Purification of Glucoamylase from Lactobacillus amylovorus ATCC 33621

James, Jennylynd; Berger, Jean-Luc

doi:10.1007/s002849900166

Cited by 23 publications

(15 citation statements)

References 34 publications

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“…The compositions of monosaccharide in EPSs were analyzed with the alditol acetate derivatization method, using GC-MS. 23) After the purified EPS (5 mg) was hydrolyzed with 2 M trifluoroacetic acid at 120°C for 1 h to obtain each monosaccharide, the reduced reaction was performed, followed by acetylation. The resulting derivative was analyzed by GC-MS, using a JMS-T100GCV AccuTOF GCv 4G gas chromatograph high-resolution time-of-flight mass spectrometer (JEOL, Tokyo, Japan) equipped with a source of ions for electron ionization (EI), using a DB-WAX capillary column (0.25 mm×0.25 µm×30 m) (Agilent, Santa Clara, CA, U.S.A.).…”

Section: Analysis Of Constitution Sugars In Epssmentioning

confidence: 99%

Characterization of Exopolysaccharides Produced by Thermophilic Lactic Acid Bacteria Isolated from Tropical Fruits of Thailand

Panthavee

Noda

Danshiitsoodol

et al. 2017

Biological & Pharmaceutical Bulletin

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In the present study, we have obtained two exopolysaccharide (EPS)-producing thermophilic lactic acid bacteria (LAB) that were isolated from tropical fruits of Thailand. The two strains, designated LY45 and PY45, were identified as Pediococcus pentosaceus and Lactobacillus amylovorus, respectively. Both plantderived LAB strains, which produce neutral EPSs together with the acidic one, can grow vigorously at 45°C and even at 50°C. Hyaluronidase (EC 3.2.1.35), which catalyzes the degradation of hyaluronic acid, activates an inflammatory reaction. Interestingly, EPSs produced by the LY45 and PY45 strains were found to inhibit hyaluronidase activity at the same order of IC 50 values as did sodium cromoglicate and dipotassium glycyrrhizinate, which are well-known as anti-inflammatory agents. The LY45-derived neutral EPS consists of glucose and mannose as monosaccharide components, whereas the acidic one contains mainly mannose, together with glucose and galactose. On the other hand, although Lactobacillus amylovorus PY45 also produces neutral and acidic EPSs, the main monosaccharide in both EPSs is mannose, and glucose is a minor component. Furthermore, the PY45 strain may be probiotically and industrially useful because the microorganism can utilize starch and glycogen as carbon sources. Key words exopolysaccharide; Lactobacillus amylovorus; Pediococcus pentosaceus; thermophilic lactic acid bacteria; anti-inflammatory substanceTraditionally, lactic acid bacteria (LAB) have been useful for producing fermented foods, such as yogurt, kimchi, and cheese. LAB-Gram-positive bacteria-are non-pathogenic and are generally recognized as safe (GRAS) microorganisms; therefore, they have been the focus of attention for their industrial importance. 1)Microorganisms that contribute to human health are called probiotics. This word is internationally defined as "living microorganisms conferring a health benefit on the host when administered in adequate amounts."2) We have isolated many kinds of LAB strains from plant sources such as vegetables, fruits, flowers, and medicinal plants to establish a plantderived LAB library with more than 600 strains. We have demonstrated that some strains stocked in the LAB library are useful for enhancing intestinal immunity, improving liver function, and preventing metabolic syndrome. [3][4][5][6] LAB fermentation often proceeds even at low temperatures, causing the overgrowth of LABs. As the result, the degradation of flavors is proceeded and affected with taste of the fermented foods. Since thermophilic LAB strains hard to ferment at lower temperatures, therefore, if the thermophile ones are successfully isolated, they will contribute to prevent over-fermentation. In the present study, we isolated 28 strains of thermophilic LAB from tropical fruits of Thailand. In the isolates, we found two exopolysaccharide (EPS)-producing LAB strains. The LY45 strain was isolated from lychee, and the PY45 strain was obtained from pineapple. These strains were identified as Pediococcus (P.) pentosaceus and Lactob...

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Section: Analysis Of Constitution Sugars In Epssmentioning

confidence: 99%

Characterization of Exopolysaccharides Produced by Thermophilic Lactic Acid Bacteria Isolated from Tropical Fruits of Thailand

Panthavee

Noda

Danshiitsoodol

et al. 2017

Biological & Pharmaceutical Bulletin

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“…᭺, ␣-amylase activity; ᭡, cell count (cfu/ ml). and became constant after 40 h. Glucoamylase activity was not detected by the method of James et al (1997) throughout the experiments, suggesting that only a minimal amount of the enzyme was synthesized to supply glucose to the cells. Production of lactic acid and pH of the culture broth followed the change in ␣-amylase activity.…”

Section: Resultsmentioning

confidence: 94%

“…Lactobacillus amylovorus, a member of the L. acidophilus-group, is one of these amylolytic species (Nakamura, 1981). ␣-Amylase and glucoamylase produced by the type strain of L. amylovorus have been characterized in detail (Giraud & Cuny, 1997;James et al, 1997). We previously compared ten strains of L. amylovorus with respect to lactic acid fermentation from starch and found that strain JCM 10628 produced lactic acid from raw cornstarch at a much higher rate than other strains (Oda et al, 2000).…”

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confidence: 99%

Utilization of Lactobacillus amylovorus as an Alternative Microorganism for Saccharifying Boiled Rice.

Oda

Ichinose

Yamauchi

2002

FSTR

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The growth condition of Lactobacillus amylovorus strain JCM 10628, an amylolytic lactic acid bacterium, was tested as a source of enzymes for saccharifying boiled rice to produce a beverage similar to Amazake, a traditional nonalcoholic beverage. The composition of a medium that should leave no unpleasant taste in the Amazake-like product was confined to 1.0% raw cornstarch, 2.0% skim milk, and 0.1% yeast extract. ␣-Amylase activity and cell growth decreased in the medium lacking any of the three ingredients. Production of ␣-amylase was stimulated by soluble starch as well as raw cornstarch, but neither by glucose nor sucrose. When 100 ml of the culture broth was mixed with 16 g of rice after boiling and incubated at 55˚C for 24 h, the obtained fluid had an acceptable sweet and sour taste.

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“…Microbial strains and culture media [4]. One unit of the enzyme activity was defined at the amount of enzyme which releases 100 μg of glucose from soluble starch for 1 hr under the above condition.…”

Section: Methodsmentioning

confidence: 99%

“…Rhizomucor pusilus and Endomyces sp. [4,5,16] In the yeast, Sacchoromyces diastaticus, three isoenzymes of the extracellular glucoamylase have been reported; glucoamylase I, glucoamylase II and glucoamylase III. These isozymes are encoded by the STA1 (DEX2), STA2 (DEX1) and STA3 (DEX3) gene, respectively [14].…”

Section: Introductionmentioning

confidence: 99%

Characterization of Sporulation-Specific Glucoamylase of Saccharomyces diastaticus

2010

Journal of Life Science

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The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and 45 o C, respectively, whereas those for the extracellular glucoamylase were 5.0 and 50 o C. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

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Purification of Glucoamylase from Lactobacillus amylovorus ATCC 33621

Cited by 23 publications

References 34 publications

Characterization of Exopolysaccharides Produced by Thermophilic Lactic Acid Bacteria Isolated from Tropical Fruits of Thailand

Characterization of Exopolysaccharides Produced by Thermophilic Lactic Acid Bacteria Isolated from Tropical Fruits of Thailand

Utilization of Lactobacillus amylovorus as an Alternative Microorganism for Saccharifying Boiled Rice.

Characterization of Sporulation-Specific Glucoamylase of Saccharomyces diastaticus

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