43.3%,胆固醇去除率为26.2%。
说明书附图
图1为本发明提供的系统发育树分析结果图;
图2为本发明提供的抗生素耐药性结果图;
图3为本发明提供的胞外多糖产量结果图。
生物保藏说明
高加索酸奶乳杆菌Lactobacillus kefiri MSR101。本菌种保藏在中国微生物菌种保藏管理委员会普通微生物中心,地址为北京市朝阳区北辰西路 1号院3号,中国科学院微生物研究所,保藏编号为CGMCC No.17506,保藏时间为2019年4月1日。
下面结合实施例和附图对本发明进一步说明。
本发明提供了一株高加索酸奶乳杆菌Lactobacillus kefiri MSR101,保藏编号为CGMCC No.17506。
高加索酸奶乳杆菌Lactobacillus kefiri MSR101是一种厌氧、革兰氏阳性菌,杆状细菌。
本发明所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101是革兰氏阳性,过氧化氢酶阴性,杆状和嗜温的。其16s rRNA基因序列如SEQ ID NO.1所示:
所述序列显示其与NCBI数据库中可获得的已知物种的序列相似99%,并且序列以登录号MK491609保藏在Gene Bank中。在抗生素敏感性,肠道条件下耐受性(低pH,胆汁盐耐受性和0.2%至0.4%酚抗性),疏水性,抗氧化能力,降低胆固醇和对HT-29的粘附等方面有有益效果。所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101对青霉素,氨苄青霉素,链霉素和四环素具有抗性。此外,它还表现出对HT-29细胞的粘附性(34.3%)和抗氧化活性(67%)。在胃肠道条件下显示出高的存活率(>80%)。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在抗高胆汁盐中的应用。本发明菌株在高胆汁盐条件下(0.3%、0.5%和1%胆汁(w/v)).菌株生长状态良好。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在苯酚耐受中的应用。本发明所述菌株在苯酚浓度为0.2%~0.4%下具有良好的存活能力。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在抗氧化中的应用。本发明所述菌株具有抗氧化活性(67%)。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiriMSR101在降低胆固醇浓度中的应用。本发明所述菌株降低胆固醇能力为26.2±0.6%。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在抗青霉素、氨苄青霉素、链霉素和四环素中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在治疗高胆固醇血症中的应用。
本发明还提供了上述技术方案所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在治疗高胆汁盐疾病中的应用。
下面结合具体实施例进一步阐明本发明,本实施例在以本发明技术方案为前提下进行实施,应理解这些方式仅用于说明本发明而不用于限制本发明的范围。
实施例1
采样
实验样品来自中国青海省西藏自治区的中国传统乳制品,共有20份,分别有牦牛奶(4)、马乳酒(5)、奶酪(4)和克菲尔谷物(7)。将样品收集在无菌管中并保存在-15℃的迷你冷冻机中。实验室收到样品后立即进行了试验。
乳酸杆菌的分离和鉴定
将样品在磷酸盐缓冲液(PBS,pH 7.2)中连续稀释,并将每批稀释液的小等分试样(50μl)涂布在补充有0.05%L-半胱氨酸(MRSc)的de Man-Rogosa-Sharpe(Merck,China)琼脂平板上。将平板在37℃下厌氧培养72h。选择具有不同形态的菌落并在新制备的MRS
c琼脂平板上重新划线数代,分离纯化单个菌落。所有分离株均经革兰染色和过氧化氢酶试验证实为乳酸菌,革兰阳性和过氧化氢酶阴性的菌株均为乳酸菌。通过对16S rDNA基因测序,进行乳杆菌分离物的鉴定。使用基因组DNA纯化试剂盒(TransGenBiotech Co.,Ltd.,Beijing,China),按照使用说明提取总基因组DNA。用于扩增16S rDNA序列的引物是正向5“-AGAGTTTGATCCTGGCTC AG-3”(SEQ ID NO.2)和反向5“-CCGTCAATTCCTTTGAGTTT-3”(SEQ ID NO.3)。在Techne-TC 512热循环仪(英国)中,在以下条件下扩增片段:95℃,1min;95℃,30s,30个循环;55℃,30秒,最后72℃,5min。将扩增的片段在琼脂糖凝胶上筛选,并由中国深圳的广州IGE生物技术有限公司测序。通过BLAST程序(https://blast.ncbi.nlm.nih.gov/Blast.cgi)测试所有获得的序列。序列保存在基因库中。通过ClustalW2 (http://www.ebi.ac.uk/Tool/mas/clustalw2/)进行序列比对,通过邻接(Tamura等,2004)和最大复合似然法构建系统发育树。使用Mega 6.0软件(http://megasoftware.net/)(Saitou和Nei,1987)。结果表明,克菲尔活菌元是乳酸杆菌的来源之一。
系统发育树分析结果如图1所示,系统发育树是在16S rRNA序列的基础上构建的。基因组概率用1000个重复数确定,并以百分比值表示。实心圆表示菌株来自NCBI,空心圆表示分离的乳酸杆菌菌株用于发育树构建。
实施例2
在低pH值和高胆汁盐条件下的生存状况
按照所(Lee等人,2011)描述的方法评估乳酸杆菌在pH2.0,pH2.5,pH3.0和pH6.5(对照组)下的存活能力。同时,使用含有0.3%,0.5%和1%胆汁(w/v)的MRS
c肉汤培养基,按照(Sabir等人,2010b)方法测定在酸性条件下存活3小时的乳酸杆菌的胆汁盐耐受性。没有胆汁盐的MRS
c肉汤培养基用作对照。根据公式(1),通过MRS
c琼脂平板上的计数评估乳酸杆菌对酸性条件以及胆汁盐条件的抗性。
增长率(%)=(N
1/N
0)×100 公式(1)
其中,N
1是处理后MRS
c肉汤中活细菌细胞的总数,N
0是处理前MRS
c肉汤中活细菌细胞的总数。结果表明,在低pH值(pH2.0、pH2.5pH3.0)和高胆汁盐条件下(0.3%、0.5%和1%胆汁(w/v)).菌株MSR101生长状态良好。
实施例3
苯酚耐受性
苯酚耐受性测定通过先前报道(Shehata等人,2016)的方法进行,对其稍作修改。将1%乳酸菌份分离物培养24h后接种于添加了0.2%苯酚和0.4%苯酚的MRsc发酵液中。将不含苯酚的MRSc肉汤培养基用作对照。在37℃下,温育24h后,按公式测定吸光度(A)OD630nm来评价酚抗性。
苯酚耐受性(%)=(A1/A0)×100
其中,A
1是处理后培养物的吸光度,A
0是处理前培养物的吸光度。结果表明,菌株MSR101在苯酚浓度为0.2%~0.4%下具有良好的存活能力。
实施例4
溶血活性
乳酸杆菌分离物以条纹状分布在有5%羊血的哥伦比亚血培养基(Sigma,中国)的表面上,并将平板在37℃下培养72h。温育后,检查平板的溶血活性。
结果表明,MSR101菌株无溶血活性,因此在溶血活性方面是安全的。
实施例5
抗生素耐药性
通过之前报道过的(Riaz Rajoka等,2017a)琼脂重叠扩散法评估乳酸杆菌分离物的抗生素耐药性。检测了所有乳酸杆菌分离物对10种抗生素的抗性,包括青霉素,红霉素,氨苄青霉素,链霉素,四环素,万古霉素,庆大霉素,卡那霉素,氯霉素和克林霉素。方法是将新鲜制备的MRS
c琼脂平板用50μl的活性乳酸杆菌培养物覆盖,并将平板在4℃孵育1h,将抗生素盘置于平板上,并将平板在37℃下温育24h后,测量抑制区的直径。抗生素耐药性结果如图2和表1所示,所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101对青霉素,氨苄青霉素,链霉素和四环素具有抗性。
表1 抗生素耐药性
*(S)=易感染and(R)=免疫
实施例6
胞外多糖的生产
乳酸杆菌培养物在新鲜制备的100ml并补充有3%(w/v)葡萄糖的 MRS
c肉汤的烧瓶中生长,并在37℃下孵育72h。通过离心(7000×g,10min)除去细菌细胞,并将两体积的预冷乙醇加入到一体积的上清液中,用于胞外多糖(EPS)沉淀。通过在4℃下离心(10000×g,35min)回收EPS沉淀,透析(6000Da至8000Da)48h,然后冻干。使用葡萄糖作为标准液,用苯酚硫酸法(Nikolic等人,2012)测量糖的总量。目前,生产EPS的L.kefiri MSR101菌株最初是从青海省西藏自治区开菲尔谷物中分离得到的,MSR101菌株显示出粘稠的外观,这表明它是一株EPS生产菌株。L.kefiri MSR101菌株培养48h后处于稳定期且EPS量达到最大值753mg/L,随后进入衰亡期。结果表明,MSR101菌株在培养的32h内生长迅速,同时培养基pH值快速下降,而48h后,该菌株生长速度下降,培养基pH值略有下降,在48h时pH值约为3.1±0.5。
实施例7
细胞表面疏水性
通过微生物对烃类化合物的粘附试验,对乳酸菌的细胞表面疏水性进行了研究(Kotzamanidis等人,2010)。简言之,通过离心(8000×g,5min)收获静止期细胞,用PBS(pH7.2)洗涤两次,最后重悬浮于PBS(pH7.2)中,在630nm处达到0.6±0.02的OD(A
0)。将1ml二甲苯与1ml细胞悬浮液混合,并在室温下静置30min以形成两相体系。小心地除去水相,测量其在630nm处的吸光度(A
1)。通过下面的公式测量细胞表面疏水性(%):
细胞表面疏水性(%)=(1-A
1/A
0)×100
Lactobacillus kefiri MSR101菌株对二甲苯显示出较高的疏水性(70%);二甲苯是测量微生物细胞疏水性的标准碳氢化合物。
实施例8
抗氧化活性
制备完整细胞
将过夜的乳酸杆菌培养物离心(7000×g,4℃下15min),用PBS(pH7.2)洗涤细胞三次,最后重悬于PBS(pH7.2)中,终浓度为1×10
6cfu/ml,进行抗氧化剂分析。
超氧阴离子清除试验
采用改进的邻苯三酚自氧化法(Re等人,2014年)进行超氧阴离子清除实验。即将PBS(pH7.2)中最终浓度为1×10
6cfu/ml的乳酸杆菌细胞悬浮液(100μl)、900μl水与2ml Tric-HCl缓冲液(pH8.1)快速混合。使用无菌蒸馏水代替乳酸杆菌细胞悬浮液作为对照。加入50μmol/ml邻苯三酚溶液,孵育10min后测定对照组和样品在330nm处的吸光度,评价其自氧化作用。通过以下公式计算测试的乳酸杆菌的超氧阴离子清除能力:
清除能力(%)=(ΔA
0-ΔA)×100/ΔA
0
其中ΔA
0和ΔA分别是加入样品和去离子水之前和之后的连苯三酚的自氧化速率。超氧化物阴离子自由基的清除率为65.5%。
DPPH自由基清除活性
通过先前报道的方法(Son等,2018)测定乳酸杆菌的DPPH自由基清除能力。即将三(3)ml乳酸杆菌细胞悬浮液和3ml 0.5mM DPPH自由基的乙醇溶液混合,并在室温下黑暗中温育30min。温育后,将反应混合物离心(10000×g,4℃下20min),测量上清液在517nm处的吸光度。通过下式计算DPPH清除能力:
清除能力(%)=(1-OD
sample/OD
control)×100
其中OD
Sample和OD
Control分别是样品和蒸馏水的吸光度,与DPPH溶液混合。Lactobacillus kefiriMSR101对DPPH自由基清除率为43.3%。
实施例9
降低胆固醇能力
分离的乳酸菌对胆固醇的吸收能力是按照对之前报道的方法(Wang et al.,2012)进行修改后测定的。简言之,将每种乳酸杆菌分离物(1%v/v)接种到10ml新鲜制备的MRS
c肉汤培养基中,该肉汤含有0.2%硫代乙酸钠(Sigma,中国),0.3%牛胆汁(Sigma,中国)和100μl/ml水溶性胆固醇(Sigma,中国),将每个试管在37℃下孵育24h。通过离心(10,000×g,4℃下35min)除去细胞。用测量胆固醇的邻苯二甲醛方法(Rudel和Morris,1973)来测定使用过的肉汤和未接种的无菌肉汤中的胆固醇量。 菌株Lactobacillus kefiri MSR101的胆固醇去除率为26.2%。
实施例10
粘附测定
使用添加有2mM L-谷氨酰胺,10%热灭活的胎牛血清,100μg链霉素/ml,1%非必需氨基酸和100Ul的Dulbecco改良Eagle培养基(DMEM),在细胞培养瓶中培养HT-29细胞。随后将HT-29细胞以2.5×10
5细胞/孔的浓度接种到24孔培养板中,培养3天,每天更换培养基。将细胞在37℃,5%CO
2气氛中温育。将乳杆菌分离物的过夜培养物离心,用PBS(0.1M,pH7.2)洗涤两次,并在相同的缓冲液中重悬至适当浓度的稀释液(吸光度OD6300.2,约2×10
8CFU/ml)。然后将细菌细胞加入到每个细胞孔中,37℃孵育4h,孵育后用PBS(0.1M,pH7.2)冲洗细胞,0.1%Triton X-100溶液裂解。细胞裂解液经连续稀释后涂于MRS
c琼脂平板上。37℃孵育3天。计算细菌粘附率:
黏附率%=(附着菌/总添加菌)×100
菌株Lactobacillus kefiri MSR101在HT-29细胞中的最高的粘附率为34.4%。
实施例11
统计分析
数值以平均值和标准偏差给出,平行三次测定。Tukey多重比较检验在所有试验中均有显著性差异(p<0.05),有统计学意义。
技术效果:
乳酸杆菌kefiri MSR101及其益生菌潜力
从中国青海省西藏自治区收集的克非尔产品样品中分离出乳酸杆菌MSR101。它是革兰氏阳性,过氧化氢酶阴性,杆状和嗜温的。基于16s rRNA基因序列(GCTTGGCGTCGTGCTATACATGCAAGTCGAACGCGTTTCCGTTATTGATTTTAGAGTGTTGCATTTGAATGATTTAACACGAAACGAGTGGCGAACTGGTGAGTAACACGTGGGTACCTGCCCTTGAAGTAGGGGATAACACTTGGAAACAGGTGCTAATACCGTATAACAACCAAAACCACATGG TTTTGGTTTAAAAGATGGCTTCGGCTATCACTTTAGGATGGACCCGGGCGTATTAGCTTGTTGGTAAGGTAATGGCCTACCAAGGCAATGATACGTAGCCGACCTAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGATGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTGGAGAAGAACAGGTGTCAGAGTAACTGTTGACATCTTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTCTTAGGTCTGATGTGAAAGCCTTCGGCTTAACCGGAGAAGTGCATCGGAAACCAGGAGACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGAGCATGTGTTTAATTCGATGCTACGCGAAGACCTTACCAGGTCTTGACATCTTTCTCCTATTTCTGTCACCTTAGACGGCTGGTCCCCGAAGGTTA)鉴定分离物,并且获得的序列显示其与NCBI数据库中可获得的已知物种的序列相似99%,并且序列以登录号MK491609保藏在Gene Bank中。在抗生素敏感性,肠道条件下耐受性(低pH,胆汁盐耐受性和0.2%至0.4%酚抗性),疏水性,抗氧化能力,降低胆固醇和对HT-29的粘附等方面测试益生菌性质。本发明菌株对青霉素,氨苄青霉素,链霉素和四环素具有抗性。此外,它还表现出对HT-29细胞的粘附性(34.3%)和抗氧化活性(67%)。在胃肠道条件下显示出高的存活率(>80%),表明它们在益生菌应用方面有潜力(如表2所示)。
表2 益生菌潜力
以上实施例的说明只是用于帮助理解本发明的方法及其核心思想。应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以对本发明进行若干改进和修饰,这些改进和修饰也落入本发明权利要求的保护范围内。对这些实施例的多种修改对本领域的专业技术人员来说是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
Claims (8)
- 一株高加索酸奶乳杆菌Lactobacillus kefiri MSR101,其特征在于,保藏编号为CGMCC No.17506。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在抗高胆汁盐中的应用。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在苯酚耐受中的应用。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在抗氧化中的应用。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在降低胆固醇浓度中的应用。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在抗青霉素、氨苄青霉素、链霉素和四环素中的应用。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在治疗高胆固醇血症中的应用。
- 权利要求1所述高加索酸奶乳杆菌Lactobacillus kefiri MSR101在治疗高胆汁盐疾病中的应用。
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CN104073455A (zh) * | 2014-06-18 | 2014-10-01 | 兰州大学 | 一株具有降低胆固醇能力的植物乳杆菌 |
CN104818230A (zh) * | 2015-05-05 | 2015-08-05 | 广西科技大学 | 一株具有降解胆固醇功能的植物乳杆菌l01及其应用 |
CN107058161A (zh) * | 2016-12-29 | 2017-08-18 | 石家庄君乐宝乳业有限公司 | 具有抗氧化功能的高加索酸奶乳杆菌jmcc0101、其筛选方法及应用 |
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104073455A (zh) * | 2014-06-18 | 2014-10-01 | 兰州大学 | 一株具有降低胆固醇能力的植物乳杆菌 |
CN104818230A (zh) * | 2015-05-05 | 2015-08-05 | 广西科技大学 | 一株具有降解胆固醇功能的植物乳杆菌l01及其应用 |
CN107058161A (zh) * | 2016-12-29 | 2017-08-18 | 石家庄君乐宝乳业有限公司 | 具有抗氧化功能的高加索酸奶乳杆菌jmcc0101、其筛选方法及应用 |
CN110106113A (zh) * | 2019-05-10 | 2019-08-09 | 深圳大学 | 一株高加索酸奶乳杆菌msr101及其应用 |
Non-Patent Citations (4)
Title |
---|
CHEN, XIAO-YONG , LI JIAN , ZHAO XIN , QIAN YU , CHEN LIAN-HONG , CHEN JUAN , DUO HUA-YI: "In vitro screening of lactic acid bacteria with probiotic properties from traditional fermented Yak yogurt", FOOD AND FERMENTATION INDUSTRIES, vol. 42, no. 4, 3 March 2016 (2016-03-03), pages 85 - 90, XP055753416, ISSN: 0253-990X, DOI: 10.13995/j.cnki.11-1802/ts.201604016 * |
TONG ZHAO , YONG-XIA WANG, YI-PING XUN, LIAN-ZHONG AI, LIN-SEN ZHAO, LI-FENG WANG, HONG ZHU, SHI-JIE WANG: "Analysis of Probiotic Characteristics of Lactobacillus kefiri KL22", SCIENCE AND TECHNOLOGY OF FOOD INDUSTRY, vol. 40, no. 9, 29 December 2019 (2019-12-29), pages 115 - 120, XP055753400, ISSN: 1002-0306, DOI: 10.13386/j.issn1002-0306.2019.09.021 * |
ZHAO TONG , WANG YONG-XIA , XUN YI-PING , ZHANG DONG , FENG LI-LI , ZHU HONG ,WANG SHI-JIE: "Study on the characteristics of L. kefir KL22", CHINA FOOD ADDITIVES, no. 10, 31 October 2018 (2018-10-31), pages 45 - 49, XP055753409, ISSN: 1006-2513 * |
ZHUQING XING, WEI TANG, WEITAO GENG, YONGNA ZHENG, YANPING WANG: "In vitro and in vivo evaluation of the probiotic attributes of Lactobacillus kefiranofaciens XL10 isolated from Tibetan kefir grain", APPLIED MICROBIAL AND CELL PHYSIOLOGY, vol. 101, no. 6, 1 December 2016 (2016-12-01), pages 2467 - 2477, XP036164209, ISSN: 0175-7598, DOI: 10.1007/s00253-016-7956-z * |
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CN110106113A (zh) | 2019-08-09 |
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